



@article {3273,
	title = {Characterization of hLF1-11 immobilization onto chitosan ultrathin films, and its effects on antimicrobial activity},
	journal = {Acta Biomaterialia},
	volume = {10},
	year = {2014},
	note = {Cited By (since 1996):1Export Date: 21 November 2014},
	month = {2014},
	pages = {3513 - 3521},
	abstract = {hLF1-11 (GRRRRSVQWCA) is an antimicrobial peptide (AMP) with high activity against methicillin-resistant Staphylococcus aureus (MRSA), the most prevalent species in implant-associated infection. In this work, the effect of the surface immobilization on hLF1-11 antimicrobial activity was studied. Immobilization was performed onto chitosan thin films as a model for an implant coating due to its reported osteogenic and antibacterial properties. Chitosan thin films were produced by spin-coating on gold surfaces. hLF1-11 was immobilized onto these films by its C-terminal cysteine in an orientation that exposes the antimicrobial activity-related arginine-rich portion of the peptide. Two levels of exposure (with and without a polyethylene glycol (PEG) spacer) were analyzed. Covalent immobilization was further compared with the AMP physical adsorption onto chitosan films. Surfaces were characterized using ellipsometry, contact angle measurements, atomic force microscopy, infrared and X-ray photoelectron spectroscopies and using a fluorimetric assay for hLF1-11 quantification. Surface antimicrobial activity was assessed through surface adhesion and viability assays using an MRSA (S. aureus ATCC 33591). The incorporation of hLF1-11 increased significantly bacterial adhesion to chitosan films. However, the presence of hLF1-11, namely when immobilized through a PEG spacer, decreased the viability of adherent bacteria with regard to the control surface. These results demonstrated that hLF1-11 after covalent immobilization by its cysteine can maintain activity, particularly if a spacer is applied. However, further studies, exploring the opposite orientation or the same C-terminal orientation, but non-cysteine related, can help to clarify the potential of the hLF1-11 immobilization strategy. {\textcopyright} 2014 Acta Materialia Inc. Published by Elsevier Ltd. All rights reserved.},
	keywords = {Antimicrobial peptides (AMP), Bacterial adhesion, chitosan, Surface characterization, Surface immobilization},
	url = {http://www.scopus.com/inward/record.url?eid=2-s2.0-84903724424\&partnerID=40\&md5=e8009cd480b49c37a991753cd145cf34},
	author = {Costa, F. and Maia, S. and Gomes, J. and Gomes, P. and Martins, M.C.L.}
}
@article {3300,
	title = {Chitosan cross-linked docetaxel loaded EGF receptor targeted nanoparticles for lung cancer cells},
	journal = {International Journal of Biological Macromolecules},
	volume = {69},
	year = {2014},
	note = {Cited By (since 1996):1Export Date: 21 November 2014},
	month = {2014},
	pages = {532 - 541},
	abstract = {Lung cancer, associated with the up-regulated epidermal growth factor receptor (EGFR) led to the development of EGFR targeted anticancer therapeutics. The biopolymeric nanoparticles form an outstanding system for the targeted delivery of therapeutic agents. The present work evaluated the in vitro effects of chitosan cross-linked γ-poly(glutamic acid) (γ-PGA) nanoparticles (Nps) loaded with docetaxel (DTXL) and decorated with Cetuximab (CET), targeted to EGFR over-expressing non-small-cell-lung-cancer (NSCLC) cells (A549). CET-DTXL-γ-PGA Nps was prepared by ionic gelation and CET conjugation via EDC/NHS chemistry. EGFR specificity of targeted Nps was confirmed by the higher uptake rates of EGFR +ve A549 cells compared to that of EGFR -ve cells (NIH3T3). The cytotoxicity of Nps quantified using cell based (MTT/LDH) and flowcytometry (Cell-cycle analysis, Annexin V/PI and JC-1) assays showed superior antiproliferative activity of CET-DTXL-γ-PGA Nps over DTXL-γ-PGA Nps. The A549 cells treated with CET-DTXL-γ-PGA NPs underwent a G2/M phase cell cycle arrest followed by reduction in mitochondrial membrane potential of A549 cells, inducing apoptosis and necrosis resulting in enhanced cancer cell death. CET-DTXL-γ-PGA Nps exhibited enhanced cellular internalization and therapeutic activity, by actively targeting EGFR on NSCLC cells and hence could be an effective alternative to non-specific, conventional chemotherapy by increasing its efficiency by many folds. {\textcopyright} 2014 Elsevier B.V.},
	keywords = {Cetuximab, chitosan, Epidermal growth factor receptor, Poly(γ-glutamic acid) nanoparticles, Targeted nanoparticles},
	url = {http://www.scopus.com/inward/record.url?eid=2-s2.0-84903523289\&partnerID=40\&md5=728cbd3502e7922319f5f88fa8d6ec6d},
	author = {Maya, S. and Sarmento, B. and Lakshmanan, V.-K. and Menon, D. and Seabra, V. and Jayakumar, R.}
}
@article {3322,
	title = {Chitosan-modified porous silicon microparticles for enhanced permeability of insulin across intestinal cell monolayers},
	journal = {Biomaterials},
	volume = {35},
	year = {2014},
	note = {Cited By (since 1996):1Export Date: 21 November 2014},
	month = {2014},
	pages = {7172 - 7179},
	abstract = {Porous silicon (PSi) based particulate systems are emerging as an important drug delivery system due to its advantageous properties such as biocompatibility, biodegradability and ability to tailor the particles{\textquoteright} physicochemical properties. Here, annealed thermally hydrocarbonized PSi (AnnTHCPSi) and undecylenic acid modified AnnTHCPSi (AnnUnTHCPSi) microparticles were developed as a PSi-based platform for oral delivery of insulin. Chitosan (CS) was used to modify the AnnUnTHCPSi microparticles to enhance the intestinal permeation of insulin. Surface modification with CS led to significant increase in the interaction of PSi microparticles with Caco-2/HT-29 cell co-culture monolayers. Compared to pure insulin, the CS-conjugated microparticles significantly improved the permeation of insulin across the Caco-2/HT-29 cell monolayers, with ca. 20-fold increase in the amount of insulin permeated and ca. 7-fold increase in the apparent permeability (Papp) value. Moreover, among all the investigated particles, the CS-conjugated microparticles also showed the highest amount of insulin associated with the mucus layer and the intestinal Caco-2 cells and mucus secreting HT-29 cells. Our results demonstrate that CS-conjugated AnnUnTHCPSi microparticles can efficiently enhance the insulin absorption across intestinal cells, and thus, they are promising microsystems for the oral delivery of proteins and peptides across the intestinal cell membrane. {\textcopyright} 2014 Elsevier Ltd.},
	keywords = {Cell monolayers, chitosan, Diabetes, Insulin, Microparticles, Porous silicon},
	url = {http://www.scopus.com/inward/record.url?eid=2-s2.0-84902079022\&partnerID=40\&md5=4de20e5572512006f72e11cb61a2035c},
	author = {Shrestha, N. and Shahbazi, M.-A. and Ara{\'u}jo, F. and Zhang, H. and M{\"a}kil{\"a}, E.M. and Kauppila, J. and Sarmento, B. and Salonen, J.J. and Hirvonen, J.T. and Santos, H.A.}
}
@article {3264,
	title = {The impact of nanoparticles on the mucosal translocation and transport of GLP-1 across the intestinal epithelium},
	journal = {Biomaterials},
	volume = {35},
	year = {2014},
	note = {Export Date: 21 November 2014},
	month = {2014},
	pages = {9199 - 9207},
	abstract = {Glucagon like peptide-1 (GLP-1) is an incretin hormone that is in the pipeline for type 2 diabetes mellitus (T2DM) therapy. However, oral administration of GLP-1 is hindered by the harsh conditions of the gastrointestinal tract and poor bioavailability. In this study, three nanosystems composed by three different biomaterials (poly(lactide-co-glycolide) polymer (PLGA), Witepsol E85 lipid (solid lipid nanoparticles, SLN) and porous silicon (PSi) were developed and loaded with GLP-1 to study their permeability invitro. All the nanoparticles presented a size of approximately 200nm. The nanoparticles{\textquoteright} interaction with the mucus and the intestinal cells were enhanced after coating with chitosan (CS). PSi nanosystems presented the best association efficiency (AE) and loading degree (LD), even though a high AE was also observed for PLGA nanoparticles and SLN. Among all the nanosystems, PLGA and PSi were the only nanoparticles able to sustain the release of GLP-1 in biological fluids when coated with CS. This characteristic was also maintained when the nanosystems were in contact with the intestinal Caco-2 and HT29-MTX cell monolayers. The CS-coated PSi nanoparticles showed the highest GLP-1 permeation across the intestinal invitro models. In conclusion, PLGA+CS and PSi+CS are promising nanocarriers for the oral delivery of GLP-1. {\textcopyright} 2014 Elsevier Ltd.},
	keywords = {chitosan, Diabetes, Glucagon like peptide-1, nanoparticles, Oral delivery systems, Triple co-culture},
	url = {http://www.scopus.com/inward/record.url?eid=2-s2.0-84906788144\&partnerID=40\&md5=38c557a51eb271561ef59bfe12421842},
	author = {Ara{\'u}jo, F. and Shrestha, N. and Shahbazi, M.-A. and Fonte, P. and M{\"a}kil{\"a}, E.M. and Salonen, J.J. and Hirvonen, J.T. and Granja, P.L. and Santos, H.A. and Sarmento, B.}
}
@article {3289,
	title = {The potential utility of chitosan micro/nanoparticles in the treatment of gastric infection},
	journal = {Expert Review of Anti-Infective Therapy},
	volume = {12},
	year = {2014},
	note = {Export Date: 21 November 2014},
	month = {2014},
	pages = {981 - 992},
	abstract = {Gastric infections are mainly caused by Helicobacter pylori (H. pylori), a bacterium that colonizes the gastric mucosa of over 50\% of the world{\textquoteright}s population. Chronic H. pylori infection has been associated with gastric diseases such as chronic gastritis, peptic ulcer and gastric adenocarcinoma. Current eradication treatment relies on antibiotic-based therapies that are unsuccessful in approximately 20\% of the patients. Chitosan, a natural and cationic polysaccharide has been investigated in the treatment of H. pylori infection. Due to its mucoadhesive properties, it has been used in the form of micro/nanoparticles, polyelectrolyte complexes or coatings as antibiotic encapsulation systems for gastric delivery, but alternative molecules may also be incorporated. It has been recently proposed that chitosan can also be used for H. pylori binding and scavenging from the host stomach due to its antimicrobial/binding properties. In this manuscript, a brief description of the use of chitosan in H. pylori treatment is reviewed. {\textcopyright} Informa UK, Ltd.},
	keywords = {Biomaterials, chitosan, Crosslinking, Drug delivery, Gastric cancer, Helicobacter pylori, Microspheres, mucoadhesion, nanoparticles, Polyelectrolytes},
	url = {http://www.scopus.com/inward/record.url?eid=2-s2.0-84904003586\&partnerID=40\&md5=d8a682d7ce6be8633b1505d0a7b8adae},
	author = {Gon{\c c}alves, I.C. and Henriques, P.C. and Seabra, C.L. and Martins, M.C.L.}
}
@article {2203,
	title = {Biosynthesis of highly pure poly-γ-glutamic acid for biomedical applications},
	journal = {Journal of Materials Science: Materials in MedicineJ. Mater. Sci. Mater. Med.},
	volume = {23},
	year = {2012},
	note = {Export Date: 21 November 2012Source: ScopusCODEN: JSMMEdoi: 10.1007/s10856-012-4639-xPubMed ID: 22532096Language of Original Document: EnglishCorrespondence Address: Pereira, C.L.; INEB-Instituto de Engenharia Biom{\'e}dica, Universidade Do Porto, Rua do Campo Alegre, 823, 4150-180 Porto, Portugal; email: Catarina.pereira@ineb.up.ptChemicals/CAS: chitosan, 9012-76-4; glutamic acid, 11070-68-1, 138-15-8, 56-86-0, 6899-05-4; polyglutamic acid, 24991-23-9, 25513-46-6; Biocompatible Materials; Polyglutamic Acid, 25513-46-6; poly(gamma-glutamic acid)References: Ratner, B.D., Hoffman, A.S., Shoen, F.J., Lemons, J.E., (2004) Natural Materials. Biomaterials Science-an Introduction to Materials in Medicine, , 2nd ed. San Diego, CA: Academic Press;Buescher, J.M., Margaritis, A., Microbial biosynthesis of polyglutamic acid biopolymer and applications in the biopharmaceutical, biomedical and food industries (2007) Critical Reviews in Biotechnology, 27 (1), pp. 1-19. , DOI 10.1080/07388550601166458, PII 773236840; 
Kang, H.-S., Park, S.-H., Lee, Y.-G., Son, T.-I., Polyelectrolyte complex hydrogel composed of chitosan and poly(γ-glutamic acid) for biological application: Preparation, physical properties, and cytocompatibility (2007) Journal of Applied Polymer Science, 103 (1), pp. 386-394. , DOI 10.1002/app.24623; 
Lin, Y.-H., Chung, C.-K., Chen, C.-T., Liang, H.-F., Chen, S.-C., Sung, H.-W., Preparation of nanoparticles composed of chitosan/poly-γ-glutamic acid and evaluation of their permeability through Caco-2 cells (2005) Biomacromolecules, 6 (2), pp. 1104-1112. , DOI 10.1021/bm049312a; 
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Park, C., Choi, J.-C., Choi, Y.-H., Nakamura, H., Shimanouchi, K., Horiuchi, T., Misono, H., Sung, M.-H., Synthesis of super-high-molecular-weight poly-γ-glutamic acid by Bacillus subtilis subsp. chungkookjang (2005) Journal of Molecular Catalysis B: Enzymatic, 35 (4-6), pp. 128-133. , DOI 10.1016/j.molcatb.2005.06.007; 
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Kubota, H., Matsunobu, T., Uotani, K., Takebe, H., Satoh, A., Tanaka, T., Production of poly(c-glutamic acid) by Bacillus subtilis F-2-01 (1993) Biosci Biotechnol Biochem, 57 (7), pp. 1212-1213; 
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},
	month = {2012///},
	pages = {1583 - 1591},
