Involvement of RAMP1/p38MAPK signaling pathway in osteoblast differentiation in response to mechanical stimulation: a preliminary study.

OBJECTIVE: The present study aimed to investigate the underlying mechanism of mechanical stimulation in regulating osteogenic differentiation. MATERIALS AND METHODS: Osteoblasts were exposed to compressive force (0-4 g/cm2) for 1-3 days or CGRP for 1 or 3 days. Expression of receptor activity modifying protein 1 (RAMP1), the transcription factor RUNX2, osteocalcin, p38 and p-p38 were analyzed by western blotting. Calcium mineralization was analyzed by alizarin red straining. RESULTS: Using compressive force treatments, low magnitudes (1 and 2 g/cm2) of compressive force for 24 h promoted osteoblast differentiation and mineral deposition whereas higher magnitudes (3 and 4 g/cm2) did not produce osteogenic effect. Through western blot assay, we observed that the receptor activity-modifying protein 1 (RAMP1) expression was upregulated, and p38 mitogen-activated protein kinase (MAPK) was phosphorylated during low magnitudes compressive force-promoted osteoblast differentiation. Further investigation of a calcitonin gene-related peptide (CGRP) peptide incubation, a ligand for RAMP1, showed that CGRP at concentration of 25 and 50 ng/ml could increase expression levels of RUNX2 and osteocalcin, and percentage of mineralization, suggesting its osteogenic potential. In addition, with the same conditions, CGRP also significantly upregulated RAMP1 and phosphorylated p38 expression levels. Also, the combination of compressive forces (1 and 2 g/cm2) with 50 ng/ml CGRP trended to increase RAMP1 expression, p38 activity, and osteogenic marker RUNX2 levels, as well as percentage of mineralization compared to compressive force alone. This suggest that RAMP1 possibly acts as an upstream regulator of p38 signaling during osteogenic differentiation. CONCLUSION: These findings suggest that CGRP-RAMP1/p38MAPK signaling implicates in osteoblast differentiation in response to optimal magnitude of compressive force. This study helps to define the underlying mechanism of compressive stimulation and may also enhance the application of compressive stimulation or CGRP peptide as an alternative approach for accelerating tooth movement in orthodontic treatment.

Light orthodontic force with high-frequency vibration accelerates tooth movement with minimal root resorption in rats.

OBJECTIVES: To determine and compare the effects of high-frequency mechanical vibration (HFV) with light force and optimal force on the tooth movement and root resorption in rat model. MATERIALS AND METHODS: Seventy-two sites in 36 male Wistar rats were randomly assigned using a split-mouth design to control (no force/no vibration) or experimental groups: HFV (125 Hz), light force (5 g), optimal force (10 g), light force with HFV, and optimal force with HFV for 14 and 21 days. The amount of tooth movement, 3D root volume, and root resorption area were assessed by micro-computed tomography and histomorphometric analysis. RESULTS: Adjunction of HFV with light force significantly increased the amount of tooth movement by 1.8-fold (p = 0.01) and 2.0-fold (p = 0.01) at days 14 and 21 respectively. The HFV combined with optimal force significantly increased the amount of tooth movement by 2.1-fold (p = 0.01) and 2.2-fold (p = 0.01) at days 14 and 21 respectively. The root volume in control (distobuccal root (DB): 0.60 ± 0.19 mm3, distopalatal root (DPa): 0.60 ± 0.07 mm3) and HFV (DB: 0.60 ± 0.08 mm3, DPa: 0.59 ± 0.11 mm3) were not different from the other experimental group (range from 0.44 ± 0.05 to 0.60 ± 0.1 mm3) with the lowest volume in optimal force group. CONCLUSIONS: Adjunction of HFV with orthodontic force significantly increased tooth movement without causing root resorption. CLINICAL RELEVANCE: Using light force with HFV could help to identify alternative treatment option to reduce the risk of root resorption.

Collagen-Based Biomaterials in Periodontal Regeneration: Current Applications and Future Perspectives of Plant-Based Collagen.

Collagen is the most widely distributed protein in human body. Within the field of tissue engineering and regenerative medical applications, collagen-based biomaterials have been extensively growing over the past decades. The focus of this review is mainly on periodontal regeneration. Currently, multiple innovations of collagen-based biomaterials have evolved, from hemostatic collagen sponges to bone/tissue regenerative scaffolds and injectable collagen matrices for gene or cell regenerative therapy. Collagen sources also differ from animal to marine and plant-extracted recombinant human type I collagen (rhCOL1). Animal-derived collagen has a number of substantiated concerns such as pathogenic contamination and transmission and immunogenicity, and rhCOL1 is a potential solution to those aforementioned issues. This review presents a brief overview of periodontal regeneration. Also, current applications of collagen-based biomaterials and their mechanisms for periodontal regeneration are provided. Finally, special attention is paid to mechanical, chemical, and biological properties of rhCOL1 in pre-clinical and clinical studies, and its future perspectives in periodontal regeneration are discussed.

