Rat Incisor Research Articles

Quantity and diameter of dentinal tubules of human teeth and teeth of experimental animals according to scanning electron microscopy data

Insight into the interrelation between the structure and mechanical properties of dentin of a tooth is crucial for prevention and treatment of oral cavity diseases. The study of the structure of dental dentin is important for understanding the pathogenetic mechanisms of the spread of cariogenic microflora in progressive dental caries to the pulp and periodontium, as well as for the interpretation of the results of studies of dental dentin-adhesive materials. The in vitro research presented here involved the examination of coronal dentin in 46 intact teeth from rats, cats, dogs, and humans at various distances from the pulp chamber, with a focus on the quantity and diameter of dentinal tubules. The calculations were based on the analysis of 155 scanning electron microscope photographs of the enamel surface and pulp chamber wall. The researchers calculated the quantity and diameter of dentinal tubules per unit area, which they measured at various distances from the pulp chamber. It was found that dentinal tubules are the widest near the pulp chamber wall, where the average diameter ranged from 1.7 to 2.8 µm. The highest values were found in premolars of young humans and the lowest - in the incisors of rats. In the middle portion of the dentin, the diameter of dentinal tubules ranged from 1.0 to 13.3 µm, while at the periphery, it was 0.6–0.9 µm. The highest density of dentinal tubules per unit area was found closest to the pulp horn, and the lowest was in the enamel region. Near the pulp chamber wall, the average density of dentinal tubules ranged from 50 000 to 90 000/mm², in the middle of the dentin, it ranged from 37 000 to 50 000/mm², and at the periphery, it ranged from 10 000 to 25 000/mm². All examined teeth exhibited straight dentinal tubules, except for the rats’ incisors, which showed local irregularities. A comparison was made based on the quantity and width of dentinal tubules in the examined animals and humans.

Effects of obesity on the rat incisor enamel and dentine thickness, as well as on the hemimandible shape over generations

Obesity has several effects on the general body metabolism. However, little is known about the impact of obesity on the growth and shape of mineralized tissues like mandibles and teeth, as well as if it effects are passed down from generation to next. Therefore, in this study, we aimed to evaluate, over nine generations using the consanguineous mating (inbreeding), the effect of the obesity condition produced by the reduction in the number of rats per litter during the lactation period on the hemimandible shape, dentine, and enamel of the rat incisor. Litters were reduced to two males and two females after birth, and were consanguinity mated in adulthood for nine generations. For all evaluations performed in this investigation, only males were used. The control group was formed by a non-consanguineous litter containing eight males. The parameters evaluated were food consumption, body weight, Lee Index, and bone density of the hemimandible bone. Incisor enamel and dentine thickness were also evaluated. The hemimandible shape was evaluated using geometric morphometry. The results show a significant and progressive increase in food intake, Lee Index, body weight, hemimandible weight, and enamel thickness, and a decrease in dentine thickness. The linear measurements of the length of the ramus ascending hemimandibular segment were found to be shorter, while its height was increased. In contrast, the geometric morphometry shows that the general hemimandible shape changed over the consanguineous obesity generations. We conclude that over generations, obesity increases and maintains the parameters evaluated with significant changes in hemimandible shape as well as in the dimensions of enamel and dentine of incisors, suggesting that enamel and dentine could be used as phenotype biomarkers to detect changes in tooth and craniofacial development related to obesity effects.

Effect of ELVAX polymer subgingival implants with echistatin on extracted and reimplanted rats' teeth.

To investigate the effect of ELVAX polymer subgingival implants incorporated with echistatin peptide on incisor reimplanted tooth in rats. Forty-two male Wistars rats were divided into two groups: echistatin-treated rats (E) and control rats (C). The animals had their right maxillary incisors extracted and treated according to the International Association of Dental Traumatology replantation protocol. The extra-alveolar dry period was 30 and 60min, and the post-surgical experimental periods were 15, 60, and 90days. The samples were stained with H&E and analyzed for the presence of an inflammatory response, incidence of resorptions, and dental ankylosis. Results were statistically analyzed (p < 0.05). The presence of inflammatory resorption was significantly higher in group C at 30 and 60min extra-alveolar time, in the 15-day postoperative period as compared with the E group (p < 0.05). Dental ankylosis was significantly more prevalent in group E in 30min extra-alveolar time and 15days postoperative period (p < 0.05). However, in 60min extra-alveolar time and 60days postoperative period, dental ankylosis was more prevalent in C group (p < 0.05). The use of ELVAX subgingival implants with echistatin demonstrated therapeutic potential in preventing the experimental resorption process after replantation of maxillary incisors in rats.

