聚乳酸-壳聚糖纤维/羟基磷灰石-硅酸钙复合支架材料的细胞相容性

背景：实践证明，有机和无机材料单独应用都不是理想的支架材料。聚乳酸具有良好的生物相容性、生物降解性和生物吸收性，聚乳酸类复合材料将成为21世纪最重要的生物复合材料之一。目的：观察复合支架材料聚乳酸-壳聚糖纤维/羟基磷灰石-硅酸钙对成骨细胞黏附、增殖、分化的影响。方法：取新生24 h内Wistar大鼠的颅盖骨，采用改良胶原酶消化法进行成骨细胞原代培养。通过倒置相差显微镜、苏木 精-伊红染色、碱性磷酸酶染色、钙结节茜素红染色对获得的细胞进行生物学特性的观察与鉴定。然后将第3代细胞与聚乳酸/壳聚糖纤维、聚乳酸-壳聚糖纤维/硅酸钙、聚乳酸-壳聚糖纤维/羟基磷灰石-硅酸钙三种支架材料体外复合培养。培养3，6，9 d后，采用倒置相差显微镜观察材料周边的细胞形态，通过碱性磷酸酶活性测定法和MTT法观察3种支架材料对细胞分化、增殖的影响。结果与结论：三种材料均有利于成骨细胞的黏附、生长、分化、增殖，而聚乳酸-壳聚糖纤维/羟基磷灰石-硅酸钙复合支架材料较聚乳酸/壳聚糖纤维、聚乳酸-壳聚糖纤维/硅酸钙支架材料促进成骨细胞的分化增殖效果更好，证实其生物相容性好，有望成为一种新型的骨组织工程支架材料。关键词：复合支架；细胞相容性；生物材料；成骨细胞；组织工程支架材料；骨组织工程doi:10.3969/j.issn.1673-8225.2010.08.016

Compatibility of osteoblasts on the polylactic acid-chitosan fiber/hydroxyapatite-calcium silicate composite scaffold material

BACKGROUND: Practice has proved that organic material and inorganic materials used alone are not ideal scaffold materials. Polylactic acid (PLA) possessing excellent biocompatibility, degradability and absorbability, PLAs composites will be one of the most important biocomposite in the 21st century. OBJECTIVE: To observe the effects of PLA-chitosan fiber (CF)/ hydroxyapatite-calcium silicate (HA-CS) on adhesion, proliferation and differentiation of osteoblasts.METHODS: The rat osteoblasts were obtained from the cranium of newborn Wistar rats within 24 hours, and primarily cultured using modified collagenase digestion. The cells were generated and their biological characteristic was examined by inverted phase-contrast microscope, hematoxylin-eosin staining, alkaline phosphatase (ALP) staining and mineralized nodules staining. Then the cells at passage 3 were co-cultured with PLA-CF, PLA-CF/CS and PLA-CF/HA-CS in vitro. At 3, 6 and 9 days of the culture, cell morphology was observed by inverted phase contrast microscopy. In addition, MTT assay and ALP activity test were used to observe the effects of three kinds of materials on cell differentiation and proliferation. RESULTS AND CONCLUSION: Osteoblasts attached on all three scaffold materials can adhere, grow, differentiate and proliferate. The effect of three materials on cell activity was PLA-CF/HA-CS>PLA-CF/CS>PLA-CF. The composite scaffold PLA-CF/HA-CS has a good compatibility, indicating that the material has a great potential for application in bone tissue engineering.