



@article {3298,
	title = {Effect of cell density on mesenchymal stem cells aggregation in RGD-alginate 3D matrices under osteoinductive conditions},
	journal = {Macromolecular Bioscience},
	volume = {14},
	year = {2014},
	note = {Cited By (since 1996):1Export Date: 21 November 2014},
	month = {2014},
	pages = {759 - 771},
	abstract = {Cellular activities in 3D are differentially affected by several matrix-intrinsic and extrinsic factors. This study highlights the relevance of optimizing initial cell densities when establishing 3D cultures for specific applications. Independently of the entrapping density, MSCs cultured within RGD-alginate hydrogels showed steady-state levels of metabolic activity and were in a nearly non-proliferative state, but recovered "normal" activity levels when retrieved from 3D matrices and re-cultured as monolayers. Importantly, high-densities promoted the establishment of cell-cell contacts with formation of multicellular clusters stabilized by endogenous ECM, and also stimulated MSCs osteogenic differentiation. These MSC-ECM microtissues may be used as building blocks for tissue engineering. {\textcopyright} 2014 WILEY-VCH Verlag GmbH \& Co. KGaA, Weinheim.},
	keywords = {alginate hydrogels, encapsulation, Self-assembly, Tissue engineering},
	url = {http://www.scopus.com/inward/record.url?eid=2-s2.0-84902536382\&partnerID=40\&md5=d6659c40247c561753aca566f938d788},
	author = {Maia, F.R. and Louren{\c c}o, A.H. and Granja, P.L. and Gon{\c c}alves, R.M. and Barrias, C.C.}
}
@article {3296,
	title = {Hydrogel depots for local co-delivery of osteoinductive peptides and mesenchymal stem cells},
	journal = {Journal of Controlled Release},
	volume = {189},
	year = {2014},
	note = {Export Date: 21 November 2014},
	month = {2014},
	pages = {158 - 168},
	abstract = {The outcome of cell-based therapies can benefit from carefully designed cell carriers. A multifunctional injectable vehicle for the co-delivery of human mesenchymal stem cells (hMSCs) and osteoinductive peptides is proposed, to specifically direct hMSCs osteogenic differentiation. The osteogenic growth peptide (OGP) inspired the design of two peptides, where the bioactive portion of OGP was flanked by a protease-sensitive linker, or its scrambled sequence, to provide faster and slower release rates, respectively. Peptides were fully characterized and chemically grafted to alginate. Both OGP analogs released bioactive fragments in vitro, at different kinetics, which stimulated hMSCs proliferation and osteogenesis. hMSCs-laden OGP-alginate hydrogels were tested at an ectopic site in a xenograft mouse model. After 4 weeks, OGP-alginate hydrogels were more degraded and colonized by vascularized connective tissue than the control (without OGP). hMSCs were able to proliferate, migrate outward the hydrogels, produce endogenous extracellular matrix and mineralize it. Moreover, OGP-groups stimulated hMSCs osteogenesis, as compared with the control. Overall, the ability of the proposed platform to direct the fate of transplanted hMSCs in loco was demonstrated, and OGP-releasing hydrogels emerged as a potentially useful system to promote bone regeneration. {\textcopyright} 2014 Elsevier B.V.},
	keywords = {Bone Regeneration, Co-delivery systems, Guided differentiation, Injectable alginate hydrogels, Mesenchymal Stem Cells, Osteoinductive peptides},
	url = {http://www.scopus.com/inward/record.url?eid=2-s2.0-84904318888\&partnerID=40\&md5=5f671d3584c3b7fc785ffaf40c2f0b1d},
	author = {Maia, F.R. and Barbosa, M. and Gomes, D.B. and Vale, N. and Gomes, P. and Granja, P.L. and Barrias, C.C.}
}
@article {3266,
	title = {Injectable alginate hydrogels for cell delivery in tissue engineering},
	journal = {Acta Biomaterialia},
	volume = {10},
	year = {2014},
	note = {Export Date: 21 November 2014},
	month = {2014},
	pages = {1646 - 1662},
	abstract = {Alginate hydrogels are extremely versatile and adaptable biomaterials, with great potential for use in biomedical applications. Their extracellular matrix-like features have been key factors for their choice as vehicles for cell delivery strategies aimed at tissue regeneration. A variety of strategies to decorate them with biofunctional moieties and to modulate their biophysical properties have been developed recently, which further allow their tailoring to the desired application. Additionally, their potential use as injectable materials offers several advantages over preformed scaffold-based approaches, namely: Easy incorporation of therapeutic agents, such as cells, under mild conditions; minimally invasive local delivery; and high contourability, which is essential for filling in irregular defects. Alginate hydrogels have already been explored as cell delivery systems to enhance regeneration in different tissues and organs. Here, the in vitro and in vivo potential of injectable alginate hydrogels to deliver cells in a targeted fashion is reviewed. In each example, the selected crosslinking approach, the cell type, the target tissue and the main findings of the study are highlighted. {\textcopyright} 2013 Acta Materialia Inc.},
	keywords = {Alginate, Cell delivery, Injectable, Regeneration, Tissue engineering},
	url = {http://www.scopus.com/inward/record.url?eid=2-s2.0-84896403229\&partnerID=40\&md5=fd56573baeefa91edfc45ba61cf0573a},
	author = {Bidarra, S.J. and Barrias, C.C. and Granja, P.L.}
}
@article {3280,
	title = {Injectable MMP-sensitive alginate hydrogels as hMSC delivery systems},
	journal = {Biomacromolecules},
	volume = {15},
	year = {2014},
	note = {Cited By (since 1996):3Export Date: 21 November 2014},
	month = {2014},
	pages = {380 - 390},
	abstract = {Hydrogels with the potential to provide minimally invasive cell delivery represent a powerful tool for tissue-regeneration therapies. In this context, entrapped cells should be able to escape the matrix becoming more available to actively participate in the healing process. Here, we analyzed the performance of proteolytically degradable alginate hydrogels as vehicles for human mesenchymal stem cells (hMSC) transplantation. Alginate was modified with the matrix metalloproteinase (MMP)-sensitive peptide Pro-Val-Gly-Leu-Iso-Gly (PVGLIG), which did not promote dendritic cell maturation in vitro, neither free nor conjugated to alginate chains, indicating low immunogenicity. hMSC were entrapped within MMP-sensitive and MMP-insensitive alginate hydrogels, both containing cell-adhesion RGD peptides. Softer (2 wt \% alginate) and stiffer (4 wt \% alginate) matrices were tested. When embedded in a Matrigel layer, hMSC-laden MMP-sensitive alginate hydrogels promoted more extensive outward cell migration and invasion into the tissue mimic. In vivo, after 4 weeks of subcutaneous implantation in a xenograft mouse model, hMSC-laden MMP-sensitive alginate hydrogels showed higher degradation and host tissue invasion than their MMP-insensitive equivalents. In both cases, softer matrices degraded faster than stiffer ones. The transplanted hMSC were able to produce their own collagenous extracellular matrix, and were located not only inside the hydrogels, but also outside, integrated in the host tissue. In summary, injectable MMP-sensitive alginate hydrogels can act as localized depots of cells and confer protection to transplanted cells while facilitating tissue regeneration. {\textcopyright} 2013 American Chemical Society.},
	url = {http://www.scopus.com/inward/record.url?eid=2-s2.0-84892606344\&partnerID=40\&md5=835c11a6460fa62bc6da4a189fd06b25},
	author = {Fonseca, K.B. and Gomes, D.B. and Lee, K. and Santos, S.G. and Sousa, A. and Silva, E.A. and Mooney, D.J. and Granja, P.L. and Barrias, C.C.}
}
@article {3297,
	title = {Matrix-driven formation of mesenchymal stem cell-extracellular matrix microtissues on soft alginate hydrogels},
	journal = {Acta Biomaterialia},
	volume = {10},
	year = {2014},
	note = {Cited By (since 1996):2Export Date: 21 November 2014},
	month = {2014},
	pages = {3197 - 3208},
	abstract = {Mesenchymal stem cells (MSCs) can be made to rearrange into microtissues in response to specific matrix cues, a process that depends on a balance between cell-matrix and cell-cell interactions. The effect of such cues, and especially their interplay, is still not fully understood, particularly in three-dimensional (3-D) systems. Here, the behaviour of human MSCs cultured within hydrogel matrices with tailored stiffness and composition was evaluated. MSC aggregation occurred only in more compliant matrices (G' <= 120 Pa), when compared to stiffer ones, both in the presence and in the absence of matrix-bound arginine-glycine-aspartic acid cell-adhesion ligands (RGD; 0, 100 and 200 μM). Fibronectin assembly stabilized cell-cell contacts within aggregates, even in non-adhesive matrices. However, MSCs were able to substantially contract the artificial matrix only when RGD was present. Moreover, compliant matrices facilitated cell proliferation and provided an environment conducive for MSC osteogenic differentiation, even without RGD. Cell interactions with the original matrix became less important as time progressed, while the de novo-produced extracellular matrix became a more critical determinant of cell fate. These data provide further insights into the mechanisms by which MSCs sense their microenvironment to organize into tissues, and provide new clues to the design of cell-instructive 3-D matrices. {\textcopyright} 2014 Published by Elsevier Ltd. on behalf of Acta Materialia Inc.},