MicroRNA-146b-5p Suppresses Pro-Inflammatory Mediator Synthesis via Targeting TRAF6, IRAK1, and RELA in Lipopolysaccharide-Stimulated Human Dental Pulp Cells.

MicroRNA-146b-5p (miR-146b-5p) is up-regulated during and to suppress the inflammation process, although mechanisms involved in the action of miR-146b-5p have not been fully elucidated. This study examined the anti-inflammation effects of miR-146b-5p in lipopolysaccharide (LPS)-stimulated human dental pulp cells (hDPCs). An increase in human miR-146b-5p (hsa-miR-146b-5p) expression following the mRNA expression of pro-inflammatory cytokines was observed in LPS-stimulated hDPCs. The expression of hsa-miR-146b-5p and pro-inflammatory cytokines was down-regulated by a nuclear factor-kappa B (NF-κB) inhibitor, and the expression of hsa-miR-146b-5p was also decreased by a JAK1/2 inhibitor. Enforced expression of hsa-miR-146b-5p abolished phosphorylation of NF-κB p65 and down-regulated the expression of pro-inflammatory cytokines and NF-κB signaling components, such as interleukin-1 receptor-associated kinase 1 (IRAK1), tumor necrosis factor receptor-associated factor 6 (TRAF6), and REL-associated protein involved in NF-κB (RELA). Expression of rat miR-146b-5p (rno-miR-146b-5p) and pro-inflammatory cytokine mRNA was also up-regulated in experimentally-induced rat pulpal inflammation in vivo, and rno-miR-146b-5p blocked the mRNA expression of pro-inflammatory mediators and NF-κB signaling components in LPS-stimulated ex vivo cultured rat incisor pulp tissues. These findings suggest that the synthesis of miR-146b-5p is controlled via an NF-κB/IL6/STAT3 signaling cascade, and in turn, miR-146b-5p down-regulates the expression of pro-inflammatory mediators by targeting TRAF6, IRAK1, and RELA in LPS-stimulated hDPCs.

Immunomodulatory Effect of Novel Electrospun Nanofibers Loaded with Doxycycline as an Adjuvant Treatment in Periodontitis.

The immunomodulatory effect of a novel biomaterial obtained through electrospinning, based on polylactic acid (PLA) and nano-hydroxyapatite (nano-HAP), loaded with doxycycline (doxy) was evaluated in an animal model. The treatment capabilities as a local non-surgical treatment of periodontitis was investigated on the lower incisors of Wistar rats, after the induction of localized periodontitis using the ligature technique. Following the induction of the disease, the non-surgical treatment of scaling and root planing was applied, in conjunction with the application of the new material. The results of the treatment were evaluated clinically, using the tooth mobility and gingival index scores, as well as histologically. The salivary concentrations of matrix metalloproteinase 8 (MMP-8) and plasmatic concentrations of interleukin 1 (IL-1), interleukin 6 (IL-6) and tumor necrosis factor alpha (TNF-α) were also monitored. Two weeks after the ligature application, the periodontal disease was successfully induced in rats. The application of the novel biomaterial obtained through electrospinning was proven to be more effective in improving the clinical parameters, while decreasing the salivary MMP-8 and plasmatic IL-1 and TNF-α concentrations, compared to the simple scaling and root planing. Thus, the novel electrospun biomaterial could be a strong candidate as an adjuvant to the non-surgical periodontal therapy.

