Microstructure Of Alveolar Bone Research Articles

Effect of estrogen depression on alveolar bone microarchitecture and periodontal ligament cells during orthodontic movement.

This study aimed to evaluate the effects of the estrogen depression during orthodontic tooth movement on alveolar bone microarchitecture and periodontal ligament. Female Wistar rats were divided into two groups, one consisting of non-ovariectomized animals subjected to orthodontic tooth movement, and one comprising ovariectomized animals subjected to orthodontic tooth movement. Micro-CT assessment of bone volume to total volume (BV/TV), total porosity, trabecular thickness (Tb.Th), trabecular number (Tb.N), and trabecular separation (Tb.Sp) in the alveolar bone of the orthodontically moved tooth was performed. Histomorphometric analyses were made in the periodontal ligament, and immunoexpression of RANK, RANKL, OPG, and TUNEL were quantified. Orthodontic tooth movement in the group of ovariectomized rats was faster than in non-ovariectomized animals. The alveolar bone area showed lower values of BV/TV and trabecular thickness, and higher bone porosity and trabeculae numbers in the ovariectomized rats. Histological analyses in the ovariectomized group revealed an increase in collagen fibers in the periodontal ligament. The apoptotic cell counts in the periodontal ligament were higher in the group of ovariectomized rats than in the sham-operated rats. Ovariectomy resulted in an increase in tooth movement and alteration of the alveolar bone microstructure in the first 7day of orthodontic tooth movement, and in the presence of apoptotic cells in the periodontal ligament.

Local E-rhBMP-2/β-TCP Application Rescues Osteocyte Dendritic Integrity and Reduces Microstructural Damage in Alveolar Bone Post-Extraction in MRONJ-like Mouse Model.

The pathology of medication-related osteonecrosis of the jaw (MRONJ), often associated with antiresorptive therapy, is still not fully understood. Osteocyte networks are known to play a critical role in maintaining bone homeostasis and repair, but the exact condition of these networks in MRONJ is unknown. On the other hand, the local application of E-coli-derived Recombinant Human Bone Morphogenetic Protein 2/β-Tricalcium phosphate (E-rhBMP-2/β-TCP) has been shown to promote bone regeneration and mitigate osteonecrosis in MRONJ-like mouse models, indicating its potential therapeutic application for the treatment of MRONJ. However, the detailed effect of BMP-2 treatment on restoring bone integrity, including its osteocyte network, in an MRONJ condition remains unclear. Therefore, in the present study, by applying a scanning electron microscope (SEM) analysis and a 3D osteocyte network reconstruction workflow on the alveolar bone surrounding the tooth extraction socket of an MRONJ-like mouse model, we examined the effectiveness of BMP-2/β-TCP therapy on the alleviation of MRONJ-related bone necrosis with a particular focus on the osteocyte network and alveolar bone microstructure (microcrack accumulation). The 3D osteocyte dendritic analysis showed a significant decrease in osteocyte dendritic parameters along with a delay in bone remodeling in the MRONJ group compared to the healthy counterpart. The SEM analysis also revealed a notable increase in the number of microcracks in the alveolar bone surface in the MRONJ group compared to the healthy group. In contrast, all of those parameters were restored in the E-rhBMP-2/β-TCP-treated group to levels that were almost similar to those in the healthy group. In summary, our study reveals that MRONJ induces osteocyte network degradation and microcrack accumulation, while application of E-rhBMP-2/β-TCP can restore a compromised osteocyte network and abrogate microcrack accumulation in MRONJ.

Thermogenic preworkout supplement induces alveolar bone loss in a rat model of tooth movement via RANK/RANKL/OPG pathway.