	keywords = {cell aggregation, Cell traction, Cell-matrix interactions, Hydrogels, Viscoelastic properties},
	url = {http://www.scopus.com/inward/record.url?eid=2-s2.0-84901785690\&partnerID=40\&md5=3d3a11adce17a7f1be2a2bb5d4e51089},
	author = {Maia, F.R. and Fonseca, K.B. and Rodrigues, G. and Granja, P.L. and Barrias, C.C.}
}
@article {2871,
	title = {Advanced biofabrication strategies for skin regeneration and repair},
	journal = {Nanomedicine},
	volume = {8},
	year = {2013},
	note = {Cited By (since 1996):3Export Date: 12 December 2013Source: Scopus},
	month = {2013///},
	pages = {603 - 621},
	url = {http://www.scopus.com/inward/record.url?eid=2-s2.0-84875943184\&partnerID=40\&md5=6ad3f1ffd0f6a7e567b16f0fd9b31121},
	author = {Pereira, R.F. and Barrias, C.C. and Granja, P.L. and Bartolo, P.J.}
}
@article {2876,
	title = {Enzymatic, physicochemical and biological properties of MMP-sensitive alginate hydrogels},
	journal = {Soft Matter},
	volume = {9},
	year = {2013},
	note = {Cited By (since 1996):1Export Date: 12 December 2013Source: Scopus},
	month = {2013///},
	pages = {3283 - 3292},
	url = {http://www.scopus.com/inward/record.url?eid=2-s2.0-84874428389\&partnerID=40\&md5=39d83a72fc608cc7f1c875e612421ff5},
	author = {Fonseca, K.B. and Maia, F.R. and Cruz, F.A. and Andrade, D. and Juliano, M.A. and Granja, P.L. and Barrias, C.C.}
}
@article {2838,
	title = {Functionalization of biomaterials with small osteoinductive moieties},
	journal = {Acta Biomaterialia},
	volume = {9},
	year = {2013},
	note = {Export Date: 12 December 2013Source: Scopus},
	month = {2013///},
	pages = {8773 - 8789},
	url = {http://www.scopus.com/inward/record.url?eid=2-s2.0-84885024380\&partnerID=40\&md5=a9ee12bc1c5789cd64861fd8201fc645},
	author = {Maia, F.R. and Bidarra, S.J. and Granja, P.L. and Barrias, C.C.}
}
@article {2235,
	title = {Molecularly designed alginate hydrogels susceptible to local proteolysis as three-dimensional cellular microenvironments},
	journal = {Acta BiomaterialiaActa Biomater.},
	volume = {7},
	year = {2011},
	note = {Cited By (since 1996): 9Export Date: 21 November 2012Source: Scopusdoi: 10.1016/j.actbio.2010.12.029PubMed ID: 21193068Language of Original Document: EnglishCorrespondence Address: Barrias, C. C.; 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: ccbarrias@ineb.up.ptChemicals/CAS: alginic acid, 28961-37-7, 29894-36-8, 9005-32-7, 9005-38-3; gelatin, 9000-70-8; gelatinase A, 146480-35-5; Alginates; Glucuronic Acid, 576-37-4; Hexuronic Acids; Hydrogels; Matrix Metalloproteinases, 3.4.24.-; Peptides; alginic acid, 9005-32-7References: Lukashev, M.E., Werb, Z., ECM signalling: Orchestrating cell behaviour and misbehaviour (1998) Trends in Cell Biology, 8 (11), pp. 437-441. , DOI 10.1016/S0962-8924(98)01362-2, PII S0962892498013622;Bosman, F.T., Stamenkovic, I., Functional structure and composition of the extracellular matrix (2003) Journal of Pathology, 200 (4), pp. 423-428. , DOI 10.1002/path.1437; 
Lutolf, M.P., Hubbell, J.A., Synthetic biomaterials as instructive extracellular microenvironments for morphogenesis in tissue engineering (2005) Nature Biotechnology, 23 (1), pp. 47-55. , DOI 10.1038/nbt1055; 
Lutolf, M.P., Gilbert, P.M., Blau, H.M., Designing materials to direct stem-cell fate (2009) Nature, 462 (7272), pp. 433-441; 
Tibbitt, M.W., Anseth, K.S., Hydrogels as extracellular matrix mimics for 3D cell culture (2009) Biotechnol Bioeng, 103 (4), pp. 655-663; 
Augst, A.D., Kong, H.J., Mooney, D.J., Alginate hydrogels as biomaterials (2006) Macromolecular Bioscience, 6 (8), pp. 623-633. , DOI 10.1002/mabi.200600069; 
Evangelista, M.B., Hsiong, S.X., Fernandes, R., Sampaio, P., Kong, H.-J., Barrias, C.C., Salema, R., Granja, P.L., Upregulation of bone cell differentiation through immobilization within a synthetic extracellular matrix (2007) Biomaterials, 28 (25), pp. 3644-3655. , DOI 10.1016/j.biomaterials.2007.04.028, PII S014296120700347X; 
Grellier, M., Granja, P.L., Fricain, J.C., Bidarra, S.J., Renard, M., Bareille, R., The effect of the co-immobilization of human osteoprogenitors and endothelial cells within alginate microspheres on mineralization in a bone defect (2009) Biomaterials, 30 (19), pp. 3271-3278; 
Bidarra, S.J., Barrias, C.C., Barbosa, M.A., Soares, R., Granja, P.L., Immobilization of human mesenchymal stem cells within RGD-grafted alginate microspheres and assessment of their angiogenic potential (2010) Biomacromolecules, 11 (8), pp. 1956-1964; 
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Page-Mccaw, A., Ewald, A.J., Werb, Z., Matrix metalloproteinases and the regulation of tissue remodelling (2007) Nat Rev Mol Cell Biol, 8 (3), pp. 221-233; 
Murphy, G., Nagase, H., Progress in matrix metalloproteinase research (2008) Mol Aspects Med, 29 (5), pp. 290-308; 
Chau, Y., Luo, Y., Cheung, A.C.Y., Nagai, Y., Zhang, S., Kobler, J.B., Incorporation of a matrix metalloproteinase-sensitive substrate into self-assembling peptides - A model for biofunctional scaffolds (2008) Biomaterials, 29 (11), pp. 1713-1719; 
Chau, Y., Tan, F.E., Langer, R., Synthesis and characterization of dextran-peptide-methotrexate conjugates for tumor targeting via mediation by matrix metalloproteinase II and matrix metalloproteinase IX (2004) Bioconjugate Chemistry, 15 (4), pp. 931-941. , DOI 10.1021/bc0499174; 
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Oliveira, S.M., Barrias, C.C., Almeida, I.F., Costa, P.C., Pena Ferreira, M.R., Bahia, M.F., Injectability of a bone filler system based on hydroxyapatite microspheres and a vehicle with in situ gel-forming ability (2008) J Biomed Mater Res B Appl Biomater, 87 (1), pp. 49-58; 
Kavimandan, N.J., Losi, E., Wilson, J.J., Brodbelt, J.S., Peppas, N.A., Synthesis and characterization of insulin-transferrin conjugates (2006) Bioconjugate Chemistry, 17 (6), pp. 1376-1384. , DOI 10.1021/bc050344k; 
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Dent, A., Aslam, M., The functional chemistry of proteins and protein coupling (1998) Bioconjugation: Protein Coupling Techniques for the Biomedical Sciences, pp. 50-100; 
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Drury, J.L., Boontheeku, T., Mooney, D.J., Cellular cross-linking of peptide modified hydrogels (2005) Journal of Biomechanical Engineering, 127 (2), pp. 220-228. , DOI 10.1115/1.1865194; 
Boontheekul, T., Kong, H.-J., Mooney, D.J., Controlling alginate gel degradation utilizing partial oxidation and bimodal molecular weight distribution (2005) Biomaterials, 26 (15), pp. 2455-2465. , DOI 10.1016/j.biomaterials.2004.06.044, PII S0142961204006064; 
Ries, C., Egea, V., Karow, M., Kolb, H., Jochum, M., Neth, P., MMP-2, MT1-MMP, and TIMP-2 are essential for the invasive capacity of human mesenchymal stem cells: Differential regulation by inflammatory cytokines (2007) Blood, 109 (9), pp. 4055-4063. , http://bloodjournal.hematologylibrary.org/cgi/reprint/109/9/4055, DOI 10.1182/blood-2006-10-051060; 
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Will, H., Atkinson, S.J., Butler, G.S., Smith, B., Murphy, G., The soluble catalytic domain of membrane type 1 matrix metalloproteinase cleaves the propeptide of progelatinase A and initiates autoproteolytic activation. Regulation by TIMP-2 and TIMP-3 (1996) Journal of Biological Chemistry, 271 (29), pp. 17119-17123. , DOI 10.1074/jbc.271.29.17119; 
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Barbolina, M.V., Adley, B.P., Ariztia, E.V., Liu, Y., Stack, M.S., Microenvironmental regulation of membrane type 1 matrix metalloproteinase activity in ovarian carcinoma cells via collagen-induced EGR1 expression (2007) Journal of Biological Chemistry, 282 (7), pp. 4924-4931. , http://www.jbc.org/cgi/reprint/282/7/4924, DOI 10.1074/jbc.M608428200
},
	month = {2011///},
	pages = {1674 - 1682},
	abstract = {The development of sophisticated three-dimensional (3-D) cell culture microenvironments that recreate some of the complexity of the natural extracellular matrix (ECM) remains a challenging task. Here, the modification of alginate through partial crosslinking with a matrix metalloproteinase (MMP) cleavable peptide (proline-valine-glycine-leucine-isoleucine-glycine, PVGLIG) is described, and its use in the preparation of injectable, in situ crosslinkable hydrogel-like matrices is proposed. PVGLIG-grafted alginates were synthesized by carbodiimide chemistry and characterized. Their biological performance was evaluated by comparing the response of 3-D cultured mesenchymal stem cells (MSCs) to alginate hydrogels containing only cell-adhesion peptides (RGD-alginate) or both peptides (PVGLIG/RGD-alginate). After 1 week, cells remained essentially round within RGD-alginate, while they exhibited an elongated morphology within PVGLIG/RGD-alginate hydrogels, forming cellular networks. This suggests that cells were able to structurally reorganize the matrix, through enzymatic hydrolysis of PVGLIG residues, overcoming biophysical hydrogel resistance. As shown by gelatine-zymography, MSC presented higher activity of MMP-2 when cultured within alginate functionalized with MMP-sensitive peptide, suggesting that the cell{\textquoteright}s proteolytic phenotype was modulated by the matrix composition. Additionally, PVGLIG/RGD-alginate hydrogels were clearly degraded in cell culture. Taken together, the results demonstrate that the co-incorporation of MMP-labile peptides in cell-adhesive RGD-alginate hydrogels improved their performance as ECM analogues, providing a more dynamic and physiological 3-D cellular microenvironment. {\textcopyright} 2010 Acta Materialia Inc. Published by Elsevier Ltd. All rights reserved.},