The correlation between biotribological function and structural characteristics of bamboo rat teeth

Chinese bamboo rats consume a diet that consists mainly mechanically demanding bamboo. The ability of the rats to process food depends on the capacity of their teeth to resist wear during biting and chewing. The mechanisms of the bamboo rat teeth were determined by investigating and correlating the structure and tribological properties of the teeth. The rat's incisor has a cutting-edge cusp and outer enamel iron pigmentation, making the teeth harder, more resistant to acid attacks, and improving the resistance to cracking. Additionally, the incisor is designed to self-sharpen. The complex morphology at the macroscale, and the three-dimensional fabric body formed by densely packed rods and inter-rods at the microscale of molar enamel benefit molar rapid capture and effective breaking of bamboo, resisting the high stresses required to process bamboo without wear. The results of the structure–function relations of bamboo rat teeth in this study may provide new ideas for improving plant cutting and grinding tools.

The Role of Lead Toxicity on Eruption Rate of Hypofunctional Incisors in Albino Wistar Rats

OBJECTIVES This objective of this study was to evaluate the role of a heavy metal- lead acetate in the eruption rate of hypo functional incisors in albino Wistar rats. METHODOLOGY An experimental study was done in animal house of Post Graduate Medical Institute, Lahore since March 2019 to March 2020. 34 adult albino Wistar rats were randomly divided into two groups (n=17 for each group) i.e., control and lead acetate group. Right mandibular incisors were selected for this study. Selected incisors were marked 1mm above the level of gingival papillae. The incisors were cut above this mark to make it hypo-functional. The readings were measured by digital Vernier caliper. This was considered as day 0. Incisors length was measured at day 0, 3, 6, 12 and 15 and eruption was calculated. The data was analyzed using SPSS version 22. RESULTS Eruption rate was similar throughout the study except last follow up. At the end of this study eruption of incisors in albino Wistar rats in control was 03.30±0.72mm, in lead 02.43±1.19mm. At day 15, the difference between control and lead group was statistically significant (p-value 0.033). CONCLUSION These results reveal that besides other causes of delayed tooth eruption excessive lead intoxication are also acausative factor of delayed tooth eruption.

Effects of immunoglobulin Y-loaded amorphous calcium phosphate on dentinal tubules occlusion and antibacterial activity.

Aim: This study aimed to evaluate the effects of immunoglobulin Y (IgY)-loaded amorphous calcium phosphate (ACP) (IgY@ACP) on dentinal tubule occlusion and antibacterial activity. Methodology: IgY@ACP was synthesized based on a biomimetic mineralization strategy. The structure was examined by transmission electron microscopy and Fourier transform infrared spectroscopy. The IgY release property was assessed in vitro. The cell biocompatibility of IgY@ACP was evaluated by CCK-8. The dentin disks were prepared using healthy human molars, and their dentinal tubules were exposed to EDTA. Subsequently, they were randomly selected and treated with or without IgY@ACP for 7 days. The tubule occlusion morphologies and newly formed layers were observed by scanning electron microscopy (SEM) and x-ray diffraction, respectively. To evaluate the acid resistance and abrasion resistance of IgY@ACP, dentin disks that were treated for 1 day were immersed in acid solution or subjected to a toothbrush. The antibacterial effects against Streptococcus mutans (S. mutans) were evaluated by colony-forming unit (CFU) counting, adhesion property assessment, and crystal violet and live/dead bacterial staining. Finally, the occlusion effect was evaluated in rat incisors in vivo. One-way analysis of variance (ANOVA) was performed for statistical analysis. The level of significance was set at 0.05. Results: IgY@ACP presented an amorphous phase with a nanosize (60–80 nm) and sustained release of protein within 48 h. The CCK-8 results showed that IgY@ACP had good biocompatibility. After treatment with IgY@ACP for 1 day, the majority of dentinal tubules were occluded by a 0.3-μm-thick mineralized layer. Seven days later, all dentinal tubules were occluded by mineralization with a thickness of 1.4 μm and a depth of 16 μm. The newly mineralized layer showed hydroxyapatite-like diffraction peaks. In addition, IgY@ACP had good acid and abrasion resistance. After treatment with IgY@ACP, the CFU counting and adhesion rate of S. mutans were significantly reduced, the crystal violet staining was lighter, and the S. mutans staining revealed more dead cells. Most importantly, IgY@ACP had a certain occluding property in rat incisors in vivo. Conclusion: IgY@ACP can effectively occlude dentinal tubules with acid-resistant stability and has prominent anti-S. mutans effects, rendering it a potentially suitable desensitization material in the clinic.

Investigating the Effects of Dehydrated Human Amnion-Chorion Membrane on Periodontal Healing.

