Ex vivo and in vivo modulatory effects of umbilical cord Wharton’s jelly stem cells on human oral mucosa stroma substitutes
C.A. Alfonso-Rodríguez1,2, E. González-Andrades1, B.D. Jaimes-Parra1, R. Fernández-Valadés1,3, A. Campos1, M.C. Sánchez-Quevedo1, M. Alaminos1 and I. Garzón1
1Department of Histology (Tissue Engineering Group). University of Granada, Spain and Instituto de Investigación Biosanitaria ibs. Granada, 2PhD program in Biomedicine, University of Granada and 3Craniofacial Malformations and Cleft Lip and Palate Management Unit, Division of Pediatric Surgery, University Hospital Virgen de las Nieves, Granada, Spain
Offprint requests to: Dr. I. Garzón, Department of Histology. Faculty of Medicine. University of Granada. Avenida de Madrid 11, E18012, Granada, Spain. e-mail address: ingt@correo.ugr.es
Summary. Novel oral mucosa substitutes have been developed in the laboratory using human umbilical cord Wharton’s jelly stem cells -HWJSC- as an alternative cell source. In the present work, we have generated human oral mucosa substitutes with oral mucosa keratinocytes and HWJSC to determine the influence of these cell sources on stromal differentiation. First, acellular and cellular stroma substitutes and bilayered oral mucosa substitutes with an epithelial layer consisting of oral mucosa keratinocytes -OM samples- or HWJSC -hOM- were generated. Then, tissues were analyzed by light and electron microscopy, histochemistry and immunohistochemistry to quantify all major extracellular matrix components after 1, 2 and 3 weeks of ex vivo development, and OM and hOM were also analyzed after in vivo grafting. The results showed that bioengineered oral mucosa stromas displayed an adequate fibrillar mesh. Synthesis of abundant collagen fibers was detected in OM and hOM after 3 weeks, and in vivo grafting resulted in an increased collagen synthesis. No elastic or reticular fibers were found. Glycoprotein synthesis was found at the epithelial-stromal layer when samples were grafted in vivo. Finally, proteoglycans, decorin, versican and aggrecan were strongly dependent on the in vivo environment and the presence of a well-structured epithelium on top. The use of HWJSC was associated to an increased synthesis of versican. These results confirm the usefulness of fibrin-agarose biomaterials for the generation of an efficient human oral mucosa stroma substitute and the importance of the in vivo environment and the epithelial-mesenchymal interaction for the adequate differentiation of the bioengineered stroma. Histol Histopathol 30, 1321-1332 (2015)
Key words: Human Wharton’s jelly stem cells, Bioengineered oral mucosa, Extracellular matrix, Epithelial-mesenchymal transition, Tissue engineering
DOI: 10.14670/HH-11-628