	abstract = {The remarkable properties of poly-aminoacids, mainly their biocompatibility and biodegradability, have prompted an increasing interest in these polymers for biomedical applications. Poly-γ-glutamic acid (γ-PGA) is one of the most interesting poly-aminoacids with potential applications as a biomaterial. Here we describe the production and characterization of γ-PGA by Bacillus subtilis natto. The γ-PGA was produced with low molecular weight (10-50 kDa), high purity grade (>99 \%) and a D-/L-glutamate ratio of 50-60/50-40 \%. To evaluate the feasibility of using this γ-PGA as a biomaterial, chitosan (Ch)/γ-PGA nanoparticles were prepared by the coacervation method at pH ranging from 3.0 to 5.0, with dimensions in the interval 214-221 nm with a poly-dispersion index of ca. 0.2. The high purity of γ-PGA produced by this method, which is firstly described here, renders this biopolymer suitable for biomedical applications. Moreover, the Ch/γ-PGA nanocomplexes developed in this investigation can be combined with biologically active substances for their delivery in the organism. The fact that the assembly between Ch and γ-PGA relies on electrostatic interactions enables addition of other molecules that can be released into the medium through changes from acidic to physiological pH, without loss in biological activity. {\textcopyright} Springer Science+Business Media, LLC 2012.},
	keywords = {Amino acids, animal cell, article, Bacillus subtilis, Bacillus subtilis natto, Bioactivity, Biocompatibility, Biocompatible Materials, biodegradability, Biodegradable polymers, Biodegradation, Biologically active substances, biomaterial, Biomedical applications, biomedicine, biosynthesis, chitosan, circular dichroism, complex formation, controlled study, dispersion, Electrophoresis, Polyacrylamide Gel, feasibility study, Functional polymers, gamma polyglutamic acid, Glutamic acid, High purity, Low molecular weight, Medical applications, molecular interaction, molecular weight, Nanocomplexes, nanoparticle, nonhuman, pH, Physiological pH, polyglutamic acid, Potential applications, priority journal, purification, Spectroscopy, Fourier Transform Infrared, static electricity, Stereoisomerism, unclassified drug},
	isbn = {09574530 (ISSN)},
	url = {http://www.scopus.com/inward/record.url?eid=2-s2.0-84863314791\&partnerID=40\&md5=d0d618bb2684c8e80e984cfe35f3eb77},
	author = {Pereira, C.L. and Antunes, J.C. and Gon{\c c}alves, R.M. and Ferreira-da-Silva, F. and Barbosa, M.A.}
}
@article {2180,
	title = {Chitosan drives anti-inflammatory macrophage polarisation and pro-inflammatory dendritic cell stimulation.},
	journal = {European cells \& materialsEur Cell Mater},
	volume = {24},
	year = {2012},
	note = {Export Date: 21 November 2012Source: ScopusPubMed ID: 22828991Language of Original Document: EnglishCorrespondence Address: Oliveira, M.I.Chemicals/CAS: chitosan, 9012-76-4; Antigens, CD86; Biocompatible Materials; Chitosan, 9012-76-4; Histocompatibility Antigens Class II; IL10 protein, human; Interleukin-10, 130068-27-8; Transforming Growth Factor beta1; Tumor Necrosis Factor-alpha},
	month = {2012///},
	pages = {136 - 152; discussion 152-153},
	abstract = {Macrophages and dendritic cells (DC) share the same precursor and play key roles in immunity. Modulation of their behaviour to achieve an optimal host response towards an implanted device is still a challenge. Here we compare the differentiation process and polarisation of these related cell populations and show that they exhibit different responses to chitosan (Ch), with human monocyte-derived macrophages polarising towards an anti-inflammatory phenotype while their DC counterparts display pro-inflammatory features. Macrophages and DC, whose interactions with biomaterials are frequently analysed using fully differentiated cells, were cultured directly on Ch films, rather than exposed to the polymer after complete differentiation. Ch was the sole stimulating factor and activated both macrophages and DC, without leading to significant T cell proliferation. After 10 d on Ch, macrophages significantly down-regulated expression of pro-inflammatory markers, CD86 and MHCII. Production of pro-inflammatory cytokines, particularly TNF-α, decreased with time for cells cultured on Ch, while anti-inflammatory IL-10 and TGF-β1, significantly increased. Altogether, these results suggest an M2c polarisation. Also, macrophage matrix metalloproteinase activity was augmented and cell motility was stimulated by Ch. Conversely, DC significantly enhanced CD86 expression, reduced IL-10 secretion and increased TNF-α and IL-1β levels. Our findings indicate that cells with a common precursor may display different responses, when challenged by the same biomaterial. Moreover, they help to further comprehend macrophage/DC interactions with Ch and the balance between pro- and anti-inflammatory signals associated with implant biomaterials. We propose that an overall pro-inflammatory reaction may hide the expression of anti-inflammatory cytokines, likely relevant for tissue repair/regeneration.},
	keywords = {antibody specificity, Antigens, CD86, article, Biocompatible Materials, biomaterial, biosynthesis, CD86 antigen, Cell culture, cell differentiation, cell motion, Cell Movement, Cells, Cultured, chitosan, cytology, dendritic cell, Dendritic Cells, drug effect, flow cytometry, Histocompatibility Antigens Class II, HLA antigen class 2, human, Humans, IL10 protein, human, immunology, interleukin 10, Interleukin-10, macrophage, Macrophages, metabolism, monocyte, Monocytes, Organ Specificity, transforming growth factor beta1, tumor necrosis factor alpha, Tumor Necrosis Factor-alpha},
	isbn = {14732262 (ISSN)},
	url = {http://www.scopus.com/inward/record.url?eid=2-s2.0-84868253478\&partnerID=40\&md5=5a496411709ccbe5d3d70de13b58b08a},
	author = {Oliveira, M.I. and Santos, S.G. and Oliveira, M.J. and Torres, A.L. and Barbosa, M.A.}
}
@booklet {2186,
	title = {Effect of gastric environment on Helicobacter pylori adhesion to a mucoadhesive polymer},
	howpublished = {Acta BiomaterialiaActa Biomater.},
	year = {2012},
	note = {Export Date: 21 November 2012Source: ScopusArticle in Pressdoi: 10.1016/j.actbio.2012.09.011Language of Original Document: EnglishCorrespondence Address: Martins, M.C.L.; INEB-Instituto de Engenharia Biom{\'e}dica, Universidade do Porto, Rua do Campo Alegre, 823, 4150email: cmartins@ineb.up.pt},
	month = {2012///},
	abstract = {Helicobacter pylori infection has been associated with several gastric diseases. This bacterium colonizes the gastric mucosa of half of the world{\textquoteright}s population, and available treatments are unsuccessful in practically one in every five patients. Mucoadhesive polymers, such as chitosan, are being investigated as gastric drug delivery systems. However, since chitosan is also known for its antimicrobial properties, this work aims to evaluate H. pylori interactions with chitosan under simulated gastric environments, namely using various pHs (2.6, 4 and 6), pepsin and urea. To enable the visualization of adherent bacteria, ultrathin chitosan films were produced by spin-coating on gold/glass surfaces, cross-linked with genipin and characterized by Fourier transform infrared reflection absorption spectroscopy, ellipsometry and electrokinetic analysis. Films with homogeneous thickness of 11.7 {\textpm} 0.6 nm were produced, and were stable and protonated at all the pHs used. Furthermore, they adsorbed pepsin in all these pHs, in contrast to urea, of which a small adsorption was only observed at pH 6. H. pylori binding to chitosan was higher at pH 2.6 although most of adherent bacteria were dead. The presence of pepsin decreased bacterial adhesion, but increased its viability while in a more stressed morphology (coccoid form). The presence of urea did not affect the amount, morphology or viability of chitosan-adherent bacteria. In suspension, the decrease in pH changed H. pylori zeta potential from negative to positive. Moreover, bacteria were only culturable when incubated in pH 6 with and without urea (without pepsin). This work demonstrates that chitosan has the capacity to bind and kill H. pylori in a range of pHs independently of urea. This opens new perspectives for the application of chitosan-based materials to the elimination of H. pylori gastric colonization, though pepsin might appear to be an obstacle. {\textcopyright} 2012 Acta Materialia Inc.},
	keywords = {Bacterial adhesion, Biomaterials, chitosan, Helicobacter pylori, Thin films},
	isbn = {17427061 (ISSN)},
	url = {http://www.scopus.com/inward/record.url?eid=2-s2.0-84867507239\&partnerID=40\&md5=defd903a9833b81e1d4e9e377550ce07},
	author = {Nogueira, F. and Gon{\c c}alves, I.C. and Martins, M.C.L.}
}
@article {2211,
	title = {Enhanced mesenchymal stromal cell recruitment via natural killer cells by incorporation of inflammatory signals in biomaterials},
	journal = {Journal of the Royal Society InterfaceJ. R. Soc. Interface},
	volume = {9},
	year = {2012},
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},
	month = {2012///},
	pages = {261 - 271},
	abstract = {An exacerbated inflammatory response questions biomaterial biocompatibility, but on the other hand, inflammation has a central role in the regulation of tissue regeneration. Therefore, it may be argued that an {\textquoteright}ideal{\textquoteright} inflammatory response is crucial to achieve efficient tissue repair/regeneration. Natural killer (NK) cells, being one of the first populations arriving at an injury site, can have an important role in regulating bone repair/regeneration, particularly through interactions with mesenchymal stem/stromal cells (MSCs). Here, we studied how biomaterials designed to incorporate inflammatory signals affected NK cell behaviour and NK cell-MSC interactions. Adsorption of the pro-inflammatory molecule fibrinogen (Fg) to chitosan films led to a 1.5-fold increase in adhesion of peripheral blood human NK cells, without an increase in cytokine secretion. Most importantly, it was found that NK cells are capable of stimulating a threefold increase in human bone marrow MSC invasion, a key event taking place in tissue repair, but did not affect the expression of the differentiation marker alkaline phosphatase (ALP). Of significant importance, this NK cell-mediated MSC recruitment was modulated by Fg adsorption. Designing novel biomaterials leading to rational modulation of the inflammatory response is proposed as an alternative to current bone regeneration strategies. {\textcopyright} 2011 The Royal Society.},
	keywords = {Adhesion, Adsorption, alkaline phosphatase, article, Biocompatibility, biomaterial, Biomaterials, blood cell, Bone, bone marrow cell, Bone Regeneration, Bone repair, cell adhesion, Cell culture, cell differentiation, cell function, cell interaction, cell invasion, Cells, chitosan, Chitosan film, cytokine, Cytokine secretions, Differentiation markers, fibrinogen, gamma interferon, human, Human bone marrow, human cell, Inflammation, Inflammatory response, Mesenchymal Stem Cells, mesenchymal stroma cell, Mesenchymal stromal cells, natural killer cell, Natural killer cells, NK cells, ossification, Pathology, Peripheral blood, Phosphatases, protein expression, Tissue, Tissue regeneration, tissue repair},
	isbn = {17425689 (ISSN)},
	url = {http://www.scopus.com/inward/record.url?eid=2-s2.0-84856583891\&partnerID=40\&md5=8a6b58ebb9ee4c20099877008acde805},
	author = {Almeida, C.R. and Vasconcelos, D.P. and Gon{\c c}alves, R.M. and Barbosa, M.A.}
}
@article {2214,
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	volume = {23},
	year = {2012},
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},
	month = {2012///},
	pages = {249 - 261},
	abstract = {Human mesenchymal stem cells (hMSCs) have an enormous potential for tissue engineering and cell-based therapies. With a potential of differentiation into multiple lineages and immune-suppression, these cells play a key role in tissue remodelling and regeneration. Here a method of hMSC recruitment is described, based on the incorporation of a chemokine in Chitosan (Ch)/ Poly(γ-glutamic acid) (γ-PGA) complexes. Ch is a nontoxic, cationic polysaccharide widely investigated. γ-PGA is a hydrophilic, non-toxic, biodegradable and negatively charged poly-amino acid. Ch and γ-PGA, being oppositely charged, can be combined through electrostatic interactions. These biocompatible structures can be used as carriers for active substances and can be easily modulated in order to control the delivery of drugs, proteins, DNA, etc. Using the layer-by-layer method, Ch and γ-PGA were assembled into polyelectrolyte multilayers films (PEMs) with thickness of 120 nm. The chemokine stromal-derived factor-1 (SDF-1) was incorporated in these complexes and was continuously released during 120 h. The method of SDF-1 incorporation is of crucial importance for polymers assembly into PEMs and for the release kinetics of this chemokine. The Ch/γ-PGA PEMs with SDF-1 were able to recruit hMSCs, increasing the cell migration up to 6 fold to a maximum of 16.2 {\textpm} 4.9 cells/mm                        2. The controlled release of SDF-1 would be of great therapeutic value in the process of hMSC homing to injured tissues. This is the first study suggesting Ch/γ-PGA PEMs as SDF-1 reservoirs to recruit hMSCs, describing an efficient method of chemokine incorporation that allows a sustained released up to 5 days and that can be easily scaled-up.
