Histological and histochemical evaluation of human oral mucosa constructs developed by tissue engineering
M.C. Sanchez-Quevedo1, M. Alaminos1,2, L.M. Capitan3, G. Moreu4, I. Garzon1, P.V. Crespo1 and A. Campos1
1Department of Histology, University of Granada, Spain, 2Fundación Hospital Clínico, Granada, Spain, 3Division of Oral and Maxillofacial Surgery, University Hospital Virgen de las Nieves, Granada, Spain and 4Department of Stomatology, University of Granada, Spain
Offprint requests to: Prof. M.C. Sanchez-Quevedo, Department of Histology, University of Granada, Avenida de Madrid 11, E-18012, Granada, Spain. e-mail: firstname.lastname@example.org
This work is dedicated to Prof. J. Gómez on his 85th anniversary.
Summary. Reconstruction of large oral mucosa defects is often challenging, since the shortage of healthy oral mucosa to replace the excised tissues is very common. In this context, tissue engineering techniques may provide a source of autologous tissues available for transplant in these patients. In this work, we developed a new model of artificial oral mucosa generated by tissue engineering using a fibrin-agarose scaffold. For that purpose, we generated primary cultures of human oral mucosa fibroblasts and keratinocytes from small biopsies of normal oral mucosa using enzymatic treatments. Then we determined the viability of the cultured cells by electron probe quantitative X-ray microanalysis, and we demonstrated that most of the cells in the primary cultures were alive and had high K/Na ratios. Once cell viability was determined, we used the cultured fibroblasts and keratinocytes to develop an artificial oral mucosa construct by using a fibrin-agarose extracellular matrix and a sequential culture technique using porous culture inserts. Histological analysis of the artificial tissues showed high similarities with normal oral mucosa controls. The epithelium of the oral substitutes had several layers, with desmosomes and apical microvilli and microplicae. Both the controls and the oral mucosa substitutes showed high suprabasal expression of cytokeratin 13 and low expression of cytokeratin 10. All these results suggest that our model of oral mucosa using fibrin-agarose scaffolds show several similarities with native human oral mucosa. Histol Histopathol 22, 631-640 (2007)
Key words: Fibrin-agarose, Constructs, Tissue engineering