双层胶原支架与人牙周膜细胞的体外生物相容性

Biocompatibility of a Double-layer Collagen Scaffold and Human Periodontal Ligament Cells in Vitro

【目的】研究双层胶原支架与人牙周膜细胞的体外生物相容性。【方法】组织块培养法培养原代人牙周膜细胞。MTT法测胶原支架不同浓度浸提液的细胞毒性。将人牙周膜细胞与支架材料三维培养,通过扫描电镜、组织切片观察其复合情况,并检测三维培养后细胞碱性磷酸酶(ALP)、羟脯氨酸(HYP)的分泌情况。【结果】不同浓度材料浸提液毒性评级为0~1级。细胞与支架复合培养后,细胞在支架上黏附、增殖良好,支架材料内部细胞分布均匀,且致密层可起屏障膜作用,阻挡细胞进入支架内部。细胞与胶原支架复合培养24 h后,细胞的ALP活性与对照组无明显差异(P>0.05),复合培养48、72 h后,细胞的ALP活性高于对照组(P<0.05)。复合培养24、48、72 h后,细胞的HYP活性均高于对照组(P<0.05)。【结论】双层胶原支架具有良好的生物相容性,可进一步应用于牙周组织工程的研究。

【Objective】 To investigate biocompatibility between a double-layer collagen scaffold and human periodontal ligament cells(hPDLC).【Methods】 The hPDLC were obtained by explant culture.The cytotoxicity of different concentrations of an extract of the collagen scaffold was evaluated by an MTT assay.The morphology of hPDLC after seeding on the scaffolds was evaluated by SEM and histological examination.Alteration of alkaline phosphatase(ALP) and hydroxyproline(HYP) activity of the hPDLC after seeding on the scaffolds was also evaluated.【Results】 The grade of the cytotoxicity of the collagen scaffold was from 0 to 1.The hPDLC proliferated well on the scaffold and a uniform distribution of cells in the scaffold could be observed.The dense layer prevented cells from migrating into the scaffold.ALP activity between the scaffold group and the control group showed no significantly difference(P > 0.05) at 24 h after cell seeding,however,at 48 and 72 h after cell seeding,ALP activity of the scaffold group was significantly higher than that of the control group(P < 0.05).HYP activity of the scaffold group was significantly higher than that of the control group(P < 0.05) at 24,48,and 72 h after cell seeding.【Conclusion】 The double-layer collagen scaffold which showed good biocompatibility may be a promising biomaterial for periodontal tissue engineering.