},
	keywords = {adsorption kinetics, article, artificial membrane, Biocompatible Materials, biomaterial, cell differentiation, cell migration, cell motion, Cell Movement, Cell recruitment, chemistry, Chemokine CXCL12, chitosan, controlled study, CXCL12 protein, human, cytology, drug adsorption, drug delivery system, Drug Delivery Systems, drug derivative, drug effect, ellipsometry, enzyme linked immunosorbent assay, human, human cell, human tissue, Humans, in vitro study, isotope labeling, Layer-by-layer, Membranes, Artificial, Mesenchymal stem cell, Mesenchymal Stem Cells, metabolism, methodology, poly(gamma glutamic acid), poly(gamma-glutamic acid), Poly(γ-glutamic acid), polyglutamic acid, Protein delivery, static electricity, stromal cell derived factor 1, sustained drug release},
	isbn = {14732262 (ISSN)},
	url = {http://www.scopus.com/inward/record.url?eid=2-s2.0-84864582297\&partnerID=40\&md5=2fe4a066582cc820d2a8f9dd64fde21b},
	author = {Gon{\c c}alves, R.M. and Antunes, J.C. and Barbosa, M.A.}
}
@article {2196,
	title = {A novel nanoparticle delivery system for in vivo targeting of the sciatic nerve: Impact on regeneration},
	journal = {NanomedicineNanomedicine},
	volume = {7},
	year = {2012},
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},
	month = {2012///},
	pages = {1167 - 1180},
	abstract = {Aim: Innovative solutions in the development of drug delivery systems targeting the nerve tissue are awaited. In this regard, a novel system for the delivery of drugs to the sciatic nerve was created using nanomedical principles. Materials \& methods: Chitosan was the vehicle material used in the experiment. Heparin bound to growth factors has been administered to enhance peripheral nerve regeneration, and since heparin possesses the appropriate charge to be able to form nanoparticles with chitosan, it appears to be a good candidate to base this new delivery system on. Results: Maximal absorption took place throughout the extracellular matrix at day 15. No major inflammatory response was observed, indicating that this is a safe and biocompatible system for drug delivery to nerves. Sensorimotor performance and nerve regeneration of mice receiving these nanoparticles were superior as compared with controls. Conclusion: Our work demonstrates a versatile nanoparticle delivery system that successfully targets drugs {\textquoteright}in vivo to the sciatic nerve, opening novel avenues in the field of nanomedicine to the design of therapeutic strategies that enhance axonal regeneration. {\textcopyright} 2012 Future Medicine Ltd.},
	keywords = {chitosan, drug delivery system, heparin, nanomedicine, nanoparticles, peripheral nerve regeneration, sciatic nerve},
	isbn = {17435889 (ISSN)},
	url = {http://www.scopus.com/inward/record.url?eid=2-s2.0-84868111412\&partnerID=40\&md5=5db0c76b3a921ca70565c9f3f63cf004},
	author = {Gon{\c c}alves, N.P. and Oliveira, H. and P{\^e}go, A.P. and Saraiva, M.J.}
}
@article {2212,
	title = {Synthesis of an O-alkynyl-chitosan and its chemoselective conjugation with a PEG-like amino-azide through click chemistry},
	journal = {Carbohydrate PolymersCarbohydr Polym},
	volume = {87},
	year = {2012},
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},
	month = {2012///},
	pages = {240 - 249},
	abstract = {N-Phthaloyl-chitosan O-prop-2-ynyl carbamate was prepared as a biopolymer amenable to undergo chemoselective conjugation by azide-alkyne coupling, while allowing upturn of chitosan{\textquoteright}s amines after dephthaloylation. N-phthaloylchitosan was prepared according to previously described methods and, due to its low solubility in current organic media, subsequent modifications were run in heterogeneous conditions. Activation of hydroxyls with carbonyl-1,1'- diimidazole and coupling to propargylamine yielded N-phthaloyl-chitosan O-prop-2-ynyl carbamate, then coupled to a model PEG-like azide by azide-alkyne coupling, giving the expected triazolyl conjugate. N-Dephthaloylation allowed recovery of the free amines, responsible for chitosan{\textquoteright}s bioadhesion and tissue-regeneration properties. The structures of all polymers were confirmed by Fourier-transformed infra-red (FT-IR) and X-ray photoelectron (XPS) spectroscopies, as well as by solid-state nuclear magnetic resonance (SSNMR). All chitosan derivatives were poorly soluble in both aqueous and organic media, which makes them suitable for topical applications or for removal of toxic substances from either the gastric intestinal tract or environmental sources. {\textcopyright} 2011 Elsevier Ltd. All Rights Reserved.},
	keywords = {1 ,3-Dipolarcycloaddition, Acetylene, Amines, Azide-alkyne coupling, Bio-adhesion, Bioadhesion, Biomaterials, chitosan, click chemistry, Cycloaddition, FTIR, Huisgen{\textquoteright}s 1,3-dipolar cycloaddition, Nuclear magnetic resonance, Nuclear magnetic resonance spectroscopy, Photoelectron spectroscopy, Resonance, Size exclusion chromatography, Size-exclusion chromatography, Solid state NMR, Solid-state NMR, Tissue, X ray photoelectron spectroscopy, XPS},
	isbn = {01448617 (ISSN)},
	url = {http://www.scopus.com/inward/record.url?eid=2-s2.0-80054771159\&partnerID=40\&md5=3760f41563573759f0ce676fe813d6c6},
	author = {Oliveira, J.R. and Martins, M.C.L. and Mafra, L. and Gomes, P.}
}
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	volume = {12},
	year = {2011},
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},
	month = {2011///},
	pages = {4183 - 4195},
	abstract = {Chitosan (Ch) is a nontoxic and biocompatible polysaccharide extensively used in biomedical applications. Ch, as a polycation, can be combined with anionic polymers by layer-bylayer (LbL) self-assembly, giving rise to multilayered complexed architectures. These structures can be used in tissue engineering strategies, as drug delivery systems, or artificial matrices mimicking the extracellular microenvironment. In this work, Ch was combined with poly(γ-glutamic acid) (γ-PGA). γ-PGA is a polyanion, which was microbially produced, and is known for its low immunogenic reaction and low cytotoxicity. Multilayered ultrathin films were assembled by LbL, with a maximum of six layers. The interaction between both polymers was analyzed by: ellipsometry, quartz crystal microbalance with dissipation, Fourier transform infrared spectroscopy, atomic force microscopy, and zeta potential measurements. Ch/γ-PGA polyelectrolyte multilayers (PEMs) revealed no cytotoxicity according to ISO 10993-5. Overall, this study demonstrates that Ch can interact electrostatically with γ-PGA forming multilayered films. Furthermore, this study provides a comprehensive characterization of Ch/γ-PGA PEM structures, elucidating the contribution of each layer for the nanostructured films. These model surfaces can be useful substrates to study cell-biomaterial interactions in tissue regeneration. {\textcopyright} 2011 American Chemical Society.},
	keywords = {Amino acids, Anionic polymers, article, Atomic force microscopy, Atomic spectroscopy, Biocompatible Materials, Biocompatible polysaccharides, Biomaterials, Biomedical applications, Cell-biomaterial interactions, chemical interaction, chitosan, controlled study, Cytotoxicity, Drug delivery, drug delivery system, Drug Delivery Systems, Electrolytes, ellipsometry, enzyme activity, Extracellular, Fourier transform infrared spectroscopy, Glutamic acid, immunogenicity, infrared spectroscopy, Layer by layer self assembly, Medical applications, microenvironment, Microenvironments, Microscopy, Atomic Force, Model surface, Models, Molecular, Multi-layered, Multi-layered films, Multilayers, nanofilm, Nanostructured Films, polyanion, Polyanions, Polycations, polyelectrolyte, Polyelectrolyte complexes, Polyelectrolyte multilayer, Polyelectrolytes, Polyethylenes, polyglutamic acid, Polymers, priority journal, Programmed control systems, Quartz, quartz crystal microbalance, Quartz crystal microbalance with dissipation, Self assembly, Tissue, Tissue engineering, Tissue regeneration, Ultrathin films, Viscoelastic Substances, Zeta potential, Zeta potential measurements},
	isbn = {15257797 (ISSN)},
	url = {http://www.scopus.com/inward/record.url?eid=2-s2.0-84856258191\&partnerID=40\&md5=8ba447b7a45e8004e5b3aa3d5cff2404},
	author = {Antunes, J.C. and Pereira, C.L. and Molinos, M. and Ferreira-da-Silva, F. and Dessi, M. and Gloria, A. and Ambrosio, L. and Gonca̧lves, R.M. and Barbosa, M.A.}
}
@article {2244,
	title = {Chitosan-based gene delivery vectors targeted to the peripheral nervous system},
	journal = {Journal of Biomedical Materials Research - Part AJ. Biomed. Mater. Res. Part A},
	volume = {95},
	year = {2010},
	note = {Cited By (since 1996): 5Export Date: 21 November 2012Source: ScopusCODEN: JBMRCdoi: 10.1002/jbm.a.32874PubMed ID: 20734332Language of Original Document: EnglishCorrespondence Address: P{\^e}go, A. P.; INEB, Instituto de Engenharia Biom{\'e}dica, Universidade Do Porto, Rua do Campo Alegre, 823, 4150-180 Porto, Portugal; email: apego@ineb.up.ptChemicals/CAS: DNA, 9007-49-2; chitosan, 9012-76-4; imidazole, 1467-16-9, 288-32-4; macrogol, 25322-68-3; Cations; Chitosan, 9012-76-4; DNA, 9007-49-2; Polyethylene GlycolsTradenames: Chimarin, medicarb, SwedenManufacturers: medicarb, SwedenReferences: Rodriguez, F.J., Valero-Cabre, A., Navarro, X., Regeneration and functional recovery following peripheral nerve injury (2004) Drug Discov Today, 1, pp. 177-185;Apfel, S.C., Neurotrophic factors in peripheral neuropathies: Therapeutic implications (1999) Brain Pathol, 9, pp. 393-413; 
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Oliveira, H., Fernandez, R., Pires, L.R., Sim{\~o}es, S., Martins, M.C.L., Barbosa, M.A., P{\^e}go, A.P., Targeted gene delivery into peripheral sensorial neurons mediated by self-assembled vectors composed of poly(ethilenimine) and tetanus toxin fragment c (2010) J Control Release, 143, pp. 350-358; 
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},
	month = {2010///},
	pages = {801 - 810},
	abstract = {A non-toxic, targeted, simple and efficient system that can specifically transfect peripheral sensorial neurons can pave the way towards the development of new therapeutics for the treatment of peripheral neuropathies. In this study chitosan (CH), a biodegradable polymer, was used as the starting material in the design of a multicomponent vector targeted to the peripheral nervous system (PNS). Polycation-DNA complexes were optimized using imidazole-and thiol-grafted CH (CHimiSH), in order to increase transfection efficiency and allow the formation of ligand conjugated nanocomplexes, respectively. The 50 kDa non-toxic fragment from the tetanus toxin (HC), shown to interact specifically with peripheral neurons and undergo retrograde transport, was grafted to the binary complex via a bi-functional poly(ethylene glycol) (HC-PEG) reactive for the thiol moieties present in the complex surface. The targeting of the developed nanocomplexes was assessed by means of internalization and transfection studies in the ND7/23 (neuronal) vs. NIH 3T3 (fibroblast) cell lines. Targeted transfection was further confirmed in dorsal root ganglion dissociated primary cultures. A versatile, multi-component nanoparticle system that successfully targets and transfects neuronal cell lines, as well as dorsal root ganglia (DRG) primary neuron cultures was obtained for the 1.0 (w/w) HC-PEG/DNA formulation. {\textcopyright} 2010 Wiley Periodicals, Inc.},
	keywords = {animal cell, Animals, article, Binary complexes, Biodegradable polymers, Carbohydrate Conformation, Carbohydrate Sequence, Cations, Cell culture, Cell lines, cell population, Cells, Cultured, Chitin, chitosan, Complex surface, DNA, DNA complex, Dorsal root ganglia, Efficient systems, embryo, Ethylene, Ethylene Glycol, Female, Ganglia, Spinal, Gene delivery, gene delivery system, Gene delivery vectors, Gene therapy, Gene transfer, Gene Transfer Techniques, genetic transfection, Genetic Vectors, Grafting (chemical), Humans, imidazole, macrogol, materials testing, Mice, Molecular Sequence Data, Molecular Structure, mouse, Multicomponents, Nanocomplexes, Nanoparticle systems, nanoparticles, nerve cell culture, Neuron culture, Neuronal cell, NIH 3T3 Cells, nonhuman, Nontoxic fragment of tetanus toxin, Peripheral nervous system, Peripheral neuropathy, Polycations, Polyethylene Glycols, Polyethylene oxides, Pregnancy, Primary culture, rat, Rats, Rats, Wistar, spinal ganglion, Starting materials, Targeted, Tetanus toxin, Toxic materials, Transfection, Transfection efficiency},
	isbn = {15493296 (ISSN)},
	url = {http://www.scopus.com/inward/record.url?eid=2-s2.0-78249240032\&partnerID=40\&md5=af9c3ca5ca712d53abb5e80f45ed09c0},
	author = {Oliveira, H. and Pires, L.R. and Fernandez, R. and Martins, M.C.L. and Sim{\~o}es, S. and P{\^e}go, A.P.}
}
@article {2250,
	title = {Chitosan/siRNA nanoparticles biofunctionalize nerve implants and enable neurite outgrowth},
	journal = {Nano LettersNano Lett.},
	volume = {10},
	year = {2010},
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},
	month = {2010///},
	pages = {3933 - 3939},
	abstract = {Microstructured 20 μm thick polymer filaments used as nerve implants were loaded with chitosan/siRNA nanoparticles to promote nerve regeneration and ensure local delivery of nanotherapeutics. The stable nanoparticles were rapidly internalized by cells and did not affect cell viability. Target mRNA was successfully reduced by 65-75\% and neurite outgrowth was enhanced even in an inhibitory environment. This work, thus, supports the application of nanobiofunctionalized implants as a novel approach for spinal cord and nerve repair. {\textcopyright} 2010 American Chemical Society.},
	keywords = {animal, Animals, article, Cell Line, cell viability, chemistry, Chitin, chitosan, cytology, Drug delivery, genetic transfection, genetics, Local delivery, metabolism, Microstructured, nanobiofunctionalized implants, nanoparticle, nanoparticles, nerve cell, nerve regeneration, Nerve repair, neurite, Neurite outgrowth, Neurites, Neurons, Polymer filaments, Prostheses and Implants, prostheses and orthoses, rhoA GTP-Binding Protein, RhoA guanine nucleotide binding protein, RNA, RNA, Small Interfering, siRNA, small interfering RNA, Spinal cords, Transfection, ultrastructure},
