| Objective To investigate the effect of fibroblast(FBs)composited with acellular dermal matrix of sheep(sADM)on wound healing in nude mice Methods (1)Isolated from abandoned prepuce,the hFBs(homo Fibroblasts)we obtained that can cultured in vitro.Identified by immunofluorescence.The expression of VEGF and bFGF was detected by RT-PCR;(2)sADM was prepared and detected whether it was toxic to cell proliferation and growth by MTS method.(3)Prepare sADM composite Fbs,and the combination of Fbs and sADM was identified by HE staining;(4)Selecting 45 nude mice of7 weeks old,adaptive feeding for 1 week,preparing a full-thickness skin injury model,randomly divided into Group A of FBs and sADM composite group,group B of sADM control group,group C of gauze control group.The wound healing rate and blood vessel were calculated at 7d,14 d and 21 d after operation.The expression of VEGF,Bax and bcl-2 genes were detected by qRT-PCR.The experimental data of the obtained data was expressed by`x±s,and statistical analysis was performed using SPSS 17.0.Results The expression of vimentin was positive by cellular immunity,and the expression of VEGF and bFGF was detected by cell RT-PCR.MTS assay showed that it had no toxicity to cell proliferation and growth.The results showed that the OD value of the control group and the OD value of the experimental group P<0.05 were significantly poor at the 3rd and 5th day.There was no significant difference in the OD values between the experimental group and the control group.Image J statistically calculated the wound healing rate: the wound healing rate of sADM composite FBs group and sADM group on the 7th day was(45.55±7.16)%,(26.76±1.56)%,P<0.01,the difference was statistically significant.On the 14 th day,the wound healing rate was(88.71±2.03)%,(79.00±1.48)%,P<0.05),the difference was statistically significant,and on the 21 th day,the wound healing rate was(99.67±0.08)%.(97.25±1.44)%,P<0.05,the difference was statistically significant.The wound healing rate of sADM combined with FBs group was higher than that of sADM group;the wound healing rate of sADM composite FBs group and gauze group,on the 7th day,the wound healing rate was(45.55±7.16)%,(18.31±4.61)%,P<0.001,the difference was statistically significant The wound healing rate at 14 days was(88.71±2.03)%,(72.74±6.35)%,P<0.01,the difference was statistically significant.The wound healing rate at 21 days was(99.67±0.08%%,(95.11±1.52)%,P<0.05,and the difference was statistically significant.sADM combined with FBs group the healing rate is higher than the gauze group.The results of qRT-PCR showed that at 7th,14 th and 21 st day,the expression of Bax in sADM complex Fbs group,ADM group and gauze group showed a downward trend.sADM combined with FBs group and sADM group: Qn the 7th day,the relative expression of genes was(0.382±0.021),(0.394±0.040),P>0.05,and the difference was not statistically significant.On the 14 th day,the relative expression of genes was(0.226±0.014),(0.272±0.014),P<0.01 The difference was statistically significant.On the 21 st day,the relative expression of genes was(0.023±0.013),(0.030±0.005),P>0.05,and the difference was not statistically significant;sADM complex FBs group,gauze group: The relative expression of genes at 7 days was(0.382±0.021),(0.495±0.036),P<0.01,the difference was statistically significant.On the 14 st day,the relative expression of the genes was(0.226±0.014),(0.363±0.016),P<0.001,the difference was statistically significant.On the 21 st day,the relative expression of the gene was(0.023±0.013),(0.054±0.003),P<0.01,and the difference was statistically significant.The expression of bcl-2 was up-regulated.sADM combined with FBs group and sADM group:On the 7th day,the relative expression levels of were(0.067±0.013),(0.048±0.017),P>0.05,the difference was not statistically significant.On the 14 th day,the relative expression levels of the genes were(0.169±0.007),(0.140±0.004),P<0.01,the difference was statistically significant..On the 21 st day,the relative expression of the genes was(0.406±0.018),(0.322±0.015),P<0.001,with significant differencethe difference was statistically significant.;sADM complex FBs group,gauze group: on the 7th day,the relative expression of genes were(0.067±0.013),(0.055±0.030),P>0.05 The difference was not statistically significant.On the 14 th day,the relative expression of genes was(0.169±0.007),(0.123±0.005),P<0.001,the difference was statistically significant.On the 21 st day,the relative expression of genes were(0.406±0.018),(0.233±0.011),P<0.001,and the difference was statistically significant.The expression of VEGF was down-regulated.sADM in FBs group and sADM group: On the 7st day,the relative expression levels of were(1.191±0.051),(1.092±0.027),P<0.05,and the difference was statistically significant.On the 14 st day,the relative expression levels of were(0.632±0.023),(0.553±0.046),P>0.05,and the difference was not statistically significant.On the 21 st day,the relative expression levels of were(0.135±0.045),(0.319±).0.017),P<0.01,the difference was statistically significant.;sADM complex FBs group,gauze group: the relative expression of genes on the 7th day were(1.191±0.051),(1.032±0.025),P<0.01,the difference was statistically significant,the relative expression of genes on day 14 was(0.632±0.023),(0.491±0.064),P<0.05,the difference was statistically significant.On the 21 st day,the relative expression of genes was0.135±0.045),(0.292±0.038),P<0.01,the difference was statistically significant.Conclusions (1)FBs with high purity and proliferative ability were isolated and cultured from the foreskin tissue discarded by circumcision,and the sADM functional dressing of composite FBs was successfully prepared.(2)The composite was prepared.The sADM functional biological dressing of FBs promotes wound healing. |
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