Research on the construction of corneal endothelium transplantation with acellular amniotic membrane as a scaffold
This study aimed to fabricate a human acellular amniotic membrane (HAAM) by utilizing sequential chemical treatments (including trypsin/EDTA, Triton X-100, sodium deoxycholate, and peracetic acid/ethanol) with physical agitation. Following lyophilization, the HAAM was sterilized, coated with fibronectin and chondroitin sulfate (FNC), and then seeded with immortalized human corneal endothelial cells (HCECs) to evaluate its biocompatibility. The biocompatibility of the scaffold was assessed through various methods, including cell adhesion (observed via microscopy), viability (measured using CCK-8/EdU assays), functional gene expression (analyzed by qPCR/immunofluorescence) and transcriptome sequencing. The results demonstrated that the HAAM was successfully prepared with an intact collagen fiber structure. The corneal endothelial cells adhered closely to the HAAM scaffold and formed a continuous monolayer. Furthermore, the HAAM promoted cell viability and proliferation, exhibited positive expression of tight junction proteins, and upregulated key functional genes. Transcriptome analysis also identified genes regulating proliferation and matrix synthesis. In conclusion, the HAAM scaffold exhibits excellent transparency, mechanical properties, and biocompatibility, making it suitable for the attachment and proliferation of human corneal endothelial cells. It also effectively maintains the expression levels of key functional genes, thereby mimicking the characteristics of the natural corneal endothelial layer functionally and providing experimental evidence for its potential clinical application.