In vitro bioartificial skin culture model of tissue rejection and inflammatory/immune mechanisms

We hypothesized that an in vitro bioartificial skin rejection model using living LSEs grown in tissue culture could be developed for the study of autologous, allogenic, and/or xenogeneic inflammatory/immune mechanisms and topical immunosuppressive drugs. Human fibroblasts were mixed with type 1 rat-tail collagen to form a matrix (4 to 5 days), on which human keratinocytes were seeded. After a keratinocyte monolayer formed, CT cultures were raised to the air-liquid interface for continued growth. In the REJ LSE model, immunocytes isolated from human blood were seeded on top of the NHEK monolayer at the time of air-lifting. Thickness measurements of the acellular keratin and keratinocyte layers, and nuclear/cytoplasmic ratios, in both CT and REJ were made using digital image analysis. Immunostaining with anticytokeratin demonstrated a viable, keratin-producing epidermal layer; staining with anti-TGF-beta suggested a role for this cytokine in the rejection or wound-healing process. The LSE appeared histologically similar to normal human epidermis. Immunocytes added to the REJ cultures caused an obvious rejection response and were clearly identifiable in the gels as CD45+ staining cells. The LSE model appears promising for the study of immune/inflammatory mechanisms, thermal injury, screening antirejection agents that might be applied topically and as an in vitro replacement for skin graft studies in animals.     

Intralesional bleomycin-mediated electrochemotherapy in 20 patients with basal cell carcinoma

BACKGROUND: A new anticancer therapy, electrochemotherapy (ECT), has been introduced that entails exposing cancerous tissues to short pulses of electricity during chemotherapy. This enhances cell membrane permeability and has been shown to have potent antitumor effects in vitro in animal models and in several clinical trials, including nevoid basal cell carcinoma (BCC). OBJECTIVE: We report the effects of ECT on 20 patients with primary BCC. METHODS: Electrical pulses were delivered to 54 tumors after administration of intralesional bleomycin sulfate. RESULTS: Complete responses were observed in 53 (98%), and in the majority of these (94%) after a single treatment. No recurrences have been recorded with a mean of 18 months of observation. CONCLUSION: Although these are preliminary results, ECT appears to be an effective alternative to surgical excision for the treatment of primary BCC.