	keywords = {3-D Cell culture, Alginate, Alginates, alginic acid, Amino Acid Sequence, article, cell adhesion, cell composition, Cell Count, Cell culture, Cell Culture Techniques, Cell Shape, cell structure, cell survival, cell viability, Cells, Cultured, controlled study, Enzyme Activation, enzyme activity, extracellular matrix, Extracellular matrix (ECM), gelatin, gelatinase A, Glucuronic Acid, Hexuronic Acids, human, human cell, Humans, hydrogel, Hydrogels, hydrolysis, matrix metalloproteinase, Matrix Metalloproteinases, Matrix metalloproteinases (MMPs), Mesenchymal stem cell, Mesenchymal Stem Cells, Molecular Sequence Data, morphology, peptide derivative, Peptides, phenotype, priority journal, prolylvalylglycylleucylisoleucylglycine, Protease-sensitive, protein cleavage, protein degradation, Protein Processing, Post-Translational, Subcellular Fractions, tumor microenvironment, unclassified drug, zymography},
	isbn = {17427061 (ISSN)},
	url = {http://www.scopus.com/inward/record.url?eid=2-s2.0-79952189303\&partnerID=40\&md5=191596b948bd4cc2569264a992052b88},
	author = {Fonseca, K.B. and Bidarra, S.J. and Oliveira, M.J. and Granja, P.L. and Barrias, C.C.}
}
@article {2274,
	title = {Characterization of polymeric solutions as injectable vehicles for hydroxyapatite microspheres},
	journal = {AAPS PharmSciTechAAPS PharmSciTech},
	volume = {11},
	year = {2010},
	note = {Cited By (since 1996): 1Export Date: 21 November 2012Source: Scopusdoi: 10.1208/s12249-010-9447-3PubMed ID: 20490958Language of Original Document: EnglishCorrespondence Address: Oliveira, S. M.; Dep. de Eng. Mec{\^a}nica e Gest{\~a}o Industrial, Escola Superior de Tecnologia e Gest{\~a}o de Viseu, Campus Polit{\'e}cnico de Repeses, 3504-510, Viseu, Portugal; email: soliveira@estv.ipv.ptChemicals/CAS: alginic acid, 28961-37-7, 29894-36-8, 9005-32-7, 9005-38-3; carboxymethylcellulose, 8050-38-2, 9000-11-7, 9004-32-4, 9050-04-8; hydroxyapatite, 1306-06-5, 51198-94-8; hydroxypropylmethylcellulose, 9004-65-3; Biocompatible Materials; Capsules; Durapatite, 1306-06-5; Excipients; Polymers; Solutions; VehiclesReferences: Verlaan, J.J., Oner, F.C., Dhert, W.J., Anterior spinal column augmentation with injectable bone cements (2006) Biomat, 27 (3), pp. 290-301;Chenite, A., Chaput, C., Wang, D., Combes, C., Buschmann, M.D., Hoemann, C.D., Novel injectable neutral solutions of chitosan form biodegradable gels in situ (2000) Biomat, 21 (21), pp. 2155-2161; 
Hoemann, C.D., Sun, J., Legare, A., McKee, M.D., Buschmann, M.D., Tissue engineering of cartilage using an injectable and adhesive chitosan-based cell-delivery vehicle (2005) Osteoarthritis Cart., 13 (4), pp. 318-329; 
Balakrishnan, B., Jayakrishnan, A., Self-cross-linking biopolymers as injectable in situ forming biodegradable scaffolds (2005) Biomat, 26 (18), pp. 3941-3951; 
Shu, X.Z., Ghosh, K., Liu, Y., Palumbo, F.S., Luo, Y., Clark, R.A., Attachment and spreading of fibroblasts on an RGD peptidemodified injectable hyaluronan hydrogel (2004) J. Biomed. Mater. Res., 68 A (2), pp. 365-375; 
Shu, X.Z., Ahmad, S., Liu, Y., Prestwich, G.D., Synthesis and evaluation of injectable, in situ crosslinkable synthetic extracellular matrices for tissue engineering (2006) J. Biomed. Mater. Res. A, 79 (4), pp. 902-912; 
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Virto, M.R., Frutos, P., Torrado, S., Frutos, G., Gentamicin release from modified acrylic bone cements with lactose and hydroxypropylmethylcellulose (2003) Biomat, 24 (1), pp. 79-87; 
Bodic, F., Amouriq, Y., Gayet-Delacroix, M., Gauthier, O., Bouler, J.-M., Daculsi, G., M{\'e}thode nom invasive d{\textquoteright}evaluation d{\textquoteright}un substitut osseux injectable/Non-invasive evaluation of an injectable bone substitute (2002) C R Biologies, 325, pp. 345-353; 
Andrews, G.P., Gorman, S.P., Jones, D.S., Rheological characterisation of primary and binary interactive bioadhesive gels composed of cellulose derivatives designed as ophthalmic viscosurgical devices (2005) Biomat, 26 (5), pp. 571-580; 
He, S., Yaszemski, M.J., Yasko, A.W., Engel, P.S., Mikos, A.G., Injectable biodegradable polymer composites based on poly (propylene fumarate) crosslinked with poly (ethylene glycol) dimethacrylate (2000) Biomat, 21 (23), pp. 2389-2394; 
Temenoff, J.S., Mikos, A.G., Injectable biodegradable materials for orthopedic tissue engineering (2000) Biomat, 21 (23), pp. 2405-2412; 
Iooss, P., Le Ray, A.M., Grimandi, G., Daculsi, G., Merle, C., A new injectable bone substitute combining poly (epsilon-caprolactone) microparticles with biphasic calcium phosphate granules (2001) Biomat, 22 (20), pp. 2785-2794; 
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},
	month = {2010///},
	pages = {852 - 858},
	abstract = {A polymeric solution and a reinforcement phase can work as an injectable material to fill up bone defects. However, the properties of the solution should be suitable to enable the transport of that extra phase. Additionally, the use of biocompatible materials is a requirement for tissue regeneration. Thus, we intended to optimize a biocompatible polymeric solution able to carry hydroxyapatite microspheres into bone defects using an orthopedic injectable device. To achieve that goal, polymers usually regarded as biocompatible were selected, namely sodium carboxymethylcellulose, hydroxypropylmethylcellulose, and Na-alginate (ALG). The rheological properties of the polymeric solutions at different concentrations were assessed by viscosimetry before and after moist heat sterilization. In order to correlate rheological properties with injectability, solutions were tested using an orthopedic device applied for minimal invasive surgeries. Among the three polymers, ALG solutions presented the most suitable properties for our goal and a non-sterile ALG 6\% solution was successfully used to perform preliminary injection tests of hydroxyapatite microspheres. Sterile ALG 7.25\% solution was found to closely match non-sterile ALG 6\% properties and it was selected as the optimal vehicle. Finally, sterile ALG 7.25\% physical stability was studied at different temperatures over a 3-month period. It was observed that its rheological properties presented minor changes when stored at 25{\textdegree}C or at 4{\textdegree}C. {\textcopyright} 2010 American Association of Pharmaceutical Scientists.},
	keywords = {Alginate, alginic acid, article, Biocompatibility, Biocompatible Materials, Capsules, Carboxymethylcellulose, Drug Compounding, Durapatite, Excipients, hydroxyapatite, Hydroxypropylmethylcellulose, Injectability, injection, Injections, instrument sterilization, microsphere, Microspheres, minimally invasive surgery, molecular weight, orthopedic equipment, Polymers, priority journal, Solutions, Vehicles, viscometry},
	isbn = {15309932 (ISSN)},
	url = {http://www.scopus.com/inward/record.url?eid=2-s2.0-77954819260\&partnerID=40\&md5=e8503462fa39dcc285aef7e07f765eef},
	author = {Oliveira, S.M. and Almeida, I.F. and Costa, P.C. and Barrias, C.C. and Ferreira, M.R.P. and Bahia, M.F. and Barbosa, M.A.}
}
@article {2264,
	title = {Immobilization of human mesenchymal stem cells within RGD-grafted alginate microspheres and assessment of their angiogenic potential},
	journal = {BiomacromoleculesBiomacromolecules},
	volume = {11},
	year = {2010},
	note = {Cited By (since 1996): 18Export Date: 21 November 2012Source: ScopusCODEN: BOMAFdoi: 10.1021/bm100264aPubMed ID: 20690708Language of Original Document: EnglishCorrespondence Address: Granja, P. L.; INEB-Instituto de Engenharia Biom{\'e}dica, Rua do Campo Alegre No. 823, 4150-180 Porto, Portugal; email: pgranja@ineb.up.ptMolecular Sequence Numbers: GENBANK: BC021289, NM_000088, NM_002046;Chemicals/CAS: alginic acid, 28961-37-7, 29894-36-8, 9005-32-7, 9005-38-3; alkaline phosphatase, 9001-78-9; arginylglycylaspartic acid, 99896-85-2; Alginates; Alkaline Phosphatase, 3.1.3.1; Collagen Type I; DNA Primers; Oligopeptides; arginyl-glycyl-aspartic acid, 99896-85-2References: Temenoff, J.S., Mikos, A.G., Injectable biodegradable materials for orthopedic tissue engineering (2000) Biomaterials, 21, pp. 2405-2412;Barralet, J.E., Wang, L., Lawson, M., Triffitt, J.T., Cooper, P.R., Shelton, R.M., Comparison of bone marrow cell growth on 2D and 3D alginate hydrogels (2005) J. Mater. Sci.: Mater. Med., 16, pp. 515-519; 
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},
	month = {2010///},
	pages = {1956 - 1964},
	abstract = {In this work, human mesenchymal stem cells (hMSC) immobilized in RGD-coupled alginate microspheres, with a binary composition of high and low molecular weight alginate, were investigated. Cells immobilized within RGD-alginate microspheres (during 21 days) showed metabolic activity, with an overall viability higher than 90\%, short cell extensions, and, when induced, they were able to differentiate into the osteogenic lineage. In osteogenic conditions (comparing to basal conditions), immobilized cells presented alkaline phosphatase (ALP) activity and an upregulation of ALP, collagen type I, and Runx 2 expression. Moreover, mineralization was also detected in immobilized cells under osteogenic stimulus. In addition, it was demonstrated for the first time that MSCs immobilized in this 3D matrix were able to enhance the ability of neighboring endothelial cells to form tubelike structures. Overall, these findings represent a step forward in the development of injectable stem cell carriers for bone tissue engineering. {\textcopyright} 2010 American Chemical Society.},
