An investigation of preferential fibroblast wound repopulation using a novel in vitro wound model

Abstract

To overcome the difficulties of studying wounding and wound repopulation in monolayer systems, a 3-dimensional model of wound repopulation has been developed which allows the in vitro investigation of fibroblast migration in response to experimental wounding. This model was utilized to determine whether fibroblasts derived from sites which demonstrate preferential healing (child and oral mucosal fibroblasts) possessed an increased ability to repopulate experimental wounds when compared to adult dermal fibroblasts. Fibroblasts were established from specimens derived from healthy donors undergoing minor elective surgery. Standard wounds were created in fibroblast populated collagen lattices (FPCLs) which were then overlaid upon an extracellular wound matrix. Fibroblast repopulation of the wounds was studied over 12 days using light- and scanning electron microscopy and quantified using computerized image analysis. Wound repopulation by fibroblasts derived from child donors (n = 3) was significantly (P < 0.001) more rapid than their adult tissue-matched counterparts (n = 3). Wound repopulation by oral mucosal fibroblasts (n = 3) was significantly greater than that exhibited by age-matched dermal fibroblasts (n = 3; P < 0.05). These differences were not reflected in differences in DNA synthesis (P > 0.5) or cell number (P > 0.5) within similar attached FPCL systems. These findings further support the concept of a gradual transition from the fetal to adult phenotype in wound healing. The potential applications of the model are discussed.