Experimental Periodontitis In Rats Research Articles

HA2-FimA DNA Vaccine Treats Experimental Periodontitis.

To study the therapeutic effect of hemagglutinin-2 and fimbrial (HA2-FimA) vaccine on experimental periodontitis in rats. The first batch of rats was divided into two groups and immunised with pure water or pVAX1-HA2-FimA at the age of 6, 7, and 9 weeks. After sacrificing the animals, total RNA was extracted from the spleens for RNA high-throughput sequencing (RNA-Seq) analysis. The second batch of rats was divided into four groups (A, B, C, D), and an experimental periodontitis rat model was established by suturing silk thread around the maxillary second molars of rats in groups B, C, and D for 4 weeks. The rats were immunised with pure water, pVAX1-HA2-FimA vaccine, empty pVAX1 vector, and pure water at 10, 11, and 13 weeks of age, respectively. Secretory immunoglobulin A (SIgA) antibodies and cathelicidin antimicrobial peptide (CAMP) levels in saliva were measured by enzyme-linked immunosorbent assay (ELISA). All rats were euthanised at 17 weeks of age, and alveolar bone loss was examined using micro-computed tomography (Micro-CT). Through sequencing analysis, six key genes, including Camp, were identified. Compared with the other three groups, the rats in the periodontitis+pVAX1-HA2-FimA vaccine group showed higher levels of SIgA and CAMP (p < 0.05). Micro-CT results showed significantly less alveolar bone loss in the periodontitis+pVAX1-HA2-FimA vaccine group compared to the periodontitis+pVAX1 group and periodontitis+pure water group (p < 0.05). HA2-FimA DNA vaccine can increase the levels of SIgA and CAMP in the saliva of experimental periodontitis model rats and reduce alveolar bone loss.

Preventive and Therapeutic Potential of Streptococcus cristatus CA119 in Experimental Periodontitis in Rats.

Periodontitis is an inflammatory condition of the oral cavity caused by a mixed infection of various bacteria, which not only severely affects the alveolar bone and connective tissues but also displays potential correlations with distal intestinal inflammation. In this study, we aimed to elucidate the therapeutic effects of Streptococcus cristatus CA119 on experimental periodontitis in rats and its impact on intestinal morphology. The results demonstrate that CA119 is capable of colonizing the oral cavity and exerting antagonistic effects on Porphyromonas gingivalis and Fusobacterium nucleatum, thus leading to a significant reduction in the oral pathogen load. Following CA119 intervention, there was a significant alleviation of weight loss in rats induced by periodontitis (P < 0.001). CA119 also regulated the expression of IL-6 (P < 0.05), IL-1β (P < 0.001), IL-18 (P < 0.001), COX-2 (P < 0.001), iNOS (P < 0.001), and MCP-1 (P < 0.01) in the gingival tissue. Additionally, CA119 reduced oxidative stress levels in rats and enhanced their antioxidant capacity. Microcomputed tomography (micro-CT) and histological analysis revealed that CA119 significantly reduced alveolar bone loss and reversed the downregulation of OPG/RANKL (P < 0.001). Furthermore, CA119 exhibited a significant protective effect against intestinal inflammation induced by periodontal disease and improved the colonic morphology in rats. In conclusion, this study demonstrates the role of CA119 as a potential oral probiotic in the prevention and treatment of experimental periodontitis, underscoring the potential of probiotics as a complementary approach to traditional periodontal care.

Impact of circadian clock protein Bmal1 on experimentally-induced periodontitis-associated renal injury.

To investigate the mechanism of circadian clock protein Bmal1 (Bmal1) on renal injury with chronic periodontitis, we established an experimental rat periodontitis model. Twelve male Wistar rats were randomly divided into control and periodontitis groups (n=6, each group). The first maxillary molars on both sides of the upper jaw of rats with periodontitis were ligated by using orthodontic ligature wires, whereas the control group received no intervention measures. After 8 weeks, clinical periodontal parameters, including probing depth, bleeding index, and tooth mobility, were evaluated in both groups. Micro-CT scanning and three-dimensional image reconstruction were performed on the maxillary bones of the rats for the assessment of alveolar bone resorption. Histopatholo-gical observations of periodontal and renal tissues were conducted using hematoxylin-eosin (HE) and periodic acid-Schiff (PAS) staining. Renal function indicators, such as creatinine, albumin, and blood urea nitrogen levels, and oxidative stress markers, including superoxide dismutase, glutathione, and malondialdehyde levels, were measured using biochemical assay kits. MitoSOX red staining was used to detect reactive oxygen species (ROS) content in the kidneys. The gene and protein expression levels of Bmal1, nuclear factor erythroid 2-related factor 2 (Nrf2), and heme oxygenase-1 (HO-1) in rat renal tissues were assessed using real-time quantitative polymerase chain reaction (RT-qPCR) and immunohistochemical staining. Micro-CT and HE staining results showed significant bone resorption and attachment loss in the maxillary first molar region of the periodontitis group. Histological examination through HE and PAS staining revealed substantial histopathological damage to the renal tissues of the rats in the periodontitis group. The findings of the assessment of renal function and oxidative stress markers indicated that the periodontitis group exhibited abnormal levels of oxidative stress, whereas the renal function levels showed abnormalities without statistical significance. MitoSOX Red staining results showed that the content of ROS in the renal tissue of the periodontitis group was significantly higher than that of the control group, and RT-qPCR and immunohistochemistry results showed that the expression levels of Bmal1, Nrf2, and HO-1 in the renal tissues of the rats in the periodontitis group showed a decreasing trend. Circadian clock protein Bmal1 plays an important role in the oxidative damage process involved in the renal of rats with periodontitis.