The aim of this study was to investigate the effect of thermogenic supplementation on the bone tissue of rats subjected to orthodontic movement. A total of 38 male Wistar rats underwent orthodontic movement of the left permanent maxillary first molars for 21 days. The rats were assigned to three groups: Control group: water; Thermogenic 1: C4 Beta Pump thermogenic; or Thermogenic 2: PRE-HD/Pre-workout. Micro-computed tomography (micro-CT) was used to investigate the dynamic changes in the microstructure of alveolar bone during orthodontic tooth movement in rats. Histopathologic analysis was performed by hematoxylin and eosin (H&E) staining, whereas tartrate-resistant acid phosphatase (TRAP) was employed for osteoclast count. Maxillary tissue was collected and evaluated by immunohistochemistry for receptor activator of NF-κB (RANK), receptor activator of NF-κB ligand (RANKL), and osteoprotegerin (OPG). The Thermogenic 2 group exhibited a significantly lower percentage of bone volume fraction (BV/TV) (68.21% ± 17.70%) compared to the control (86.84% + 12.91%) and Thermogenic 1 groups (86.84% + 15.94%) (p < 0.05). The control group had a significantly higher mean orthodontic movement in the mesial direction (0.2143 mm + 0.1513 mm) than the Thermogenic 2 group (0.0420 mm + 0.05215 mm) (p < 0.05). The Thermogenic 2 and Thermogenic 1 groups showed a stronger immunostaining for RANKL when compared to the control group (p < 0.05). The supplementation used in the Thermogenic 2 group (PRE-HD/Pre-workout) induced alveolar bone loss in rats subjected to orthodontic movement, which can be related to the regulation of the RANK/RANKL/OPG signaling pathway. This suggests the influence of thermogenic supplements on bone metabolism seems to depend on their composition.

Alveolar Bone Microstructure Surrounding Orthodontic Anchor Screws with Plasma Surface Treatment in Rats.

A lateral load was applied to anchor screws that had undergone surface treatment, and the structure, cellular dynamics, and quality of the bone surrounding anchor screws were analyzed to investigate the effect of this surface treatment on the peri-implant jawbone. In addition, bone microstructural characteristics were quantitatively evaluated for each site of loading on the bone around the anchor screw. Rats were euthanized after observation on days 3, 5, or 7, and bone quality analyses were performed. Bone-implant contact rate increased more rapidly at an early stage in the treated surface group than in the untreated surface group. Bone lacuna morphometry showed that the measured values adjacent to the screw at the screw neck on the compressed side (A) and at the screw tip on the uncompressed side (D) were significantly lower than those at the screw tip on the compressed side (B) and at the screw neck on the uncompressed side (C). Collagen fiber bundle diameter showed that the measured values adjacent to regions A and D were significantly higher than those at regions B and C. Anchor screw surface activation facilitates initial bone contact of the screw, suggesting that early loading may be possible in clinical practice.

Effects of Hederin (Hed) on Alveolar Bone Microstructure During Tooth Movement in Rats

Purpose: To discuss effects of Hederin (Hed) to tooth movement process in rats. Materials and methods: 54 rats were divided into Model and Hed groups at 7 d, 14 d and 21 d, establishment of rat tooth movement model, the Hed and Model group injected Hed [5 mg/(kg· d)] and equal volume normal saline intraperitoneally respectively, Take the medicine regularly every night. After 14 days, 9 rats in each group were killed, BV/TV, Tb. SP and trabecular number (Tb. N) were measured by Mirco CT. Using TRAP staining and HE staining to observe osteoclasts number and pathology change. The relative protein expressions were measured by SP staining. Results: Compared with Model group, BV/TV and Tb.N were significantly down-regulation and Tb.Sp was significantly up-regulation in Hed group (P &lt; 0.05, respectively); meanwhile, tooth movement and osteoclast number were significantly improved in Hed groups at 7 d, 14 d and 21 d (P &lt;0.05, respectively). By SP staining, compared with Model group, ADRB2 and RANKL proteins expression were significantly enhanced at 7 d, 14 d and 21 d (P &lt;0.05, respectively). Conclusion: Hed could promote alveolar bone resorption and increase the expression of ADRB2 and RANKL during orthodontic tooth movement.

