人牙髓干细胞在不同HA含量的PLGA/HA复合支架材料上的粘附研究

目的:构建不同质量构成比的PLGA/HA复合支架并评价材料机械性能;探讨不同比例PLGA/HA复合生物学支架对牙髓干细胞粘附能力的影响。方法:采用溶液浇筑/颗粒沥析技术构建出分别含有10%HA、20%HA及30%HA的PLGA/HA复合支架,对照组采用单纯PLGA支架。应用电子万能测验机检测支架的拉伸强度,扫描电镜观察表面结构。将牙髓干细胞与不同比例的PLGA/HA支架复合培养,应用DAPI染色细胞计数法检测细胞粘附能力。结果:在对3组实验组的拉伸强度测试中,10% HA组拉伸强度最高,20% 组次之,两组均高于对照组,而30% HA组拉伸强度低于对照组(P<0.05)。扫描电镜观察支架表面,显示支架孔径在100~250 μm之间,孔隙间 连通性良好,孔隙率较高。细胞接种于支架2、6、12 h后,荧光染色显示细胞与3种支架紧密贴合,粘附良好,细胞计数结果显示各实验组细胞粘附数量均多于对照组,其中在30%HA组中细胞粘附性能最佳(P<0.05)。结论:3种不同构成比的HA/PLGA复合支架均为良好的组织工程支架材料,其中10%HA组具有良好的机械性能,30%HA组细胞粘附性能更佳。

Adhesion of Human Dental Pulp Stem Cells on Poly Lactic-co-glycolic Acid/hydroxylapatite (PLGA/HA) Composite Scaffolds with Different Proportions.

Objective: To prepare and evaluate the Poly (lactide-co-glycolide)/Hydroxylapatite (PLGA/HA) composite scaffold with different proportions. Methods: PLGA/HA composite scaffolds with different HA contents (10%, 20% and 30%) were fabricated by solvent casting/particulate leaching technology, while PLGA without HA was used as control group. The tensile strength and microstructure were detected by using electro-mechanical universal testing machines and scanning electron microscopy (SEM). Human dental pulp stem cells were isolated, cultured and identified. The cells were seeded in the PLGA/HA scaffolds with different proportions. The adhesion properties were measured by DAPI staining. Results: The tensile strength of 10% HA was higher than that of 20% HA, and both of them showed a higher tensile strength than the control group. However, that of 30% HA was lower than control group (P<0.05). Scaffolds with different proportions of PLGA/HA showed high porosity and pore connectivity and the pore size was 100-250 μm investigated by SEM. Cell attachment and number in PLGA/HA after the incubation of 2h, 6h and 12h were significantly higher than the control group (P<0.05), and the cell adhesion ability in 30% HA group was the highest (P<0.05). Conclusions: The present study shows that PLGA/HA composite scaffolds with HA contents may be suitable for tissue engineering. In addition, 10% HA shows a higher tensile strength, while the adhesion ability of hDPSCs in 30% HA is better.