The role of 25-hydroxyvitamin-D3 and vitamin D receptor gene in human periodontal ligament fibroblasts as response to orthodontic compressive strain: an in vitro study

Abstract

Background This study aimed to investigate, if different physiological concentrations of vitamin D (25(OH)D3) and single nucleotide polymorphisms in vitamin D receptor (VDR) gene have an impact on gene expression in human periodontal ligament (hPDL) fibroblasts induced by simulated orthodontic compressive strain. Methods A pool of hPDL fibroblasts was treated in absence or presence of 25(OH)D3 in 3 different concentrations (10, 40 and 60 ng/ml). In order to evaluate the role of single nucleotide polymorphisms in the VDR gene, hPDL fibroblasts from 9 patients were used and treated in absence or presence of 40 ng/ml 25(OH)D3. Each experiment was performed with and without simulated orthodontic compressive strain. Real-time PCR was used for gene expression and allelic discrimination analysis. Relative expression of dehydrocholesterol reductase (DHCR7), Sec23 homolog A, amidohydrolase domain containing 1 (AMDHD1), vitamin D 25-hydroxylase (CYP2R1), Hydroxyvitamin D-1-α hydroxylase, receptor activator of nuclear factor-κB ligand (RANKL), osteoprotegerin (OPG), cyclooxygenase-2 (COX-2) and interleukin-6 (IL6) was assessed. Three single nucleotide polymorphisms in VDR were genotyped. Parametric or non-parametric tests were used with an alpha of 5%. Results RANKL, RANKL:OPG ratio, COX-2, IL-6, DHCR7, CYP2R1 and AMDHD1 were differentially expressed during simulated orthodontic compressive strain (p < 0.05). The RANKL:OPG ratio was downregulated by all concentrations (10 ng/ml, 40 ng/ml and 60 ng/ml) of 25(OH)D3 (mean = 0.96 ± 0.68, mean = 1.61 ± 0.66 and mean = 1.86 ± 0.78, respectively) in comparison to the control (mean 2.58 ± 1.16) (p < 0.05). CYP2R1 gene expression was statistically modulated by the different 25(OH)D3 concentrations applied (p = 0.008). Samples from individuals carrying the GG genotype in rs739837 presented lower VDR mRNA expression and samples from individuals carrying the CC genotype in rs7975232 presented higher VDR mRNA expression (p < 0.05). Conclusions Simulated orthodontic compressive strain and physiological concentrations of 25(OH)D3 seem to regulate the expression of orthodontic tooth movement and vitamin-D-related genes in periodontal ligament fibroblasts in the context of orthodontic compressive strain. Our study also suggests that single nucleotide polymorphisms in the VDR gene regulate VDR expression in periodontal ligament fibroblasts in the context of orthodontic compressive strain.

The relationship between vitamin D receptor gene and TREM‐1 gene polymorphisms and the susceptibility and prognosis of neonatal sepsis

“…In this study, we found that the serum 25(OH)D level of patients with sepsis was significantly lower than that of the control group, and the VDR rs739837 G > T allelic variation was correlated with the increase of serum 25(OH)D. Küchler et al 24 found that the VDR mRNA expression level in samples of GG genotype in rs739837 was lower, which was consistent with the results of this study. 25(OH)D levels are closely related to neonatal sepsis.…”

“…In addition to the classic regulation of calcium and phosphorus metabolism, when combined with VDR, vitamin D can also inhibit proliferation, promote differentiation, regulate immunity, inhibit cell necrosis, inhibit tumor infiltration and metastasis, and other non-calcium regulatory effects. [21][22][23] In this study, we found that the serum 25(OH)D level of patients with sepsis was significantly lower than that of the control group, and the VDR rs739837 G > T allelic variation was correlated with the increase of serum 25(OH)D. Küchler et al 24 with sepsis and bronchial bubble lavage fluid of patients with pneumonia, sTREM-1 is significantly increased. 30 In this study, we found that the serum sTREM-1 level of neonatal sepsis was significantly higher than that of the control group.…”

Radiographic Bone Loss and Its Relation to Patient-Specific Risk Factors, LDL Cholesterol, and Vitamin D: A Cross-Sectional Study

“…These results are in line with a recently published systematic review and meta-analysis concluding that periodontitis is associated with lower 25OHD levels [15]. This concurs with the known bone-protective effect of higher 25OHD levels, which have been shown to decrease the ratio of RANKL to OPG expression by periodontal ligament fibroblasts controlling osteoclastogenesis [60]. Ketharanathan et al focused on radiographic bone loss in their cohort study to investigate the impact of 25OHD levels in periodontitis patients.…”

Association between genetic variants in key vitamin‐D‐pathway genes and external apical root resorption linked to orthodontic treatment

“…Even though this situation would not necessarily occur in vivo, this could explain why ff subjects for rs2228570, associated with higher vitamin D levels, would show more external apical root resorption during orthodontic treatment. Besides, it was shown that other SNPs in VDR influence VDR mRNA expression under conditions of simulated pressure and 25(OH)D 3 supply [54]. These findings would provide new insights into the molecular mechanisms of how SNPs in VDR would influence the amount of external apical root resorption related to orthodontic treatment.…”