基于聚己内酯纳米纤维膜的组织工程化视网膜色素上皮的研究

Research of tissue-engineered retinal pigment epithelium based on nanofiber membrane of polycaprolactone

目的 探讨聚己内酯(PCL)纳米纤维膜在构建组织工程化视网膜色素上皮(RPE)的应用前景。方法参考有关文献采用自发分化法将人胚胎干细胞(ESC)诱导为视网膜色素上皮(ESC-RPE)细胞;采用静电纺丝法制备PCL纳米纤维膜,经扫描电镜检测PCL纳米纤维膜的表面形貌;利用CCK-8法检测PCL纳米纤维膜浸提液对于ESC-RPE细胞活性的影响;将ESC-RPE细胞培养在PCL纳米纤维膜上,通过透射电镜和免疫荧光法分别检测组织工程化RPE的超微结构和RPE特异性标记物的表达,通过与大鼠视网膜共培养检测组织工程化RPE吞噬感光细胞外节的能力。结果ESC-RPE细胞表达RPE特异性标记物MITF和PAX6,PCL纳米纤维膜呈纤维多孔状,具有良好的生物相容性。基于PCL纳米纤维膜的组织工程化RPE具有典型的RPE超微结构,表达ZO-1、NaKATPase呈极性表达,并且具有吞噬感光细胞外节的能力。结论PCL纳米纤维膜支持RPE细胞的生长,可用于构建组织工程化RPE。

Objective To explore the potential of nanofiber membrane of polycaprolactone(PCL)to constitute tissue-engineered retinal pigment epithelium(RPE).Methods The embryonic stem cells(ESC)were differentiated into retinal pigment epithelium(ESC-RPE)cells by spontaneous differentiation methods.The nanofiber membrane of PCL was prepared by electrospinning,and the topography of PCL nanofiber membrane was observed by scanning electron microscope.The viability of ESC-RPE cells treated by the extracts of PCL nanofiber membrane was studied by CCK-8.After the ESC-RPE cells were cultured on the PCL nanofiber membrane,transmission electron microscope and immunostaining were used to detect the ultrastructure and the expression of RPE markers of tissue-engineered RPE,respectively.Moreover,neural retinas of rats were co-cultured with tissue-engineered RPE to detect the capacity to phagocytize photoreceptor outer segment.Results ESC-RPE cells were positive for MITF and PAX6.The PCL nanofiber membrane was porous with high biocompatibility.The tissue-engineered RPE based on PCL nanofiber membrane showed typical ultrastructure of RPE cells.The tissue-engineered RPE was positive for ZO-1 and showed polarized expression of NaKATPase.Besides,the tissue-engineered RPE was able to phagocytize photoreceptor outer segment.Conclusion PCL nanofiber membrane is favor for the growth of RPE cells,and it can be used as a scaffold to constitute tissue-engineered RPE.