Oscillatory perfusion seeding and culturing of osteoblast-like cells on porous beta-tricalcium phosphate scaffolds

被引:33
|
作者
Du, Dajiang [1 ]
Furukawa, Katsuko [1 ,2 ]
Ushida, Takashi [3 ]
机构
[1] Univ Tokyo, Grad Sch Engn, Dept Mech Engn, Biomed Engn Lab,Bunkyo Ku, Tokyo 1138656, Japan
[2] Univ Tokyo, Grad Sch Engn, Dept Bioengn, Bunkyo Ku, Tokyo 1138656, Japan
[3] Univ Tokyo, Sch Med, Ctr Dis Biol & Integrat Med, Div Biomed Mat & Syst,Bunkyo Ku, Tokyo 1130033, Japan
关键词
flow perfusion bioreactor; bone tissue engineering; oscillatory fluid flow; cell culture techniques; porous ceramic scaffolds;
D O I
10.1002/jbm.a.31641
中图分类号
R318 [生物医学工程];
学科分类号
0831 ;
摘要
Perfusion culture systems have proven to be effective bioreactors for constructing tissue engineered bone in vitro, but existing circuit-based perfusion systems are complicated and costly for conditioned culture due to the large medium volume required. A compact perfusion system for artificial bone fabrication using oscillatory flow is described here. Mouse osteoblast-like MC 3T3-E1 cells were seeded at 1.5 X 10(6) cells/100 mu L and cultured for 6 days in porous ceramic beta-tricalcium phosphate scaffolds (10 mm in diameter, 8 mm in height) by only 1.5 mL culture media per scaffold. The seeding efficiency, cell proliferation, distribution and viability, and promotion of early osteogenesis by both a static and an oscillatory perfusion method were evaluated. The oscillatory perfusion method generated higher seeding efficiency, alkaline phosphatase activity, and scaffold cellularity (by DNA content) after 6 days of culture. Stereo-microscopic observation of 3(4,5-dimethylthiazol-2-yl)-2,5-diphenyltetrazolium bromide staining and Calcein-AM/ propidium iodide double, staining also demonstrated homogeneous seeding, proliferation, and viability of cells throughout the scaffolds in the oscillatory perfusion system. By contrast, the static culture yielded polarized seeding and proliferation favoring the outer and upper scaffold surfaces, with only dead cells in the center of the scaffolds. Thus, these results suggest that the oscillatory flow condition not only allow a better seeding efficiency and homogeneity, but also facilitates uniform culture and early osteogenic differentiation. The oscillatory perfusion system could be a simple and effective bioreactor for bone tissue engineering. (c) 2007 Wiley Periodicals, Inc.
引用
收藏
页码:796 / 803
页数:8
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