Cultivo in vitro con colágeno y fibroblastos humanos de un equivalente de mucosa oral de espesor total

objectives. the objective of this study was to obtain, by in vitro culture, sheets of oral tissue in which complete oral mucosa structures can be identified. clinical application of the findings of this study will allow the replacement of free skin grafts or autologous oral mucosa grafts by this technique in certain cases. material and method. primary keratinocyte cultures were prepared from small biopsy samples of oral mucosa. secondary cultures were prepared from these primary cultures on an artificial submucosa constituted by collagen and human fibroblasts. the cell cultures were analyzed histologically in vitro and then used for graft implants in athymic mice to study their behavior in vivo. results. the primary cultures were confluent within a minimum period of 10 days and maximum of 12 days, which is similar to the period that the secondary cultures required to reach confluence. the time from sampling to achieving a complete artificial mucosa ranged from 20 to 22 days. the artificial mucosa showed histologic characteristics of a normal mucosa. after 17 days of graft implantation in immunoincompetent mice without any clinical contingency, histologic and immunohistochemical characterization (cytokeratins 19 and 13, collagen iv, and laminin) confirmed the similarity of the mucosa in vitro to healthy oral mucosa. conclusion. a complete oral mucosa equivalent can be prepared with collagen and fibroblasts using in vitro culture techniques. although this mucosa shows considerable retraction, its clinical handling is very favorable.