改良明胶/聚己内酯复合纳米纤维电纺膜构建复层皮肤的研究

Use of modified gelatin/PCL electrospun membranes in engineering bilayered skin graft

目的:探讨改良明胶/聚己内酯复合纳米纤维电纺膜成功构建组织工程表皮的可行性。方法:2013年1月至2019年12月,于上海交通大学医学院附属第九人民医院整复外科组织工程实验室分别制备3个组不同配比的明胶/聚己内酯纳米纤维电纺膜(70∶30、50∶50、30∶70),体外测试细胞相容性,构建复层皮肤修复裸鼠皮肤缺损。结果:细胞接种实验显示,随着膜片中明胶含量的增加,成纤维细胞和表皮细胞在材料上的黏附和增殖都明显增强。组织学染色可见,经过21 d体外培养构建的复层皮肤中,明胶/聚己内酯(70∶30)组能形成相对较好的皮肤结构。而裸鼠皮肤缺损修复试验发现,回植术后14 d时,所有组材料未降解,移植物均脱落,提示3种配比材料虽具有较好的生物相容性,但材料降解过慢不适于构建复层皮肤。结论:改良的明胶/聚己内酯纳米纤维电纺膜随明胶比例增高,细胞相容性逐渐增强,可促进皮肤愈合,但最终会被自体组织替代。

Objective For severe skin defects which are deep to dermis,engineered skin with epidermis and dermis(bilayered)is required.Based on the success of engineering epidermis with GT/PCL electrospun membranes,our study was to investigate whether this membrane could be also used for engineering bilayered skin graft.Methods From 2013 to 2019,we first prepared three GT/PCL electrospun membranes with different proportion(70∶30;50∶50;30∶70)in our laboratory;the biocompatibility of the membrane was evaluated in vitro by seeding fibroblasts or keratinocytes on the membranes.Then the outcome of GT/PCL membranes repairing skin defects in the nude mouse was investigated.Results Cell attachment and proliferation were significantly improved with increase of gelatin.Histological analyses showed that bilayered skin engineered with GT/PCL(70∶30)group could form relatively better structure after 3 weeks of cultivation in vitro.Further in vivo transplantation studies revealed that scaffolds were not degraded in all three groups,indicating that these materials were not suitable for engineering bilayered skin although they had good biocompatibility.Conclusions The higher gelatin membranes possess better biocompatibility.Further in vivo transplantation studies reveal that bilayered skin engineered with GT/PCL membranes is able to repair skin defects in the nude mouse.