Evaluation of Alveolar Bone Microstructure around Impacted Maxillary Canines Using Fractal Analysis in Dravidian Population: A Retrospective CBCT Study

The aim of this study was to assess alveolar bone microstructure around impacted maxillary canines derived from fractal analysis. The present study was a retrospective cone-beam computed tomography (CBCT) study. In total, 61 scans (25 males, 36 females; age range - 12-28 years) were analyzed. About 64 × 64-pixel regions of interest (ROI) in the maxillary alveolar process containing trabecular bone between the premolars were selected. ImageJ software was utilized to process images and bone density was assessed by determining bone surface area (BSA) and bone marrow surface area (BMSA) for the impacted and nonimpacted sides separately. Selected fractals were plotted in a histogram using box-counting method and the results were tabulated. Paired t-tests were used to determine significant differences between the groups and gender differences. For both buccally and palatally impacted maxillary canines, BSA was increased, BMSA decreased in the region of the impacted canine and the difference was statistically significant (p <0.05) in both genders. Bone fractal dimension (FD) values were greater in the region of the impacted canine for both buccally (1.47 > 1.21) and palatally (1.53 > 1.43) displaced canines, while bone marrow FD values were greater in the region of the nonimpacted canine for both buccally (1.37 > 1.28) and palatally displaced canines (1.41 > 1.33). Females had significantly higher BMSA than males around impacted canines (p <0.05). Denser bone microstructure was noted around impacted canines when compared with fully erupted canines. No gender-related differences were noted for BSA, whereas BMSA was higher in females implying lower bone density when compared with males. Retrospective evaluation of bone microstructure surrounding unerupted/impacted canines can provide analytical information about treatment prognosis and anchorage considerations. With FD analysis of CBCT images, BSA and BMSA can be measured and bone density estimated in a reliable manner.

Fluoride exposure duringintrauterine and lactation periods promotes changes in the offspring rats' alveolar bone

The importance of fluoride (F) for oral health is well established in the literature. However, evidence suggests that excessive exposure to this mineral is associated with adverse effects at different life stages and may affect many biological systems, especially mineralized tissues. The purpose of this study was to investigate the effects of F exposure during pregnancy and breastfeeding on the alveolar bone of the offspring since the alveolar bone is one of the supporting components of the dental elements. For this, the progeny rats were divided into three groups: control, 10 mg F/L, and 50 mg F/L for 42 (gestational and lactation periods). Analysis of the quantification of F levels in the alveolar bone by particle-induced gamma emission; Raman spectroscopy to investigate the physicochemical aspects and mineral components; computed microtomography to evaluate the alveolar bone microstructure and analyses were performed to evaluate osteocyte density and collagen quantification using polarized light microscopy. The results showed an increase in F levels in the alveolar bone, promoted changes in the chemical components in the bone of the 50 mg F/L animals (p < 0.001), and had repercussions on the microstructure of the alveolar bone, evidenced in the 10 mg F/L and 50 mg F/L groups (p < 0.001). Furthermore, F was able to modulate the content of organic bone matrix, mainly collagen; thus, this damage possibly reduced the amount of bone tissue and consequently increased the root exposure area of the exposed groups in comparison to a control group (p < 0.001). Our findings reveal that Fcan modulate the physicochemical and microstructural dimensions and reduction of alveolar bone height, increasing the exposed root region of the offspring during the prenatal and postnatal period. These findings suggest that F can modulate alveolar bone mechanical strength and force dissipation functionality.

Therapeutic Potential of Liraglutide for Diabetes-Periodontitis Comorbidity: Killing Two Birds with One Stone.

Background The relationship between diabetes and periodontitis is bidirectional, and there is now consensus that periodontitis and diabetes are comorbid. There is a quest for a drug that can be used to treat both conditions simultaneously. This study evaluated the anti-inflammatory and osteoprotective effects of liraglutide (LIRA) on periodontitis in diabetic rats. Methods Male Wistar rats (n = 46) were randomly divided into four groups: control group (n = 8), LIRA group (n = 8), diabetes-associated periodontitis+0.9% saline group (diabetic periodontitis (DP)+NaCl group, n = 15), and diabetes-associated periodontitis+LIRA group (DP+LIRA group, n = 15). LIRA treatment lasted for 4 weeks (300 μg/kg/d) after establishment of a rat model of DP. The expression of IL-6, TNF-α, and IL-1β was detected by enzyme-linked immunosorbent assay. The morphological changes of periodontal tissues were observed by hematoxylin–eosin staining. The absorption of alveolar bone and its ultrastructural changes were observed by histomorphometry and microcomputed tomography. The expression of receptor activator of NF-κB ligand (RANKL) and osteoprotegerin (OPG) in alveolar bone was detected by immunohistochemistry. The levels of Runx2 mRNA and ALP mRNA in the gingival epithelium were examined by quantitative real-time polymerase chain reaction. Results LIRA decreased alveolar bone resorption, improved the microstructure of alveolar bone, and reduced periodontal inflammation and damage (P < 0.05). LIRA also reduced blood glucose level and inhibited the secretion of serum IL-6, TNF-α, and IL-1β (P < 0.05). In addition, after treatment with LIRA, the ratio of RANKL/OPG was reduced, and the expression levels of ALP mRNA and Runx2 mRNA were upregulated (P < 0.05). Conclusions LIRA not only controls blood glucose level but also reduces inflammation and bone loss and enhances osteogenic differentiation in diabetes-associated periodontitis. Those indicate that LIRA may be used as a potential medicine for the adjunctive therapy of diabetes-periodontitis comorbidity.

