三维支架材料外消旋聚乳酸复合藻酸钙的生物相容性观察

Biocompatibility investigation of three-dimensional porous Poly-D- Lactide/Calcium Alginate Gel scaffolds

目的:观察外消旋聚乳酸(PDLLA)复合藻酸钙(CAG)制备的三维支架材料的组织相容性、细胞相容性,为进一步改善材料性能提供实验依据。方法:体外大鼠成骨细胞与PDLLA、PDLLA/CAG支架材料联合培养,倒置显微镜、环境扫描电镜观察细胞黏附情况,MTT法分析细胞增殖情况;将支架材料植入大鼠肌袋中,观察局部伤口情况,术后2,4,8,12周行组织学观察。结果:复合支架材料在细胞贴壁时间和细胞活性方面均较单纯支架优良;两种支架材料在体内均无明显炎症反应。MTT法测定结果显示:随着培养时间的延长,各组细胞数量都明显增加,各时间点PDLLA组细胞数高于对照组,差异有显著性意义(t=-285.091,-154.681,-72.794,-81.306,P<0.01);PDLLA/CAG组细胞数高于PDLLA组,差异有显著性意义(t=-297.156,-167.687,-84.900,-103.103,P<0.01)。结论:PDLLA/CGA是一种具有良好生物相容性的支架材料,有望在骨组织工程中得到广泛应用。

AIM:To investigate the tissue compatibility and cell compatibility of the three dimensional porous Poly DL Lactide (PDLLA)/Calcium Alginate Gel (CAG) scaffold and provide an experimental reference for improving the constitution of composite material. METHODS:The materials of PDLLA and PDLLA/CAG were cultured in vitro with rat osteoblasts. The cell morphology and adhesion was examined by inverted microscope and environmental scanning electron microscope, and the cell proliferation was detected by MTT reduction assay. The scaffolds were embedded in rat's muscle tissue and incisions were observed.The specimens were harvested in 2, 4, 8 and 12 weeks respectively and examined by histology to evaluate the tissue response. RESULTS:The results showed that the cells on PDLLA/CAG were more favorable to the adhesion and activity than those on PDLLA. The PDLLA/CAG and PDLLA had no apparent inflammatory reaction in vivo. MTT assay showed that the number of cells in each group was significantly increased along with the cultured time prolonged; the number of cells in the PDLLA group was higher than that of the control group at different intervals, with a significant difference (t=- 285.091,- 154.681,- 72.794,- 81.306,P< 0.01); the number of cells in the PDLLA/CAG group was higher than that of the PDLLA group at different intervals, with a significant difference( t=- 297.156,- 167.687,- 84.900,- 103.103,P< 0.01) .