Therapeutic effect of adipose derived mesenchymal stem cell conditioned medium thermosensitive hydrogel on skin scald
10.3969/j.issn.2095-4344.2017.30.017
- VernacularTitle:脂肪间充质干细胞条件培养基制备温敏凝胶外用治疗皮肤烫伤
- Author:
Pei LIU
1
;
sheng Zhen HU
;
Ling MA
;
huan Huan WANG
;
Dong LI
Author Information
1. 山东大学齐鲁医院
- From:
Chinese Journal of Tissue Engineering Research
2017;21(30):4852-4859
- CountryChina
- Language:Chinese
-
Abstract:
BACKGROUND: Studies have shown that adipose derived mesenchymal stem cells are involved in the skin repair after scald, but the hydrogel made of the excreta by adipose derived mesenchymal stem cells is rarely reported in the treatment of skin scald. OBJECTIVE: To analyze the therapeutic effect of human adipose derived mesenchymal stem cell conditioned medium hydrogel in a mouse model of skin scald. METHODS: Adipose derived mesenchymal stem cells were obtained from adipose tissues by enzyme digestion combined with adherent culture method. Morphological and flow cytometry were used to identify phenotype and induce differentiation. Secondly, the stable proliferative phase cells were harvested to obtain the conditioned medium, and chitosan, mannitol, beta-glycerol phosphate sodium and hyaluronic acid were added to prepare the thermosensitive hydrogel. Then the 95 ℃ aluminum block was used to rapidly establish a model of degree III skin scald on the left (experimental group) and right (control group) sides of the back of 24 C57BL/6 mice. In the experimental group, adipose derived stem cell conditioned medium hydrogel was applied twice a day on the right side of the mouse back, and in the control group, fresh medium hydrogel was applied twice a day on the left side of the mouse back. The treatment period lasted for 7 days. Healing time and healing process were observed to calculate the healing rate. Histopathological changes were observed by hematoxylin-eosin staining at paraffin sections at 4, 14, 28 days after skin scald. RESULTS AND CONCLUSION: (1) The human adipose derived mesenchymal stem cells had fibroblast-like morphology and proliferated vigorously, and the average doubling time was 55 hours. These cells could be induced to differentiate into osteoblasts, chondrocytes and adipocytes. High expression of CD29, CD44, CD90 and CD105 were observed on these cells with low expression of CD31 and CD34, which met the standard of mesenchymal stem cells. (2) Thethermosensitive hydrogel prepared by the conditioned medium was cool and transparent viscous liquid at 4-20 ℃, and was changed into semi-solid gel at( 37 ℃ after 15 minutes. (3) The normal structure) subcutaneous fat and muscle tissue (of 95 ℃ aluminum block scalded mice wer) standards of degree III burns. The wound area was roughly 3 cm2. (4) In the repair process, shorter wound healing time, less scar and better dermis structure were observed in the experimental group compared with the control group. (5) Inflammatory infiltration, thickness of granulation tissue, epidermal thickness, fibroblasts and vascular density were significantly improved in the experimental group as compared with the control group (P < 0.05). To conclude, human adipose derived mesenchymal stem cells conditioned medium hydrogel can promote the wound healing and promote the quality of regenerated skin after skin scald.