	isbn = {15306984 (ISSN)},
	url = {http://www.scopus.com/inward/record.url?eid=2-s2.0-77958033530\&partnerID=40\&md5=6862e82e01c3aedf5423ebb14248c6ec},
	author = {Mittnacht, U. and Hartmann, H. and Hein, S. and Oliveira, H. and Dong, M. and P{\^e}go, A.P. and Kjems, J. and Howard, K.A. and Schlosshauer, B.}
}
@article {2280,
	title = {Evaluation of the effect of the degree of acetylation on the inflammatory response to 3D porous chitosan scaffolds},
	journal = {Journal of Biomedical Materials Research - Part AJ. Biomed. Mater. Res. Part A},
	volume = {93},
	year = {2010},
	note = {Cited By (since 1996): 3Export Date: 21 November 2012Source: ScopusCODEN: JBMRCdoi: 10.1002/jbm.a.32499PubMed ID: 19484769Language of Original Document: EnglishCorrespondence Address: Barbosa, J. N.; INEB - Instituto de Engenharia Biom{\'e}dica, Divis{\~a}o de Biomateriais, Universidade do Porto, Rua do Campo Alegre 823, 4150-180 Porto, Portugal; email: judite@ineb.up.ptChemicals/CAS: chitosan, 9012-76-4; Chitosan, 9012-76-4References: Sefton, M.V., Babensee, J.E., Woodhouse, K.A., Innate and adaptative immune responses in tissue engineering (2008) Semin Immunol, 20, pp. 83-85;VandeVord, P.J., Matthew, H.W.T., DeSilva, S.P., Mayton, L., Wu, B., Wooley, P.H., Evaluation of the biocompatibility of a chitosan scaffold in mice (2002) J Biomed Mater Res, 59, pp. 585-590; 
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},
	month = {2010///},
	pages = {20 - 28},
	abstract = {The effect of the degree of acetylation (DA) of 3D chitosan (Ch) scaffolds on the inflammatory reaction was investigated. Chitosan porous scaffolds with DAs of 4 and 15\% were implanted using a subcutaneous air-pouch model of inflammation. The initial acute inflammatory response was evaluated 24 and 48 h after implantation. To characterize the initial response, the recruitment and adhesion of inflammatory cells to the implant site was studied. The fibrous capsule formation and the infiltration of inflammatory cells within the scaffolds were evaluated for longer implantation times (2 and 4 weeks). Chitosan with DA 15\% attracted the highest number of leukocytes to the implant site. High numbers of adherent inflammatory cells were also observed in this material. For longer implantation periods Ch scaffolds with a DA of 15\% induced the formation of a thick fibrous capsule and a high infiltration of inflammatory cells within the scaffold. Our results indicate that the biological response to implanted Ch scaffolds was influenced by the DA. Chitosan with a DA of 15\% induce a more intense inflammatory response when compared with DA 4\% Ch. Because inflammation and healing are interrelated, this result may provide clues for the relative importance of acetyl and amine functional groups in tissue repair and regeneration. {\textcopyright} 2009 Wiley Periodicals, Inc.},
	keywords = {acetic acid derivative, Acetylation, Acute inflammatory response, amine, Amine functional groups, animal cell, animal experiment, animal tissue, Animals, article, Biological materials, Biological response, Biomaterials, cell adhesion, cell infiltration, Chitin, chitosan, Chitosan scaffold, Degree of acetylation, Fibrous capsule, Fibrous capsule formation, freeze drying, Functional groups, Hydrogels, implant, implantation, Implantation periods, Implants, Experimental, Inflammation, Inflammatory cells, Inflammatory reaction, Inflammatory response, leukocyte count, Leukocytes, male, Mice, Mice, Inbred BALB C, Microscopy, Electron, Scanning, mouse, nonhuman, Pathology, porosity, Porous scaffold, Prosthesis Implantation, Relative importance, Scaffolds, Subcutaneous Tissue, Surface Properties, Three dimensional, Tissue regeneration, tissue repair, Tissue response, Tissue responses, Tissue Scaffolds},
	isbn = {15493296 (ISSN)},
	url = {http://www.scopus.com/inward/record.url?eid=2-s2.0-77649219383\&partnerID=40\&md5=58781af29b82f3108ded649c4776e428},
	author = {Barbosa, J.N. and Amaral, I.F. and {\'A}guas, A.P. and Barbosa, M.A.}
}
@article {2316,
	title = {Engineering endochondral bone: In vitro studies},
	journal = {Tissue Engineering - Part ATissue Eng. Part A},
	volume = {15},
	year = {2009},
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Oliveira, S.M., Turner, G., Mijares, D., Amaral, I.F., Barbosa, M.A., Teixeira, C.C., (2007) Engineering endochondral bone: In vivo studies
},
	month = {2009///},
	pages = {625 - 634},
	abstract = {Chitosan scaffolds have been shown to possess biological and mechanical properties suitable for tissue engineering and clinical applications. In the present work, chitosan sponges were evaluated regarding their ability to support cartilage cell proliferation and maturation, which are the first steps in endochondral bone formation. Chitosan sponges were seeded with chondrocytes isolated from chicken embryo sterna. Chondrocyte/chitosan constructs were cultured for 20 days, and treated with retinoic acid (RA) to induce chondrocyte maturation and matrix synthesis. At different time points, samples were collected for microscopic, histological, biochemical, and mechanical analyses. Results show chondrocyte attachment, proliferation, and abundant matrix synthesis, completely obliterating the pores of the sponges. RA treatment caused chondrocyte hypertrophy, characterized by the presence of type X collagen in the extracellular matrix and increased alkaline phosphatase activity. In addition, hypertrophy markedly changed the mechanical properties of the chondrocyte/chitosan constructs. In conclusion, we have developed chitosan sponges with adequate pore structure and mechanical properties to serve as a support for hypertrophic chondrocytes. In parallel studies, we have evaluated the ability of this mature cartilage scaffold to induce endochondral ossification. {\textcopyright} Copyright 2009, Mary Ann Liebert, Inc. 2009.},
	keywords = {alkaline phosphatase, Alkaline phosphatase activity, animal, animal cell, animal tissue, Animals, article, biomechanics, Body fluids, Bone, Bone and Bones, Bone formation, Cartilage, cartilage cell, Cell culture, cell damage, cell hypertrophy, cell isolation, cell maturation, Cell membranes, cell proliferation, Cells, Cultured, chick embryo, Chicken embryo, Chitin, chitosan, Chitosan scaffold, Chondrocyte, Chondrocyte maturation, Chondrocytes, Clinical application, collagen type 10, Collagen Type X, controlled study, cytology, DNA, drug effect, embryo, enchondral ossification, Endochondral ossification, enzyme activity, enzymology, Extracellular matrices, extracellular matrix, gene expression profiling, Hypertrophic chondrocytes, in vitro study, In-vitro, Ligaments, Matrix synthesis, Mechanical analysis, Mechanical properties, metabolism, methodology, nonhuman, Pathology, physiology, Porifera, porosity, priority journal, retinoic acid, Retinoic acids, Scaffolds, sponge (Porifera), Sterna, sternum, synthesis, Time points, Tissue engineering, tissue scaffold, Tissue Scaffolds, Tretinoin, Type X collagen},
	isbn = {19373341 (ISSN)},
	url = {http://www.scopus.com/inward/record.url?eid=2-s2.0-64549143844\&partnerID=40\&md5=310f8e62d4dd51ccb9399db171440148},
	author = {Oliveira, S.M. and Amaral, I.F. and Barbosa, M.A. and Teixeira, C.C.}
}
@article {2317,
	title = {Engineering endochondral bone: In vivo studies},
	journal = {Tissue Engineering - Part ATissue Eng. Part A},
	volume = {15},
	year = {2009},
	note = {Cited By (since 1996): 14Export Date: 21 November 2012Source: Scopusdoi: 10.1089/ten.tea.2008.0052Language of Original Document: EnglishCorrespondence Address: Teixeira, C. C.; Department of Basic Science and Craniofacial Biology, College of Dentistry, New York University, 345 East. 24th St., New York, NY 10010; email: ct40@nyu.eduChemicals/CAS: chitosan, 9012-76-4; Chitosan, 9012-76-4; MineralsReferences: Yelin, E., Callahan, L.F., The economic cost and social and psychological impact of musculoskeletal conditions. National Arthritis Data Work Groups (1995) Arthritis Rheum, 38, p. 1351;Service, R.F. Tissue engineers build new bone. Science 289,1498, 2000Dupraz, A., Delecrin, J., Moreau, A., Pilet, P., Passuti, N., Long-term bone response to particulate injectable ceramic (1998) J Biomed Mater Res, 42, p. 368; 
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Deckers, M.M., Karperien, M., van der Bent, C., Yamashita, T., Papapoulos, S.E., Lowik, C.W., Expression of vascular endothelial growth factors and their receptors during osteoblast differentiation (2000) Endocrinology, 141, p. 1667; 
Petersen, W., Tsokos, M., Pufe, T., Expression of VEGF121 and VEGF165 in hypertrophic chondrocytes of the human growth plate and epiphyseal cartilage (2002) J Anat, 201, p. 153; 
Nakagawa, M., Kaneda, T., Arakawa, T., Morita, S., Sato, T., Yomada, T., Hanada, K., Hakeda, Y., Vascular endothelial growth factor (VEGF) directly enhances osteoclastic bone resorption and survival of mature osteoclasts (2000) FEBS Lett, 473, p. 161; 
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Oliveira, S.M., Amaral, I.F., Barbosa, M.A., Teixeira, C.C., (2007) Engineering endochondral bone: In vitro studies; 
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},
	month = {2009///},
	pages = {635 - 643},
	abstract = {The use of biomaterials to replace lost bone has been a common practice for decades. More recently, the demands for bone repair and regeneration have pushed research into the use of cultured cells and growth factors in association with these materials. Here we report a novel approach to engineer new bone using a transient cartilage scaffold to induce endochondral ossification. Chondrocyte/chitosan scaffolds (both a transient cartilage scaffold- experimental-and a permanent cartilage scaffold-control) were prepared and implanted subcutaneously in nude mice. Bone formation was evaluated over a period of 5 months. Mineralization was assessed by Faxitron, micro computed tomography, backscatter electrons, and Fourier transform infrared spectroscopy analyses. Histological analysis provided further information on tissue changes in and around the implanted scaffolds. The deposition of ectopic bone was detected in the surface of the experimental implants as early as 1 month after implantation. After 3 months, bone trabeculae and bone marrow cavities were formed inside the scaffolds. The bone deposited was similar to the bone of the mice vertebra. Interestingly, no bone formation was observed in control implants. In conclusion, an engineered transient cartilage template carries all the signals necessary to induce endochondral bone formation in vivo. {\textcopyright} Copyright 2009, Mary Ann Liebert, Inc. 2009.},
	keywords = {animal, animal experiment, animal tissue, Animals, article, Biological materials, Biomaterials, Bone, Bone and Bones, Bone formation, bone marrow, bone mineralization, Bone repair, bone transplantation, Calcification, Physiologic, Cartilage, cartilage cell, chick embryo, chitosan, Chondrocytes, Computerized tomography, controlled study, Cultured cell, cytology, drug effect, Ectopic bones, ectopic tissue, electron, enchondral ossification, Endochondral ossification, Fourier transform infrared spectroscopy, Growth (materials), Growth factor, Histological analysis, histology, implantation, in vivo study, In-control, In-vivo, infrared spectroscopy, Ligaments, male, metabolism, methodology, Mice, Mice, Nude, micro-computed tomography, Microcomputed tomography, Microscopy, Electron, Scanning, mineral, Minerals, mouse, Mus, Mus musculus, nonhuman, Nude mice, nude mouse, ossification, physiology, priority journal, Prosthesis Implantation, Scaffolds, Scanning electron microscopy, Spectroscopic analysis, Spectroscopy, Spectroscopy, Fourier Transform Infrared, Tissue changes, Tissue engineering, tissue scaffold, Tissue Scaffolds, Tomography, trabecular bone, vertebra},
	isbn = {19373341 (ISSN)},
	url = {http://www.scopus.com/inward/record.url?eid=2-s2.0-64549096039\&partnerID=40\&md5=d389cb8b10be8945007a5b6996b8896f},
	author = {Oliveira, S.M. and Mijares, D.Q. and Turner, G. and Amaral, I.F. and Barbosa, M.A. and Teixeira, C.C.}
}
@article {2297,
	title = {Fibronectin-mediated endothelialisation of chitosan porous matrices},
	journal = {BiomaterialsBiomaterials},
	volume = {30},
	year = {2009},
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},
	month = {2009///},
	pages = {5465 - 5475},
	abstract = {Chitosan (Ch) porous matrices were investigated regarding their ability to be colonized by human microvascular endothelial cells (HPMEC-ST1.6R cell line) and macrovascular endothelial cells namely HUVECs. Specifically we assessed if previous incubation of Ch in a fibronectin (FN) solution was effective in promoting endothelial cell (EC) adhesion to Ch matrices with different degrees of acetylation (DAs). Upon FN physiadsorption, marked differences were found between the two DAs investigated, namely DA 4\% and 15\%. While cell adhesion was impaired on Ch with DA 15\%, ECs were able to not only adhere to Ch with DA 4\%, but also to spread and colonize the scaffolds, with retention of the EC phenotype and angiogenic potential. To explain the observed differences between the two DAs, protein adsorption studies using                        125I-FN and immunofluorescent labelling of FN cell-binding domains were carried out. In agreement with the higher cell numbers found, scaffolds with DA 4\% revealed a higher number of exposed FN cell-binding domains as well as greater ability to adsorb FN and to retain and exchange adsorbed FN in the presence of competitive proteins. These findings suggest that the DA is a key parameter modulating EC adhesion to FN-coated Ch by influencing the adsorbed protein layer. {\textcopyright} 2009 Elsevier Ltd. All rights reserved.
