成骨细胞与BC/PLA/HA复合支架材料体外培养的初步结果

目的探讨成骨细胞在细菌纤维素/聚乳酸/羟基磷灰石复合支架中的生长和增殖情况。方法将成骨细胞以5×104/ml的密度接种于细菌纤维素(BC)、细菌纤维素/聚乳酸(BC/PLA)、细菌纤维素/聚乳酸/羟基磷灰石(BC/PLA/HA)三种支架中,进行体外培养。采用MTT比色法测定1、3、5、7 d的成骨细胞增殖情况;碱性磷酸酶活性测定法检测3、6、9 d的碱性磷酸酶活性变化,并通过扫描电镜观察细胞在复合支架中的形态变化。结果实验组与对照组比较1、3、5、7 d细胞的增殖量(OD值)有统计学差异(p<0.05),其中BC/PLA/HA组的OD值高于其他实验组;第3 d至第9 d三组细胞的ALP分泌量均呈上升趋势,各组间比较均有统计学差异(p<0.05);复合培养6d SEM观察可见三组材料中均有细胞的粘附,BC/PLA/HA组的细胞生长状态好于其他两组材料。结论 BC/PLA/HA较BC/PLA和BC有利于成骨细胞的增殖和生长。

Preliminary results of culture in vitro of osteoblasts on BC/PLA/HA composite scaffold materials

Objective To explore the growth and proliferation of osteoblasts in the bacterial cellulose/polylactic acid/hydroxyapatite composite scaffolds.Methods Osteoblasts growing to a density of 5×104/ml were seeded on BC,BC/PLA,BC/PLA/HA composite scaffolds,cultured in vitro.By MTT colorimetry assay of osteoblast proliferation in 1,3,5,7 days;Alkaline phosphatase activity assay to measure the change of alkaline phosphatase activity in 3,6,9 days.By Scanning electron microscopy to observe the change of osteoblasts morphology on the scaffold materials.Results Experimental group and control group compare the amount of cell proliferation（OD） in 1,3,5,7 days.have significant differences.BC/PLA/HA group OD value higher than the other experimental groups.All cells of the three groups showed an increasing tendency of ALP secretion from 3 days to 9 days of culture（P0.05）.Electron microscopic observation showed an adhesion of the cultured osteoblasts in all the 3 groups to the scaffold materials on the 6th day of culture,but the growth of osteoblasts cultured on the BC/PLA/HA composite material was better than that of the other two groups.