Vascular endothelial growth factor gene modified human amnion-derived mesenchymal stem cells promote the survival of ultra-long random skin flap

doi:10.3969/j.issn.2095-4344.1743

Abstract

Abstract:

BACKGROUND: Studies have confirmed that vascular endothelial growth factor can induce angiogenesis, and human amnion-derived mesenchymal stem cells improve the repair of ischemia-reperfusion injury using flap transplantation via various mechanisms. Therefore, the modification of human amnion-derived mesenchymal stem cells using vascular endothelial growth factor gene promotes the survival of skin flaps.
OBJECTIVE: To investigate the effect of human amnion-derived mesenchymal stem cells modified by vascular endothelial growth factor gene on the survival of ultra-long random skin flaps.
METHODS: Thirty healthy adult Sprague-Dawley rats were randomly divided into group I, group II and group III, 10 rats in each group. Two ultra-long ischemic random skin flap models were constructed at the left and right back of each rat, and the skin flap pedicle was located at iliac spine level. These skin flaps received the following treatment immediately after removal: The flaps were divided into group I-left, I-right; II-left, II-right; III-left and III-right. 0.5 mL of LG-DMEM was injected in the skin flaps in I-left and II-left groups (control group); 0.5 mL of 1×10⁹/L human amnion-derived mesenchymal stem cells was injected into the skin flaps in I-right and III-left groups (experimental group 1); 0.5 mL of 1×10⁹/L vascular endothelial growth factor-transfected human amnion-derived mesenchymal stem cells was injected into the flaps in II-right and III-right groups (experimental group 2). There were five injection sites for each flap, 0.1 mL per each site. The blood perfusion in the middle and pedicle of each skin flap was monitored using laser Doppler flow meter immediately, 1, 2, 4 and 7 days after transplantation. The survival rate of skin flaps in all groups was measured at 7 days after transplantation. The capillary density in survived skip flaps was histologically observed. The distribution and survival rate of CM-Dil labeled human amnion-derived mesenchymal stem cells were observed by fluorescent microscope. The vascular endothelial growth factor protein level was detected by western blot.
RESULTS AND CONCLUSION: (1) The survival rate of skin flaps, blood perfusion, and capillary density showed significantly difference among groups (P < 0.05) and ranked as follows: experimental group 2 > experimental group 1 > control group. (2) Under fluorescent microscope, CM-Dil labeled human amnion-derived mesenchymal stem cells were visible in the flaps in the experimental groups 1 and 2. (3) Vascular endothelial growth factor positively expressed in all the flaps, while the protein expression of vascular endothelial growth factor was ranked as follows: experimental group 2 > experimental group 1 > control group and there were significant differences among groups (P < 0.05). To conclude, the application of human amnion-derived mesenchymal stem cells in the middle and distal parts of ultra-long random skin flap can remarkably improve the survival rate of skin flaps, and moreover transfection with vascular endothelial growth factor can increase the secretion of vascular endothelial growth factor, further improving the effects of human amnion-derived mesenchymal stem cells promoting angiogenesis and survival rate of ultra-long random skip flaps.

Key words: ultra-long ischemic random flap model, human amnion-derived mesenchymal stem cells, vascular endothelial growth factor, gene transfection, flap survival rate

CLC Number:

Jin Wenhu, Zhou Aiting, Wu Zhonghuan, Fan Zhenhai, Wang Dali, Wei Zairong. Vascular endothelial growth factor gene modified human amnion-derived mesenchymal stem cells promote the survival of ultra-long random skin flap[J]. Chinese Journal of Tissue Engineering Research, 2019, 23(21): 3349-3356.

Figures/Tables 5

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