Each growth factor (GF) has different effects and targets, and plays a critical role in periodontal healing. Dehydrated human amnion-chorion membrane (dHACM) contains various GFs and has been used to enhance wound healing. The purpose of this study was to evaluate the effects of dHACM on periodontal healing, using in vitro and in vivo experimental approaches. Standardized periodontal defects were created in rats. The defects were randomly divided into three groups: Unfilled, filled with hydroxypropyl cellulose (HPC), and dHACM+HPC. At 2 and 4 weeks postoperatively, periodontal healing was analyzed by microcomputed tomography (micro-CT), and histological and immunohistochemical analyses. In vitro, periodontal ligament-derived cells (PDLCs) isolated from rat incisors were incubated with dHACM extract. Cell proliferation and migration were evaluated by WST-1 and wound healing assay. In vivo, micro-CT examination at 2 weeks revealed enhanced formation of new bone in the dHACM+HPC group. At 4 weeks, the proportions of vascular endothelial growth factor (VEGF)-positive cells and α-smooth muscle actin (α-SMA)-positive blood vessels in the dHACM+HPC group were significantly greater than those in the Unfilled group. In vitro, dHACM extracts at 100 µg/mL significantly increased cell proliferation and migration compared with control. These findings suggest that GFs contained in dHACM promote proliferation and migration of PDLCs and angiogenesis, which lead to enhanced periodontal healing.

Role of Garlic (Allium Sativum) on Lead Induced Delayed Eruption of Incisors in Albino Wistar Rats

Role of Garlic (Allium Sativum) on Lead Induced Delayed Eruption of Incisors in Albino Wistar Rats

Impact of dams' diet on the levels of magnesium and zinc in the incisors of offspring rats

Impact of dams' diet on the levels of magnesium and zinc in the incisors of offspring rats

Craniofacial Bones and Teeth in Spacefarers: Systematic Review and Meta-analysis.

Estimating the risk of dental problems in long-duration space missions to the Moon and Mars is critical for avoiding dental emergencies in an environment that does not support proper treatment. Previous risk estimates were constructed based on the experience in short-duration space missions and isolated environments on Earth. However, previous estimates did not account for potential changes in dental structures due to space travel, even though bone loss is a known problem for long-duration spaceflights. The objective of this study was to systematically analyze the changes in hard tissues of the craniofacial complex during spaceflights. Comprehensive search of Medline, Embase, Scopus, the NASA Technical Report Server, and other sources identified 1,585 potentially relevant studies. After screening, 32 articles that presented quantitative data for skull in humans (6/32) and for calvariae, mandible, and lower incisors in rats (20/32) and mice (6/32) were selected. Skull bone mineral density showed a significant increase in spacefaring humans. In spacefaring rodents, calvariae bone volume to tissue volume (BV/TV) demonstrated a trend toward increasing that did not reach statistical significance, while in mandibles, there was a significant decrease in BV/TV. Dentin thickness and incisor volume of rodent incisors were not significantly different between spaceflight and ground controls. Our study demonstrates significant knowledge gaps regarding many structures of the craniofacial complex such as the maxilla, molar, premolar, and canine teeth, as well as small sample sizes for the studies of mandible and incisors. Understanding the effects of microgravity on craniofacial structures is important for estimating risks during long-duration spaceflight and for formulating proper protocols to prevent dental emergencies. Avoiding dental emergencies in long-duration spaceflights is critical since this environment does not support proper treatment. Prior risk estimates did not account for changes in dental structures due to space travel. We reviewed and synthesized the literature for changes in craniofacial complex associated with spaceflight. The results of our study will help clinicians and scientists to better prepare to mitigate potential oral health issues in space travelers on long-duration missions.

