



@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; 
Khor, E., (2001) Chitin: Fulfilling a Biomaterials Promise, , Oxford, UK: Elsevier Science Ltd; 
Vandevord, P.J., Hwt, M., 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; 
Tomihata, K., Ikada, Y., In vitro and in vivo degradation of films of chitin and its deacetylated derivatives (1997) Biomaterials, 18, pp. 567-575; 
Rao, S.B., Sharma, C.P., Sterilization of chitosan: Implications (1995) J Biomater Appl, 10, pp. 136-143; 
Muzzarelli, R.A.A., Zucchini, C., Ilari, P., Pugnaloni, A., Belmonte, M.M., Biagini, G., Castaldini, C., Osteoconductive properties of methylpyrrolidinone chitosan in an animal-model (1993) Biomaterials, 14, pp. 925-929; 
Lee, J.Y., Nam, S.H., Im, S.Y., Park, Y.J., Lee, Y.M., Seol, Y.J., Chung, C.P., Lee, S.J., Enhanced bone formation by controlled growth factor delivery from chitosan-based biomaterials (2002) J Control Release, 78, pp. 187-197; 
Zhao, F., Yin, Y.J., Lu, W.W., Leong, J.C., Zhang, W.J., Zhang, J.Y., Zhang, M.F., Yao, K.D., Preparation and histological evaluation of biomimetic three-dimensional hydroxyapatite/chitosan-gelatin network composite scaffolds (2002) Biomaterials, 23, pp. 3227-3234; 
Nettles, D.L., Elder, S.H., Gilbert, J.A., Potential use of chitosan as a cell scaffold material for cartilage tissue engineering (2002) Tissue Eng, 8, pp. 1009-1016; 
Zhang, Y., Zhang, M.Q., Calcium phosphate/chitosan composite scaffolds for controlled in vitro antibiotic drug release (2002) J Biomed Mater Res, 62, pp. 378-386; 
Prasitsilp, M., Jenwithisuk, R., Kongsuwan, K., Damrongchai, N., Watts, P., Cellular responses to chitosan in vitro: The importance of deacetylation (2000) J Mater Sci Mater Med, 11, pp. 773-778; 
Chatelet, C., Damour, O., Domard, A., Influence of the degree of acetylation on some biological properties of chitosan films (2001) Biomaterials, 22, pp. 261-268; 
Dhiman, H.K., Ray, A.R., Panda, A.K., Characterization and evaluation of chitosan matrix for in vitro growth of MCF-7 breast cancer cell lines (2004) Biomaterials, 25, pp. 5147-5154; 
Amaral, I.F., Lamghari, M., Sousa, S.R., Sampaio, P., Barbosa, M.A., Rat bone marrow stromal cells osteogenic differentiation and fibronectin adsorption on chitosan membranes: The effect of the degree of acetylation (2005) J Biomed Mater Res Part A, 75 A, pp. 387-397; 
Cukierman, E., Pankov, R., Stevens, D.R., Yamada, K.M., Taking cell-matrix adhesions to the third dimension (2001) Science, 294, pp. 1708-1712; 
Brugnerotto, J., Lizardi, J., Goycoolea, F.M., Arguelles-Monal, W., Desbrieres, J., Rinaudo, M., An infrared investigation in relation with chitin and chitosan characterization (2001) Polymer, 42, pp. 3569-3580; 
Mima, S., Miya, M., Iwamoto, R., Yoshikawa, S., Highly deacetylated chitosan and its properties (1983) J Appl Polym Sci, 28, pp. 1909-1917; 
Aiba, S., Studies on Chitosan: 1. Determination of the degree of N-acetylation of chitosan by ultraviolet spectrophotometry and gel-permeation chromatography (1986) Int J Biol Macromol, 8, pp. 173-176; 
Hirano, S., Ohe, Y., Ono, H., Selective N-acylation of chitosan (1976) Carbohydr Res, 47, pp. 315-320; 
Baxter, A., Dillon, M., Taylor, K.D.A., Roberts, G.A.F., Improved method for Ir determination of the degree of N-acetylation of chitosan (1992) Int J Biol Macromol, 14, pp. 166-169; 
Terbojevich, M., Cosani, A., Muzzarelli, R.A.A., Molecular parameters of chitosans depolymerized with the aid of papain (1996) Carbohydr Polym, 29, pp. 63-68; 
Zhang, R.Y., Ma, P.X., Poly(alpha-hydroxyl acids) hydroxyapatite porous composites for bone-tissue engineering. I. Preparation and morphology (1999) J Biomed Mater Res, 44, pp. 446-455; 
Mosmann, T., Rapid colorimetric assay for cellular growth and survival: Application to proliferation and cytotoxicity assays (1983) J Immunol Methods, 65, pp. 55-63; 
Ashton, B.A., Abdullah, F., Cave, J., Williamson, M., Sykes, B.C., Couch, M., Poser, J.W., Characterization of cells with high alkaline-phosphatase activity derived from human-bone and marrow - Preliminary assessment of their osteogenicity (1985) Bone, 6, pp. 313-319; 
