Effects of tacrolimus on morphology, proliferation and differentiation of mesenchymal stem cells derived from gingiva tissue

Abstract

Tacrolimus is a 23-membered macrolide lactone with potent immunosuppressive activity that is effective in the prophylaxis of organ rejection following kidney, heart and liver transplantation. Tacrolimus also exerts a variety of actions on bone metabolism. The aim of the present study was to evaluate the effects of different concentrations of tacrolimus on the morphology and viability of human stem cells derived from the gingiva. Gingival-derived stem cells were grown in the presence of tacrolimus at final concentrations ranging from 0.001 to 100 µg/ml. The morphology of the cells was viewed under an inverted microscope and the cell viability was analyzed using Cell Counting kit-8 (CCK-8) on days 1, 3, 5 and 7. Alizarin Red S staining was used to assess mineralization of treated cells. The control group showed spindle-shaped, fibroblast-like morphology and the shapes of the cells in 0.001, 0.01, 0.1, 1 and 10 µg/ml tacrolimus were similar to those of the control group. All groups except the 100 µg/ml group showed increased cell proliferation over time. Cultures grown in the presence of tacrolimus at 0.001, 0.01, 0.1, 1 and 10 µg/ml were not identified to be significantly different compared with the control at days 1, 3 and 5 using the CCK-8 assays. Increased mineralized deposits were noted with increased incubation time. Treatment with tacrolimus from 0.001 to 1 µg/ml led to an increase in mineralization compared with the control group. Within the limits of this study, tacrolimus at the tested concentrations (ranging from 0.001 to 10 µg/ml) did not result in differences in the viability of stem cells derived from gingiva; however it did enhance osteogenic differentiation of the stem cells.

Evaluation of the effects of dimethylsulphoxide on morphology, cellular viability, mRNA, and protein expression of stem cells culture in growth media

“…In this study, a CCK-8 assay, which is based on the evaluation of mitochondrial activity, is used for the evaluation of cellular viability ( 17 ). A qPCR and a western blot analysis were performed to detect the mRNA and protein expression of collagen I and Runx2 to achieve information on the possible mechanisms.…”

The effects of on osteogenic differentiation and mineralization of human stem cells derived from the gingiva

“…1) and Alizarin red S staining assay measures the presence of calcium in cellular deposits which is used for the evaluation of early matrix mineralization. 17) Alkaline phosphatase activities increased between 3 and 7 days and mineralization increased between 3 and 7 Days. The highest alkaline phosphatase activity was achieved with 0.01% PPT on Day 3 and 0.01% group on Day 7.…”

“…The cellular viability was evaluated on Day 1 based on the previous report using the Counting Kit-8 (CCK-8, Dojindo, Tokyo, Japan) assay. 17) In short, cells were incubated with tetrazolium monosodium salt for 2 hours at 37 ˚C. Spectrophotometric absorbance at 450 nm was (417) measured using a microplate reader (BioTek Instruments Inc., Winooski, VT, USA).…”

Evaluation of the effects of Bambusa tulda on osteogenic differentiation and mineralization of human stem cells

“…Previously, differentiation effects of individual herbal extracts have been tested on human mesenchymal stem cells mainly derived from bone marrow and elaborated on the plausible underlying mechanisms of action [5]. Mesenchymal stem cells may be isolated from various tissues including gingival, as used in this study [16][17][18][19][20]. Gingiva can be easily accessible and obtainable [10,[21][22][23][24][25].…”