Abstract
Introduction The aim of the present study was to determine the effects of vitamins D and E on the proliferation, morphology, and differentiation of human dental pulp stem cells (hDPSCs). Methods In this in vitro experimental study, hDPSCs were isolated, characterized, and treated with vitamins D and E, individually and in combination, utilizing different doses and treatment periods. Changes in morphology and cell proliferation were evaluated using light microscopy and the resazurin assay, respectively. Osteoblast differentiation was evaluated with alizarin red S staining and expression of RUNX2, Osterix, and Osteocalcin genes using real-time RT-PCR. Results Compared with untreated cells, the number of cells significantly reduced following treatment with vitamin D (49%), vitamin E (35%), and vitamins D + E (61%) after 144 h. Compared with cell cultures treated with individual vitamins, cells treated with vitamins D + E demonstrated decreased cell confluence, with more extensive and flatter cytoplasm that initiated the formation of a significantly large number of calcified nodules after 7 days of treatment. After 14 days, treatment with vitamins D, E, and D + E increased the transcription of RUNX2, Osterix, and Osteocalcin genes. Conclusions Vitamins D and E induced osteoblastic differentiation of hDPSCs, as evidenced by the decrease in cell proliferation, morphological changes, and the formation of calcified nodules, increasing the expression of differentiation genes. Concurrent treatment with vitamins D + E induces a synergistic effect in differentiation toward an osteoblastic lineage.
Highlights
In the field of orthodontics, the study of mechanisms accelerating dental movement is being highlighted to enable a shortened treatment time and reduce side effects resulting from long periods of orthodontic treatment, such as tooth decay, periodontal disease, and root resorption [1].Several methods have been developed to accelerate orthodontic tooth movement, including surgeries, flaps, periodontal invasion, infections, and surgical site interventions, such as osteotomy and corticotomy
HDPSC treatment with vitamin E significantly reduced cell proliferation at all doses after 120 h of treatment, with a maximal decline of 31% observed with the highest concentration (12 μM) (Figure 2(a))
After 120 h, all treatment concentrations induced a significant decrease in cell number, with the greatest decline in cell proliferation (53%) observed in cells treated with vitamin D at 1 × 10−7 M when compared to proliferation in the untreated group (Figure 2(b))
Summary
In the field of orthodontics, the study of mechanisms accelerating dental movement is being highlighted to enable a shortened treatment time and reduce side effects resulting from long periods of orthodontic treatment, such as tooth decay, periodontal disease, and root resorption [1].Several methods have been developed to accelerate orthodontic tooth movement, including surgeries, flaps, periodontal invasion, infections, and surgical site interventions, such as osteotomy and corticotomy. In the field of orthodontics, the study of mechanisms accelerating dental movement is being highlighted to enable a shortened treatment time and reduce side effects resulting from long periods of orthodontic treatment, such as tooth decay, periodontal disease, and root resorption [1]. Ese substances are proposed to increase dental movement mainly through the induction of osteoclast formation; side effects such as toxicity, root resorption, and recurrence, as well as the mechanisms of action of these approaches, remain unclear [2]. Vitamin D demonstrates immunomodulatory actions by stimulating osteoblasts that generate bone mineralization. Erefore, vitamin D induces osteoclasts to produce bone resorption that facilitates dental movement, as well as bone formation that allows tooth stabilization. The biological mechanism that induces osteoblast and osteoclast activities mediated by vitamin D remains elusive [3]
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