Kurita, K., Controlled functionalization of the polysaccharide chitin (2001) Prog Polym Sci, 26, pp. 1921-1971; 
Wang, W., Bo, S.Q., Li, S.Q., Qin, W., Determination of the Mark-Houwink equation for chitosans with different degrees of deacetylation (1991) Int J Biol Macromol, 13, pp. 281-285; 
Roberts, G.A.F., (1992) Chitin Chemistry, , Houndmills: The Macmillan Press Ltd; 
Ueda, H., Hong, L., Yamamoto, M., Shigeno, K., Inoue, M., Toba, T., Yoshitani, M., Shimizu, Y., Use of collagen sponge incorporating transforming growth factor-beta 1 to promote bone repair in skull defects in rabbits (2002) Biomaterials, 23, pp. 1003-1010; 
Hanthamrongwit, M., Grant, M.H., Wilkinson, R., Confocal laser-scanning microscopy (Clsm) for the study of collagen sponge microstructure (1994) J Biomed Mater Res, 28, pp. 213-216; 
Zinger, O., Anselme, K., Denzer, A., Habersetzer, P., Wieland, M., Jeanfils, J., Hardouin, P., Landolt, D., Time-dependent morphology and adhesion of osteoblastic cells on titanium model surfaces featuring scale-resolved topography (2004) Biomaterials, 25, pp. 2695-2711; 
Eggli, P.S., Muller, W., Schenk, R.K., Porous hydroxyapatite and tricalcium phosphate cylinders with 2 different pore-size ranges implanted in the cancellous bone of rabbits - A comparative histomorphometric and histologic-study of bony ingrowth and implant substitution (1988) Clin Orthop, 232, pp. 127-138; 
Madihally, S.V., Matthew, H.W.T., Porous chitosan scaffolds for tissue engineering (1999) Biomaterials, 20, pp. 1133-1142; 
Madihally, S.V., Frattaroli, T.L., Matthew, H.W.T., (1999) In Vivo Colonization of Chitosan Based Porous Scaffolds, p. 172. , Providence, RI: Transactions from the 25th Annual Meeting of the Society for Biomaterials, Providence, RI; 
Zhang, Y., Zhang, M.Q., Synthesis and characterization of macroporous chitosan/calcium phosphate composite scaffolds for tissue engineering (2001) J Biomed Mater Res, 55, pp. 304-312; 
Chung, T.W., Yang, J., Akaike, T., Cho, K.Y., Nah, J.W., Kim, S.I., Cho, C.S., Preparation of alginate/galactosylated chitosan scaffold for hepatocyte attachment (2002) Biomaterials, 23, pp. 2827-2834; 
Masi, L., Franchi, A., Santucci, M., Danielli, D., Arganini, L., Giannone, V., Formigli, L., Beghe, F., Adhesion, growth, and matrix production by osteoblasts on collagen substrata (1992) Calcif Tissue Int, 51, pp. 202-212; 
Wiesmann, H.P., Nazer, N., Klatt, C., Szuwart, T., Meyer, U., Bone tissue engineering by primary osteoblast-like cells in a monolayer system and 3-dimensional collagen gel (2003) J Oral Maxillofac Surg, 61, pp. 1455-1462; 
Henriksen, I., Green, K.L., Smart, J.D., Smistad, G., Karlsen, J., Bioadhesion of hydrated chitosans: An in vitro and in vivo study (1996) Int J Pharmaceut, 145, pp. 231-240; 
Lebaron, R.G., Athanasiou, K.A., Extracellular matrix cell adhesion peptides: Functional applications in orthopedic materials (2000) Tissue Eng, 6, pp. 85-103; 
Cuy, J.L., Beckstead, B.L., Brown, C.D., Hoffman, A.S., Giachelli, C.M., Adhesive protein interactions with chitosan: Consequences for valve endothelial cell growth on tissue-engineering materials (2003) J Biomed Mater Res Part A, 67 A, pp. 538-547; 
Keselowsky, B.G., Collard, D.M., Garcia, A.J., Surface chemistry modulates fibronectin conformation and directs integrin binding and specificity to control cell adhesion (2003) J Biomed Mater Res Part A, 66 A, pp. 247-259; 
Webb, K., Hlady, V., Tresco, P.A., Relationships among cell attachment, spreading, cytoskeletal organization, and migration rate for anchorage-dependent cells on model surfaces (2000) J Biomed Mater Res, 49, pp. 362-368; 
Anselme, K., Bigerelle, M., Noel, B., Dufresne, E., Judas, D., Lost, A., Hardouin, P., Qualitative and quantitative study of human osteoblast adhesion on materials with various surface roughnesses (2000) J Biomed Mater Res, 49, pp. 155-166; 
Domard, A., Chitosan interactions (1997) Advances in Chitin Science, pp. 410-420. , Domard A, Roberts GAF, Varum KM, editors. Lyon, France: Jacques Andr{\'e} Publisher;
},
	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.}
}
