新型胶原膜体外生物相容性研究

Study on biocompatibility of a new bovine tendon collagen membrane in vitro

目的评估新型胶原膜的体外生物相容性,包括浸提液对细胞毒性影响和细胞与胶原膜表面的生物相容性。方法培养人牙周成纤维细胞(HPLFs),采用细胞计数试剂盒CCK-8实验评价新型胶原膜4种浸提液浓度(100%、50%、25%、12.5%)的细胞毒性;并将HPLFs与新型胶原膜复合培养,倒置相差显微镜、扫描电子显微镜及组织学观察细胞在胶原膜上的形态学变化。结果新型胶原膜不同浓度浸提液第1、3、5、7天的CCK-8检测值与空白对照组差异无统计学意义(P>0.05),细胞毒级评分为0级或1级。胶原膜上HPLFs黏附数目低于培养板上的黏附数,差异有统计学意义(P<0.01)。HPLFs在胶原膜上增殖、迁移,分泌大量的细胞外基质,复层生长,形成纤维组织结构。结论新型胶原膜无明显细胞毒性,具有良好的细胞相容性,初步具备了生物屏障膜的基本性能。

Objective To evaluate the biocompatibility of a new type of collagen membrane prepared from bovine tendon in vitro by cytotoxicity and cytocompatibility.Methods The cyctotoxic effects of four different concentrations of leaching liquor(100%,50%,25%,and 12.5%) prepared from collagen membrane on human periodontal ligament fibroblasts(HPLFs) were evaluated by Cell Counting Kit-8(CCK-8) assay.The fibroblasts were seeded on the membranes and the composites were observed by inverted phase contrast microscope,SEM and HE staining to explore morphological changes of cells.Results According to CCK-8 assay,the absorbance values among each tested groups and the blank control group were no significantly different(P > 0.05) at 1,3,5 and 7 d after cells seeded,and the levels of toxicity ranged from 0 to 1.The cell adhesion cultured on membrane was less than that on culture plate(P < 0.01).The proliferation,migration,secreting extracellular matrix,stratified cells and reticular collagen fibers of HPLFs cultured on the collagen membrane could be observed.Conclusion The new collagen membrane from bovine tendon has a good biocompatibility and potential as biomembrane and scaffold for tissue engineering.