N-acetylcysteine promotes cyclic mechanical stress-induced osteogenic differentiation of periodontal ligament stem cells by down-regulating Nrf2 expression

Background/purposeMechanical stress plays a vital role in osteogenic differentiation of periodontal ligament stem cells (PDLSCs). Cyclic mechanical stress may up-regulate reactive oxygen species (ROS) level. N-acetylcysteine (NAC) possesses powerful antioxidant capacity. However, it is undefined the impact of NAC on osteogenic differentiation stimulated by cyclic mechanical stress in PDLSCs. The aim of our research was to study the effect of NAC on PDLSCs during osteogenic differentiation under cyclic mechanical stress. Materials and methodsThe expression levels of osteogenesis markers were used to examine the osteogenic differentiation of PDLSCs. ROS production were measured by flow cytometry. The levels of reduced glutathione (GSH) and oxidized glutathione (GSSG) were analyzed. We also examined the changes of alveolar bone and periodontal ligament (PDL) tissues in orthodontic rats by micro-computed tomography (micro-CT) system and immunohistochemistry (IHC) staining. The nuclear factor erythroid-2-related factor-2 (Nrf2) expression was examined. ResultsNAC could enhance the osteogenic differentiation and up-regulate the GSH level as well as the ratio of GSH/GSSG, while down-regulate ROS generation and Nrf2 expression induced by cyclic mechanical stress in PDLSCs. NAC had beneficial effects on the microstructure of alveolar bone and enhanced the expression levels of osteogenesis markers, such as alkaline phosphatase (ALP) and collagen type 1 (COL1) in PDL in orthodontic rats at the tension side. ConclusionNAC could improve the osteogenic differentiation stimulated by cyclic mechanical stress in PDLSCs and in orthodontic rats, suggesting a potential therapeutic approach for alveolar bone remodeling in orthodontics.

Chronic exposure to lead acetate promotes changes in the alveolar bone of rats: microstructural and physical-chemical characterization.

There are a few data relating to the effects of lead (Pb) exposure on the alveolar bone, which has very distinct morphophysiological characteristics and is of great importance in the oral cavity. In this context, the aim of this study was to investigate the changes promoted after long-term exposure to Pb in the microstructure of the alveolar bone of rats. Twenty adult Wistar rats were exposed to 50 mg/kg/day of lead acetate for 55 days. These animals were euthanized and had their mandible removed. Each mandible was divided into hemimandibles, and the alveolar bone was used for bone lead quantification, crystallinity analysis, microstructure evaluation by the percentage of bone volume (BV/TV), number of trabeculae (Tb.N), thickness of the trabecular (Tb.Th), and trabecular space (Tb.Sp). Morphometric analysis of the exposed root area was also performed. Long-term exposure to Pb resulted in high levels of Pb in the alveolar bone but showed no changes in the organization of crystallinity. The microstructural analyses showed a reduction of BV/TV, Tb.Th, and Tb.N and increase of Tb.Sp parameters, resulting in an increase in the exposed root area and an alveolar bone loss in height. The findings of this study reveal the ability of Pb to alter the alveolar bone microstructure after long-term exposure to the metal, possibly due to changes in tissue homeostasis, contributing to the reduction of bone quality.

