多聚乳酸/乙醇酸多管道支架的构建及其三维结构与雪旺细胞生物相容性研究

Biocompatibility of poly (lactic -co -glycolic acid)multichannel three -dimensional scaffold with schwann cells

目的将雪旺细胞(SCs)移植到聚乳酸羟基乙酸(PLGA)多通道支架材料中,观察SCs在支架中的黏附、增殖和迁移,探讨细胞和支架的生物相容性。方法将SCs种植在PLGA多通道支架中,继续培养7 d和14 d,利用钙黄绿素-AM(Calcein-AM)4 μl和胡米胺二聚体(EthD-Ⅲ)试剂盒检测细胞在支架中的活性,活细胞被Calcein-AM染成绿色,死细胞被EthD-Ⅲ染成红色。细胞死亡率由计算EthD-1阳性细胞百分比获得。每组每张切片至少5个区域被照相。最终的细胞死亡率取于3组的平均值。扫描电镜及光镜观察细胞形态和S-100蛋白免疫鉴定细胞的迁移状况。结果倒置显微镜下显示SCs在多管道支架中于7 d表现出良好的黏附能力,增殖14 d达到高峰。Viability/Cytotoxicity Assay Kit检测细胞活性发现7 d后支架中细胞死亡率为(1.39±0.46)%。14 d后支架中细胞死亡率为(1.43±0.51)%。免疫荧光切片及扫描电镜均显示,SCs不仅在管道壁贴附并生长,并且能在14 d迁移到通道之间的空泡结构中。结论 SCs和PLGA多管道支架的三维结构之间具有良好的生物相容性。

Objective After seeding schwann cells (SCs) into poly (lactic-co-glycolic acid) (PLGA) multichannel scaffolds, the biocompatibility between SCs and the scaffold was evaluated by adhesion, proliferation and survival of SCs.Methods After seeding into PLGA scaffolds, SCs were cultured for 7 days and 14 days respectively. Then, Viability/Cytotoxicity Assay Kits were used to evaluate the activity of SCs. The living cells were dyed green by Calcein-AM and dead cells were dyed red by EthD-Ⅲ. Cell death rate was calculated by measuring the percentage of EthD-1 positive cells. At least 5 areas of each slice were photographed. Cellular morphological changes were observed under the scanning electron microscopy (SEM) and light microscopy. In addition, immumofluorescence method was used to estimate the migration of SCs in the scaffold.Results Under an inverted microscope, SCs displayed satisfactory characteristics of adhesion and proliferation on the 7th day and reached the peak on the 14th day. Cell viability was detected by Viability/Cytotoxicity Assay Kits at 7th day, and the cell death rate in the stent was (1.39±0.46)%. At 14th day, the cell death rate was (1.43±0.51)%. Immumofluorescence method and SEM also showed that SCs could migrate into the gradient porous structure of the scaffold.Conclusion SCs have a good biocompatibility with PLGA multichannel scaffolds.