Human acellular amniotic membrane and acellular subamniotic membrane in repairing skin defects

doi:10.3969/j.issn.2095-4344.2355

Abstract

Abstract:

BACKGROUND: Amniotic membrane and subamniotic membrane are homologous and belong to medical waste. They have low immunogenicity and certain toughness. Amniotic membrane as tissue-engineered scaffold material has been reported more, but the research of subamniotic membrane as tissue-engineered scaffold is less.

OBJCETIVE: To compare the difference between acellular amniotic membrane and acellular subamniotic membrane in combination with bone marrow mesenchymal stem cells for skin repair.

METHODS: The acellular subamniotic membrane materials were prepared by sodium dodecyl sulfate + nuclease method, and the acellular amniotic membrane materials were prepared by Triton X-100 + pancreatin method. Bone marrow mesenchymal stem cells were cultured with two kinds of acellular materials. The cytotoxicity of the two kinds of acellular materials was detected by CCK-8 method. Bone marrow mesenchymal stem cells were seeded on the surfaces of two kinds of acellular materials. The growth and adhesion of cells were observed under light microscope and scanning electron microscope. Skin defects deep into the dermis were made on the back of one spine of each Sprague-Dawley rat, and 36 rats were randomly divided into three groups (n=12). No materials were implanted in the blank group. Acellular amniotic membrane and bone marrow mesenchymal stem cell complex were implanted in the control group. Acellular subamniotic membrane and bone marrow mesenchymal stem cell complex were implanted in the observation group. Wound healing rate, wound pathology, wound gene and protein were detected at 7, 14 and 21 days after operation, respectively. This study was approved by the Animal Ethics Committee of Jiangnan University.

RESULTS AND CONCLUSION: (1) In the 3-7 days of culture, two kinds of acellular materials could promote the proliferation of bone marrow mesenchymal stem cells. (2) The results of light microscope and scanning electron microscope showed that bone marrow mesenchymal stem cells grew well on the surfaces of two kinds of acellular materials, and the morphology of bone marrow mesenchymal stem cells was the same as that of cultured in culture bottle. (3) The wound healing rate of the observation and control groups was higher than that of the blank group at various time points after surgery (P < 0.05); and that in the observation group was higher than that in the control group (P < 0.05). (4) At 14 days after surgery, pathological observation showed that the skin repair effect of the observation group was better than that of the blank group, and the skin repair of the observation group was better than that in the control group. (5) The expression of type III collagen, CK18, type I collagen and vascular endothelial growth factor gene in the observation and control groups was higher than that in the blank group at various time points after surgery (P < 0.05); and the expression was higher in the observation group than that in the control group (P < 0.05). (6) The expression of CK18 and vascular endothelial growth factor protein in the observation and control groups was higher than that in the blank group at various time points (P < 0.05); and the expression was higher in the observation group than that in the control group (P < 0.05). (7) Compared with acellular amniotic membrane, the lower layer of acellular subamniotic membrane can promote skin repair.

Key words:

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CLC Number:

Wang Dan, Yin Jing, Wang Yuying, Du Yong, Li Jingling, Yu Shun. Human acellular amniotic membrane and acellular subamniotic membrane in repairing skin defects[J]. Chinese Journal of Tissue Engineering Research, 2020, 24(34): 5570-5576.

Figures/Tables 8

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