Effect of Curcumin on Alterations of Alveolar Bone Remodeling and Expression of Receptor Activator of Nuclear Factor-κ B Ligand in Rat Tooth During Tooth Movement

Objective: To evaluate the effect of Curcumin on Alterations of Alveolar Bone Remodeling and Expression of RANKL in Rat Tooth during Tooth Movement. Methods: 64 SD rats were randomly divided into 4 groups, Model, Adrb2, Cur and Cur + Pro groups. The rat orthodontic teeth movement models were established.The rats were injected corresponding reagents according to weight and were sacrificed on day 0, 7, 14 and 21. The movement distance of first molar of rats was measured by Vernier caliper.The numbers of osteoclasts were observed through TRAP staining. The change of micro-structure of alveolar bone was evaluated by Micro-CT. Results: The trends of the distance of teeth movement and numbers of osteoclast were the same: Cur group β Adrb2 group &gt; Model groups Cur+Pro group (P &lt; 0.05). Micro-CT scan showed that curcumin could reduce the bone volume fraction (BV/TV), bone trabecular density (MTPD), and increase the trabecular resolution (TB. SP). When propranolol was given at the same time, the effect of curcumin disappeared. Conclusion: Curcumin could promote the resorption of alveolar bone at the pressure side and increase the osteoclast numbers so that the alveolar bone became looser which was beneficial to the movement of orthodontic tooth.

Effects of irradiation in the mandibular bone loaded with dental implants. An experimental study with a canine model

ABSTRACT Radiation therapy may compromise the quality of bone around dental implants, and its ability to regenerate, remodel, and revascularize. This study aimed to describe the irradiation effect on the bone microstructure of the mandible using dental implants in a canine model. Five beagle dogs were exposed to 40 Gy fractionated radiation. In total, 20 dental implants were inserted, two in the irradiated and two in the non-irradiated side. The mandible bone blocks were subjected to 3D micro-computed tomography (µCT) imaging, later evaluated histomorphometrically by light microscopy and scanning electron microscopy. Alterations in irradiated bone were observed under µCT imaging showing an increased anisotropy, porosity, and pore volume. Bone surface-to-bone volume decreased. The bone to implant contact index was significantly reduced in the irradiated bone (75.6% ± 5.8%) as compared to the non-irradiated bone (85.1% ± 6.8%). In the irradiated mandible, osteocytes with their filopodial processes, the bone beneath the periosteum, and subperiosteal veins showed structural differences but were not significant, whereas the diameter of Haversian canals were smaller statistical significant as compared to the control side. The study highlights that radiation dosage of fractioned 40 Gy causes alterations in the alveolar bone microstructure with compatible osseointegration and clinically stable dental implants.

Biomechanical Role of Trabecular Microstructure on Peri-implant Strain Based on Micro-CT Finite Element Modeling of Aging Mice.

To evaluate the influence of age and trabecular microstructure on peri-implant strain in aging and young mice models under compressive load. Eighteen 4-week-old female C57BL/6 mice (n = 6) were subjected to a 1.2% calcium content diet (young normal calcium group), and 7-month-old mice (n = 12) were randomly subjected to 0.01% and 1.2% calcium content diets (aging low and normal calcium groups, respectively) for 3 weeks. Histomorphometric and microcomputed tomography (micro-CT) analyses were used to investigate local alveolar bone microstructure. One maxilla segment from each group was reconstructed using micro-CT images to highlight the trabecular microstructure. A finite element analysis based on a computational model of the maxilla segment was performed to investigate peri-implant strain. Implants with three different diameters (0.3, 0.4, and 0.5 mm) were analyzed in these models. The aging low calcium group showed worse cancellous microstructure in hematoxylin and eosin (HE) staining, significantly increased osteoclast numbers (P < .05), and reduced bone volume fraction and trabecular thickness compared with the aging normal calcium group (P < .05). However, the young normal calcium group presented no difference in trabecular microstructure and osteoclast numbers compared with the aging normal calcium group. The aging low calcium group demonstrated increased strain intensity compared with the aging normal calcium group, whereas the young normal calcium and aging normal calcium groups showed comparable strain magnitude. The strain intensity of peri-implant bone increased with worse cancellous microstructure. When the diameter increased from 0.3 mm to 0.4 mm, the percentages of pathologic overload decreased regardless of bone microstructure. Deteriorated bone microstructure induced by a low calcium diet determined higher strain intensity, whereas, whenever age had no significant effect on trabecular microstructure, consequently, there was no substantial influence on strain. An increase of implant diameters can improve the strain distribution. Clinical decision-making should take into consideration the patient-specific and site-specific trabecular microstructure in preoperative assessment.

