Direct induction of the differentiation of bone mesenchymal stem cell for enhanced wound regeneration by Wnt3a in rats
CAO Hua-ping
YE Tao
LI Yong-zhong
YU Lu
MIN Hua
Abstract：Objective To explore the in vitro and in vivo inductive process of Wnt 3a on the differentiation of BMSCs into the crucial cells for wound regeneration.Methods Eighty SD male rats of 40-50 grams were select-ed.BMSCs from the bone marrow of femur and tibia were isolated from 3-week male SD rats and separated from the non-adherent cells and then cultured until the three passages.After expansion in vitro,they were divided into control group and experimental group.Next,BMSCs were cultured with DMEM +Wnt3a(200μg/L) for the experimental group, while only DMEM were used for control group.qRT-PCR and immunofluorescence staining were used for an-alyzing the expression of NG2,VEGF-CD31and α-SMA in vitro and in vivo to verify the induced differentiation effect of Wnt3a in vitro on BMSCs.The effect of BMSC and Wnt3a on wound repair by using full-thickness skin defect model of rats as the research object was also observed.Results The results of RT-PCR showed that the expression of BMSCs transcription factors (Oct4 and Klf4) was decreased in Wnt3a group (0.732 ±0.048,0.591 ±0.097, P＜0.001)and their differentiation into the angiogenic cells and pericytes were also observed ,which were indispen-sable for wound regeneration.In vivo,the findings also demonstrated that BMSC +Wnt3a were able to accelerate the wound healing by forming more active and robust granulation tissue and stimulating the angiogenesis process of the wound(P＜0.001).Expression of angiogenesis markers (NG2,VEGF,CD31 and α-SMA) was also upregulated in wounds treated with combined BMSCs +Wnt3a ( P＜0.001).Conclusion These findings verify the capacity of Wnt3a to enhance BMSC angiogenic property for cutaneous wound healing potentially by preserving the viability of the cells, stimulating, and inducing them to differentiate into the desired cells that are beneficial for angiogenesis within wounds.This combinatory strategy is superior to either of cells transplantation and Wnt 3a therapy alone,lead- ing to the accelerated wound healing and providing new idea for massive skin defect repair with BMSCs .
Keywords：soft tissue defectbone marrow mesenchymal stem cellsproteindirectional differentiationwound repairrats
Publication Date：2018-01-01
Online Publishing Date：2025-08-15(First online date of this platform, not the publication date of the document)
Pages：7( 450-456 )