	keywords = {Alginate, Alginates, alginic acid, alkaline phosphatase, Alkaline phosphatase activity, angiogenesis, arginylglycylaspartic acid, article, Basal conditions, Base Sequence, Binary composition, bone development, Bone tissue engineering, Cell culture, cell differentiation, Cell extension, Cell immobilization, cell metabolism, cell structure, cell viability, collagen type 1, Collagen type I, controlled study, DNA Primers, Endothelial Cells, endothelium cell, enzyme activity, human, human cell, Human mesenchymal stem cells, Humans, immobilized cell, Immobilized cells, Low molecular weight, matrix, Mesenchymal stem cell, Mesenchymal Stem Cells, Metabolic activity, Microscopy, Electron, Scanning, microsphere, Microspheres, molecular weight, Neovascularization, Physiologic, nucleotide sequence, Oligopeptides, osteoblast, Osteogenic, Phosphatases, priority journal, protein expression, quantitative analysis, Reverse Transcriptase Polymerase Chain Reaction, stem cell, Stem cells, three dimensional imaging, Tissue engineering, transcription factor RUNX2, umbilical vein, Up-regulation, upregulation},
	isbn = {15257797 (ISSN)},
	url = {http://www.scopus.com/inward/record.url?eid=2-s2.0-77955575798\&partnerID=40\&md5=d2d378c63dc49de9324e8dddb24ff2e9},
	author = {Bidarra, S.J. and Barrias, C.C. and Barbosa, M.A. and Soares, R. and Granja, P.L.}
}
@article {2323,
	title = {The correlation between the adsorption of adhesive proteins and cell behaviour on hydroxyl-methyl mixed self-assembled monolayers},
	journal = {BiomaterialsBiomaterials},
	volume = {30},
	year = {2009},
	note = {Cited By (since 1996): 41Export Date: 21 November 2012Source: ScopusCODEN: BIMADdoi: 10.1016/j.biomaterials.2008.09.048PubMed ID: 18952279Language of Original Document: EnglishCorrespondence Address: Barrias, C.C.; INEB - Instituto de Engenharia Biom{\'e}dica, Divis{\~a}o de Biomateriais, Universidade do Porto, R. Campo Alegre 823, 4150-180 Porto, Portugal; email: ccbarrias@ineb.up.ptChemicals/CAS: fibronectin, 86088-83-7; gold, 7440-57-5; thiol derivative, 13940-21-1; Biocompatible Materials; Fibronectins; VitronectinReferences: Wilson, C.J., Clegg, R.E., Leavesley, D.I., Pearcy, M.J., Mediation of biomaterial-cell interactions by adsorbed proteins: a review (2005) Tissue Eng, 11, pp. 1-18;Garc{\'\i}a, A.J., Vega, M.D., Boettiger, D., Modulation of cell proliferation and differentiation through substrate-dependent changes in fibronectin conformation (1999) Mol Biol Cell, 10, pp. 785-798; 
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},
	month = {2009///},
	pages = {307 - 316},
	abstract = {The objective of this study was to compare the biological effects of two key cell-adhesive proteins, fibronectin (FN) and vitronectin (VN), upon adsorption onto molecularly-designed model surfaces. Single-component and mixed self-assembled monolayers (SAMs) of alkanethiols on gold with OH and CH                        3 terminal groups were prepared at 100\%, 65\%, 36\% and 0\% of OH at the surface, to generate a range of surfaces with a simple chemistry and a wettability gradient. FN and VN were adsorbed under non-competitive (single-protein solutions) and competitive (multi-protein solutions) conditions, and compared at different levels: adsorbed amount (radiolabelling), elution, functional presentation of cell-binding domains (ELISA), and role in mediating cell adhesion (antibody-based assay). The observed trends were related to mesenchymal stem cell response in terms of adhesion and overall cell morphology. Under non-competitive conditions, adsorption of both proteins increased with surface hydrophobicity. The presence of competitive proteins significantly decreased the adsorbed amounts, although both proteins were still detected in all SAMs. Adsorption of FN followed a trend similar to that of non-competitive conditions, while adsorption of VN was higher on 100\%OH-SAMs. Concerning elution, retention of adsorbed VN was always higher than that of FN. For both proteins, functional presentation of cell-binding domains was more effective on the more hydrophilic 100\%OH-SAMs. This fact, coupled to the ability of this type of SAMs to selectively recruit and retain VN in the presence of competitive serum proteins, seems to correlate with the better cell response observed on these surfaces, as compared with hydrophobic 0\%OH(100\%CH
                        3)-SAMs. {\textcopyright} 2008 Elsevier Ltd. All rights reserved.
},
	keywords = {Adhesion, Adhesiveness, Adsorption, alkane, Animals, article, Biocompatible Materials, Biological Assay, bone marrow, Cattle, cell adhesion, Cell culture, Cell Movement, Cells, Cells, Cultured, cytology, cytoskeleton, Electric currents, Electron energy levels, enzyme linked immunosorbent assay, Fibronectin, Fibronectins, Focal adhesions, Functional groups, Functionalization, gold, human, human cell, Humans, hydrophilicity, hydrophobicity, Hydroxylation, isotope labeling, Mesenchymal stem cell, Mesenchymal Stem Cells, Microscopy, Confocal, Missile bases, Molecular interactions, monoclonal antibody, priority journal, Protein adsorption, protein analysis, protein binding, Protein Structure, Tertiary, Proteins, Self assembled monolayers, Self-assembled monolayers, stem cell, Stem cells, Surface Properties, thiol derivative, Vitronectin},
	isbn = {01429612 (ISSN)},
	url = {http://www.scopus.com/inward/record.url?eid=2-s2.0-55249093830\&partnerID=40\&md5=132fa4efe300c4f76764942bfe3bce30},
	author = {Barrias, C.C. and Martins, M.C.L. and Almeida-Porada, G. and Barbosa, M.A. and Granja, P.L.}
}
@book {2320,
	title = {Morphology and mechanical properties of injectable ceramic microspheres},
	series = {Key Engineering MaterialsKey Eng Mat},
	volume = {396-398},
	year = {2009},
	note = {Cited By (since 1996): 1Export Date: 21 November 2012Source: ScopusCODEN: KEMAEdoi: 10.4028/0-87849-353-0.691Language of Original Document: EnglishCorrespondence Address: Oliveira, S. M.; ESTV, Departamento de Engenharia Mec{\^a}nica, Instituto Polit{\'e}cnico de Viseu, Viseu, PortugalReferences: Barrias, C.C., Ribeiro, C.C., Lamghari, M., Miranda, C.S., Barbosa, M.A., Proliferation, activity, and osteogenic differentiation of bone marrow stromal cells cultured on calcium titanium phosphate microspheres (2005) Journal of Biomedical Materials Research, 72 A (1), pp. 57-66;Barrias, C.C., Ribeiro, C.C., Barbosa, M.A., Adhesion and proliferation of human osteoblastic cells seeded on injectable hydroxyapatite microspheres (2004) Bioceramics, 16, pp. 254-262,877-880; 
Ribeiro, C.C., Barrias, C.C., Barbosa, M.A., Calcium phosphate-alginate microspheres as enzyme delivery matrices (2004) Biomaterials, 25 (18), pp. 4363-4373; 
Silva, R.V., Camilli, J.A., Bertran, C.A., Moreira, N.H., The use of hydroxyapatite and autogenous cancellous bone grafts to repair bone defects in rats (2005) International Journal of Oral and Maxillofacial Surgery, 34 (2), pp. 178-184; 
Boyan, B.D., Lossd{\"o}rfer, S., Wang, L., Zhao, G., Lohmann, C.H., Cochran, D.L., Schwartz, Z., Osteoblasts generate an osteogenic microenvironment when grown on surfaces with rough microtopographies (2003) Eur Cell Mater, 6, pp. 22-27; 
Cardoso, O.N., Sotto Mayor, T., Pinto, A.M.F.R., Campos, J.B.L.M., Axial dispersion of particles in a slugging column - the role of the laminar wake of the bubbles (2003) Chemical Engineering Science, 58 (18), pp. 4159-4172; 
Gauthier, O., Bouler, J.M., Weiss, P., Bosco, J., Aguado, E., Daculsi, G., Short-term effects of mineral particle sizes on cellular degradation activity after implantation of injectable calcium phosphate biomateriais and the consequences for bone substitution (1999) Bone, 25 (2), pp. 71S-74S; 
Xu, H.H., Weir, M.D., Burguera, E.F., Fraser, A.M., Injectable and macroporous calcium phosphate cement scaffold (2006) Biomaterials, 27 (24), pp. 4279-4287; 
Ginebra, M.P., Rilliard, A., Fernandez, E., Elvira, C., San Roman, J., Planell, J.A., Mechanical and rheological improvement of a calcium phosphate cement by the addition of a polymeric drug (2001) J Biomed Mater Res, 57 (1), pp. 113-118
},
	month = {2009///},
	pages = {691 - 694},
	abstract = {The aim of this study was to analyze the effect of starting powder granulometry and sintering conditions on the morphological structure and mechanical properties of injectable hydroxyapaptite (HAp) microspheres. The mechanical properties of the microspheres were evaluated, to investigate if their integrity could be maintained during the injection process. To obtain microspheres, HAp powders were dispersed in a sodium alginate solution and spherical particles were prepared by droplet extrusion under a co-axial air stream, coupled with ionotropic gelation in the presence of Ca                        2+. This was followed by a sintering process at various temperatures and times. The morphology of microspheres was observed under SEM, diameter measurements were performed in an optical microscope and the compression strength was evaluated using a texture analyzer. Finally, microspheres prepared using lower granulometry HAp powders and sintered at 1200{\textdegree}C for 1 hour presented the best properties and were selected as the most suitable for the envisaged application.
