RECONSTITUTED HUMAN GINGIVAL EPITHELIUM - NONSUBMERGED IN-VITRO MODEL

Many studies have shown that human gingival keratinocytes grown in sub merged culture fail to attain optimal differentiation. This study repo rts an in vitro culture system for oral gingival epithelial cells, in which they are grown at the air-liquid interface, on polycarbonate ins erts, in the presence of an NIH-3T3 feeder layer. This model was compa red with two submerged culture methods for gingival keratinocytes, on type I collagen gel and on an NIH-3T3 feeder layer. Transmission elect ron microscopy showed an advanced level of stratification (over six la yers of cells) for cultures grown at the air-liquid interface. Immunof luorescence and electrophoretic patterns showed the presence of cytoke ratins 10 and 11 in cytoskeletal protein extracts of these cultured ke ratinocytes. In this air-liquid interface culture model in the presenc e of NIH-3T3 feeder cells, keratinocytes can achieve an advanced level of stratification and differentiation and a resemblance to in vivo gi ngiva. The obtention of a highly differentiated epithelium will permit in vitro pharmacological studies and studies on the biocompatability of certain alloys with the superficial periodontium; it will also prov ide grafts for patients undergoing periodontal iodontal surgery.