Effects of porcine acellular dermal matrix combined with sodium carboxymethylcellulose on the ex-pression of vimentin and zinc finger proteins secreted by keratinocytes
Objective To investigate the effect of porcine acellular dermal matrix(pADM)com-bined with sodium carboxymethyl cellulose(CMC)on epithelial-mesenchymal transformation(EMT)of ke-ratinocytes and its mechanism.Methods Rat hair follicle keratinocytes were established in vitro and di-vided into four groups:group A(blank control),group B(2％CMC),group C(5％pADM)and group D(5％pADM+2％CMC).The effect of gel on the viability of keratinocytes was detected by cell counting kit(CCK-8)assay.Transwell assay was used to detect the migration ability of keratinocytes.Polymerase chain reaction(PCR)and Western blotting were used to detect the expression of EMT-related proteins in each group.Non-paired t-test was used for comparison between the two groups,and single factor analysis of variance was used for comparison between multiple groups.Results Results The results of CCK-8 assay showed that keratinocyte activity was the strongest in group D(group A:100.0±8.70,group B:124.1±8.10,group C:130.4±7.30,group D:152.3±5.50),and the difference was statistically significant a-mong all groups(F=189.2,P＜0.01).In Transwell experiment,the number of migrating keratinocytes was 95.889±8.638,149.556±7.091,172.778±7.710,and 236.667±9.695 in groups A,B,C and D,respectively.There was a statistically significant difference between groups B,C and D and group A(F=354.9,P＜0.01).The number of migrating keratinocytes in group D was the greatest.PCR results showed that the expression levels of keratin 10(CK10),Vimentin(Vim)and zinc finger protein(Snail)in group D were the highest(4.69,4.25,4.52;F=125.3,146.8,100.3.all P＜0.05)and the expres-sion of CK14 was the lowest(0.18,F=96.5,P＜0.05).Western blotting results showed that the expres-sion levels of CK10,Vim and Snail in group D were the highest(0.43,0.54,0.69;F=187.5,168.9,151.3.all P＜0.05),while the expression level of CK14 was the lowest(0.10,F=151.3,F=167.4,P＜0.05).Conclusion Both pPADM and CMC can improve the biological activity of keratinocytes.pADM combined with CMC can promote the expression of CK10,Vim and Snail,and inhibit the expression of CK14.
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