},
	keywords = {Acetylation, Adhesion, Adsorption, angiogenesis, article, Biocompatible Materials, Blood vessel prostheses, cell adhesion, Cell culture, cell invasion, Cell Line, Cell lines, Cell membranes, Cell number, cell proliferation, Cell size, cell spreading, cell survival, Cell-binding domain, Chitin, Chitin/chitosan, chitosan, colony formation, controlled study, Degrees of acetylation, Electric conductivity measurement, Electric currents, Endothelial cell, Endothelial Cells, Endothelialisation, endothelium, endothelium cell, epithelization, Fibronectin, Fibronectins, human, human cell, Human microvascular endothelial cells, Humans, immunofluorescence test, Key parameters, Labels, materials testing, Matrix algebra, Neovascularization, Physiologic, phenotype, porosity, Porous matrixs, priority journal, Protein adsorption, Protein layers, Scaffold, Scaffolds, Self assembly, Tissue engineering, tissue scaffold, umbilical vein},
	isbn = {01429612 (ISSN)},
	url = {http://www.scopus.com/inward/record.url?eid=2-s2.0-68549107574\&partnerID=40\&md5=9babb57c1a4f2ae6eb7e7e0643bffe48},
	author = {Amaral, I.F. and Unger, R.E. and Fuchs, S. and Mendon{\c c}a, A.M. and Sousa, S.R. and Barbosa, M.A. and P{\^e}go, A.P. and Kirkpatrick, C.J.}
}
@article {2296,
	title = {Improving chitosan-mediated gene transfer by the introduction of intracellular buffering moieties into the chitosan backbone},
	journal = {Acta BiomaterialiaActa Biomater.},
	volume = {5},
	year = {2009},
	note = {Cited By (since 1996): 17Export Date: 21 November 2012Source: Scopusdoi: 10.1016/j.actbio.2009.04.021PubMed ID: 19427930Language of Original Document: EnglishCorrespondence Address: P{\^e}go, A.P.; INEB-Instituto de Engenharia Biom{\'e}dica, Divis{\~a}o de Biomateriais, Universidade do Porto, Rua do Campo Alegre 823, 4150-180 Porto, Portugal; email: apego@ineb.up.ptChemicals/CAS: DNA, 9007-49-2; chitosan, 9012-76-4; imidazole, 1467-16-9, 288-32-4; Biocompatible Materials; Buffers; Chitosan, 9012-76-4; DNA, 9007-49-2; Drug CarriersReferences: Kirkpatrick, C.J., Fuchs, S., Peters, K., Brochhausen, C., Hermanns, M.I., Unger, R.E., Visions for regenerative medicine: interface between scientific fact and science fiction (2006) Artif Organs, 30, pp. 822-827;Sim{\~o}es, S., Filipe, A., Faneca, H., Mano, M., Penacho, N., Duzgunes, N., Cationic liposomes for gene delivery (2005) Expert Opin Drug Deliv, 2, pp. 237-254; 
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},
	month = {2009///},
	pages = {2995 - 3006},
	abstract = {Chitosan was functionalized with imidazole moieties (CHimi) with the aim of improving its buffering capacity and promoting the endosomal escape ability of chitosan-DNA complexes, ultimately increasing their transfection efficiency. 5.6\%, 12.9\% and 22.1\% of the glucosamine residues of chitosan were substituted. Complexes with different molar ratios of primary amines to DNA phosphate anion (N/P) were prepared by a coacervation method. For an N/P \&gt; 3, CHimi polymers are able to complex electrostatically with DNA and condense it into positively charged nanostructures (average size 260 nm and zeta potential +16 mV at pH 5.5). In the concentration range 2.5-100 μg ml},
	keywords = {article, beta galactosidase, Biocompatible Materials, Buffers, Cell Line, cell transport, Chemical structure, chitosan, coacervation, complex formation, controlled study, Cytotoxicity, Diffusion, DNA, Drug Carriers, Drug Compounding, endosome, Gene delivery, Gene transfer, genetic transfection, human, human cell, Humans, imidazole, Kidney, materials testing, nanomaterial, nanoparticle, priority journal, protein expression, proton transport, Tissue engineering, Transfection},
	isbn = {17427061 (ISSN)},
	url = {http://www.scopus.com/inward/record.url?eid=2-s2.0-70349162856\&partnerID=40\&md5=fb6672dd0cf06d6ced21eec115b597d2},
	author = {Moreira, C. and Oliveira, H. and Pires, L.R. and Sim{\~o}es, S. and Barbosa, M.A. and P{\^e}go, A.P.}
}
@article {2324,
	title = {Physical, chemical and in vitro biological profile of chitosan hybrid membrane as a function of organosiloxane concentration},
	journal = {Acta BiomaterialiaActa Biomater.},
	volume = {5},
	year = {2009},
	note = {Cited By (since 1996): 10Export Date: 21 November 2012Source: Scopusdoi: 10.1016/j.actbio.2008.07.022PubMed ID: 18722166Language of Original Document: EnglishCorrespondence Address: Hayakawa, S.; Biomaterial Laboratory, Graduate School of Natural Science and Technology, Okayama University, Tsushima, Okayama-shi, 700-8530, Japan; email: satoshi@cc.okayama-u.ac.jpChemicals/CAS: (3 glycidyloxypropyl)trimethoxysilane, 2530-83-8; calcium phosphate, 10103-46-5, 13767-12-9, 14358-97-5, 7758-87-4; chitosan, 9012-76-4; silicate, 12627-13-3; Chitosan, 9012-76-4; Cross-Linking Reagents; gamma-glycidoxypropyltrimethoxysilane, 2530-83-8; Silanes; SiloxanesTradenames: AldrichReferences: Lai, J.Y., Lin, F.C., Wang, C.C., Wang, D.M., Effect of nonsolvent additives on the porosity and morphology of asymmetric TPX membranes [J] (1996) J Membrane Sci, 118, pp. 49-61;Wang, D.M., Lin, F.C., Chiang, J.C., Lai, J.Y., Control of porosity of asymmetric TPX membranes [J] (1998) J Membrane Sci, 141, pp. 1-12; 
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Dupoirieux, L., Pourquier, D., Picot, M.C., Neves, M., Comparative study of three different membranes for guided bone regeneration of rat cranial defects (2001) Int J Oral Maxillofac Surg, 30 (1), pp. 58-62; 
Zhang, R., Ma, P.X., Porous poly(l-lactic acid)/apatite composites created by biomimetic process (1999) J Biomed Mater Res, 45, pp. 285-293; 
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Muzzareli, R.A.A., Mattioli-Belmonte, M., Tietz, C., Biagini, R., Ferioli, G., Brunelli, M.A., Stimulatory effect on bone formation exerted by a modified chitosan (1994) Biomaterials, 15, pp. 1075-1081; 
Shirosaki, Y., Tsuru, K., Hayakawa, S., Osaka, A., Lopes, M.A., Santos, J.D., In vitro cytocompatibility of MG63 cells on chitosan-organosiloxane hybrid membranes (2005) Biomaterials, 26, pp. 485-493; 
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Costa, M.A., Gutierres, M., Almeida, L., Lopes, M.A., Santos, J.D., Fernandes, M.H., In vitro mineralization of human bone marrow cells cultured on Bonelike {\textregistered} (2004) Key Eng Mater, 254-256, pp. 821-824
},
	month = {2009///},
	pages = {346 - 355},
	abstract = {We attempted to prepare chitosan-silicate hybrid for use in a medical application and evaluated the physico-chemical properties and osteocompatibility of the hybrids as a function of γ-glycidoxypropyltrimethoxysilane (GPTMS) concentration. Chitosan-silicate hybrids were synthesized using GPTMS as the reagent for cross-linking of the chitosan chains. Fourier transform infrared spectroscopy,                        29Si CP-MAS NMR spectroscopy and the ninhydrin assay were used to analyze the structures of the hybrids, and stress-strain curves were recorded to estimate their Young{\textquoteright}s modulus. The swelling ability, contact angle and cytocompatibility of the hybrids were investigated as a function of the GPTMS concentration. A certain fraction of GPTMS in each hybrid was linked at the epoxy group to the amino group of chitosan, which was associated with the change in the methoxysilane group of GPTMS due to hybridization. The cross-linking density was around 80\% regardless of the volume of GPTMS. As the content of GPTMS increased, the water uptake decreased and the hydrophilicity of the hybrids increased except when the content exceeded amolar ratio of 1.5, when it caused a decrease. The values of the mechanical parameters assessed indicated that significant stiffening of the hybrids was obtained by the addition of GPTMS. The adhesion and proliferation of the MG63 osteoblast cells cultured on the chitosan-GPTMS hybrid surface were improved compared to those on the chitosan membrane, regardless of the GPTMS concentration. Moreover, human bone marrow osteoblast cells proliferated on the chitosan-GPTMS hybrid surface and formed a fibrillar extracellular matrix with numerous calcium phosphate globular structures, both in the presence and in the absence of dexamethasone. Therefore, the chitosan-GPTMS hybrids are promising candidates for basic materials that can promote bone regeneration because of their controllable composition (chitosan/GPTMS ratio). {\textcopyright} 2008 Acta Materialia Inc.
