Cell Scaffolds Complex Promote Survival And Vascularization Of Prefabricated Flap In Rat

Objective: To investigate the survival effect of complex of collagen sponge seededamniotic epithelial cells on prefabricated flap after the complex was transplanted on bothsides of vascular bundle of the prefabricated flap.Methods: To investigate the growth of rat amniotic epithelial cells (AECs) in thecollagen sponge when the cells implant to the scaffolds after they became3rd generationapart were cultured in vitro. To observe the growth and proliferation of cells before andafter planting with Light microscopy and method of fluorescence labeling. ELISA wastaken to detect the angiogenesis factors VEGF, bFGF, TGF in the supernatant fluid of theAECs complex. The method of Fluorescently labeled was taken to observe the survival ofthe cells after transplantation. SD rat cartilage cells were implanted to the collagen spongescaffold after they became3rd generation apart. The SD rats were divided into normalcontrol group, blank control group, cartilage cell group and amniotic epithelial cells group.In the first phase of operation, the blank collagen sponge scaffolds, the collagen spongescaffolds seeded the corresponding cells were implanted to the both sides of neurovascularbundle of prefabricated flaps. On the7th day and14th day after first-stage surgery ELISAwas taken to test angiogenesis factors in the flaps for each group. In the second phase ofoperation, four sides of the flap were set off and then sutured. The survival of the flaps andthe number and density of vas capillary were assessed on one week after surgery. Anothersix rats as normal control group does not undergo surgery, when the other three groups ofrats after two operations, the corresponding position of the abdominal skin and soft tissuewere taken to test relevant indicators.Results: Rat AECs in collagen sponge scaffolds grown well, VEGF, TGFβ1, bFGFin the supernatant fluid of amniotic epithelial cells complex increased gradually.Cells growwell in the collagen sponges after the cell scaffold complex was transplanted in vivo.Content of the factors was the highest in the flap tissue of amniotic epithelial cells group, then the content in normal control group, blank control group and cartilage cells groupdecreased one by one. Additionally, the content of the factors on7th day after surgery wasclosed to the data on14th day after surgery. There was no significant difference betweenthe blank control group and cartilage cell group of the survival flap percentage, and thesurvival percentage of amniotic epithelial cells group was higher significantly.Subcutaneous microvascular density was significantly higher in the amniotic epithelialcells groups than that in the other groups in which the density was closed.Conclusion: AECs grow well in the collagen sponges and transplanting theAECs/collagen sponge complex in vivo, which lay the foundation of cell therapy. Thecomplex of collagen sponge seeded amniotic epithelial cells can promote the survival ofprefabricated flap evidently, because some of the angiogenesis factors secreted by theamniotic epithelial cells can promote the neovascularization in the prefabricated flap.