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The effect of hyaluronan on osteoblast proliferation and differentiation in rat calvarial‐derived cell cultures
Author(s) -
Huang L.,
Cheng Y. Y.,
Koo P. L.,
Lee K. M.,
Qin L.,
Cheng J. C. Y.,
Kumta S. M.
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.10535
Subject(s) - alkaline phosphatase , osteocalcin , osteoblast , cell growth , cellular differentiation , hyaluronic acid , mesenchymal stem cell , extracellular matrix , cell adhesion , morphogenesis , microbiology and biotechnology , cell , cell culture , biology , biochemistry , chemistry , enzyme , in vitro , anatomy , gene , genetics
Hyaluronan (or hyaluronic acid, HA) is an essential component of extracellular matrices. It interacts with other macromolecules and plays a predominant role in tissue morphogenesis, cell migration, differentiation, and adhesion. The cell signaling functions of HA are mediated through the CD‐44 receptor and are dependent upon the molecular weight of the polymer. We hypothesized that an HA of appropriate molecular weight alone in optimal concentration may induce osteoblast differentiation and bone formation. Enzyme‐digested calvarial‐derived mesenchymal cells from 2‐day‐old newborn rats were cultured with the addition of HA of three different molecular weights (2300, 900, and 60 kDa). We added, 0.5, 1.0, and 2.0 mg/mL HA for each molecular weight to the medium at the first plating of cells. After 7 to 20 days in culture, cell proliferation and differentiation were evaluated by measuring thymidine incorporation, alkaline phosphatase activity, and osteocalcin gene expression. The effects of HA on bone formation were examined by using Alizarin red staining for mineralization. The results showed that low molecular weight HA (60 kDa) significantly stimulated cell growth, increased osteocalcin mRNA expression in a dose‐dependent manner, but showed no apparent effects on alkaline phosphatase activity and bone mineralization. On the other hand, high‐weight HA (900 and 2300 kDa) significantly increased all the parameters examined, particularly alkaline phosphatase activity, in a dose‐dependent manner and stimulated cell mineralization to 126% and 119% of the controls, respectively, in the 1.0 mg/mL dose. Our findings suggest that HA has a molecular weight‐specific and dose‐specific mode of action that may enhance the osteogenic and osteoinductive properties of bone graft materials and substitutes due to its stimulatory effects on osteoblasts. © 2003 Wiley Periodicals, Inc. J Biomed Mater Res 66A: 880–884, 2003