Varied temporal expression patterns of trigeminal TRPA1 and TRPV1 and the neuropeptide CGRP during orthodontic force-induced pain.

OBJECTIVE: The aim of this study was to quantify the temporal changes in inflammation and TRPA1, TRPV1 and CGRP expression in the trigeminal ganglion during force-induced orthodontic pain. DESIGN: Orthodontic force was applied to both maxillary first molars in 8-week-old Wistar rats for 12 h, 24 h, 3 d or 7 d. The rat grimace scale (RGS) score and duration of face grooming were used to measure orthodontic pain. Western blotting was performed to assess TRPA1, TRPV1 and CGRP expression in trigeminal ganglia. NF-кB levels and colocalization of TRPA1, TRPV1 and CGRP were evaluated by immunofluorescent staining. RESULTS: Application of continuous force significantly increased pain behaviours at 1 and 3 d. NF-кB significantly increased in periodontal ligament at 12 h until 3 d. TRPV1 was significantly elevated within 1 d; TRPA1 significantly increased from 1-3 d; CGRP expression significantly increased from 12 h to 3 d. The TRPV1/TRPA1 expression ratio was highest at 12 h; the TRPA1/TRPV1 ratio peaked at 3 d. The percentages of trigeminal neurons co-expressing TRPA1/TRPV1, TRPA1/CGRP, and TRPV1/CGRP significantly increased by 12 h and peaked at 24 h. CGRP expression had a stronger positive correlation with TRPV1 than TRPA1. CONCLUSIONS: Inflammation induced by application of orthodontic force sensitizes trigeminal TRPV1 and TRPA1; TRPV1 is primarily activated as an early response, whereas TRPA1 is activated as a late response. Activation of both nociceptors results in CGRP release. Thus, blocking both TRPV1 and TRPA1 may represent a primary therapeutic target for relief of orthodontic pain.

Interval Vibration Reduces Orthodontic Pain Via a Mechanism Involving Down-regulation of TRPV1 and CGRP.

BACKGROUND/AIM: The transient receptor potential vanilloid 1 (TRPV1) ion receptor is involved in the release of calcitonin gene-related peptide (CGRP), a major contributor to orthodontic pain. Approaches that attenuate expression of TRPV1 and CGRP may reduce orthodontic pain. We explored the ability of high-frequency interval vibration to reduce orthodontic pain. MATERIALS AND METHODS: Orthodontic force (50 g) was applied to both maxillary first molars in 8-week-old Wistar rats (n=72). Vibration was applied at 125 Hz for 15 min/day. Duration of face grooming was assessed as a measure of orthodontic pain. Immunofluorescence and western blotting were used to assess TRPV1 and CGRP in the trigeminal ganglia. RESULTS: Compared to orthodontic force alone, application of vibration significantly decreased the duration of face grooming at 24 h and day 3 and reduced expression of TRPV1 and CGRP at 24 h. CONCLUSION: Vibration represents a promising mechanical approach to reduce orthodontic pain.

Vibration synergistically enhances IL-1ß and TNF-α in compressed human periodontal ligament cells in the frequency-dependent manner.

OBJECTIVES: To investigate whether mechanical vibration at 30 or 60 Hz combined with compressive force alter IL-1ß and TNF-α expression in human periodontal ligament (hPDL) cells. METHODS: hPDL cells isolated from the roots of first premolar teeth extracted from four independent donors were cultured and exposed to vibration (0.3 g, 20 min per cycle, every 24 h for 3 cycles) at 30 or 60 Hz (V30 or V60), 2.0 g/cm2 compressive force for 2 days (CF), or a combination of compressive force and vibration at 30 Hz or 60 Hz (V30CF or V60CF). Quantitative real-time polymerase chain reaction (qPCR) and enzyme-linked immunosorbent assays (ELISAs) were used to determine IL-1ß and TNF-α mRNA and protein, respectively. RESULTS: The levels of IL-1ß and TNF-α did not alter in groups V30 and V60. While, they were upregulated in groups CF, V30CF and V60CF. In addition, IL-1ß mRNA and TNF-α mRNA and protein were expressed at significantly higher levels in group V30CF compared to CF group. However, IL-1ß protein levels between V30CF and CF groups did not reach statistical significance. CONCLUSIONS: 30 Hz vibration had the synergistic effects with compressive force on the upregulation of IL-1ß mRNA and TNF-α mRNA and protein in PDL cells, while 60 Hz vibration did not have this synergistic effect.