Reference genes for valid gene expression studies on rat dental, periodontal and alveolar bone tissue by means of RT-qPCR with a focus on orthodontic tooth movement and periodontitis

Kirschneck, Christian; Proff, Peter; Fanghänel, Jochen; Wolf, Michael; Roldán, Juan; Römer, Piero

doi:10.1016/j.aanat.2015.11.005

Cited by 35 publications

(51 citation statements)

References 52 publications

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“…Apart from orthodontic tooth movement, PDLF also play a major role in the pathogenesis of periodontitis , a progressive infectious multifactorial inflammatory disease characterized by an excessive inflammatory host response as a defense reaction to pathogenic microorganisms within dental plaque, which results in loss of periodontal attachment and bone . Periodontal ligament fibroblasts have been shown to react to bacterial toxins, such as lipopolysaccharides, which are recognized by Toll‐like receptors , with a similar (but uncontrolled) release of proinflammatory cytokines and RANKL, inducing excessive osteoclastogenesis, matrix degradation, and periodontal bone loss Furthermore, interactive effects between periodontitis and orthodontic strain or tooth movement have been reported previously .…”

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confidence: 99%

Sodium‐chloride‐induced effects on the expression profile of human periodontal ligament fibroblasts with focus on simulated orthodontic tooth movement

Schröder¹,

Nazet²,

Neubert

et al. 2019

European J Oral Sciences

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Increased salt (NaCl) consumption triggers chronic diseases such as hypertension or osteopenia. Its impact on orthodontic tooth movement and periodontitis, however, has not been investigated, although both processes are related to the immune system, with periodontal ligament fibroblasts (PDLFs) playing a key mediating role. Here, we investigated the impact of NaCl on the expression pattern of PDLFs in a model of simulated compressive orthodontic strain. Periodontal ligament fibroblasts were preincubated for 24 h with additional 0 or 40 mM NaCl and concurrently treated for another 48 h with or without compressive strain of 2 g cm−2. We analyzed the expression of genes and proteins involved in orthodontic tooth movement by reverse transcription quantitative polymerase chain reaction (RT‐qPCR), ELISA, and immunoblot. Co‐culture experiments were performed to observe PDLF‐mediated osteoclastogenesis. A higher (40 mM) concentration of NaCl in the culture medium resulted in increased secretion of prostaglandin, expression of alkaline phosphatase, and expression of genes involved in extracellular matrix remodeling, but decreased compression‐induced expression of the interleukin‐6 (IL6) gene. The 40 mM concentration of NaCl also enhanced receptor activator of nuclear factor kappa‐B ligand (RANKL) but reduced that of osteoprotegerin (OPG), resulting in upregulated PDLF‐mediated osteoclastogenesis. A high NaCl concentration in the periodontal ligament, corresponding to a high‐salt diet in vivo, may influence orthodontic tooth movement and periodontitis through increased secretion of prostaglandins by PDLFs and upregulated PDLF‐mediated osteoclastogenesis, possibly accelerating orthodontic tooth movement and propagating periodontitis and periodontal bone loss.

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confidence: 99%

Sodium‐chloride‐induced effects on the expression profile of human periodontal ligament fibroblasts with focus on simulated orthodontic tooth movement

Schröder¹,

Nazet²,

Neubert

et al. 2019

European J Oral Sciences

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“…Hence, this gene can be regulated by mechanical forces, as demonstrated in some previous studies. Even though no significant difference in Gapdh expression was detected among the groups in our study, peptidylprolyl isomerase B ( Ppib ) and tyrosine 3‐monooxygenase/tryptophan 5‐monooxygenase activation protein, zeta ( Ywhaz ) might be more suitable as reference genes for normalization because in a previous study they were recommended as the most stable reference genes for normalization in the combined dental, periodontal, and alveolar bone tissue of the rat . In addition, we used the incisors as the anchor for the mesial M1 movement in the rat tooth‐movement model.…”

Section: Discussionmentioning

confidence: 87%

“…Using this model, we evaluated the expression of Hmgb1 mRNA by real‐time PCR and HMGB1 by immunofluorescence. In standardized dental‐periodontal tissue , which consists of the adjacent periodontal ligament, alveolar bone ,and some dental pulp, Hmgb1 was downregulated at the transcriptional level, as determined using Gapdh as the reference gene. The 100‐g group showed increases in the translocation and extracellular expression of HMGB1, whereas a decreased mean optical density of HMGB1 in the nucleus was found in the force groups.…”

