Dose-Dependent Effect of Cordycepin on Viability, Proliferation, Cell Cycle, and Migration in Dental Pulp Stem Cells

Abstract

Objective: To examine the effect of Cordycepin on the viability, proliferation, and migratory properties of dental pulp-derived mesenchymal stem cells. Materials and methods: The pulp was derived from human premolar teeth extracted for orthodontic purposes after obtaining informed consent. The samples were transferred to the laboratory for processing. DPSCs were expanded and characterized using flow cytometry and differentiation to the bone, adipose, and cartilage cells was examined. MTT Assay was performed using various concentrations of Cordycepin. The growth curve was plotted for 13 days. Cell cycle analysis was performed by flow cytometry. Migratory ability was assessed by wound healing assay. ROS generation was detected by flow cytometry. Gene expression was quantified by RT-qPCR. Statistical analysis was performed. p < 0.05 was considered as significant and p < 0.01 was considered as highly significant (* p < 0.05, and ** p < 0.01). Results: DPSCs expressed characteristic MSC-specific markers and trilineage differentiation. Cordycepin at lower concentrations did not affect the viability of DPSCs. The growth curve of cells showed a dose-dependent increase in cell numbers till the maximum dose. DPSCs treated with 2.5 µM Cordycepin was found to have a reduced G1 phase cell percentage. DPSCs treated with 2.5 µM and 5 µM Cordycepin showed a significant decrease in G2 phase cells. No significant difference was observed for S phase cells. Cordycepin treatment affected the migratory ability in DPSCs in a concentration-dependent manner. Conclusion: Cordycepin can be used at therapeutic doses to maintain stem cells.

Effect of Different Intracanal Medicaments on the Viability and Survival of Dental Pulp Stem Cells

“…The cells obtained by this method have shown characteristics similar to those obtained by the enzymatic method [17]. They also show the ability to differentiate into different lineages similar to MSCs and the potential for differentiation demonstrated by DPSCs [9,18,19].…”

Dose-Dependent Effects of Melatonin on the Viability, Proliferation, and Differentiation of Dental Pulp Stem Cells (DPSCs)

“…Separating the dental pulp stem cells from the pulp tissue samples ( n = 12) and characterization for MSC characteristics was conducted using the explant culture system based on a previous narration [ 28 ]. Briefly, pulp tissue was cut into small pieces using a blade, positioned in 25 mm polystyrene cell culture plates, and covered completely using enough volume of fetal bovine serum (FBS) (Gibco, Baltimore, MD, USA).…”

“…The cell surface markers were examined using a previously reported protocol [ 28 ]. Confluent DPSCs were extracted using trypsin, rinsed twice with phosphate-buffered saline (PBS), and then incubated for 30 min at 4 °C with anti-human CD73-APC, anti-human CD90-APC, anti-human CD105-APC, anti-human CD34-PE, anti-human CD45-FITC, and anti-human human leukocyte antigen-DR isotype-APC antibodies (BergischGladbach, Germany).…”

“…To verify the differentiation capacity of the DPSCs, 1 × 10 4 cells were plated into a 12-well cell culture plate and treated with two specific concentrations of melatonin (1 µM and 25 µM). The number of cells was assessed on alternate days for a total duration of 13 days using an inverted microscope [ 24 , 28 ] followed by plotting the growth curve by considering cell numbers for 13 days.…”

“…DPSCs were seeded at a density of 2500 cells/cm 2 into a 24-well cell culture plate and treated with adipocyte-inducing medium (Dulbecco’s modified Eagle medium containing ten percent fetal bovine serum, 10 micromoles insulin, 1 µM dexamethasone, 0.5 millimoles 3-isobutyl-1-methylxanthine, and 200 micromoles indomethacin) (Sigma-Aldrich Corporation, St. Louis, MO, USA). At every twenty-four hours, the medium was replaced twice every seven days until 21 days [ 28 ]. Adipogenic differentiation was performed by fixation of mature adipose cells using 4% paraformaldehyde and further adding 0.3% oil red O for one hour.…”