Abstract
Background: dental pulp-derived stem cells are easy to access and collect and are an excellent source of stem cells for regenerative therapy. These cells can interact with many biomolecules and scaffolds and can pass on the instructive signals to the sites of regeneration where they are used. In this regard cordycepin, a potential biomolecule derived from medicinal mushrooms with a spectrum of bioactive properties such as antioxidant, anti-inflammatory, and anticancer has not yet been tested for its effect on human dental pulp stem cells. Objective: the objective of the present study was to assess the in vitro adipogenic and osteogenic differentiation potential of human dental pulp stem cells with or without induction after administration of cordycepin. Materials and methods: human dental pulp stem cells DPSCs were isolated from a healthy permanent tooth extracted for orthodontic purposes after obtaining informed consent. Flow cytometry technique was used to assess the surface markers of these cells such as CD73, CD90, and CD105, CD34, CD45, and HLA-DR. Further, an MTT assay was performed on the cells after subjecting them to various concentrations of cordycepin. Following this, the adipogenic and osteogenic potential of the dental pulp stem cells was assessed with or without induction under the influence/absence of 5 µM of cordycepin. The results obtained were statistically analyzed and documented. Results: it was found that the dental pulp stem cells showed strong positive expression for CD73, CD90, and CD105 and faint expression of CD34, CD45, and HLA-DR. MTT assay revealed that 5 µM was the optimum concentration of cordycepin for all the assays. Concerning adipogenesis experiments, there was a statistically significant lowering of all the 4 adipogenesis-related genes PPARγ, FABP4, LPL, and C/EBPα following cordycepin treatment in the presence of induction compared to the only induction group and untreated control cells (p < 0.05). In connection with osteogenesis, was found that there was a statistically significant increase in the expression of RUNX2, COL1A1, OSX and OCN genes along with the increase in alkaline phosphatase and alizarin red staining in the DPSC treated with cordycepin along with the presence of induction and simultaneous addition of PDTC compared to the control untreated cells and cells treated with induction and simultaneous addition of PDTC (p < 0.05). Conclusion: cordycepin can be exploited for its osteopromotive properties and can be used as a bioactive molecule alongside the administration of dental pulp stem cells in the area of regenerative biology and medicine.
Highlights
Stem cells are a capable means to mankind, can differentiate into an array of cell types, and can be potentially used in the field of regenerative medicine to reconstruct lost tissues and organ parts [1,2,3]
MTT—48 h Absorbance at 570 nm Concerning the experiments assessing adipogenesis related genes in the dental pulp stem cells (DPSC) following induction and cordycepin treatment, it was found that there was a statistically significant lowering of all four adipogenesis related genes PPARγ, FABP4, LPL, and C/EBPα following cordycepin treatment in the presence of induction compared to the only induction group and untreated control cells (p < 0.05)
In connection with the ethe DPSC following induction with and without cordycepin treated experiments assessing the osteogenesis related genes and bone-forming characteristics of nt versus control cells along with the use of Pyrrolidine dithiocarbamate (PDTC) a nuclear factor (NF) kappa B inhibitor, it was found that there was a statistically significant increase in the expression of Runt-related transcription factor 2 (RUNX2), collagen, type I, alpha 1 (COL1A1), osterix (OSX) and osteocalcin (OCN) in the DPSC treated with cordycepin along with the presence of induction and simultaneous addition of PDTC compared to the control untreated cells and cells treated with induction and simultaneous addition of PDTC (p < 0.05) (Figure 3A–D, Table 6)
Summary
Stem cells are a capable means to mankind, can differentiate into an array of cell types, and can be potentially used in the field of regenerative medicine to reconstruct lost tissues and organ parts [1,2,3]. The success of stem cell harvesting largely depends on the access to the collection site and the invasiveness of the collection method. In this regard, the dental pulp is an easy to access site for stem cell collection, and the methodology of the collection is associated with a very low degree of morbidity and invasiveness [4]. It has been demonstrated that dental pulp stem cells (DPSC) are multipotent and can be differentiated into bone-forming osteoblasts. It is noteworthy that DPSC can interact well with scaffolds and biomaterials making the differentiation process easy and well-orchestrated [4]
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