Biocompatibility of human amniotic mesenchymal stem cells with human acellular amniotic membrane scaffold / 中国组织工程研究

ABSTRACT

BACKGROUND: Anterior cruciate ligament (ACL) injury is commonly seen, and mainly treated with ACL reconstruction. Ligament tissue engineering treatment for ACL injury has become the current focus due to the extensive use of tissue engineering techniques in clinical practice. OBJECTIVE: To investigate the biocompatibility of human amniotic mesenchymal stem cells (hAMSCs) and human acellular amniotic membrane(HAAM)scaffolds in vitro. METHODS: hAMSCs were isolated using enzyme digestion and then identified. The fresh human amniotic membrane was subjected to decellularization using enzymatic digestion and chemical decontamination. Hematoxylin-eosin staining and immunofluorescence were used to detect the removal of cell components and destruction of the extracellular matrix. hAMSCs were cultured with HAAM extract (experimental group) and normal L-DMEM/F12 medium (control group). Cell proliferation was detected by cell counting kit-8. Cell adhesion and growth were observed by scanning electron microscope and inverted microscope at 14 days after co-culture with HAAM. RESULTS AND CONCLUSION: hAMSCs exhibited a spiral, adherent growth under the inverted phase contrast microscope; and the cells highly expressed vimentin and lowly expressed CK-19, with osteogenic, adipogenic, and chondrogenic potential. Hematoxylin-eosin staining showed that human acellular amniotic epithelial cells and mesenchymal stem cells were completely removed and the extracellular matrix had no obvious damage. Immunofluorescence staining showed positive expression of type I collagen and type III collagen. The HAAM extract had no cytotoxicity to hAMSCs and exerted no effect on its proliferative activity. HAAM could promote the proliferation and adhesion of hAMSCs after incubation under the inverted microscope and scanning electron microscope. These findings indicate that HAAM has good biocompatibility and is favorable for the proliferation of hAMSCs.