RESUMEN
BACKGROUND: Epidermal suction blister grafts are an effective treatment for chronic wounds or vitiligo, but this treatment is time consuming and limited to small areas. OBJECTIVES: To compare two novel strategies to create fractional epidermal grafts. METHODS: Epidermal blisters were raised from fresh human skin ex vivo at 38-40 °C, with suction of 380-510 mmHg. In Strategy 1, a 1-cm blister was micromeshed into approximately 500 pieces, transferred to elastic adhesive dressing, then pneumatically expanded to approximately nine times the original blister area. In Strategy 2, a 25-cm(2) array of 100 small blisters was raised, simultaneously harvested and captured directly onto an adhesive dressing. Measurements were taken for the pneumatic expansion limit, the release of microblisters upon hydration of the dressing adhesive, light microscopy, epidermal cell viability and positive L-3,4 dihydroxyphenylalanine melanocyte presence in blisters. RESULTS: Both strategies yielded viable fractional epidermal microblister arrays, carried on a dressing for transfer to graft recipient sites. The microblisters were gradually released upon hydration of the dressing adhesive. Strategy 2 has major advantages as only small blisters are made at the donor site, skilful dissection and physical expansion are not required and the strategy can be scaled to create large-area grafts. CONCLUSIONS: Strategy 2 is the more practical method for fractional epidermal micrografting to treat larger lesions with less donor-site trauma and has recently been commercialized.
Asunto(s)
Trasplante de Piel/métodos , Recolección de Tejidos y Órganos/métodos , Vesícula/fisiopatología , Supervivencia Celular , Enfermedad Crónica , Humanos , Melanocitos/fisiología , Succión , Colgajos Quirúrgicos , Vitíligo/cirugía , Cicatrización de HeridasRESUMEN
PURPOSE: To compare the induction and repair of radiation-induced base damage in human and rodent cell lines. MATERIAL AND METHODS: Experiments were performed with two human (normal fibroblasts HSF1 and tumour HeLa cells) and two rodent (mouse L929 and hamster CHO-K1) cell lines. Base damage was determined with the alkaline comet assay combined with the repair enzyme formamidopyrimidine-glycosylase (Fpg). Proteins were detected by Western blot. RESULTS: The induction of Fpg-sensitive sites was measured in human and rodent cell lines for doses up to 8 or 5 Gy, respectively. Comets were analysed in terms of tail moments, which were transformed into Gy-equivalents. The amount of Fpg-sensitive sites increased linearly with doses up to 4 Gy, whereby the ratio of single-strand breaks (ssb) to Fpg-sensitive sites was nearly identical for human and rodent cells with ssb:Fpg-sensitive sites=1:0.41+/-0.07 and 1:0.45+/-0.05, respectively. For doses exceeding 4 Gy, the amount of Fpg-sensitive sites did not increase further, indicating a dose limit up to which the comet assay can be used to detect Fpg-sensitive sites. Repair of Fpg-sensitive sites was studied for an X-ray dose of 4 Gy. For all four cell lines, the repair was measured to be completed 24 h after irradiation, but with pronounced differences in the kinetics. In both rodent cell lines, 50% of Fpg-sensitive sites were removed after t((1/2))=25+/-10 min in contrast to t((1/2))=80+/-20 min in the two human cell lines. The two species also differed in the level of polymerase ss with, on average, a three- to fivefold higher level in rodent cells compared with human cells. CONCLUSIONS: Repair of radiation-induced Fpg-sensitive sites was much faster in rodent than in human cells, which might result from the higher level of polymerase ss found in rodent cells.