目的：制备一种新型的聚乳酸-生物玻璃（Poly-L-Lactic Acid/Bioglass, PLLA/BG）亲水性纳米纤维膜，探讨其作为引导骨再生屏障膜的生物相容性。方法采用氧等离子处理PLLA/BG纳米纤维引导骨再生膜，改善其亲水性能，将人成骨样细胞（MG63）接种于膜表面，通过Hoechst荧光染色观察其在膜表面的生长，计算细胞粘附率及增殖率，检测MG63细胞碱性磷酸酶活性，最后通过扫描电镜观察细胞在膜上生长形态及钙结节形成，分析该膜生物相容性及促成骨性能。结果氧等离子处理的PLLA/BG膜在1、3、6 h的细胞粘附率分别为（30.570±0.96）%、（47.27±0.78）%、（66.78±0.69）%，细胞黏附率高于两组膜（P<0.01）；3组膜的细胞增殖率均随时间延长提高，在不同时间点氧等离子处理的PLLA/BG膜的细胞增殖率高于另外两组（P<0.01）；Hoechst荧光染色中可观察到氧等离子处理的PLLA/BG膜表面早期有更多细胞黏附；碱性磷酸酶活性测定实验结果显示加入BG的复合膜可在早期促进成骨细胞分泌基质，差异具有统计学意义（P<0.01）；扫描电镜下可观察到MG-63细胞呈梭形，黏附于膜表面，与复合膜相互交联，形成钙结节。结论经氧等离子处理的PLLA/BG复合膜在早期可促进细胞黏附、增殖及促进成骨细胞分泌基质，具有良好的生物相容性和促成骨性能。%Objective To prepare and characterize a nano-scale fibrous hydrophilic poly-L-lactic acid/Bioglass (PLLA/BG) composite membrane and evaluate its biocompatibility as a composite membrane for guiding bone regeneration (GBR). Methods PLLA/BG-guided bone regeneration membrane was treated by oxygen plasma to improved its hydrophilicity. The growth of MG-63 osteoblasts on the membrane was observed using Hoechst fluorescence staining, and the biocompatibility of the membrane was evaluated by calculating the cells adhesion rate and proliferation rate. Osteogenesis of MG-63 cells was assessed by detecting alkaline phosphatase (ALP), and the formation of calcified nodules and cell morphology changes were observed using scanning electron microscope (SEM). Results The cell adhesion rates of PLLA/BG-guided bone regeneration membrane treated with oxygen plasma were (30.570 ± 0.96)%, (47.27 ± 0.78)%, and (66.78 ± 0.69)%at 1, 3, and 6 h, respectively, significantly higher than those on PLLA membrane and untreated PLLA/BG membrane (P<0.01). The cell proliferation rates on the 3 membranes increased with time, but highest on oxygen plasma-treated PLLA/BG membrane (P<0.01). Hoechst fluorescence staining revealed that oxygen plasma treatment of the PLLA/BG membrane promoted cell adhesion. The membranes with Bioglass promoted the matrix secretion of the osteoblasts. Under SEM, the formation of calcified nodules and spindle-shaped cell morphology were observed on oxygen plasma-treated PLLA/BG membrane. Conclusion Oxygen plasma-treated PLLA/BG composite membrane has good biocompatibility and can promote adhesion, proliferation and osteogenesis of the osteoblasts.
展开▼
