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Mineralization of matrix vesicles isolated from a human osteosarcoma cell line in culture with water‐soluble chitosan‐containing medium
Author(s) -
Yamada Shizuka,
Ohara Naoko,
Hayashi Yoshihiko
Publication year - 2003
Publication title -
journal of biomedical materials research part a
Language(s) - English
Resource type - Journals
SCImago Journal Rank - 0.849
H-Index - 150
eISSN - 1552-4965
pISSN - 1549-3296
DOI - 10.1002/jbm.a.10009
Subject(s) - chitosan , alkaline phosphatase , collagenase , incubation , mineralization (soil science) , osteoblast , in vitro , fetal bovine serum , cell culture , vesicle , microbiology and biotechnology , biochemistry , biology , chemistry , materials science , enzyme , membrane , genetics , ecology , soil water
Chitosan is a natural bioactive material. Although it has been reported that chitosan promotes osteogenesis in bone lesions, little is known about how chitosan modulates this process. The present study was designed to investigate the effect of water‐soluble chitosan relative to initiation of biologic mineralization, especially in the matrix‐vesicles‐(MVs) mediated process in vitro . A human osteoblastic cell line (NOS‐1) was used. After 3 days of incubation, the number of cells and alkaline phosphatase (ALP) activity increased significantly in the chitosan group. RT‐PCR analysis revealed that chitosan induced an increase in the expression of bone morphogenetic protein‐2 mRNA after 7 days of incubation. MVs were isolated from NOS‐1 cells using a collagenase digestion and ultracentrifugation method. ALP activity of MVs isolated from chitosan‐supplemented cells was significantly higher than that of the control group. Furthermore, isolated MVs were incubated in medium supplemented with Na‐β‐glycerophosphate without fetal bovine serum. Needle‐like crystals were observed in association with MVs after 24 h of incubation. These needle‐like crystals were densely accumulated in the chitosan group. The present findings suggest that water‐soluble chitosan would promote osteoblast proliferation and differentiation and may be useful for the acceleration of initial biologic mineralization. © 2003 Wiley Periodicals, Inc. J Biomed Mater Res 66A: 500–506, 2003