},
	keywords = {Air streams, Apatite, Calcium, Ceramic microspheres, Co-axial, Coagulation, Compression strengths, Diameter measurements, Gelation, Granulometry, hydroxyapatite, Injectable, Injection process, Ionotropic gelations, Mechanical properties, Microspheres, Morphological structures, Morphology and mechanical properties, Optical microscopes, Powders, SEM, Sintering, Sintering conditions, Sintering process, Sodium, Sodium alginates, Spherical particles, Starting powders, Texture analyzers},
	isbn = {10139826 (ISSN)},
	url = {http://www.scopus.com/inward/record.url?eid=2-s2.0-58849147162\&partnerID=40\&md5=5bf4364653a3ad10b5ab28c1a01568ab},
	author = {Oliveira, S.M. and Barrias, C.C. and Ribeiro, C.C. and Almeida, I.F. and Bahia, M.F. and Barbosa, M.A.}
}
@article {2342,
	title = {Comparative study of nanohydroxyapatite microspheres for medical applications},
	journal = {Journal of Biomedical Materials Research - Part AJ. Biomed. Mater. Res. Part A},
	volume = {86},
	year = {2008},
	note = {Cited By (since 1996): 22Export Date: 21 November 2012Source: ScopusCODEN: JBMRCdoi: 10.1002/jbm.a.31634PubMed ID: 17975824Language of Original Document: EnglishCorrespondence Address: Mateus, A. Y. P.; INEB - Instituto de Engenharia Biom{\'e}dica, Laborat{\'o}rio de Biomateriais, Rua do Campo Alegre 823, 4150-180 Porto, Portugal; email: yovana@ibmc.up.ptChemicals/CAS: alginic acid, 28961-37-7, 29894-36-8, 9005-32-7, 9005-38-3; calcium hydroxide, 1305-62-0; calcium phosphate, 10103-46-5, 13767-12-9, 14358-97-5, 7758-87-4; calcium, 7440-70-2; dodecyl sulfate sodium, 151-21-3; neutral red, 553-24-2; phosphoric acid, 7664-38-2; Biocompatible Materials; Durapatite, 1306-06-5; PowdersReferences: Jarcho, M., Calcium phosphates ceramics as hard tissue prosthetics (1981) Clin Orthop Relat Res, 157, pp. 259-278;Bucholz, R., Carlton, A., Holmes, R., Hydroxyapatite and tricalcium phosphate bone graft substitutes (1987) Orthop Clin N Am, 18, pp. 323-334; 
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},
	month = {2008///},
	pages = {483 - 493},
	abstract = {This study concerns the preparation, physical, and in vitro characterization of two different types of hydroxyapatite (HA) microspheres, which are intended to be used as drug-delivery systems and bone-regeneration matrices. Hydroxyapatite nanoparticles (HA-1 and HA-2) were prepared using the chemical precipitation synthesis with H                        3PO
                        4, Ca(OH) 
                        2, and a surfactant, SDS (sodium dodecyl sulfate), as starting reagents. The HA powders were dispersed in a sodium alginate solution, and spherical particles were obtained by droplet extrusion coupled with ionotropic gelation in the presence of Ca
                        2+. These were subsequently sintered to produce HA-1 and HA-2 microspheres with a uniform size and interconnected microporosity. Both powders and microspheres were characterized using FTIR and X-ray diffraction. Moreover, SEM and mercury intrusion porosimetry were used to analyze the microspheres, and TEM was used to analyze the powders. Results showed that pure HA and mixtures of HA/β-TCP in the nanometer range and needlelike shape were obtained for HA-1 and HA-2 powders, respectively. Neutral Red, scanning electron microscopy and confocal microscopy were used to evaluate the behavior of osteoblastic-like MG-63 cells cultured on HA microspheres surfaces for 7 days. Results showed that good adhesion and proliferation of osteoblasts on the HA microspheres surface. Cells built bridges between adjacent microspheres, forming microspheres-cells clusters in both types of materials. {\textcopyright} 2007 Wiley Periodicals, Inc.
},
	keywords = {alginic acid, analytic method, Apatite, article, Biocompatible Materials, Biological materials, biomaterial, Bone Regeneration, Calcium, calcium hydroxide, calcium phosphate, cell adhesion, Cell Line, cell proliferation, Chemical precipitation, Chemotherapy, Comparative studies, comparative study, controlled study, dodecyl sulfate sodium, Drug delivery, Drug delivery system (DDS), Drug Delivery Systems, Drug dosage, Durapatite, Gelation, human, human cell, Humans, hydroxyapatite, Hydroxyapatite (HAP), Hydroxylation, In vitro characterization, in vitro study, infrared spectroscopy, materials testing, Medical applications, Mercury intrusion porosimetry, microsphere, Microspheres, Nano-hydroxyapatite (n-HA), Nanohydroxyapatite, nanoparticle, nanoparticles, neutral red, osteoblast, Osteoblasts, Particle Size, phosphoric acid, porosity, powder, Powders, precipitation, Precipitation (chemical), reagent, Regeneration (renovating), Scanning electron microscopy, surfactant, synthesis, Synthesis (chemical), X ray diffraction},
	isbn = {15493296 (ISSN)},
	url = {http://www.scopus.com/inward/record.url?eid=2-s2.0-47049103112\&partnerID=40\&md5=138b3e2f3a97d8b398546d3f85567a0f},
	author = {Mateus, A.Y.P. and Barrias, C.C. and Ribeiro, C. and Ferraz, M.P. and Monteiro, F.J.}
}
@article {2336,
	title = {Injectability of a bone filler system based on hydroxyapatite microspheres and a vehicle with in situ gel-forming ability},
	journal = {Journal of Biomedical Materials Research - Part B Applied BiomaterialsJ. Biomed. Mater. Res. Part B Appl. Biomater.},
	volume = {87},
	year = {2008},
	note = {Cited By (since 1996): 9Export Date: 21 November 2012Source: ScopusCODEN: JBMRGdoi: 10.1002/jbm.b.31066PubMed ID: 18437700Language of Original Document: EnglishCorrespondence Address: Oliveira, S. M.; Escola Superior de Tecnologia de Viseu (ESTV), Dep. de Eng. Mec{\^a}nica e Gest{\~a}o Industrial, 3504-510 Viseu, Portugal; email: serafim@ineb.up.ptChemicals/CAS: alginic acid, 28961-37-7, 29894-36-8, 9005-32-7, 9005-38-3; calcium carbonate, 13397-26-7, 13701-58-1, 14791-73-2, 471-34-1; gluconolactonase, 9012-44-6, 9012-73-1; hydroxyapatite, 1306-06-5, 51198-94-8; Bone Substitutes; Durapatite, 1306-06-5; GelsTradenames: Captal S, Plasma BiotalManufacturers: Plasma BiotalReferences: Ribeiro, C.C., Barrias, C.C., Barbosa, M.A., Calcium phosphate-alginate microspheres as enzyme delivery matrices (2004) Biomaterials, 25, pp. 4363-4373;Grabowski, G.A., Leslie, N., Wenstrup, R., Enzyme therapy for Gaucher disease: The first 5 years (1998) Blood Rev, 12, pp. 115-133; 
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Jasper, L.E., Deramond, H., Mathis, J.M., Belkoff, S.M., Material properties of various cements for use with vertebroplasty (2002) J Mater Sci Mater Med, 13, pp. 1-5
},
	month = {2008///},
	pages = {49 - 58},
	abstract = {The aim of this study was to test the injectability of a bone filler system based on the combination of ceramic microspheres with a gel-like vehicle, for noninvasive surgery. Porous hydroxyapatite microspheres with a uniform size and an average diameter of 535 {\textpm} 38 μm were prepared, and their compression strength and friability were tested. The sodium-alginate solution with a concentration of 7.25\% (w/v) was used as the vehicle. To promote its in situ gelation, calcium carbonate and D-gluconic-δ-lactone were added to the solution. Microspheres were mixed with the vehicle at different percentages (20-40 wt \%). Gelation times in the range of 8-20 min, were obtained, depending on the formulation. Mixtures of HAp microspheres with alginate solution at 7.25\% originating a gel in 11 min present an adequate handling time to perform an injection. Their injectability was evaluated using an injection device commonly employed in vertebroplasty surgical procedures, coupled to a texturometer in compression mode. Using an extrusion rate of 0.1 mm/s, the force required to extrude any of the mixtures tested was lower than 100 N. For an extrusion rate of 1 mm/s mixtures with 40 wt \% of microspheres were very difficult to inject. Mixtures with 35 wt \% of microspheres presented the best compromise between injectability and compression strength of the gelled system. MicroCT analysis revealed a homogeneous distribution of the microspheres inside the vehicle, as well as full interconnection of the intra-microspheres spaces. The compression strength for the gelled systems ranged from 80 kPa (gel itself) to 600 kPa (composite with 40 Wt \% of microspheres). {\textcopyright} 2008 Wiley Periodicals, Inc.},
	keywords = {Alginate, alginic acid, Apatite, article, Average diameter, bioengineering, Bone, bone filler system, Bone Substitutes, Calcium, Calcium carbonate, Ceramic, Ceramic microspheres, Coagulation, Colloids, compression, Compression mode, Compression strengths, Compressive Strength, Computerized tomography, Concentration (process), Data compression, Durapatite, Extrusion, Extrusion rates, Fillers, Forming, Gel-forming ability, Gelation, Gels, gluconolactonase, Handling time, Homogeneous distribution, hydrogel, hydroxyapatite, In situ gelation, in situ hybridization, In-situ, Injectability, injection, Injection (oil wells), Micro-CT, microsphere, Microspheres, Mixtures, non invasive procedure, Particle Size, Porous hydroxyapatite, Sodium, Surgery, Surgical procedures, Uniform size, Vehicles, Vertebroplasty},
	isbn = {15524973 (ISSN)},
	url = {http://www.scopus.com/inward/record.url?eid=2-s2.0-52449123139\&partnerID=40\&md5=cc648bad01cb2c6aa01d885acac10e8a},
	author = {Oliveira, S.M. and Barrias, C.C. and Almeida, I.F. and Costa, P.C. and Pena Ferreira, M.R. and Bahia, M.F. and Barbosa, M.A.}
}
@article {2376,
	title = {Upregulation of bone cell differentiation through immobilization within a synthetic extracellular matrix},
	journal = {BiomaterialsBiomaterials},
	volume = {28},
	year = {2007},
