| Objective: To analyze the effect of skin homogenate injection on hypertrophic scar, and to investigate whether the skin homogenate injection method can successfully transplant skin tissue, and to supplement the epidermal stem cells in hypertrophic scar tissue.Methods: 1. Female New Zealand white rabbits, weight 1.5-2.0 kg,15, were manufactured hypertrophic scar model(HSM) on the ventral side of the rabbit ears, each ear make 3 HSMs, we total harvested 90 HSMs. HSMs were randomly divided into 3 groups, that is Group A(skin homogenate injection group)、Group B(normal saline group)、Group C(control group), each group contain 30 HSMs.2. Skin homogenate production: 4×8 cm2 pre-operation skin were prepared on the back of the rabbit, excision full thickness skin. The full thickness skin were cut to strips using a surgical scissors, addition NS, and continue to trim skin strips until it tune into mash.3. Management of Hypertrophic scar models: Group A: skin homogenate was injected underneath scar. Group B: Normal saline was injected underneath scar. Group C: nothing to do special.4. General observation: To measure the thickness of scar after the treatment 3 days, 5 days, 7 days, 14 days, 21 days, 28 days and record.5. Specimen Collection: To take biopsy and reserved by embedding in paraffin, after treatment 3 days, 5 days, 7 days, 14 days, 21 days, 28 days.6. Laboratory testing: routine pathologic examination was checked for each sample, then immunohistochemical assay were conducted on each sample to detect the expression of hair follicle stem cells marks K19、P63.7. Observation and compare the thickness, tissue structure and the changes of K19, P63 positive cells percentage between each group.8. Statistics analysis: The resulting data were analyzed using SPSS 20.0, and using the analysis of variance of mean comparison of multiple samples, p<0.05 was used as the criterion to judge the difference.Results: 1. The general observation: Thickness of hyperplastic scar was measured and proceed statistics analysis. After injection, the average thickness of group A and B increased, then, the average thickness decreased over time. Group C: the average thickness of hyperplastic scar decreased over time.2. Histological observation: HE dyeing: a large number of fiber cells, inflammatory cells and blood vessels can be seen in the scar tissue, fibroblasts is concentrated around the new blood vessels, and the collagen arranged promiscuously. Group A: injection of skin homogenate can be seen in the scar tissue, the appendants of skin survive, such as hair follicles and sebaceous glands.3. Special dyeing observation: After immunohistochemical staining, the cytoplasm of the cells expressing K19 and the nucleus of the cells expressing P63 is tan, and the positive cells mainly distributed in basal layer of epidermal. A large number of tan cells can be seen in the scar tissue of group A, and the positive cell percentage of K19 and P63 is significantly higher than group B and C. Proceed statistics analysis, p < 0.05, the difference was statistically significant.Conclusion: 1.The skin homogenate injection can successfully supply the epidermal stem cells to the hyperplastic scar tissue.2. The skin homogenate injection can successfully transplanted the skin tissue, the appendants of skin survive, such as hair follicles, sebaceous glands, with minimal invasion. |
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