RESUMO
BACKGROUND: Acellular amniotic membrane scaffold is a natural scaffold with good biocompatibility, which has been widely used in tissue engineering. Scleraxis can promote the differentiation of human amniotic mesenchymal stem cells into human ligament cells and promote tendon-bone healing. OBJECTIVE: To explore whether acellular amniotic membrane scaffold combined with human amniotic mesenchymal stem cells transfected with Scleraxis can promote rabbit tendon-bone healing. METHODS: (1) Human amniotic mesenchymal stem cells were isolated and cultured in vitro. After passaged, the cell morphology was observed. (2) The Scleraxis lentivirus was constructed in vitro and then transfected into passage 3 human amniotic mesenchymal stem cells with optimal multiplicity of infection. The transfection efficiency was detected by q-PCR. (3) The acellular amniotic membrane scaffold was prepared by enzymatic digestion. Then the Scleraxis lentivirus-transfected cells were seeded on the acellular amniotic membrane scaffold in vitro. The cell growth on the scaffold was observed by phalloidin staining. (4) The New Zealand white rabbit tendon was covered with the acellular amniotic membrane scaffold combined with human amniotic mesenchymal stem cells transfected with Scleraxis lentivirus, followed by implanted into the bone tunnel. The tendon-bone healing was detected. RESULTS AND CONCLUSION: The passage 3 human amniotic mesenchymal stem cells adhered well. (2) After transfected with Scleraxis lentivirus for 96 hours, stable green fluorescence was observed. The mRNA expression level of Sclerxis was significantly increased, indicating a success transfection. The epithelial cells of the acellular amniotic membrane scaffold disappeared, indicating a relatively complete decellularization. The basal layer remained intact, and the extracellular matrix component still existed. Phalloidin staining results revealed that the cells on the acellular amniotic membrane scaffold were in good adhesion and growth, and the cell proliferation was not affected. Therefore, In vivo experimental results reveal that human acellular amniotic scaffold combined with human amniotic mesenchymal stem cells transfected with Scleraxis lentivirus can promote the tendon-bone healing.