Age-related osteogenesis on lateral force application to rat incisor – Part III: Periodontal and periosteal bone remodeling

Objectives: This study was aimed to compare the histological pattern of bone modeling on either periodontal or periosteal side induced by lateral orthodontic tooth movement in different age groups. Material and Methods: A total of 50 male Sprague-Dawley rats (25 rats in the adult group – 52 weeks and 25 rats in the young group – 10 weeks) were utilized in this study. Each age group was classified into the control, 3 days, 7 days, 14 days, and 21 days groups (five rats in each) by the duration of experimental device application. A double-helical spring was produced using 0.014” stainless steel wire to provide 40 g lateral force to the left and right incisors. Hematoxylin-eosin staining, proliferating cell nuclear antigen (PCNA) immunohistochemical staining, fibroblast growth factor receptor 2 (FGFR2) immunohistochemical staining, and Masson trichrome staining were performed; and the slides were subject to histological examination. Results: In 7 days, active bone modeling represented by the scalloped surface was observed on the periosteal side of the crestal and middle alveolus at the pressure side in the young group, while similar changes were observed only on the crestal area in the adult group. In the young group, the number of PCNA-positive cells increased significantly on the crestal area and middle alveolus on the 3, 7, and 14 day groups, with subsequent decrease at 21 days. In the adult group, PCNA-positive cells were localized on the crestal area throughout the period. In the young group, FGFR2-positive cells were observed mainly on the crestal and middle alveolus at 3, 7, and 14 days than the control group. In the adult group, these cells appeared on the crestal and middle alveolus in the 3 days group, but mainly on the crestal area at 14 days. In the young group, FGFR2-positive cells were observed on the crestal and middle alveolus on the 3, 7, and 14 days groups more than on the control group. In the adult group, these cells appeared on the crestal and middle alveolus in the 3 days group, but mainly on the crestal area in the 14 days group. In Masson trichrome stain, an increased number of type I collagen fibers were observed after helical spring activation in both age groups. Large resorption lacunae indicating undermining bone resorption were progressively present in both young and adult groups. Conclusion: According to these results, orthodontic tooth movement may stimulate cell proliferation and differentiation primarily on the periosteal side according to progressive undermining bone resorption on the periodontal side. This response may lead to prominent bone modeling during tooth movement in the young group, compared to the relatively delayed response in the adult group.

Mechanical properties, microstructure and chemical composition of naked mole rat incisors

The naked mole rat incisors (NMRI) exhibit excellent mechanical properties, which make it a good prototype for design and fabrication of bionic mechanical systems and materials. In this work, we characterised the chemical composition, microstructure and mechanical properties of NMRI, and further compared these properties with the laboratory rat incisors (LRI). We found out that (i) Enamel and dentin are composed of organic matter, inorganic matter and water. The ratio of Ca/P in NMRI enamel is higher than that of LRI enamel. (ii) The dentin has a porous structure. The enamel has a three-dimensional reticular structure, which is more complex, regular and denser than the lamellar structure of LRI enamel. (iii) Enamel has anisotropy. Its longitudinal nano-hardness is greater than that of transverse nano-hardness, and both of them are higher than that of LRI enamel. Their nano-hardness and elastic modulus increase with the increase in distance from enamel-dentin boundary. The nano-hardness of dentin is smaller than that of enamel. The chemical composition and microstructure are considered to be the reasons for the excellent properties of NMRI. The chemical composition and unique microstructure can provide inspiration and guide for the design of bionic machinery and materials.

Biomimetic Fe‐hydroxyapatite nanoparticle‐reinforced bisphenol A‐glycol methacrylate/triethyleneglycol‐dimethacrylate resins for dental restorative application

AbstractCommercial bisphenol A‐glycol methacrylate (BisGMA)/triethyleneglycol‐dimethacrylate (TEGDMA) have been widely used as dental restorative material; however, the poor mechanical and anti‐wear properties greatly limited their applied range. In this study, iron‐containing hydroxyapatite (FeHA) nanoparticles that mimic the chemical composition and microstructure of natural bamboo rat incisor enamel were synthesized and used as reinforcement in BisGMA/TEGDMA resins. The as‐prepared polycrystalline FeHA have a rod like morphology with uniformly dispersed iron. The effects of nanoparticle–resin weight ratios on the mechanical properties, tribological properties, and bioactivity of FeHA‐BisGMA/TEGDMA resins were investigated. In vitro bioactivity of the studied resin composites in artificial saliva showed that a dense and continuous apatite layer was formed on the surface of the resin composite. Findings revealed that doping a suitable content of FeHA nanoparticle, especially at 0.75 wt% FeHA, would significantly improve the tribological and mechanical properties as well as bioactivity of the dental resin composites.