Nrf2 activation is involved in osteogenic differentiation of periodontal ligament stem cells under cyclic mechanical stretch

During orthodontic treatment, mechanical stretch serves a crucial function in osteogenic differentiation of periodontal ligament stem cells (PDLSCs). Up-regulated reactive oxygen species (ROS) level is a result of cyclic mechanical stretch in many cell types. Nuclear factor erythroid-2-related factor-2 (Nrf2) is a master regulator in various antioxidants expression. However, it is not known whether cyclic mechanical stretch could induce the ROS generation in PDLSCs and whether Nrf2 participated in this process. The present study was aimed to investigate the role of Nrf2 in PDLSCs under cyclic mechanical stretch. Our results showed that cyclic mechanical stretch increased ROS level and the nuclear accumulation of Nrf2 during osteoblast differentiation. Knocking down Nrf2 by siRNA transfection increased ROS formation and suppressed osteogenic differentiation in PDLSCs. T-BHQ, a Nrf2 activator, promoted the osteogenic differentiation in PDLSCs under cyclic mechanical stretch, and improved the microstructure of alveolar bone during orthodontic tooth movement in rats by employing micro-CT system. Taken together, Nrf2 activation was involved in osteogenic differentiation under cyclic mechanical stretch in PDLSCs. T-BHQ could promote the osteogenic differentiation in vitro and in vivo, suggesting a promising option for the remodeling of the alveolar bone during orthodontic tooth movement.

Imaging Microstructural Damage and Alveolar Bone Loss in Rats Systemically Exposed to Methylmercury: First Experimental Evidence.

The alveolar bone is an important mineralized structure of the periodontal support apparatus, and information about the methylmercury (MeHg) effects on the structural integrity is scarce. Therefore, this study aimed toinvestigate whether systemic, chronic, and low-dose exposure to MeHg can change the alveolar bone microstructure of rats. Adult Wistar rats (n = 30) were exposed to 0.04mg/kg/day of MeHgor vehicle through intragastric gavage. The animals were euthanized after 60days, and blood samples were collected for trolox equivalent antioxidant capacity (TEAC), glutathione (GSH), lipid peroxidation (LPO), and comet assays. The mandible of eachanimalwas collected and separated into hemimandibles that were used to determine the total Hg level in the bone and to analyze microstructural damage and alveolar bone loss in terms of trabecular number (Tb.N), trabecular thickness (Tb.Th), bone volume fraction (BV/TV), and exposed root area of the second molars. MeHg exposure triggered oxidative stress in blood represented by lower levels of GSH and TEAC and the increase in LPO and DNA damage of the blood cells. High total Hg levels were found in the alveolar bone, and the microstructural analyses showed a reduction in Tb.N, Tb.Th, and BV/TV, which resulted in an increase in the exposed root area and a decrease in bone height. Long-term MeHg exposure promotes a systemic redox imbalance associated with microstructural changes and alveolar bone loss and may indicate a potential risk indicator for periodontal diseases.

Osteopenic effects of high-fat diet-induced obesity on mechanically induced alveolar bone remodeling.

The aim of the study was to investigate the effect of obesity on the tissue and molecular reactions of alveolar bone in response to orthodontic force and its underlying mechanisms. Sixty-four rats were randomly divided into normal diet (ND) and high-fat diet (HFD) groups for eight weeks of dietary treatment. OTM was induced using nickel-titanium springs between the upper left first molar and incisor. After 1, 3, 7, and 14days of OTM, the maxillary alveolar bone and gingival tissues were harvested and analyzed. Compared with the ND rats, the HFD rats had greater OTM distance, serum levels of tartrate-resistant acid phosphatase (TRAP), and tumor necrosis factor α (TNF-α), as well as significant alveolar bone loss and bone architecture deterioration on both the compression and tension sides (p<.05 for all). This response was linked to the increased osteoclast numbers and functional activity and decreased osteoblast activity in the periodontal ligament, gingival tissue, and alveolar bone. HFD-induced obesity promoted mechanically induced alveolar bone remodeling and detrimental changes in alveolar bone microstructure by increasing osteoclastogenesis and regulating inflammatory cytokine expression. The increased alveolar bone remodeling in the obese rats lead to an accelerated OTM.