},
	keywords = {(3 glycidyloxypropyl)trimethoxysilane, article, assay, Biocompatibility, Bone Marrow Cells, calcium phosphate, cell adhesion, Cell Line, Tumor, cell proliferation, cell survival, cell viability, Chemical analysis, chitosan, concentration (parameters), controlled study, cross linking, Cross-Linking Reagents, extracellular matrix, human, human cell, Humans, hydrophobicity, in vitro study, infrared spectroscopy, Magnetic Resonance Spectroscopy, Microscopy, Electron, Scanning, molecular hybridization, Nuclear magnetic resonance spectroscopy, Organic-inorganic hybrid, osteoblast, Osteoblasts, Osteocompatibility, physical chemistry, priority journal, Silanes, silicate, siloxane, Siloxanes, Spectrophotometry, Ultraviolet, Spectroscopy, Fourier Transform Infrared, stress strain relationship, structure analysis, surface property, Tissue engineering, unclassified drug, water transport, γ-Glycidoxypropyltrimethoxysilane (GPTMS)},
	isbn = {17427061 (ISSN)},
	url = {http://www.scopus.com/inward/record.url?eid=2-s2.0-56649083638\&partnerID=40\&md5=550d9a49b04985f38609e86d9adc5735},
	author = {Shirosaki, Y. and Tsuru, K. and Hayakawa, S. and Osaka, A. and Lopes, M.A. and Santos, J.D. and Costa, M.A. and Fernandes, M.H.}
}
@book {2361,
	title = {Biodegradable chitosan-silicate porous hybrids for drug delivery},
	series = {Key Engineering MaterialsKey Eng Mat},
	volume = {361-363 II},
	year = {2008},
	note = {Export Date: 21 November 2012Source: ScopusCODEN: KEMAELanguage of Original Document: EnglishCorrespondence Address: Shirosaki, Y.; Faculdade de Engenharia da Universidade do Porto (FEUP), Rua Dr.Roberto Frias, 4200-465, Porto, Portugal; email: yuki@ineb.up.ptReferences: Tateishi, T., GChen, Ushida, T., (2002) J Artif Organs, 5, p. 77;(2004) Biomaterials science: An introduction to materials in medicine, , edited by B.D.Ratner, A.S. Hoffman, F.J.Schoen, and J.E.Lemons, Elsevier, Academic Press; 
Sinha, V.R., Singla, A.K., Wadhawan, S., Kaushik, R., Kumria, R., Bansal, K., Dhawan, S., (2004) Inter J Pharmace, 274, p. 1; 
Shirosaki, Y., Tsuru, K., Hayakawa, S., Osaka, A., Lopes, M.A., Santos, J.D., Fernandes, M.H., (2005) Biomaterials, 26, p. 485; 
Tsuru, K., Shirosaki, Y., Hayakawa, S., Osaka, A., Lopes, M.A., Santos, J.D., Fernandes, M.H., (2005) Key Engineering Mateirals, 284-286, p. 823; 
Ning, S., Yu, N., Brown, D.M., Kanekal, S., Knox, S.J., (1999) Radiother Oncol, 50, p. 215; 
Davis, R.S., Brough, A.R., Atkinson, A., (2003) J Non-Cryst Solid, 315, p. 197; 
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Akbuga, J., Durmaz, G., (1994) Int J Pharm, 111, p. 217
},
	month = {2008///},
	pages = {1219 - 1222},
	abstract = {Porous chitosan-silicate hybrids were prepared by freeze-drying the precursor sol solutions synthesized from chitosan and 3- glycidoxypropyltrimethoxysilane (GPTMS). Degradability of and the release of cytochrome C in to phosphate buffer saline solution (PBS) were examined as a function of the GPTMS content. The hybrids were less degradable with larger GPTMS contents, and the cytochrome C release profile was so controllable as to give either burst release or slow one due to the GPTMS content. Thus, the present porous chitosan-silicate hybrids were considered applicable to drug delivery systems.},
	keywords = {biodegradability, Buffer saline solution, chitosan, Chitosan silicate hybrid, Chitosan-silicate hybrid, Drug delivery, Porous materials, Proteins, Silicates, Synthesis (chemical)},
	isbn = {10139826 (ISSN)},
	url = {http://www.scopus.com/inward/record.url?eid=2-s2.0-38149045782\&partnerID=40\&md5=520d7fffb6d19fcc96b3fa38036f9bf6},
	author = {Shirosaki, Y. and Tsuru, K. and Hayakawa, S. and Osaka, A.}
}
@book {2362,
	title = {Hybrid chitosan membranes tested in sheep for guided tissue regeneration},
	series = {Key Engineering MaterialsKey Eng Mat},
	volume = {361-363 II},
	year = {2008},
	note = {Cited By (since 1996): 1Export Date: 21 November 2012Source: ScopusCODEN: KEMAELanguage of Original Document: EnglishCorrespondence Address: Cortez, P.P.; Centro de Estudos de Ci{\^e}ncia Animal (CECA), Instituto de Ci{\^e}ncias e Tecnologias Agr{\'a}rias e Agro-Alimentares (ICETA), Campus Agr{\'a}rio de Vair{\~a}o, Rua P. Armando Quintas, 4485-661, Vair{\~a}o, Portugal; email: pcortez@mail.icav.up.ptReferences: Nasser, N.J., Friedman, A., Friedman, M., Moor, E., Mosheiff, R., (2005) Injury, 36, p. 1460;Chandy, T., Sharma, C.P., (1990) Biomater. Artif. Cells Artif. Organs, 18, p. 1; 
Domard, A., Rinaudo, M., (1983) Int J Biol Macromol, 5, p. 49; 
Shirosaki, Y., Tsuru, K., Hayakawa, S., Osaka, A., Lopes, M.A., Santos, J.D., Fernandes, M.H., (2004) Key Eng. Mat, 254-256, p. 857; 
Shirosaki, Y., Tsuru, K., Hayakawa, S., Osaka, A., Lopes, M.A., Santos, J.D., Fernandes, M.H., (2005) Biomaterials, 26, p. 485; 
Ueno, H., Yamada, H., Tanaka, I., Kaba, N., Matsuura, M., Okumura, M., Kadosawa, T., Fujinaga, T., (1999) Biomaterials, 20, p. 1407
},
	month = {2008///},
	pages = {1265 - 1268},
	abstract = {Previous in vitro studies confirmed an improved cytocompatibility of chitosan-silicate hybrid membranes over chitosan membranes. The main goal of this study was to assess the in vivo histocompatibility of both membranes through subcutaneous implantations at different time periods, 1 week, 1, 2 and 3 months, using a sheep model. Chitosan membranes elicited an exuberant inflammatory response and were consequently rejected. The hybrid chitosan membranes were not rejected and the degree of inflammatory response decreased gradually until the third month of implantation. Histological evaluation also showed that these membranes can be resorbed in vivo. This study demonstrates that the incorporation of silicate into the chitosan solution improves its histocompatibility, indicating that the hybrid chitosan-silicate membranes are suitable candidates to be used in clinical applications.},
	keywords = {biodegradability, Biodegradable membrane, Biodegradable membranes, chitosan, Chitosan-silicate hybrid, Guided tissue regeneration (GTR), histology, in vivo study, Membranes, Silicates, Tissue culture},
	isbn = {10139826 (ISSN)},
	url = {http://www.scopus.com/inward/record.url?eid=2-s2.0-38149018476\&partnerID=40\&md5=9a6734a40aa84667fe014796596e1ef3},
	author = {Cortez, P.P. and Shirosaki, Y. and Botelho, C.M. and Sim{\~o}es, M.J. and Gartner, F. and Da Costa, R.M.G. and Tsuru, K. and Hayakawa, S. and Osaka, A. and Lopes, M.A. and Santos, J.D. and Maur{\'\i}cio, A.C.}
}
@article {2356,
	title = {Synthesis and cytocompatibility of porous chitosan-silicate hybrids for tissue engineering scaffold application},
	journal = {Chemical Engineering JournalChem. Eng. J.},
	volume = {137},
	year = {2008},
	note = {Cited By (since 1996): 13Export Date: 21 November 2012Source: ScopusCODEN: CMEJAdoi: 10.1016/j.cej.2007.10.012Language of Original Document: EnglishCorrespondence Address: Osaka, A.; Graduate School of Natural Science and Technology, Okayama University, Okayama, 700-8530, Japan; email: osaka@cc.okayama-u.ac.jpReferences: Tateishi, T., Chen, G., Ushida, T., Biodegradable porous scaffolds for tissue engineering (2002) J. Artif. Organs, 5, pp. 77-83;Lee, Y.-K., Mooney, J.D., Hydrogels for tissue engineering (2001) Chem. Rev., 101, pp. 1869-1879; 
Park, S.-N., Park, J.-C., Kim, H.-O., Song, M.-J., Suh, H., Characterization of porous collagne/hyaluronic acid scaffold modified by 1-ethyl-3-(3-dimethylaminopropyl)carbodiimide cross-linking (2002) Biomaterials, 23, pp. 1205-1212; 
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Hua, H., Jie, W., Changshengh, L., Development of asymmetric gradational-changed porous chitosan membrane for guided periodontal tissue regeneration (2007) Compos. B, 38, pp. 311-316; 
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Kang, H.-W., Tabata, Y., Ikada, Y., Fabrication of porous gelatin scaffolds for tissue engineering (1999) Biomaterials, 20, pp. 1339-1344; 
Ho, M.-H., Kuo, P.-Y., Hsieh, H.-J., Hsien, T.-Y., Hou, L.-T., Lai, J.-Y., Wang, D.-M., Preparation of porous scaffolds by using freeze-extraction and freeze-gelation methods (2004) Biomaterials, 25, pp. 129-138
},
	month = {2008///},
	pages = {122 - 128},
	abstract = {Chitosan-silicate hybrids with 3D porous structures were prepared with freeze-drying precursor solutions derived from chitosan and γ-glycidoxypropyltrimethoxysilane (GPTMS). They were formed easily in any shape, such as sheets, pellets, disks, granules, and even roll-cakes. The pore size was strongly dependent on the freezing temperature: lower freezing temperature resulted smaller pores, about 110 μm for the hybrids frozen at -20 {\textdegree}C, and about 50 μm for those at -85 {\textdegree}C. The pore size was little dependent on the GPTMS content. In contrast, the GPTMS content affected porosity a littlie: \~{}80\% for chitosan, and \~{}90\% for the GPTMS-containing hybrids. Thus, their porous microstructure was controllable due to the freezing temperature and composition. MG63 osteoblastic cells were cultured up to 7 days on the porous hybrids. The cells adhered on the pore walls, proliferated, and migrated deep into the pore structure. It was thus concluded that the present chitosan-silicate hybrids were promising for tissue engineering scaffold applications. {\textcopyright} 2007 Elsevier B.V. All rights reserved.},
	keywords = {chitosan, Cytocompatibility, GPTMS, Organic-inorganic hybrid, Osteoblastic cell MG63, Osteoblasts, Pore size, Pore structure, Scaffold, Silicates, Tissue engineering, γ-Glycidoxypropyltrimethoxysilane (GPTMS)},
	isbn = {13858947 (ISSN)},
	url = {http://www.scopus.com/inward/record.url?eid=2-s2.0-38949146037\&partnerID=40\&md5=e26163ca30dda2875f5c9dd2316bdd4e},
	author = {Shirosaki, Y. and Okayama, T. and Tsuru, K. and Hayakawa, S. and Osaka, A.}
}
@article {2381,
	title = {Attachment, spreading and short-term proliferation of human osteoblastic cells cultured on chitosan films with different degrees of acetylation},
	journal = {Journal of Biomaterials Science, Polymer EditionJ. Biomater. Sci. Polym. Ed.},
	volume = {18},
	year = {2007},
	note = {Cited By (since 1996): 22Export Date: 21 November 2012Source: ScopusCODEN: JBSEEdoi: 10.1163/156856207780425068Language of Original Document: EnglishCorrespondence Address: Amaral, I.F.; INEB - Instituto de Engenharia Biom{\'e}dica, Laborat{\'o}rio de Biomateriais, Rua do Campo Alegre, 823, 4150-180 Porto, Portugal; email: iamaral@ineb.up.ptChemicals/CAS: chitosan, 9012-76-4; Biocompatible Materials; Chitosan, 9012-76-4References: Vacanti, C.A., Bonassar, L.J., Vacanti, J.P., (2000) Principles of Tissue Engineering, p. 671. , R. P. Lanza, R. Langer and J. Vacanti Eds, p, Academic Press, San Diego, CA;Bruder, S.P., Caplan, A.I., (2000) Principles of Tissue Engineering, p. 683. , R. P. Lanza, R. Langer and J. Vacanti Eds, p, Academic Press, San Diego, CA; 
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},
	month = {2007///},
	pages = {469 - 485},
	abstract = {Chitosan (Ch) is being actively investigated as a non-protein template for the growth of an increasing number of anchorage-dependent cells, including chondrocytes and bone cells. In the present work, Ch films with degrees of N-acetylation (DAs) in the range of 4 to 49\% were evaluated with respect to the attachment, spreading and short-term proliferation of osteoblasts, using human osteoblastic MG-63 cells. The films were characterized in terms of surface morphology and surface charge by atomic force microscopy and streaming potential measurements, respectively. Cell attachment was assessed after 3 and 24 h of cell culture. After 24 h of incubation, cell attachment was found to be dependent on the DA, lower DAs favouring cell adhesion. With time, cell spreading and cytoskeleton organization were only attained for DAs <= 13\%. Regarding cell proliferation, cells grown on films with the lowest DA (4\%) revealed a higher specific growth rate as compared to those grown on films with higher DAs. Films with a DA of 49\% failed to sustain cell proliferation. In addition, a longer lag-phase was observed on Ch, as compared to TCPS, in accordance to an observed delay of cell spreading. The present findings revealed that differences in the DA as small as 9\% may be critical in terms of the osteoblast response to two-dimensional Ch-based matrices. {\textcopyright} 2007 VSP.},
	keywords = {Acetylation, AFM, article, Atomic force microscopy, Biocompatible Materials, biofilm, Biofilms, biomaterial, bone cell, cartilage cell, cell adhesion, cell anchorage, Cell culture, cell growth, Cell Line, Tumor, cell proliferation, Cell Shape, cell spreading, cell survival, Chemical structure, chemistry, Chitin, chitosan, comparative study, controlled study, cytoskeleton, Degree of acetylation, drug effect, Growth kinetics, human, human cell, Humans, incubation time, metabolism, Microscopy, Atomic Force, osteoblast, Osteoblast-like cells, Osteoblasts, Pathology, priority journal, Surface charge, Surface Properties, surface property, Surface treatment, tumor cell line, Zeta potential},
	isbn = {09205063 (ISSN)},
	url = {http://www.scopus.com/inward/record.url?eid=2-s2.0-34347375992\&partnerID=40\&md5=437b9ed270e859425b76aa3655c611be},
	author = {Amaral, I.F. and Cordeiro, A.L. and Sampaio, P. and Barbosa, M.A.}
}
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	title = {Functionalization of chitosan membranes through phosphorylation: Atomic force microscopy, wettability, and cytotoxicity studies},
	journal = {Journal of Applied Polymer ScienceJ. Appl. Polym. Sci.},
	volume = {102},
	year = {2006},
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},
	month = {2006///},
	pages = {276 - 284},
	abstract = {Grafting negatively charged groups such as phosphates is a well-known strategy for inducing the deposition of apatite-like layers under simulated physiological conditions. In this investigation, chitosan was phosphorylated in an attempt to enhance its osteoconduction. Chitosan membranes were phosphorylated at 30{\textdegree}C with the H                        3PO
                        4/ Et 
                        3PO
                        4/P
                        2O
                        5/butanol reaction system for periods up to 48 h. This method is an alternative to the phosphoric acid/ urea/dimethylformamide phosphorylation method, which involves the use of much higher temperatures. In this study, the effects of the phosphorylation reaction time on the surface morphology and surface free energy of phosphorylated membranes were investigated with atomic force microscopy and static-contact-angle measurements, respectively. In addition, the modified membranes were evaluated with respect to their cytotoxiciry toward bone cells through the incubation of human osteoblastic cells with extracts of the materials for two different periods: 24 and 120 h. The results revealed a reduction of the average surface roughness at the nanometer scale with increasing phosphorylation reaction time. Wettability studies showed an increase in the polar component of the surface free energy with increasing reaction time as a result of the increase in the phosphate surface concentration. Cytotoxicity studies revealed no cytotoxic effect of phosphorylated membranes on osteoblastic cells, regardless of the incubation period. {\textcopyright} 2006 Wiley Periodicals, Inc.