Age-related osteogenesis on lateral force application to rat incisor – Part II: Bony recession and cortical remodeling

Objective: The aim of this study is to analyze the age-related changes in the bony recession and cortical bone remodeling induced by lateral orthodontic tooth movement, using a three-dimensional micro-computed tomography (CT) analysis. Material and Methods: A total of 40 male Sprague-Dawley rats were divided into two distinct age groups (young, 10 weeks and adult, 52 weeks). Double-helical springs exerting 40 g of force were applied to central incisors to analysis of changes in lateral cortical bone and tooth movement with age and time. The young and adult rats were divided into four subgroups, T0 (0 week), T1 (1 week), T2 (2 weeks), and T3 (3 weeks), depending on the period of wearing the appliance. Micro-CT image was taken on each dissected pre-maxilla specimen. In each subgroup, distance between the center of teeth, suture width, tooth displacement, bony recession, and bone volume was evaluated. Results: The changes in the distance between the center of teeth and the suture width were significantly greater in the young group. However, the change in the tooth displacement showed no significant difference between groups. In the young group, bony recession of outer cortical layer was observed at T1 (P &lt; 0.05), but the amount of recession gradually decreased at T2 and T3. In contrast, in the adult group, bony recession increased gradually over observation period (P &lt; 0.05). The bone volume decreased at T1 (P &lt; 0.05), but recovered at T2 and T3 in both groups. Conclusion: The compensatory bone formation occurs in the pressure side of cortical bone more significantly in the young group than in the adult according to the lateral displacement of incisor in rats. The reduced bone reaction in the adult is considered a limiting factor of the excessive tooth movement in the compromised treatment of skeletal malocclusion.

Optimization of decalcification techniques for histologic examination of the rat maxillary and mandibular incisors for toxicity studies

ABSTRACT The objective of this study was to provide optimized processing for examination of rat incisors in nonclinical toxicity studies that enables analysis using immunohistochemistry (IHC). Rat maxillas and mandibles were decalcified in Immunocal, a formic acid decalcifier, and Decal Stat, a hydrochloric acid decalcifier, to evaluate tissue quality when with hematoxylin and eosin (H&E) stain and an IHC. Following necropsy of 10 to 13-week-old male Sprague Dawley rats, tissues were collected, trimmed, fixed in neutral buffered formalin (NBF), and placed into the corresponding decalcifying solution. After a pilot study with multiple timepoints for both decalcifying solutions, times were selected for the definitive study. Incisors in the definitive study were decalcified for 72, 96 or 120 hours in Immunocal and 24 hours in Decal Stat, trimmed, processed, embedded in paraffin, and sectioned. The microtomy process and sections were evaluated by histotechnologists. Sections were stained withH&E or an IHC to detect vimentin. Veterinary pathologists used blinded assessment to evaluate staining and tissue quality. The H&E sections from Immunocal timepoints scored higher based on criteria such as cellular morphology. However, tissue quality decreased at 120 hours with Immunocal but was adequate after 24 hours with Decal Stat. For IHC, moderate to excellent expression of vimentin was observed at timepoints for both decalcifiers. Optimal tissue sectioning and histological quality were achieved on incisor sections decalcified for 96 hours with Immunocal and 24 hours with Decal Stat.

Secreted Frizzled-Related Protein 1 Promotes Odontoblastic Differentiation and Reparative Dentin Formation in Dental Pulp Cells.

Direct pulp capping is an effective treatment for preserving dental pulp against carious or traumatic pulp exposure via the formation of protective reparative dentin by odontoblast-like cells. Reparative dentin formation can be stimulated by several signaling molecules; therefore, we investigated the effects of secreted frizzled-related protein (SFRP) 1 that was reported to be strongly expressed in odontoblasts of newborn molar tooth germs on odontoblastic differentiation and reparative dentin formation. In developing rat incisors, cells in the dental pulp, cervical loop, and inner enamel epithelium, as well as ameloblasts and preodontoblasts, weakly expressed Sfrp1; however, Sfrp1 was strongly expressed in mature odontoblasts. Human dental pulp cells (hDPCs) showed stronger expression of SFRP1 compared with periodontal ligament cells and gingival cells. SFRP1 knockdown in hDPCs abolished calcium chloride-induced mineralized nodule formation and odontoblast-related gene expression and decreased BMP-2 gene expression. Conversely, SFRP1 stimulation enhanced nodule formation and expression of BMP-2. Direct pulp capping treatment with SFRP1 induced the formation of a considerable amount of reparative dentin that has a structure similar to primary dentin. Our results indicate that SFRP1 is crucial for dentinogenesis and is important in promoting reparative dentin formation in response to injury.

