Three-dimensional scaffold from decellularized human gingiva for cell cultures: glycoconjugates and cell behavior

We studied both the presence of some carbohydrate compounds in a three-dimensional [3D] matrix harvested from human gingiva and the cell behavior in this matrix. In this experimental research, in order to prepare 3D scaffolds, human palatal gingival biopsies were harvested and physically decellularized by freeze-thawing and sodium dodecyl sulfate [SDS]. The scaffolds were placed within the rings of blastema tissues obtained from a pinna rabbit, in vitro. We evaluated the presence of glycoconjugates and cellular behavior according to histological, histochemical and spectrophotometry techniques at one, two and three weeks after culture. One-way analysis of variance [ANOVA] compared the groups. Extracellular matrix [ECM] remained after decellularization of tissue with 1% SDS. Glycoconjugate contents decreased meaningfully at a higher SDS concentration [p<0.0001]. After culture of the ECM scaffold with blastema, we observed increased staining of alcian blue, periodic acid-Schiff [PAS] and toluidine blue in the scaffold and a number of other migrant cells which was caused by cell penetration into the scaffold. Spectrophotometry results showed an increase in glycosaminoglycans [GAGs] of the decellularized scaffolds at three weeks after culture. The present study has shown that a scaffold generated from palatal gingival tissue ECM is a suitable substrate for blastema cell migration and activity. This scaffold may potentially be useful as a biological scaffold in tissue engineering applications

[ vitro histological investigation of interactions between rat's mesenchymal stem cells and human gingival matrix]

Extracellular matrix of natural tissues can be used as a scaffold for reconstructing biological tissues and organs. In this study, decellularized human gingival matrix was used as a scaffold for investigating the interactions of rat's bone marrow mesenchymal stem cells with human gingival matrix. To reach this goal, human gingival tissues were decellularized by two detergents sodium dodecyl sulfate [SDS] and Triton X-100. After washing and sterilization procedures, scaffolds were divided into 3 groups. Low density [LD] group was cultivated with 8x104 cells /cm2, high density [HD] group was cultivated with 8x105 cells/cm2, and control [C] group, was maintained in culture medium without any cells. Microscopic sections were prepared from the scaffoldsbefore and after 1, 2 and 4 weeks of culture with mesenchymal cells and were stained with Hematoxylin-Eosin. Repeated measure ANOVA was used to study the cell density alteration significance within the matrixes and post tests of means comparisons were performed within and between the groups. Also, gingival samples before and after decellularization procedure were investigated by scanning electron microscopy. Histological study of decellularized scaffolds revealed that nuclear and cellular components of the tissues were completely removed. Scanning electron microscopy of the scaffolds indicated that collagen fibers of connective tissue remained intact. Study of the scaffolds 1, 2 and 4 weeks after culture, revealed penetration of mesenchymal stem cells in scaffold, migration of cells towards connective tissue's papilla, and moreover epithelium-like structures. Statistical analysis indicated that cell density in HD group was significantly [P<0.05] higher than LD group. Cell density in both LD and HD groups significantly increased at 2nd week and decreased after4 weeks of culture. According to the results, scaffolds prepared from human gingival matrix can be a suitable scaffold for studying In vitro cell behaviors during oral wound healing