A circRNA-miRNA-mRNA network plays a role in the protective effect of diosgenin on alveolar bone loss in ovariectomized rats

BackgroundThe present study aimed to assess the perturbation in circular RNA (circRNA)/mRNA expression profiles and a circRNA-miRNA-mRNA coexpression network involved in the potential protective effect of diosgenin (DIO) on alveolar bone loss in rats subjected to ovariectomy (OVX).MethodsThe Wistar rats (female) manipulated with sham operation were classified as the SHAM group and the grouping of OVX rats administered with DIO, estradiol valerate or vehicle for 12 weeks was DIO group, EV group and OVX group respectively. Following treatments, the plasmatic levels of osteocalcin and tumor necrosis factor-alpha and the microstructure of alveolar bone were assayed. Based on microarray analyses, we identified differentially expressed (DE) circRNAs and mRNAs in alveolar bone of rats in both OVX and DIO group. The DE circRNAs and DE mRNAs involved in the bone metabolism pathway validated by RT-qPCR were considered key circRNAs/mRNAs. On the basis of these key circRNAs/mRNAs, we predicted the overlapping relative miRNAs of key circRNAs/mRNAs, and a circRNA-miRNA-mRNA network was built.ResultsDIO showed an anti-osteopenic effect on the rat alveolar bone loss induced by OVX. In total, we found 10 DE circRNAs (6 downregulated and 4 upregulated) and 614 DE mRNAs (314 downregulated and 300 upregulated) in samples of the DIO group compared with those of the OVX group. However, only one circRNA (rno_circRNA_016717) and seven mRNAs (Sfrp1, Csf1, Il1rl1, Nfatc4, Tnfrsf1a, Pik3c2g, and Wnt9b) were validated by qRT-PCR and therefore considered key circRNA/mRNAs. According to these key circRNA/mRNAs and overlapping predicted miRNAs, a coexpression network was constructed. After network analysis, one circRNA-miRNA-mRNA axis (circRNA_016717/miR-501-5p/Sfrp1) was identified.ConclusionThe mechanism of DIO inhibiting alveolar bone loss after OVX is possibly relevant to the simultaneous inhibition of osteogenesis and osteoclastogenesis by mediating the expression of important molecules in the Wnt, PI3K, RANK/RANKL or osteoclastogenic cytokine pathways. The circRNA_016717/miR-501-5p/Sfrp1 axis may play important roles in these processes.

Micro-osteoperforations accelerate orthodontic tooth movement by stimulating periodontal ligament cell cycles.

The aim of this study was to investigate the mechanism of how micro-osteoperforations (MOPs) accelerate tooth movement. We focused on inflammation, cell proliferation, and apoptosis ofperiodontal ligament cells and performed immunostaining of MOPs exposed to tumor necrosis factor-alpha (TNF-α), proliferating cell nuclear antigen (PCNA), and terminal deoxynucleotidyl transferase dUTP nick end labeling (TUNEL) during experimental tooth movement. Eleven-week-old male Wistar rats were divided into 2 groups: (1) 10g of orthodontic force applied to the maxillary first molar (TM) and (2) force application plus 3 small perforations of the cortical plate (TM+MOPs). On days 1, 4, 7, 10, and 14 after force application, we investigated tooth movement and alveolar bone microstructure using microcomputed tomography (n=5). We also determined the expression of TNF-α and PCNA in the pressure sides of periodontal ligaments via an immunohistochemical analysis. The expression of apoptotic cells was also determined by the TUNEL method. The tooth movement in the TM+MOPs group was significantly greater on days 4 to 14 than in the TM group. The TM+MOPs group showed statistically significant decreases in bone volume/tissue volume ratio and bone mineral density compared with the TM group. The ratios of TNF-α positive cells in the TM+MOPs group were increased on days 1, 4. 7, and 10 compared with the TM group. The ratios of PCNA positive cells in the TM+MOPs group were increased on days 1, 4, and 7 compared with the TM group, and the ratios of TUNEL positive cells in the TM+MOPs group were increased on days 1 and 7 compared with the TM group. These results suggest that MOPs may accelerate tooth movement through activation of cell proliferation and apoptosis of periodontal ligament cells.