H2O2 gel bleaching induces cytotoxicity and pain conduction in dental pulp stem cells via intracellular reactive oxygen species on enamel/dentin disc.

Bleaching is widely accepted for improving the appearance of discolored teeth; however, patient compliance is affected by bleaching-related complications, especially bleaching sensitivity. This study aimed to investigate the role of reactive oxygen species (ROS) in cytotoxicity and pain conduction activated by experimental tooth bleaching. Dental pulp stem cells with or without N-acetyl-L-cysteine (NAC), an ROS scavenger, were cultured on the dentin side of the enamel/dentin disc. Subsequently, 15% (90 min) and 40% (30 min) bleaching gels were painted on the enamel surface. Cell viability, intracellular ROS, Ca2+, adenosine triphosphate (ATP), and extracellular ATP levels were evaluated using the Cell Counting Kit-8 assay, 2',7'-dichlorodihydrofluorescein diacetate, CellROX, fura-3AM fluorescence assay, and ATP measurement kit. The rat incisor model was used to evaluate in vivo effects after 0, 1, 3, 7, and 30 days of bleaching. Changes in gene and protein expression of interleukin 6 (IL-6), tumor necrosis factor-alpha (TNFα), transient receptor potential ankyrin 1 (TRPA1), and Pannexin1 (PANX1) in dental pulp stem cells and pulp tissue were detected through RT-PCR, western blotting, and immunofluorescence. The bleaching gel suppressed dental pulp stem cell viability and extracellular ATP levels and increased intracellular ROS, Ca2+, and intracellular ATP levels. The mRNA and protein expression of IL-6, TNFα, TRPA1, and PANX1 were up-regulated in vitro and in vivo. Furthermore, the 40% gel had a stronger effect than the 15% gel, and NAC ameliorated the gel effects. Our findings suggest that bleaching gels induce cytotoxicity and pain conduction in dental pulp stem cells via intracellular ROS, which may provide a potential therapeutic target for alleviating tooth bleaching nociception.

Nuclear factor-kappa B decoy oligodeoxynucleotide-loaded poly lactic-co-glycolic acid nanospheres promote periodontal tissue healing after tooth replantation in rats.

Excessive inflammation in the periodontal tissue after tooth replantation can lead to inflammatory root resorption and interrupt periodontal tissue regeneration. We tested the hypothesis that nuclear factor-κB decoy oligodeoxynucleotide-loaded poly lactic-co-glycolic acid nanospheres (NF-PLGA) inhibit excessive inflammation and promote healing of periodontal tissue after replantation in rats. The upper right incisors of rats were extracted, immersed in different specific solutions, and replanted. The rats were euthanized at 7, 14, and 28 days after replantation. Morphological evaluation with micro-CT and histological assessment with hematoxylin and eosin and tartrate-resistant acid phosphatase (TRAP) staining was performed. Additionally, we examined the expression of interleukin (IL)-1β, IL-6, transforming growth factor-β1 (TGF-β1), and fibroblast growth factor-2 (FGF-2) in the periodontal ligament (PDL) by performing immunohistological assessment. The NF-PLGA group showed significantly greater dental root thickness than the other experimental groups. Root resorption was not observed after the application of NF-PLGA on day 7. The application of NF-PLGA also resulted in a significantly lower number of TRAP-positive osteoclasts on days 7 and 14 after replantation. Significantly lower expression of IL-1β and IL-6 and higher expression of TGF-β1 and FGF-2 were observed under the application of NF-PLGA in the PDL. NF-PLGA promoted the healing process by inhibiting the initial excessive inflammatory response in the PDL, preventing root resorption, and promoting periodontal tissue regeneration. The findings also suggested that the PLGA nanospheres-mediated transfection of the decoy oligodeoxynucleotides can be useful for the clinical application of replanted tooth root surfaces.