},
	keywords = {Atomic force microscopy, Atomic force microscopy (AFM), Biocompatibility, Biological applications, Biological applications of polymers, Biomaterials, Chitin, chitosan, Chitosan membranes, grafting, Grafting (chemical), Osteoblastic cells, Phosphorylation, Polymeric membranes, polysaccharide, Polysaccharides, surface property, toxicity, Urea},
	isbn = {00218995 (ISSN)},
	url = {http://www.scopus.com/inward/record.url?eid=2-s2.0-33749478403\&partnerID=40\&md5=c365a50e3e767b1da90ad4ccd331440b},
	author = {Amaral, I.F. and Granja, P.L. and Melo, L.V. and Saramago, B. and Barbosa, M.A.}
}
@article {2421,
	title = {Three-dimensional culture of human osteoblastic cells in chitosan sponges: The effect of the degree of acetylation},
	journal = {Journal of Biomedical Materials Research - Part AJ. Biomed. Mater. Res. Part A},
	volume = {76},
	year = {2006},
	note = {Cited By (since 1996): 19Export Date: 21 November 2012Source: ScopusCODEN: JBMRCdoi: 10.1002/jbm.a.30522PubMed ID: 16270345Language of Original Document: EnglishCorrespondence Address: Amaral, I.F.; INEB-Instituto de Engenharia Biom{\'e}dica, Laborat{\'o}rio de Biomateriais, Rua do Campo Alegre, 823, 4150-180 Porto, Portugal; email: iamaral@ineb.up.ptChemicals/CAS: alkaline phosphatase, 9001-78-9; chitosan, 9012-76-4; collagen, 9007-34-5; Alkaline Phosphatase, EC 3.1.3.1; Chitosan, 9012-76-4; Collagen, 9007-34-5References: Bruder, S.P., Caplan, A.I., Bone regeneration through cellular engineering (2000) Principles of Tissue Engineering, pp. 683-696. , Lanza RP, Langer R, Vacanti J, editors. San Diego: Academic Press;Anselme, K., Osteoblast adhesion on biomaterials (2000) Biomaterials, 21, pp. 667-681; 
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},
	month = {2006///},
	pages = {335 - 346},
	abstract = {In this investigation, the effect of the degree of acetylation (DA) of chitosan on the behavior of human osteoblastic MG-63 cells cultured in three-dimensional chitosan matrices was assessed. Chitosan sponges with DAs in the range of 4 to 49\% were prepared and characterized in terms of microstructure, porosity, and pore size. Collagen sponges were used as 3D control. Cell proliferation was determined using the MTT assay while the retention of the osteoblastic phenotype was monitored by assaying alkaline phosphatase activity. Cell morphology, cytoskeletal organization, and viability were assessed using different microscopy techniques. Chitosan sponges showed a similar microstructure regardless the DA, except for the highest DA used, where a more heterogeneous pore distribution was observed. In terms of cell proliferation, alkaline phosphatase activity and cell viability, cells cultured in chitosan scaffolds performed as well as in the 3D control regardless the DA, except for the highest DA used, where an inhibitory effect on cell proliferation was found. However, while in sponges with DAs <= 13\% cells attached and spread displaying long cell filopodia and numerous cell-to-cell contacts, in sponges with higher DAs cells tended to remain spherical and grow into spheroid-like cellular aggregates. In the present study, the DA played a key role in determining the affinity of osteoblastic cells towards the substrates, possibly by influencing the nature of the initial adsorbed protein layer. {\textcopyright} 2005 Wiley Periodicals, Inc.},
	keywords = {Acetylation, Aggregates, alkaline phosphatase, article, Bone Regeneration, bone tissue, Bone tissue engineering, Cell culture, Cell Culture Techniques, Cell filopodia, cell growth, cell proliferation, Cell Shape, cell structure, cell survival, cell viability, chitosan, Collagen, confocal microscopy, controlled study, Cytoskeletal organization, cytoskeleton, Degree of acetylation (DA), enzyme activity, gel permeation chromatography, human, human cell, Humans, infrared spectroscopy, Microstructure, morphology, osteoblast, Osteoblastic cells, Osteoblasts, Phosphatase activity, porosity, Proteins, Scaffold, Scaffolds, Scanning electron microscopy, Substrates, Tissue engineering},
	isbn = {00219304 (ISSN)},
	url = {http://www.scopus.com/inward/record.url?eid=2-s2.0-33645101773\&partnerID=40\&md5=5adc6336963ec1c8a83d8ec89943159d},
	author = {Amaral, I.F. and Sampaio, P. and Barbosa, M.A.}
}
@article {2438,
	title = {Chemical modification of chitosan by phosphorylation: An XPS, FT-IR and SEM study},
	journal = {Journal of Biomaterials Science, Polymer EditionJ. Biomater. Sci. Polym. Ed.},
	volume = {16},
	year = {2005},
	note = {Cited By (since 1996): 50Export Date: 21 November 2012Source: ScopusCODEN: JBSEEdoi: 10.1163/156856205774576736Language of Original Document: EnglishCorrespondence Address: Amaral, I.F.; Laborat{\'o}rio de Biomateriais, INEB - Instituto de Engenharia Biom{\'e}dica, Rua do Campo Alegre, 823, 4150-180 Porto, Portugal; email: iamaral@ineb.up.ptChemicals/CAS: chitosan, 9012-76-4; phosphate, 14066-19-4, 14265-44-2; apatite, 64476-38-6; butanol, 35296-72-1, 71-36-3; calcium ion, 14127-61-8; Chitosan, 9012-76-4; PhosphatesReferences: Khor, E., (2001) Chitin: Fulfilling a Biomaterials Promise, , Elsevier Science, Oxford;Vande Vord, P.J., Matthew, H.W.T., DeSilva, S.P., Mayton, L., Wu, B., Wooley, P.H., (2002) J. Biomed. Mater. Res., 59, p. 585; 
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},
	month = {2005///},
	pages = {1575 - 1593},
	abstract = {In the present work, the surface of chitosan membranes was modified using a phosphorylation method carried out at room temperature. Phosphorylation may be of particular interest in materials for orthopaedic applications, due to the cation-exchange properties of phosphate functionalities. Phosphate groups chelate calcium ions, thus inducing the deposition of an apatite-like layer known to improve the osteoconduction of polymer-based implants. Additionally, the negatively charged phosphate functionalities, together with the positively charged amine groups from chitosan, are expected to provide chitosan with an amphoteric character, which may be useful as a combinatorial therapeutic strategy, by simultaneously allowing the immobilization of signalling molecules like growth factors. Phosphorylation was carried out at room temperature using the H                        3 PO
                        4/Et
                        3 PO
                        4/P
                        2O
                        5/butanol method. Surface characterization was performed by XPS, ATR-FT-IR, and SEM. Cross-sections were analyzed by SEM fitted with EDS. The phosphate content increased with the reaction time, as shown by XPS and ATR-FT-IR, a P/N atomic ratio of 0.73 being obtained after 48 h of treatment. High-resolution XPS spectra regarding C
                        1s, O
                        1s, N
                        1s and P
                        2p are discussed. The introduction of a neutralization step led to a reduction of P content, which pointed out to the presence of phosphates ionically bound to protonated amines, in addition to phosphate esters. EDS analysis of cross-sections revealed a gradual P reduction up to 50\% towards the inner part of the membrane. {\textcopyright} VSP 2005.
},
	keywords = {Acetylation, amine, animal, Animals, Apatite, article, bone conduction, butanol, calcium ion, cation exchange, cephalopod, chelate, Chemical modification, chemistry, Chitin, chitosan, comparative study, controlled study, Decapodiformes, Fourier transform infrared spectroscopy, Fourier transform infrared spectroscopy (FT-IR), Growth factor, immobilization, Implants (surgical), infrared spectroscopy, isolation and purification, Microscopy, Electron, Scanning, molecular weight, organophosphate, phosphate, Phosphate groups, Phosphates, Phosphorylation, Phosphorylation method, Plastics applications, priority journal, reflectometry, room temperature, Scanning electron microscopy, SEM-EDS, signal transduction, spectrometry, Spectrometry, X-Ray Emission, Spectroscopy, Fourier Transform Infrared, Surface chemical reaction, Surface Properties, surface property, temperature, time, Time Factors, viscosity, X ray photoelectron spectroscopy, X-rayphotoelectron spectroscopy (XPS)},
	isbn = {09205063 (ISSN)},
	url = {http://www.scopus.com/inward/record.url?eid=2-s2.0-33644875542\&partnerID=40\&md5=cb42a0fba6d2000d93b90af9724a5bc1},
	author = {Amaral, I.F. and Granja, P.L. and Barbosa, M.A.}
}
@book {2437,
	title = {In vitro biodegradability of chitosan-organosiloxane hybrid membrane},
	series = {Key Engineering MaterialsKey Eng Mat},
	volume = {284-286},
	year = {2005},
	note = {Cited By (since 1996): 2Export Date: 21 November 2012Source: ScopusCODEN: KEMAELanguage of Original Document: EnglishCorrespondence Address: Tsuru, K.; Faculty of Engineering, Biomaterial Laboratory, Okayama University, Tsushima, Okayama 700-8530, Japan; email: kanji@cc.okayama-u.ac.jpReferences: Chandy, T., Sharma, P.C., (1990) Biomat Art Cells Art Org, 18, pp. 1-24;Ren, L., Tsuru, K., Hayakawa, S., Osaka, A., (2001) J Ceram Soc Japan, 109, pp. 403-408; 
Ren, L., Tsuru, K., Hayakawa, S., Osaka, A., (2002) Biomaterials, 23, pp. 4765-4773; 
Shirosaki, Y., Tsuru, K., Hayakawa, S., Osaka, A., Lopes, M.A., Santos, J.D., Fernandes, M.H., (2004) Key Engineering Materials, 254-256, pp. 857-860; 
Shirosaki, Y., Tsuru, K., Hayakawa, S., Osaka, A., Lopes, M.A., Santos, J.D., Fernandes, M.H., (2005) Biomaterials, 26, pp. 485-493; 
S. Prochazkova, K. M. V{\r a}rum, K. {\O}stgaard, Carbohydrate Polymers, 38 (1999), p. 115-122Brinker, J.C., Scherer, G.W., (1989) Sol-Gel Science, The Physics and Chemistry of Sol-Gel Processing, , Academic Press, San Diego
},
	month = {2005///},
	pages = {823 - 826},
	abstract = {Chitosan-silicate hybrid membranes were prepared using γ- glycidoxypropyltrimethoxysilane (GPSM) through a sol-gel process. The amino groups of chitosan chains were reacted with the epoxy groups of GPSM and GPSM have a function as the agent to cross-link the chitosan chains. The cross-linking degree of the hybrid membranes was determined by ninhydrin assay. Fourier-transform infrared (FT-IR) spectroscopy and                        29Si cross-polarization (CP) magic-angle spinning (MAS)-NMR spectroscopy were used to assess the structure of the hybrid membranes. The biodegradability of the hybrid membranes in phosphate-buffered saline solution or lysozyme solution was investigated as a function of the GPSM concentration.
