Premium
Osteoblastic response to collagen scaffolds varied in freezing temperature and glutaraldehyde crosslinking
Author(s) -
Chen DaiChian,
Lai YuLin,
Lee ShyhYuan,
Hung ShanLing,
Chen HenLi
Publication year - 2007
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.30932
Subject(s) - glutaraldehyde , von kossa stain , osteocalcin , alkaline phosphatase , calcium , osteoblast , sponge , mineralization (soil science) , materials science , biophysics , microbiology and biotechnology , chemistry , biochemistry , biology , in vitro , botany , chromatography , organic chemistry , nitrogen , metallurgy , enzyme
Collagen sponges are widely used scaffolds in bone engineering. To form bone, the osteoblastic cells undergo proliferation, differentiation, and mineralization stages in the scaffold. Crosslinking and freezing temperature are two important variables in fabricating collagen sponges. The purpose of this study was to examine the osteoblastic responses to collagen sponges prepared with or without glutaraldehyde crosslinking at different freezing temperatures (−20°C or −80°C). MC3T3‐E1 osteoblastic cells were cultured in differently prepared sponges. Osteoblastic responses examined included cell numbers, osteocalcin expression, and calcium deposition. Cell numbers were measured by DNA content. Osteocalcin expression was determined by RT‐PCR and real‐time RT‐PCR. Calcium deposition was assayed by ortho‐cresophthalein complexone method and von Kossa stain. The osteoblastic cells grown in all collagen sponges did not show apparent signs of cytotoxicity. Collagen sponges differed in freezing temperatures resulted in similar osteoblastic responses. Glutaraldehyde‐crosslinked sponges demonstrated less cell‐mediated contraction and more cell numbers at day 7 ( p < 0.005). However, they showed lower osteocalcin expression at day 7 ( p < 0.05) and less calcium deposition at day 21 ( p < 0.001). In summary, different freezing temperatures played a minor role in osteoblastic responses. Glutaraldehyde crosslinking process, though improved the dimensional stability of collagen sponges, might compromise the osteoblastic differentiation and mineralization. © 2006 Wiley Periodicals, Inc. J Biomed Mater Res, 2007