Effects of recombinant human metallothionein-Ⅲ combined with wound dressing on wound healing of full-thickness skin defects in mice
Objective To investigate the effects of recombinant human metallothionein-Ⅲ(rh-MT-Ⅲ)combined with wound dressing on wound healing of full-thickness skin defects in mice.Methods This study was an experimental study.Twenty-four half male and half female 6 weeks old Institute of Cancer Research mice were taken,and two symmetrical round full-thickness skin defect wounds were prepared on the back of each mouse.The mice were stratified and randomly divided into normal saline group,dressing group,rh-MT-Ⅲ group(applying the corresponding solution on the wounds),and combined treatment group(applying a mixture of rh-MT-Ⅲ and wound dressing on the wounds)according to sex and body weight,with 6 mice in each group.From 1 to 7 d after injury,all mice were observed daily for changes in activity,diet,and fur growth,their body weights and wound areas were recorded,and the relative wound area percentages were calculated.On 7 d after injury,the wound tissue of mice was taken for hematoxylin-eosin staining to observe the newborn granulation tissue,for Masson staining to observe collagen fiber formation,and for immunofluorescence staining to detect cell proliferation(denoted as Ki67 relative fluorescence intensity)and cell apoptosis(denoted as TdT-mediated dUTP nick end labeling(TUNEL)relative fluorescence intensity).The sample size in the above experiments was 6.Results There were no abnormalities in activity,diet,or fur growth in the 4 groups of mice within 7 d after injury.There were no statistically significant differences in the overall comparison of the body weights of mice in the 4 groups from 1 to 7 d after injury(P＞0.05).The relative wound area percentages of mice in combined treatment group were significantly lower than those in normal saline group and rh-MT-Ⅲ group on 2,3,4,5,6,and 7 d after injury(P＜0.05),and the relative wound area percentages of mice in combined treatment group were significantly lower than those in dressing group on 3,4,5,6,and 7 d after injury(P＜0.05).The relative wound area percentages of mice in dressing group on 6 and 7 d after injury and in rh-MT-Ⅲ group on 7 d after injury were significantly lower than those in normal saline group(P＜0.05).On 7 d after injury,a large number of capillaries and fibroblasts could be seen in wound tissue of mice in combined treatment group,and the growth of new epithelial tissue at the upper edge of the wound was better than that of the other three groups;the collagen fibers in the wound tissue of mice in combined treatment group had higher degree of density and arrangement in a more orderly manner than those of the other three groups.On 7 d after injury,the Ki67 relative fluorescence intensity in the wound tissue of mice in dressing group,rh-MT-Ⅲ group,and combined treatment group was(289±35)％,(197±17)％,and(389±56)％,which was significantly higher than(100±15)％in normal saline group,respectively(P＜0.05),and the Ki67 relative fluorescence intensity in the wound tissue of mice in combined treatment group was significantly higher than that in dressing group and rh-MT-Ⅲ group,respectively(with both P values＜0.05).On 7 d after injury,the TUNEL relative fluorescence intensity in the wound tissue of mice in dressing group,rh-MT-Ⅲ group,and combined treatment group was(55.5±5.7)％,(66.7±8.0)％,and(20.0±2.2)％,which was significantly lower than(100.0±12.9)％in normal saline group,respectively(P＜0.05),and the TUNEL relative fluorescence intensity in the wound tissue of mice in combined treatment group was significantly lower than that in dressing group and rh-MT-Ⅲ group,respectively(with both P values＜0.05).Conclusions rh-MT-Ⅲ combined with wound dressing can promote the growth of granulation tissue around the wound as well as collagen deposition,increase the cell proliferation vitality,reduce cell apoptosis,and promote the re-epithelialization of skin at the edge of the wounds,thus accelerating the healing of full-thickness skin defect wounds in mice.
Wounds and injuriesMetallothioneinBiological dressingsCell proliferationApoptosisWound repair
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