},
	keywords = {biodegradability, Biological membranes, chitosan, Chitosan chains, Chitosan inorganic hybrid membranes, Chitosan-lnorganlc hybrid, Crosslinking, Fourier transform infrared spectroscopy, Nuclear magnetic resonance spectroscopy, R- glycidoxypropyltrimethoxysilane GPSM, Reaction kinetics, Silicates, Sol-gel process},
	isbn = {10139826 (ISSN)},
	url = {http://www.scopus.com/inward/record.url?eid=2-s2.0-34249091650\&partnerID=40\&md5=51e1b69bd7dc1b4c23b022e4ec338e8a},
	author = {Tsuru, K. and Shirosaki, Y. and Hayakawa, S. and Osaka, A. and Lopes, M.A. and Santos, J.D. and Fernandas, M.H.}
}
@article {2459,
	title = {In vitro cytocompatibility of MG63 cells on chitosan-organosiloxane hybrid membranes},
	journal = {BiomaterialsBiomaterials},
	volume = {26},
	year = {2005},
	note = {Cited By (since 1996): 55Export Date: 21 November 2012Source: ScopusCODEN: BIMADdoi: 10.1016/j.biomaterials.2004.02.056PubMed ID: 15276356Language of Original Document: EnglishCorrespondence Address: Hayakawa, S.; Biomaterial Laboratory, Faculty of Engineering, Okayama Univ., 3-1-1 T., Okayama-shi, Japan; email: satoshi@cc.okayama-u.ac.jpChemicals/CAS: calcium chloride, 10043-52-4; Alkaline Phosphatase, EC 3.1.3.1; Biocompatible Materials; Calcium, 7440-70-2; Chitosan, 9012-76-4; Cross-Linking Reagents; gamma-glycidoxypropyltrimethoxysilane, 2530-83-8; Membranes, Artificial; Neoplasm Proteins; SilanesManufacturers: Chisso, Japan; Kasei, JapanReferences: Park, J.B., Lakes, R.S., (1992) Biomaterials An Introduction, pp. 223-234. , New York: Plenum Press;Wise, D.L., (2000) Biomaterials and Bioengineering Handbook, pp. 556-559. , New York: Marcel Dekker; 
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},
	month = {2005///},
	pages = {485 - 493},
	abstract = {Chitosan-silicate hybrids were synthesized using γ- glycidoxypropyltrimethoxysilane (GPSM) as the agent for cross-linking the chitosan chains. CaCl                        2 was introduced in the hybrids in expectation that it would improve cell adhesion and differentiation of the hybrid surfaces. Fourier-transform infrared (FT-IR) spectroscopy and 
                        29Si CP-MAS NMR spectroscopy were used to analyze the structures of the hybrids. Cytocompatibility of the hybrids was investigated in terms of proliferation of an osteoblastic cell line, MG63. The adhesion and proliferation of the osteoblastic cells cultured on the surface of a chitosan-GPSM hybrid without calcium were similar to those on a control culture plate, and were better than those on a chitosan membrane. The ALP activity of the cells cultured on this hybrid was higher than that on the chitosan membrane. Contrary to expectations, the incorporation of calcium ions into the hybrids did not improve cell attachment and proliferation on their surfaces. {\textcopyright} 2004 Elsevier Ltd. All rights reserved.
},
	keywords = {alkaline phosphatase, article, artificial membrane, Biocompatibility, Biocompatible Materials, Bone Neoplasms, Calcium, calcium chloride, cell adhesion, Cell attachments, Cell culture, cell differentiation, Cell Division, Cell Line, Tumor, chitosan, chitosan derivative, controlled study, cross linking, Cross-Linking Reagents, Crosslinking, Cytocompatibility, Fourier transform infrared spectroscopy, human, human cell, Humans, hybrid, infrared spectroscopy, Magnetic Resonance Spectroscopy, materials testing, Membranes, Artificial, MG63 osteoblastic cells, Microscopy, Electron, Scanning, Neoplasm Proteins, Nuclear magnetic resonance spectroscopy, Organic-inorganic hybrid, osteoblast, Osteoblasts, Osteosarcoma, priority journal, Silanes, Silicates, siloxane, Spectroscopy, Fourier Transform Infrared, structure analysis, surface property, Synthesis (chemical), wettability, γ-Glycidoxypropyltrimethoxysilane (GPSM)},
	isbn = {01429612 (ISSN)},
	url = {http://www.scopus.com/inward/record.url?eid=2-s2.0-3242791810\&partnerID=40\&md5=a22c21444fa3d2843a8783bb42f6ebc1},
	author = {Shirosaki, Y. and Tsuru, K. and Hayakawa, S. and Osaka, A. and Lopes, M.A. and Santos, J.D. and Fernandes, M.H.}
}
@article {2454,
	title = {Polysaccharides as scaffolds for bone regeneration},
	journal = {ITBM-RBMITBM-RBM},
	volume = {26},
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},
	month = {2005///},
	pages = {212 - 217},
	abstract = {This review is concerned with the use of polysaccharides for bone regeneration. Cellulose, alginate and chitosan have been chosen as examples of the formidable potential of this class of materials. Many others could be cited but our experience and research interests dictated the choice. As in many areas of research we know how they started but we cannot see how and if they will end. Modification of cellulose was our entrance to the world of polysaccharides and a lot of what we know we owe to Charles Baquey, to whom we would like to express our immense gratitude. This chapter is dedicated to him, for his cooperation, unlimited enthusiasm and friendship. {\textcopyright} 2005 Elsevier SAS. All rights reserved.},
	keywords = {Alginate, alginic acid, Amino Acid Sequence, arginylglycylaspartic acid, biomaterial, biotechnology, bone cell, bone prosthesis, Bone Regeneration, Cellulose, chitosan, enzyme degradation, polysaccharide, Polysaccharides, review, Tissue engineering},
	isbn = {12979562 (ISSN)},
	url = {http://www.scopus.com/inward/record.url?eid=2-s2.0-20744434609\&partnerID=40\&md5=43d83907f18b3e44035dfe14ff13a24f},
	author = {Barbosa, M.A. and Granja, P.L. and Barrias, C.C. and Amaral, I.F.}
}
@article {2444,
	title = {Rat bone marrow stromal cell osteogenic differentiation and fibronectin adsorption on chitosan membranes: The effect of the degree of acetylation},
	journal = {Journal of Biomedical Materials Research - Part AJ. Biomed. Mater. Res. Part A},
	volume = {75},
	year = {2005},
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},
	month = {2005///},
	pages = {387 - 397},
	abstract = {Cell adhesion, migration, and proliferation of a few anchorage-dependent cells cultured on chitosan (Ch) matrices are influenced by the degree of N-acetylation (DA) of Ch. In the present work, we examined the influence of the DA on the attachment, spreading, proliferation, and osteogenic differentiation of rat bone marrow stromal cells (rBM-SCs). Ch membranes were characterized in terms of surface morphology, roughness, and wettability, and in terms of adsorption of an adhesive protein, fibronectin (Fn). Chs with DAs in the range of 4 to 49\% were used. Among the Ch samples, the DA of 4\% led to the highest Fn surface concentration, both from single protein solution and from diluted serum. Furthermore, the levels of Fn adsorbed from serum found for this DA were threefold higher than for the tissue culture polystyrene control, indicating that in the presence of competitive proteins Ch is more specific toward Fn adsorption than tissue culture polystyrene. rBMSCs cultured on Ch carrying a DA of 4\% were able to spread, proliferate, and differentiate, reaching a higher level of osteogenic differentiation than on the control, despite the lower cell attachment observed for all Ch samples. Because the Ch sample with a DA of 4\% showed the highest Fn adsorption from serum, we suggest that cell adhesion, spreading, and osteogenic differentiation of rBMSCs on Ch may be mediated by the adsorbed layer of Fn. {\textcopyright} 2005 Wiley Periodicals, Inc.},
	keywords = {Acetylation, Adhesion, Adsorption, animal cell, Animals, article, artificial membrane, binding affinity, binding competition, Bone, bone development, bone marrow cell, Bone Marrow Cells, Bone marrow stomal cells, Bone marrow stromal cells, cell adhesion, cell anchorage, Cell culture, cell differentiation, Cell membranes, cell migration, cell proliferation, Cell Shape, cell spreading, chitosan, Coated Materials, Biocompatible, controlled study, cytoskeleton, Degree of acetylation, Fibronectin, Fibronectins, male, materials testing, morphology, nonhuman, polystyrene, Proteins, rat, Rats, Rats, Wistar, roughness, stroma cell, Stromal Cells, Surface Properties, surface property, Tissue culture, wettability},
	isbn = {00219304 (ISSN)},
	url = {http://www.scopus.com/inward/record.url?eid=2-s2.0-26844551535\&partnerID=40\&md5=fa91e3e29d946053ae7bb231b6a2640e},
	author = {Amaral, I.F. and Lamghari, M. and Sousa, S.R. and Sampaio, P. and Barbosa, M.A.}
}
@book {2489,
	title = {In Vitro Cytocompatibility of Osteoblastic Cells Cultured on Chitosan-Organosiloxane Hybrid Membrane},
	series = {Key Engineering MaterialsKey Eng Mat},
	volume = {254-256},
	year = {2004},
	note = {Conference code: 62053Cited By (since 1996): 2Export Date: 21 November 2012Source: ScopusCODEN: KEMAELanguage of Original Document: EnglishCorrespondence Address: Shirosaki, Y.; Biomaterial Laboratory, Faculty of Engineering, Okayama University, Tsushima, Okayama-shi 700-8530, JapanReferences: Chandy, T., Sharma, P.C., (1990) Biomat Art Cells Art Org, 18, pp. 1-24;Jameela, S.R., Jayakrishnan, A., (1995) Biomaterials, 16, pp. 769-775; 
Nishi, C., Nakajima, N., Ikada, Y., (1995) J Biomed Mater Res., 29, pp. 829-834; 
Ren, L., Tsuru, K., Hayajkawa, S., Osaka, A., (2002) Biomaterials, 23, pp. 4765-4773
},
	month = {2004///},
	pages = {857 - 860},
	address = {Porto},
	abstract = {Chitosan-silicate hybrids were synthesized using γ-glycidoxypropyltrimethoxysilane (GPSM) as the agent to cross-link the chitosan chains. Fourier-transform infrared (PT-IR) spectroscopy was used to analyze the structures of the hybrids. The swelling ability and the cytocompatibility of the hybrids were investigated as a function of the GPSM concentration. The swelling of the hybrids was suppressed by addition of GPSM. MG63 human osteoblastic cells were cultured on the hybrids. The adhesion and proliferation of the MG63 cells cultured on the chitosan-GPSM hybrid surface were improved comparing to that on chitosan membrane, regardless the GPSM concentration.},
	keywords = {Biocompatibility, Biological membranes, biosynthesis, Cell culture, Cells, chitosan, Chitosan-inorganic hybrid, Crosslinking, Cytocompatibility, Fourier transform infrared spectroscopy, Nuclear magnetic resonance spectroscopy, Organic polymers, Osteoblastic cell (MG63), Structural analysis, Synthetic polymers, Tissue, Tissue engineering, γ-Glycidoxypropyltrimethoxysilane (GPSM)},
	isbn = {10139826 (ISSN)},
	url = {http://www.scopus.com/inward/record.url?eid=2-s2.0-0346885502\&partnerID=40\&md5=80d45c86bb14c57c3b1a6b94b435e45d},
	author = {Shirosaki, Y. and Tsuru, K. and Hayakawa, S. and Osaka, A. and Lopes, M.A. and Santos, J.D. and Fernandes, M.H.},
	editor = {Barbosa M.A. and Monteiro F.J. and Correia R. and Leon B.}
}