	note = {Cited By (since 1996): 39Export Date: 21 November 2012Source: ScopusCODEN: BIMADdoi: 10.1016/j.biomaterials.2007.04.028PubMed ID: 17532040Language of Original Document: EnglishCorrespondence Address: Evangelista, M.B.; INEB-Instituto de Engenharia Biom{\'e}dica, Laborat{\'o}rio de Biomateriais, Rua do Campo Alegre, 823, 4150-180 Porto, Portugal; email: mbe@ineb.up.ptChemicals/CAS: alginic acid, 28961-37-7, 29894-36-8, 9005-32-7, 9005-38-3; alkaline phosphatase, 9001-78-9; arginylglycylaspartic acid, 99896-85-2; DNA, 9007-49-2; osteocalcin, 136461-80-8; Alginates; alginic acid, 9005-32-7; Alkaline Phosphatase, EC 3.1.3.1; arginyl-glycyl-aspartic acid, 99896-85-2; Glucuronic Acid, 576-37-4; Hexuronic Acids; Oligopeptides; Osteocalcin, 104982-03-8References: de Vos, P., de Haan, B., Wolters, G.H.J., VanSchilfgaarde, R., Factors influencing the adequacy of microencapsulation of rat pancreatic islets (1996) Transplantation, 62 (7), pp. 888-893;Brissova, M., Lacik, I., Powers, A.C., Anilkumar, A.V., Wang, T., Control and measurement of permeability for design of microcapsule cell delivery system (1998) J Biomed Mater Res, 39 (1), pp. 61-70; 
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Bancroft, J.D., Gamble, M., (2002) Theory and practice of histological techniques. 5th ed, , Elsevier Science, Edinburgh
},
	month = {2007///},
	pages = {3644 - 3655},
	abstract = {There is a need for new therapeutic strategies to treat bone defects caused by trauma, disease or tissue loss. Injectable systems for cell transplantation have the advantage of allowing the use of minimally invasive surgical procedures, and thus for less discomfort to patients. In the present study, it is hypothesized that Arg-Gly-Asp (RGD)-coupled in a binary (low and high molecular weight) injectable alginate composition is able to influence bone cell differentiation in a three-dimensional (3D) structure. Viability, metabolic activity, cytoskeleton organization, ultrastructure and differentiation (alkaline phosphatase (ALP), von Kossa, alizarin red stainings and osteocalcin quantification) of immobilized cells were assessed. Cells within RGD-modified alginate microspheres were able to establish more interactions with the synthetic extracellular matrix as visualized by confocal laser scanning microscope and transmission electron microscopy imaging, and presented a much higher level of differentiation (more intense ALP and mineralization stainings and higher levels of osteocalcin secretion) when compared to cells immobilized within unmodified alginate microspheres. These findings demonstrate that peptides covalently coupled to alginate were efficient in influencing cell behavior within this 3D system, and may provide adequate preparation of osteoblasts for cell transplantation. {\textcopyright} 2007 Elsevier Ltd. All rights reserved.},
	keywords = {3T3 Cells, Alginate, Alginates, alginic acid, alkaline phosphatase, animal cell, Animals, arginylglycylaspartic acid, article, Bone, bone cell, Bone Regeneration, calvaria, cell adhesion, cell differentiation, Cell encapsulation, Cell immobilization, cell metabolism, cell proliferation, cell survival, cell ultrastructure, cell viability, confocal laser microscopy, controlled study, cytology, cytoskeleton, DNA, DNA content, ECM (extracellular matrix), enzyme activity, extracellular matrix, Glucuronic Acid, Hexuronic Acids, hydrogel, immobilized cell, metabolism, Mice, Microscopy, Confocal, Microscopy, Electron, Transmission, microsphere, Microspheres, nonhuman, Oligopeptides, osteoblast, Osteoblasts, osteocalcin, Patient treatment, priority journal, rat, RGD peptide, structure analysis, three dimensional imaging, Tissue engineering, transmission electron microscopy, Transplantation (surgical)},
	isbn = {01429612 (ISSN)},
	url = {http://www.scopus.com/inward/record.url?eid=2-s2.0-34249870130\&partnerID=40\&md5=4476a4b1ee80320328c1bf37054037bb},
	author = {Evangelista, M.B. and Hsiong, S.X. and Fernandes, R. and Sampaio, P. and Kong, H.-J. and Barrias, C.C. and Salema, R. and Barbosa, M.A. and Mooney, D.J. and Granja, P.L.}
}
@book {2414,
	title = {Calcium phosphate microspheres for localised delivery of a therapeutic enzyme},
	series = {Key Engineering MaterialsKey Eng Mat},
	volume = {309-311 II},
	year = {2006},
	note = {Export Date: 21 November 2012Source: ScopusCODEN: KEMAELanguage of Original Document: EnglishCorrespondence Address: Barrias, C.C.; INEB - Instituto de Engenharia Biom{\'e}dica, Laborat{\'o}rio de Biomateriais, R. Campo Alegre 823, 4150-180 Porto, Portugal; email: ccbarrias@ineb.up.ptReferences: Barrias, C.C., Ribeiro, C.C., Barbosa, M.A., (2004) Key Eng. Mater., 254-256, p. 877;Barrias, C.C., Ribeiro, C.C., Lamghari, M., Miranda, C.S., Barbosa, M.A., (2005) J. Biomed. Mater. Res. A, 72 A, p. 57; 
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Barrias, C.C., Martins, M.A., Miranda, M.A., Barbosa, M.A., (2005) Biomaterials, 26, p. 2695
},
	month = {2006///},
	pages = {903 - 906},
	abstract = {The ability of calcium titanium phosphate (CTP) and hydroxyapatite (HAp) microspheres to reversibly adsorb the enzyme glucocerebrosidase (GCR) while preserving its biological activity, and efficiently deliver it to Gaucher disease (GD) fibroblasts was investigated. CTP microspheres adsorbed ca. 3.6-fold more GCR than HAp microspheres. The activity of adsorbed GCR was higher than the free enzyme in the case of CTP microspheres and lower when HAp was used. GCR release from both types of microspheres was characterized by the initial elution of a large percentage of enzyme followed by a delayed release that extended for at least 30 days. Released GCR was internalized by GD fibroblasts increasing their intracellular enzymatic activity. In cells treated with the same amount of GCR-loaded CTP microspheres or free-GCR a higher intracellular activity was detected in the former case, suggesting an improved efficacy.},
	keywords = {Adsorption, Biocompatibility, biological activity, Biomaterials, Calcium Compounds, Calcium titanium phosphate, Calcium titanium phosphate (CTP), Cells, Enzyme adsorption, Enzymes, hydroxyapatite, Microspheres},
	isbn = {10139826 (ISSN)},
	url = {http://www.scopus.com/inward/record.url?eid=2-s2.0-33645396489\&partnerID=40\&md5=de3cf8530e8c690b254fed782b964340},
	author = {Barrias, C.C. and Ribeiro, C.C. and Martins, M.C.L. and Barbosa, M.A. and Rodrigues, D. and Miranda, M.C.S.}
}
@article {2447,
	title = {Biological evaluation of calcium alginate microspheres as a vehicle for the localized delivery of a therapeutic enzyme},
	journal = {Journal of Biomedical Materials Research - Part AJ. Biomed. Mater. Res. Part A},
	volume = {74},
	year = {2005},
	note = {Cited By (since 1996): 12Export Date: 21 November 2012Source: ScopusCODEN: JBMRCdoi: 10.1002/jbm.a.30348PubMed ID: 16028235Language of Original Document: EnglishCorrespondence Address: Barrias, C.C.; Instituto de Engenharia Biom{\'e}dica (INEB), Laborat{\'o}rio de Biomateriais, Rua do Campo Alegre, 823, 4150-180 Porto, Portugal; email: ccbarrias@ineb.up.ptChemicals/CAS: alginic acid, 28961-37-7, 29894-36-8, 9005-32-7, 9005-38-3; calcium alginate, 9005-35-0; glucosylceramidase, 37228-64-1; iodine 125, 14158-31-7, 22822-81-7; Alginates; alginic acid, 9005-32-7; Drug Carriers; Enzymes, Immobilized; Glucosylceramidase, EC 3.2.1.45; Glucuronic Acid, 576-37-4; Hexuronic AcidsManufacturers: GenzymeReferences: Grabowski, G.A., Leslie, N., Wenstrup, R., Enzyme therapy for Gaucher disease: The first 5 years (1998) Blood Rev, 12, pp. 115-133;Zhao, H., Grabowski, G.A., Gaucher disease: Perspectives on a prototype lysosomal disease (2002) Cell Mol Life Sci, 59, pp. 694-707; 
Friedman, B., Vaddi, K., Preston, C., Mahon, E., Cataldo, J.R., McPherson, J.M., A comparison of the pharmacological properties of carbohydrate remodeled recombinant and placental-derived beta-glucocerebrosidase: Implications for clinical efficacy in treatment of Gaucher disease (1999) Blood, 93, pp. 2807-2816; 
Rasmussen, J., Barsomian, G., Bergh, M., Enzymatically active recombinant glucocerebrosidase. U.S. patent 6451600. 17 September 2002Elstein, D., Itzchaki, M., Mankin, H.J., Skeletal involvement in Gaucher{\textquoteright}s disease (1997) Baillieres Clin Haematol, 10, pp. 793-816; 
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},
	month = {2005///},
	pages = {545 - 552},
	abstract = {Gaucher disease (GD) is caused by the decreased activity and/or stability of the lysosomal enzyme glucocerebrosidase (GCR). The available treatment consists in the intravenous administration of exogenous GCR, and is effective in reverting most of the symptoms. However, in terms of bone pathology, which is among the most disabling manifestations, a slow and incomplete response is observed, indicating that adjuvant therapies are necessary to consistently restore GCR activity in bone and accelerate regeneration. In this study, calcium alginate microspheres were analyzed as a vehicle for localized GCR delivery to bone. Results demonstrated that the entrapped enzyme retained full activity and exhibited a broader pH-dependent activity profile, compared to that of free-GCR, with improved stability at physiological pH. GCR release profile was established, and it was demonstrated that GCR could be released in a sustained manner. The biological behavior of the system was evaluated by analyzing the uptake of released GCR by GCR-deficient cells from GD patients, using different techniques: GCR activity measurements, radiolabeling, and cellulose acetate electrophoresis. Results demonstrated that GCR was internalized by cells significantly enhancing the residual enzymatic activity. To achieve an activity reconstitution level comparable to that obtained using free-GCR, only half of the dose was required with entrapped-GCR. {\textcopyright} 2005 Wiley Periodicals, Inc.},
