Fabrication of cultured oral gingiva by tissue engineering techniques without materials of animal origin.
BACKGROUND
Cultured gingival substitute has been found to be a useful graft material for treatment of gingival recession. However, such substitutes include xenograft derivative materials that involve concomitant risk of viral contamination. To eliminate this risk, we designed new gingival substitutes made of recombinant human collagen types I and III sponges and cultured these substitutes in animal-free media (HFDM-1).


METHODS
Gingival fibroblasts were seeded onto sponges of type I or III recombinant collagen. These sponges were cultured in Dulbecco's modified Eagle's medium (DMEM) with 10% fetal bovine serum (FBS), HFDM-1 with 2% human serum (HS), or HFDM-1. Fibroblast proliferation in these samples was compared using the cell-counting kit assay. Vascular endothelial growth factor (VEGF) and hepatocyte growth factor (HGF) released into the cultured media were examined by enzyme-linked immunosorbent assay.


RESULTS
The fibroblasts proliferated significantly in all six combinations of collagen and medium types. The fibroblast growth rate after 9 days of culture was equal between HFDM-1 with 2% HS and DMEM with 10% FBS. The type III collagen sponge showed a higher fibroblast growth rate than the type I sponge. VEGF concentrations in HFDM-1 with 2% HS were higher than those in other media. The highest HGF levels were detected in DMEM with 10% FBS.


CONCLUSIONS
The new cultured gingival substitute containing no animal-derived materials produced good cell proliferation and VEGF release. The results suggested that the substitute may provide a new tool for the treatment of gingival recession.
Department of Research and Development, Japan Tissue Engineering, Aichi, Japan.
Kazuto
K
Yamada
Junji
J
Yamaura
Masakazu
M
Katoh
Ken-ichiro
K
Hata
Kazuhiro
K
Okuda
Hiromasa
H
Yoshie
D000818:Animals; D021522:Bioartificial Organs; D002417:Cattle; D018929:Cell Culture Techniques; D049109:Cell Proliferation; D002478:Cells, Cultured; D024042:Collagen Type I; D024061:Collagen Type III; D017077:Culture Media, Conditioned; D005347:Fibroblasts / Q000378:metabolism / Q000637:transplantation*; D005881:Gingiva / Q000166:cytology*; D005889:Gingival Recession / Q000601:surgery*; D017228:Hepatocyte Growth Factor / Q000096:biosynthesis; D006801:Humans; D011994:Recombinant Proteins; D023822:Tissue Engineering / Q000379:methods*; D042461:Vascular Endothelial Growth Factor A / Q000096:biosynthesis
D024042:Collagen Type I; D024061:Collagen Type III; D017077:Culture Media, Conditioned; D011994:Recombinant Proteins; D042461:Vascular Endothelial Growth Factor A; D017228:Hepatocyte Growth Factor