In vitro and in vivo cytokeratin patterns of expression in bioengineered human periodontal mucosa.
OBJECTIVE
Development of human oral mucosa substitutes by tissue engineering may provide new therapeutic tools for the management of periodontal diseases. In this study we evaluated a fibrin-agarose human oral mucosa substitute both in vitro and in vivo.


METHODS
In vitro bioengineered oral mucosa substitutes were developed from irrelevant biopsy samples of human oral gingiva. In vivo evaluation of the constructed tissues was performed by implantation into athymic nude mice. The expression of several epithelial markers was assessed by microarray analysis and immunohistochemistry.


RESULTS
Bioengineered oral mucosa samples kept in vitro developed a multilayered epithelium that expressed several cytokeratins, including some markers of simple epithelia (cytokeratins 7, 8 and 18), along with markers of stratified epithelia (cytokeratins 5 and 13) and of cell proliferation (proliferating cell nuclear antigen). Bioengineered tissues grafted in vivo onto nude mice exhibited very good biointegration with the host, showing a cytokeratin expression pattern that was very similar to that of normal native oral mucosa controls. Histological analysis of the artificial tissues demonstrated that oral mucosa substitutes evaluated in vivo were structurally mature, showing some typical structures of human native oral mucosa such as rete ridges and chorial papillae, along with numerous blood vessels at the fibrin-agarose stromal substitute. These structures were absent in samples evaluated in vitro.


CONCLUSIONS
The results indicate that this model of human oral mucosa, constructed using fibrin-agarose scaffolds, shows similarities to native oral mucosa controls and imply that bioengineered oral mucosa substitutes could eventually be used clinically.
Department of Histology, Tissue Engineering Group, University of Granada, Granada, Spain.
I
I
Garzón
M C
MC
Sánchez-Quevedo
G
G
Moreu
M
M
González-Jaranay
M
M
González-Andrades
A
A
Montalvo
A
A
Campos
M
M
Alaminos
D000818:Animals; D015415:Biomarkers / Q000032:analysis; D062109:Dermatologic Surgical Procedures; D004848:Epithelium / Q000033:anatomy & histology; D005337:Fibrin; D005347:Fibroblasts / Q000166:cytology; D005881:Gingiva / Q000033:anatomy & histology / Q000166:cytology* / Q000637:transplantation; D006085:Graft Survival; D006801:Humans; D053535:Keratin-13 / Q000032:analysis; D053538:Keratin-18 / Q000032:analysis; D053555:Keratin-5 / Q000032:analysis; D053552:Keratin-7 / Q000032:analysis; D053533:Keratin-8 / Q000032:analysis; D015603:Keratinocytes / Q000166:cytology; D007633:Keratins / Q000032:analysis*; D051379:Mice; D008819:Mice, Nude; D020411:Oligonucleotide Array Sequence Analysis; D018809:Proliferating Cell Nuclear Antigen / Q000032:analysis; D012685:Sepharose; D046509:Tissue Culture Techniques; D023822:Tissue Engineering; D054457:Tissue Scaffolds
D015415:Biomarkers; D053535:Keratin-13; D053538:Keratin-18; D053555:Keratin-5; D053552:Keratin-7; D053533:Keratin-8; D018809:Proliferating Cell Nuclear Antigen; D007633:Keratins; D005337:Fibrin; D012685:Sepharose