	keywords = {Alginates, alginic acid, article, Bone, bone disease, Bone Regeneration, Calcium alginate, Calcium Compounds, catalysis, Cell culture, Cells, Cultured, controlled study, Drug Carriers, drug delivery system, enzyme activity, enzyme assay, enzyme replacement, Enzymes, Enzymes, Immobilized, fibroblast culture, Fibroblasts, Gaucher disease, Gaucher disease (GD), Glucocerebrosidase (GCR), Glucosylceramidase, Glucuronic Acid, Hexuronic Acids, human, human cell, Humans, iodine 125, Localized delivery, lysosome enzyme, lysosome storage disease, microsphere, Microspheres, Pathology, pH effects, physiology, recombinant enzyme, Therapeutic enzyme},
	isbn = {00219304 (ISSN)},
	url = {http://www.scopus.com/inward/record.url?eid=2-s2.0-24044465170\&partnerID=40\&md5=bbcee8568e0c0223975853a8177439c8},
	author = {Barrias, C.C. and Lamghari, M. and Granja, P.L. and S{\'a} Miranda, M.C. and Barbosa, M.A.}
}
@book {2426,
	title = {Effect of calcium phosphate addition to alginate microspheres: modulation of enzyme release kinetics and improvement of cell adhesion},
	series = {Key Engineering MaterialsKey Eng Mat},
	volume = {284-286},
	year = {2005},
	note = {Cited By (since 1996): 4Export Date: 21 November 2012Source: ScopusCODEN: KEMAELanguage of Original Document: EnglishCorrespondence Address: Barrias, C.C.; INEB - Institute de Engenharia Biom{\'e}dica, Laborat{\'o}rio de Biomaterials, R. Campo Alegre 823, 4150-180 Porto, Portugal; email: ccbarrias@ineb.up.ptReferences: Graopwski, G.A., Leslie, N., Wenstrup, R., (1998) Blood Rev, 12, p. 115;Rowley, J.A., Madlambayan, G., Mooney, D.J., (1999) Biomaterials, 20, p. 45; 
Dalby, M.J., Di Silvio, L., Harper, E.J., Bonfield, W., (2002) Biomaterials, 23, p. 569
},
	month = {2005///},
	pages = {689 - 692},
	abstract = {In this study, the addition of calcium phosphate powders to an alginate matrix was evaluated as a strategy to modulate enzyme release-kinetics from alginate microspheres and, simultaneously, to improve cell adhesion to the polymer. Pre-adsorption of the enzyme to the ceramic powders resulted in a more adequate release pattern. The ratio of ceramic-to-polymer had a pronounced effect on osteoblast adhesion to microspheres. Cells were only able to spread on microspheres with the highest percentage of ceramic (0.4 w/w using a 1.5\% w/v alginate solution).},
	keywords = {Alginate, Bioceramics, calcium phosphate, Calcium titanium phosphate, cell adhesion, Enzyme kinetics, Enzyme release, hydroxyapatite, Microspheres, Osteoblasts},
	isbn = {10139826 (ISSN)},
	url = {http://www.scopus.com/inward/record.url?eid=2-s2.0-20744433251\&partnerID=40\&md5=30ce8fc67d1e3e37074cc056f650b798},
	author = {Barrias, C.C. and Ribeiro, C.C. and Rodrigues, D. and Miranda, M.C.S. and Barbosa, M.A.}
}
@article {2450,
	title = {In vitro degradation behavior of a novel bioresorbable composite material based on PLA and a soluble CaP glass},
	journal = {Acta BiomaterialiaActa Biomater.},
	volume = {1},
	year = {2005},
	note = {Cited By (since 1996): 24Export Date: 21 November 2012Source: Scopusdoi: 10.1016/j.actbio.2005.03.004PubMed ID: 16701822Language of Original Document: EnglishCorrespondence Address: Ginebra, M.P.; Reference Centre for Bioengineering of Catalonia (CREBEC), Department of Materials Science, Universitat Polit{\`e}cnica de Catalunya, Av. Diagonal 647, 08028 Barcelona, Spain; email: maria.pau.ginebra@upc.esChemicals/CAS: calcium phosphate, 10103-46-5, 13767-12-9, 14358-97-5, 7758-87-4; polylactic acid, 26100-51-6; Bone Substitutes; calcium phosphate, 10103-46-5; Calcium Phosphates; Lactic Acid, 50-21-5; poly(lactic acid), 26100-51-6; PolymersManufacturers: Purac, NetherlandsReferences: Rokkanen, P., Bioabsorbable fixation in orthopaedic surgery and traumatology (2000) Biomaterials, 21, pp. 2607-2613;Middleton, J.C., Tipton, A.J., Synthetic biodegradable polymers as orthopedic devices (2000) Biomaterials, 21 (23), pp. 2335-2346; 
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},
	month = {2005///},
	pages = {411 - 419},
	abstract = {Poly(α-hydroxy acids), and in particular polylactic acid (PLA), are nowadays amongst the most used bioabsorbable materials. However, this polymer may not meet some application requirements due to inadequate mechanical properties and or its degradation characteristics. A possible strategy to tackle this problem is the incorporation of an inorganic phase into the polymeric matrix. In this work a new fully biodegradable composite material made with PLA and calcium phosphate soluble glass particles has been developed. The behaviour of the PLA/ glass composite has been analysed during its degradation in simulated physiological conditions by means of weight loss, molecular weight and thermal properties analysis and electron microscopy observation. The results showed that the incorporation of phosphate glass particles into the polymer significantly accelerated the degradation of the PLA and induced the formation of calcium phosphate precipitates at the composite surface. {\textcopyright} 2005 Acta Materialia Inc. Published by Elsevier Ltd. All rights reserved.},
	keywords = {Absorbable Implants, Absorption, article, biodegradability, Biodegradable composite, Biomimetic materials, Body fluids, Bone Regeneration, Bone Substitutes, calcium phosphate, Calcium phosphates, composite material, Degradation, electron microscopy, glass, In vitro degradation, in vitro study, Lactic Acid, materials testing, molecular weight, observation, Phosphate glass, polylactic acid, Polymers, precipitation, priority journal, reaction analysis, simulation, Solubility, Surface Properties, surface property, temperature, weight reduction},
	isbn = {17427061 (ISSN)},
	author = {Navarro, M. and Ginebra, M.P. and Planell, J.A. and Barrias, C.C. and Barbosa, M.A.}
}
@article {2454,
	title = {Polysaccharides as scaffolds for bone regeneration},
	journal = {ITBM-RBMITBM-RBM},
	volume = {26},
	year = {2005},
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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 {2460,
	title = {Proliferation, activity, and osteogenic differentiation of bone marrow stromal cells cultured on calcium titanium phosphate microspheres},
	journal = {Journal of Biomedical Materials Research - Part AJ. Biomed. Mater. Res. Part A},
	volume = {72},
	year = {2005},
	note = {Cited By (since 1996): 34Export Date: 21 November 2012Source: ScopusCODEN: JBMRCdoi: 10.1002/jbm.a.30217PubMed ID: 15543603Language of Original Document: EnglishCorrespondence Address: Barrias, C.C.; INEB, Instituto de Engenharia Biomedica, Laboratorio de Biomateriais, Rua do Campo Alegre, 823, 4150-180 Porto, Portugal; email: ccbarrias@ineb.up.ptChemicals/CAS: alkaline phosphatase, 9001-78-9; calcium phosphate, 10103-46-5, 13767-12-9, 14358-97-5, 7758-87-4; osteocalcin, 136461-80-8; titanium, 7440-32-6; Alkaline Phosphatase, EC 3.1.3.1; Calcium, 7440-70-2; Osteocalcin, 104982-03-8; titanium phosphate, 17017-57-1; Titanium, 7440-32-6References: Senuma, Y., Franceschin, S., Hilborn, J.G., Tissieres, P., Bisson, I., Frey, P., Bioresorbable microspheres by spinning disk atomization as injectable cell carrier: From preparation to in vitro evaluation (2000) Biomaterials, 21 (11), pp. 1135-1144;Hsu, F.Y., Chueh, S.C., Wang, Y.J., Microspheres of hydroxyapatite/ reconstituted collagen as supports for osteoblast cell growth (1999) Biomaterials, 20 (20), pp. 1931-1936; 
McGlohorn, J.B., Grimes, L.W., Webster, S.S., Burg, K.J., Characterization of cellular carriers for use in injectable tissue-engineering composites (2003) J Biomed Mater Res, 66 A (3), pp. 441-449; 
Wu, T.J., Huang, H.H., Lan, C.W., Lin, C.H., Hsu, F.Y., Wang, Y.J., Studies on the microspheres comprised of reconstituted collagen and hydroxyapatite (2004) Biomaterials, 25 (4), pp. 651-658; 
Barrias, C.C., Ribeiro, C.C., Barbosa, M.A., Adhesion and proliferation of human osteoblastic cells seeded on injectable hydroxyapatite microspheres (2004) Key Eng Mater, 254-256, pp. 877-880; 
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Ribeiro, C.C., Barrias, C.C., Barbosa, M.A., A novel route for the preparation of injectable ceramic porous microspheres for bone tissue engineering (2003) Ceramics, Cells and Tissues Annual Conferences: Bioceramic Surfaces Behavior in Vitro and in Vivo, pp. 228-232. , Ravaglioli A, Krajewski A, editors. Faenza: ISTEC-CNR editions; 
Ribeiro, C.C., Barrias, C.C., Barbosa, M.A., (2004) Preparation and Characterisation of Calcium-phosphate Microspheres for Biomedical Applications, , Submitted for publication; 
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Shu, R., McMullen, R., Baumann, M.J., McCabe, L.R., Hydroxyapatite accelerates differentiation and suppresses growth of MC3T3-E1 osteoblasts (2003) J Biomed Mater Res, 67 A (4), pp. 1196-1204; 
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Knabe, C., Berger, G., Gildenhaar, R., Klar, F., Zreiqat, H., The modulation of osteogenesis in vitro by calcium titanium phosphate coatings (2004) Biomaterials, 25, pp. 4911-4919
},
	month = {2005///},
	pages = {57 - 66},
