Graft of autologous fibroblasts in gingival tissue in vivo after culture in vitro preliminary study on rats

Several grafting techniques and guided tissue regeneration techniques (GTR) have been well-developed in periodontal surgery. However, these techniques could induce pain and side effects, such as a gingival recession during th e healing period following the therapy. The graft of a small autologous con nective tissue, using non-invasive surgical techniques could yield several benefits for the patients. Our preliminary study explores the feasibility o f collecting healthy gingival tissues, culturing them in vitro to amplify r at gingival fibroblasts (RGF) and inoculating the obtained cells into autol ogous rat gingival tissues in vivo. Gingival tissues samples were cultured as explants as described by Freshney et al. and Adolphe. Confluent cells su rrounding explants were detached after 7 d of culture from Petri dishes usi ng 0.05% trypsin and designated "first transferred cells" (T1). At the thir d passage (T3), cells cultured as monolayer were either examined under micr oscopy - phase contrast, scanning, or transmission electron - or numerated after trypan blue exclusion test. Autologous RGF labelled with fluorochrome were inoculated at the vestibular and palatine site of gingival tissue clo se to the superior incisors. In this preliminary study, 12 Wistar rats were used; for each, 2 biopsies were dissected and fixed for phase contrast or fluorescence microscopy. On d 1, 3 and 7 after injection in rat gingival ti ssues, fluorochrome-labelled cells could be detected in all these.