壳聚糖-明胶网络/羟基磷灰石复合材料支架的研究--成骨细胞培养

目的　研究大鼠颅骨成骨细胞在壳聚糖 -明胶网络 /羟基磷灰石 (CS- Gel/ HA)复合材料支架上的生长情况。方法　将原代培养大鼠颅骨成骨细胞第 3代 ,密度为 1.0 1× 10 6 / m l悬液 ,种植于孔隙率分别为85 .2 0 %、90 .4 0 %和 95 .80 %的 CS- Gel/ HA支架材料中 ,利用细胞计数法检测种植后 3天 ,1、2及 3周的细胞增殖曲线 ,采用 HE和 von Kossa染色方法观察细胞生长、骨样组织形成和矿化沉积情况。结果　大鼠颅骨成骨细胞在孔隙率为 85 .2 0 %的支架中增殖较慢 ,在孔隙率为 90 .4 0 %和 95 .80 %的 CS- Gel/ HA复合材料支架上生长良好 ,增殖较快 ,周围分泌有大量细胞外基质 ,3周时局部已出现骨样组织 ,且细胞 /支架结构物有利于钙质沉积。结论　CS- Gel/ HA复合材料支架有望成为培养自体成骨细胞的材料 ,以重建新的骨组织

TISSUE ENGINEERING STUDY ON CHITOSAN-GELATIN / HYDROXYAPATITE COMPOSITE SCAFFOLDS--OSTEOBLASTS CULTURE

OBJECTIVE: To investigate the behavior of rat calvarial osteoblasts cultured on chitosan-gelatin/hydroxyapatite (CS-Gel/HA) composite scaffolds. METHODS: The rat calvarial osteoblasts (the 3rd passage) were seeded at a density of 1.01 x 10(6) cells/ml onto the CS-Gel/HA composite scaffolds having porosity 85.20%, 90.40% and 95.80%. Cell number was counted after cultured for 3 days, 1 week, 2 weeks and 3 weeks. Cell proliferation, bone-like tissue formation, and mineralization were separately detected by HE, von Kossa histological staining techniques. RESULTS: The CS-Gel/HA composite scaffolds supported the attachment of seeded rat calvarial osteoblasts. Cells proliferated faster in scaffold with higher porosity 90.40% and 95.80% than scaffold with lower porosity 85.20%. The osteoblasts/scaffold constructs were feasible for mineral deposition, and bone-like tissue formation in 3 weeks. CONCLUSION: This study suggests the feasibility of using CS-Gel/HA composite scaffolds for bone tissue engineering.