Fibrin u tkivnom inženjerstvu kože – proizvodnja i kliničko iskustvo

The purpose of this study was to create a fibrin-based human skin substitute in vitro with epidermal and dermal component and to assess its healing potential in deep partial and full thickness burns. Fibrin scaffolds were prepared from commercial fibrin glue kits. Human fibroblasts were cultured in fibrin gel. Human keratinocytes were seeded on the top of the gel. Viability of cells was determined fluorimetrically. Scanning electron microscope and immunocytochemistry analysis of cultured cells were performed. After hydrosurgical preparation of deep burn necrotic tissue, wound bed was prepared for skin substitutes. Progress of healing was documented using visual estimation and photos. Scanning electron microscope images showed good cell attachment and colony spreading of keratinocytes and fibroblasts on fibrin scaffold. Immunofluorescent staining of cell cultures on fibrin scaffold showed expression of vimentin, a marker of fibroblast cells, cytokeratin 19, a marker of epithelial stem cells, as well as involucrin, a marker of differentiated keratinocytes. Clinical results clearly showed that appearance of the skin did not differ significantly from the areas of transplanted skin using split-thickness skin graft techniques. In conclusion, using these fibrin-cultured autografts on massive full-thickness burn resulted in good healing.Cilj ovoga istraživanja bio je kreirati in vitro nadomjestak ljudske kože s epidermalnim i dermalnim dijelom. Pritom smo koristili fibrinski gel kao nosač stanica. Procijenili smo učinak nadomjestka kože na cijeljenje opeklina drugog i trećeg stupnja. Fibrinski gel načinjen je od komercijalnog kirurškog fibrinskog ljepila. Ljudski fibroblasti kultivirani su u samom fibrinskom nosaču. Ljudski keratinociti zasijani su na gornju površinu nosača. Vijabilnost uzgojenih stanica određena je fluorimetrijski. Nadomjestak kože analiziran je elektronskim skenirajućim mikroskopom. Napravljena je imunocitokemijska analiza kultiviranih stanica. Skenirajući mikroskop pokazao je dobro prianjanje i proliferaciju stanica kože na nosaču. Imunocitokemijska analiza pokazala je prisutnost vimentina, biljega fibroblastnih stanica; citokeratina 19, biljega epitelnih matičnih stanica; te involukrina, biljega diferenciranih keratinocita. Klinička primjena ovih nadomjestaka kože pokazala je slične rezulatate cijeljenja kao i dijelovi rana koji su prekriveni autolognim presatcima kože djelomične debljine. Zaključujemo da nadomjestci kože bazirani na fibrinskom nosaču imaju potencijala u liječenju teških opeklina