	abstract = {In this study, the behavior of bone marrow stromal cells cultured on calcium titanium phosphate (CTP) microspheres was analyzed. Cell adhesion and proliferation were estimated by the neutral red assay and by total DNA quantification. Morphology and deposition of extracellular matrix were assessed by confocal laser scanning microscopy and/or scanning electron microscopy. The expression of the osteoblastic phenotype was evaluated by monitoring alkaline phosphatase activity and osteocalcin secretion. Results revealed that cells were able to attach and spread on the surface of CTP microspheres, and gradually grow into nearly confluent monolayers. Moreover, cells were able to bridge adjacent microspheres forming microsphere-cell clusters. Cells produced an abundant amount of fibrillar extracellular matrix that covered the substrate surface. Alkaline phosphatase activity peaked around days 7-14 and then decreased until day 21. Cells secreted osteocalcin, with higher levels being detected at day 14 than at day 21. Taken together, these results suggest that CTP microspheres are appropriate scaffolds for the growth and differentiation of cells along the osteoblastic lineage. {\textcopyright} 2004 Wiley Periodicals, Inc.},
	keywords = {alkaline phosphatase, animal cell, Animals, article, Bioassay, Bone, bone marrow cell, Bone Marrow Cells, bone marrow stromal cell, Bone marrow stromal cells, Calcium, Calcium Compounds, calcium phosphate, Calcium titanium phosphate, cell adhesion, Cell culture, cell differentiation, cell growth, cell lineage, cell proliferation, cell structure, Confluent monolayers, confocal laser microscopy, Confocal laser scanning microscopy, controlled study, Deposition, DNA determination, enzyme activity, Enzyme kinetics, extracellular matrix, Microscopy, Electron, microsphere, Microspheres, nonhuman, osteoblast, osteocalcin, Osteogenesis, Osteogenic differentiation, phenotype, protein expression, protein secretion, rat, Rats, Scanning electron microscopy, Stromal Cells, surface property, Tissue culture, Tissue engineering, Titanium, unclassified drug},
	isbn = {00219304 (ISSN)},
	url = {http://www.scopus.com/inward/record.url?eid=2-s2.0-10444223137\&partnerID=40\&md5=ab1b84ba6dd86274c0102567dffe1418},
	author = {Barrias, C.C. and Ribeiro, C.C. and Lamghari, M. and S{\'a} Miranda, C. and Barbosa, M.A.}
}
@article {2443,
	title = {Recombinant glucocerebrosidase uptake by Gaucher disease human osteoblast culture model},
	journal = {Blood Cells, Molecules, and DiseasesBlood Cells Mol. Dis.},
	volume = {35},
	year = {2005},
	note = {Cited By (since 1996): 3Export Date: 21 November 2012Source: ScopusCODEN: BCMDFdoi: 10.1016/j.bcmd.2005.07.009PubMed ID: 16125984Language of Original Document: EnglishCorrespondence Address: Lamghari, M.; INEB-Instituto de Engenharia Biom{\'e}dica, Laborat{\'o}rio de Biomateriais, R. Campo Alegre 823, 4150-180 Porto, Portugal; email: lamghari@ineb.up.ptChemicals/CAS: conduritol b epoxide, 6090-95-5; imiglucerase, 154248-97-2; iodine 125, 14158-31-7, 22822-81-7; conduritol epoxide, 6090-95-5; Glucosylceramidase, EC 3.2.1.45; Inositol, 6917-35-7; Recombinant ProteinsTradenames: cerezyme, GenzymeManufacturers: GenzymeReferences: Beutler, E., Grabowski, G., Gaucher disease (1995) The Metabolic and Molecular Bases of Inherited Disease, 2, pp. 2641-2661. , C.R. Scriber A.L. Beaudet W.S. Sly 7th ed. McGraw-Hill New York, NY;Beutler, E., Enzyme replacement in Gaucher disease (2004) PLoS Med., 1 (2), p. 21; 
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Barrias, C.C., Ribeiro, C.C., Lamghari, M., S{\'a} Miranda, C., Barbosa, M.A., Proliferation, activity, and osteogenic differentiation of bone marrow stromal cells cultured on calcium titanium phosphate microspheres (2005) J. Biomed. Mater. Res., 72 A (1), pp. 57-66; 
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},
	month = {2005///},
	pages = {348 - 354},
	abstract = {Bone lesions are a major cause of morbidity in Gaucher disease (GD) type I. Enzyme replacement therapy (ERT) has been successful in treating many symptoms of type I GD but skeletal response lags behind. Local exogenous glucocerebrosidase supplementation in bone lesions via a drug delivery system may overcome this limitation. Although local enzyme supplementation aims to target lipid-engorged macrophages (Gaucher Cells) in bone compartment, enzyme uptake by osteoblasts is not excluded. To investigate the ability of human osteoblasts to internalize recombinant glucocerebrosidase (rGCR), we have used an artificial GD human osteoblasts cell culture system. MG63 human osteoblasts were treated with conduritol B epoxide (CBE) to induce complete and prolonged inhibition of endogenous glucocerebrosidase activity of cells. rGCR uptake by glucocerebrosidase-inactivated osteoblasts was examined using                        125I-radiolabelling, Western blot analysis and measurement of glucocerebrosidase activity. Analysis of radiolabeled enzyme uptake by CBE treated osteoblasts showed 67.9\% of internalized protein in cell extract. Enzyme internalization was also observed by Western blot analysis where the amount of mature form of glucocerebrosidase protein recognized by the glucocerebrosidase antibody was increased following the administrations of rGCR. Moreover, enzymatic activity measurement showed 23.9\% of glucocerebrosidase activity of control cells. The rGCR internalization by MG63 osteoblast seems to be partially mediated by mannose receptors. These data provide evidence that MG63 human osteoblasts are able to internalize rGCR. {\textcopyright} 2005 Elsevier Inc. All rights reserved.
},
	keywords = {article, Cell culture, cell extract, Cells, Cultured, conduritol b epoxide, controlled study, drug activity, drug uptake, enzyme activity, enzyme antibody, Enzyme inhibition, Enzyme uptake, epoxide, Gaucher disease, Glucocerebrosidase, Glucosylceramidase, human, human cell, Humans, imiglucerase, Inositol, internalization, iodine 125, isotope labeling, mannose receptor, osteoblast, Osteoblasts, priority journal, Recombinant Proteins, unclassified drug, Western blotting},
	isbn = {10799796 (ISSN)},
	url = {http://www.scopus.com/inward/record.url?eid=2-s2.0-27744461413\&partnerID=40\&md5=575b4d8cc3f6e5b8e846cf6de71852b3},
	author = {Lamghari, M. and Barrias, C.C. and S{\'a} Miranda, C. and Barbosa, M.A.}
}
@article {2474,
	title = {Calcium phosphate-alginate microspheres as enzyme delivery matrices},
	journal = {BiomaterialsBiomaterials},
	volume = {25},
	year = {2004},
	note = {Cited By (since 1996): 92Export Date: 21 November 2012Source: ScopusCODEN: BIMADdoi: 10.1016/j.biomaterials.2003.11.028PubMed ID: 15046927Language of Original Document: EnglishCorrespondence Address: Ribeiro, C.C.; Laborat{\'o}rio de Biomateriais, INEB, Inst. de Engenharia Biom{\'e}dica, Rua do Campo Alegre 823, Porto 4150-180, Portugal; email: cribeiro@ineb.up.ptChemicals/CAS: Alginates; alginic acid, 9005-32-7; calcium phosphate, 10103-46-5; Calcium Phosphates; Drug Carriers; Enzymes, Immobilized; Glucosylceramidase, EC 3.2.1.45; Glucuronic Acid, 576-37-4; Hexuronic Acids; PowdersReferences: Griffith, L.G., Naughton, G., Tissue engineering: Current challenges and expanding opportunities (2002) Science, 295, pp. 1009-1014;Hench, L.L., Polak, J.M., Third-generation biomedical materials (2002) Science, 295, pp. 1014-1017; 
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},
	month = {2004///},
	pages = {4363 - 4373},
	abstract = {The present study concerns the preparation and initial characterisation of novel calcium titanium phosphate-alginate (CTP-alginate) and hydroxyapatite-alginate (HAp-alginate) microspheres, which are intended to be used as enzyme delivery matrices and bone regeneration templates. Microspheres were prepared using different concentrations of polymer solution (1\% and 3\%w/v) and different ceramic-to-polymer solution ratios (0.1, 0.2 and 0.4w/w). Ceramic powders were characterised using X-ray diffraction, laser granulometry, Brunauer, Emmel and Teller (BET) method for the determination of surface area, zeta potential and Fourier transform infrared spectroscopy (FT-IR). Alginate was characterised using high performance size exclusion chromatography. The methodology followed in this investigation enabled the preparation of homogeneous microspheres with a uniform size. Studies on the immobilisation and release of the therapeutic enzyme glucocerebrosidase, employed in the treatment of Gaucher disease, were also performed. The enzyme was incorporated into the ceramic-alginate matrix before gel formation in two different ways: pre-adsorbed onto the ceramic particles or dispersed in the polymeric matrix. The two strategies resulted in distinct release profiles. Slow release was obtained after adsorption of the enzyme to the ceramic powders, prior to preparation of the microspheres. An initial fast release was achieved when the enzyme and the ceramic particles were dispersed in the alginate solution before producing the microspheres. The latter profile is very similar to that of alginate microspheres. The different patterns of enzyme release increase the range of possible applications of the system investigated in this work. {\textcopyright} 2003 Elsevier Ltd. All rights reserved.},
	keywords = {Absorption, Adsorption, Alginate, Alginates, article, Biomaterials, Bone, Calcium Compounds, Calcium phosphates, Calcium titanium phosphate, calcium titanium phosphate alginate, ceramics, Drug Carriers, Drug Delivery Systems, Enzyme delivery, Enzyme-delivery, Enzymes, Enzymes, Immobilized, Fourier transform infrared spectroscopy, Glucosylceramidase, Glucuronic Acid, Hexuronic Acids, High performance liquid chromatography, hydroxyapatite, hydroxyapatite alginate, laser, methodology, microsphere, Microspheres, Molecular Conformation, Particle Size, Powders, priority journal, surface property, unclassified drug, X ray diffraction},
	isbn = {01429612 (ISSN)},
	url = {http://www.scopus.com/inward/record.url?eid=2-s2.0-1642331315\&partnerID=40\&md5=217590793385cbaff039a6b260d3d85c},
	author = {Ribeiro, C.C. and Barrias, C.C. and Barbosa, M.A.}
}