Section: Discussionmentioning

confidence: 99%

“…The tooth‐movement model used was established as previously described by Kirschneck et al . , with some modifications. Briefly, a nickel‐titanium (Ni‐Ti) coil spring (1 mm diameter; 4 mm length; 0.2 mm wire size) (Smart Technology, Beijing, China) with an activating force of approximately 50 or 100 g (Correx Tension Gauge; Haag‐Streit, Koeniz, Switzerland), which are heavy forces for rats , was connected between the right first molar (M1) and the incisors to induce mesial migration of the M1.…”

Section: Methodsmentioning

confidence: 99%

“…In addition, according to the method described by Kirschneck et al . , standardized tissue samples were obtained, and total RNA from the resulting powder was extracted using 1 ml of TRIzol reagent (Invitrogen, Carlsbad, CA, USA), following the manufacturer's recommended protocol (Fig. ).…”

Section: Methodsmentioning

confidence: 99%

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Stress overload‐induced periodontal remodelling coupled with changes in high mobility group protein B1 during tooth movement: an in‐vivo study

Zou

Lin

2019

European J Oral Sciences

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High mobility group protein B1 (HMGB1), a bone‐active cytokine and an osteocyte alarmin, might have dual functions in bone metabolism that could benefit bone formation and accelerate osteoclastogenic activity. High mobility group protein B1 was recently shown to be involved in tooth movement. Here, we investigated the expression of HMGB1, which remains poorly elucidated, under stress overload‐induced periodontal remodelling conditions in vivo. Thirty‐six Sprague‐Dawley rats (male, 180–200 g) were randomly divided into three groups: two experimental groups, in which 50 or 100 g of force was applied to the first molars for 7 d to induce movement; and one control group, in which no force was applied. These stresses induced tooth movement over significantly different distances, and marked morphological changes were consistently observed in the periodontal tissues of the experimental rats, as demonstrated by histological staining. A real‐time PCR analysis showed upregulation of the receptor activator of nuclear factor‐kappaB ligand‐to‐osteoprotegerin ratio and downregulation of the Hmgb1 gene. Changes in both location and expression of the HMGB1 were observed through immunofluorescence analysis. Our data suggest that HMGB1 expression during orthodontic tooth movement might be regulated in a time‐ and force‐dependent manner that is substantially more complex than anticipated.

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Effects of sodium chloride on the gene expression profile of periodontal ligament fibroblasts during tensile strain

et al. 2020

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Purpose During orthodontic tooth movement, pressure and tension zones develop in the periodontal ligament, and periodontal ligament fibroblasts (PDLF) become exposed to mechanical strain. Enhanced salt (NaCl) concentrations are known to modulate responses of PDLF and immune cells to different stimuli like mechanical strain. Here, we investigated the impact of tensile strain on the gene expression profile of PDLF under normal (NS) and high salt (HS) conditions. Methods After preincubation under NS or HS (+40 mM NaCl in medium) conditions for 24 h, PDLF were stretched 16% for 48 h using custom-made spherical cap silicone stamps using an established and published setup. After determination of cell number and cytotoxicity, we analyzed expression of genes involved in extracellular matrix reorganization, angiogenesis, bone remodeling, and inflammation by quantitative real-time polymerase chain reaction (RT-qPCR). Results Tensile strain did not affect the expression of genes involved in angiogenesis or extracellular matrix reorganization by PDLF, which however modulate inflammatory responses and bone remodeling in reaction to 16% static tensile strain. Salt (NaCl) treatment triggered enhanced extracellular matrix formation, expression of cyclooxygenase 2 and bone metabolism in PDLF during tensile strain. Conclusions Salt (NaCl) consumption may influence orthodontic tooth movement and periodontal bone loss via modulation of extracellular matrix and bone metabolism. Excessive salt intake during orthodontic therapy may cause adverse effects regarding periodontal inflammation and bone resorption.

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Reference genes for valid gene expression studies on rat dental, periodontal and alveolar bone tissue by means of RT-qPCR with a focus on orthodontic tooth movement and periodontitis

Cited by 35 publications

References 52 publications

Sodium‐chloride‐induced effects on the expression profile of human periodontal ligament fibroblasts with focus on simulated orthodontic tooth movement

Sodium‐chloride‐induced effects on the expression profile of human periodontal ligament fibroblasts with focus on simulated orthodontic tooth movement

Stress overload‐induced periodontal remodelling coupled with changes in high mobility group protein B1 during tooth movement: an in‐vivo study

Effects of sodium chloride on the gene expression profile of periodontal ligament fibroblasts during tensile strain

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