Diosgenin protects against alveolar bone loss in ovariectomized rats via regulating long non-coding RNAs.

The present study assessed the changes in long non-coding (lnc)RNA and mRNA expression profiles when diosgenin (DIO) exerted a potential osteoprotective effect on the alveolar bone of ovariectomized (OVX) rats. Female Wistar rats underwent a sham operation (SHAM group) or ovariectomy. OVX rats were treated using vehicle (OVX group), DIO (DIO group) or estradiol valerate (EV group) for 12 weeks. After treatment, the biomarkers of bone turnover in plasma and the microstructure of alveolar bone were assessed. lncRNA microarrays were applied to assess lncRNA and mRNA expression profiles in alveolar bone in the OVX and DIO group rats. Subsequently, the differentially expressed mRNAs associated with the comprehensive bone metabolism pathway in Ingenuity Pathway Analysis (IPA) were identified and regarded as key mRNAs. Based on some of the key mRNAs and all the differentially expressed lncRNAs, a coexpression network was established and this network was further analyzed to identify the top 6 lncRNAs with the highest closeness scores (pivotal lncRNAs). Finally, 6 modules showing interactions between pivotal lncRNAs and key mRNAs were constructed. All of the pivotal lncRNAs and key mRNAs were validated with reverse transcription-quantitative polymerase chain reaction. The present findings demonstrated that DIO suppressed the loss of alveolar bone in OVX rats, and the changes to the expression of some lncRNAs or mRNAs occurred in the alveolar bone of the rats in the DIO group. Twenty-four key mRNAs were identified during pathway analysis. Furthermore, 8/24 key mRNAs (Ctnnb1, Smad4, Tcf2, Sp7, Il1b, Il1r1, Tnf and Tnfrsf1a) were used to establish a coexpression network, which included 1,656 nodes and 5,341 edges. During network analysis, 6 pivotal lncRNAs (XR_008346, MRuc007iji, MRAK157089, MRAK076413, MRAK143591 and AB036696) were obtained, and 6 modules illustrating pivotal lncRNA-key mRNA interactions were identified. These results revealed that the anti-osteoporotic effect of DIO on alveolar bone may be associated with the promotion of a bone formation process through increasing the signaling of the Wnt and BMPs pathways and the inhibition of the bone resorption process through decreasing stimulators of osteoclastogenesis. To conclude, several pivotal lncRNAs may serve important roles in these processes via regulating some key mRNAs in the bone metabolism pathway.

Influence of human parathyroid hormone during orthodontic tooth movement and relapse in the osteoporotic rat model: A preliminary study.

To investigate the effects of parathyroid hormone (PTH) on tooth movement in ovariectomized (OVX) rats by comparing the tooth movement distance and relapse and by examining the alveolar bone microstructure. Thirty 8-week-old female rats were classified into 3 groups: sham-operated, OVX and ovariectomized rats injected with PTH (PTH). Eight weeks later, a closed-coil spring appliance was placed between the maxillary incisor and the first molar and then activated with 50cN of force. During tooth movement, 30μg/kg of PTH was administered 3 times per week in the PTH group. Tooth movement distances were measured weekly. Five rats in each group were killed after 3weeks for microcomputerized tomographic analysis, and the remaining 5 rats in each group were killed at an additional 3weeks after the removal of the appliance to measure relapsed distance. The OVX group showed significantly greater tooth movement compared to those in the other 2 groups at 2 and 3weeks (P<.05). The relapse distance and relapse percentage for the OVX group were higher; however, it did not differ significantly from the PTH group. On micro-CT analysis, bone volume/tissue volume ratio and bone mineral density in the PTH group were significantly greater than in the OVX group (P<.05). Application of PTH did not promote tooth movement in OVX rat, however, did lead to decrease in relapse tendency. Therefore, the application of PTH during orthodontic treatment of patients with osteoporosis should be carefully considered.