Reuterin isolated from the probiotic Lactobacillus reuteri promotes periodontal tissue regeneration by inhibiting Cx43-mediated the intercellular transmission of endoplasmic reticulum stress.

The present study aimed to evaluate the effects of reuterin, a bioactive isolated from the probiotic Lactobacillus reuteri (L. reuteri) on periodontal tissue regeneration, and provide a new strategy for periodontitis treatment in the future. Data discussing the present state of the field: Probiotics are essential for maintaining oral microecological balance. Our previous study confirmed that probiotic L. reuteri extracts could rescue the function of mesenchymal stem cells (MSCs) and promote soft tissue wound healing by neutralizing inflammatory Porphyromonas gingivalis-LPS. Periodontitis is a chronic inflammatory disease caused by bacteria seriously leading to tooth loss. In this study, we isolated and purified reuterin from an extract of L. reuteri to characterize from the extracts of L. reuteri to characterize its role in promoting periodontal tissue regeneration and controlling inflammation in periodontitis. Chromatographic analysis was used to isolate and purify reuterin from an extract of L. reuteri, and HNMR was used to characterize its structure. The inflammatory cytokine TNFα was used to simulate the inflammatory environment. Periodontal ligament stem cells (PDLSCs) were treated with TNFα and reuterin after which their effects were characterized using scratch wound cell migration assays to determine the concentration of reuterin, an experimental periodontitis model in rats was used to investigate the function of reuterin in periodontal regeneration and inflammation control invivo. Real-time PCR, dye transfer experiments, image analysis, alkaline phosphatase activity, Alizarin red staining, cell proliferation, RNA-sequencing and Western Blot assays were used to detect the function of PDLSCs. In vivo, local injection of reuterin promoted periodontal tissue regeneration of experimental periodontitis in rats and reduced local inflammatory response. Moreover, we found that TNFα stimulation caused endoplasmic reticulum (ER) stress in PDLSCs, which resulted in decreased osteogenic differentiation. Treatment with reuterin inhibited the ER stress state of PDLSCs caused by the inflammatory environment and restored the osteogenic differentiation and cell proliferation functions of inflammatory PDLSCs. Mechanistically, we found that reuterin restored the functions of inflammatory PDLSCs by inhibiting the intercellular transmission of ER stress mediated by Cx43 in inflammatory PDLSCs and regulated osteogenic differentiation capacity. Our findings identified reuterin isolated from extracts of the probiotic L. reuteri, which improves tissue regeneration and controls inflammation, thus providing a new therapeutic method for treating periodontitis.

Investigating the anti-inflammatory and bone repair-promoting effects of an injectable porous hydrogel containing magnesium ions in a rat periodontitis mode

Periodontitis is associated with several systemic diseases, and advanced periodontitis is often linked to an extensive inflammatory microenvironment and irregularly shaped alveolar bone defects. However, eliminating periodontal inflammation in a minimally invasive manner while repairing irregularly shaped bone defects is clinically challenging. In comparison to traditional bone grafts, a thermo-sensitive hydrogel can be injected into deep periodontal pockets, forming and filling the alveolar bone defects in situ. In this study, porous injectable thermo-sensitive hydrogels containing magnesium ions were prepared by adding magnesium particles (MPs) to a glycerophosphate solution and combining this mixture with a chitosan solution. The incorporation of MPs created interconnected pores in the hydrogel, exhibiting high cytocompatibility and maintaining cell viability, proliferation, spreading, and osteogenesis in vitro. Evaluation on an experimental periodontitis rat model, using micro-computed tomography and histological analyses, demonstrated that this Mg2+-containing hydrogel effectively reduced periodontal inflammation, inhibited osteoclast activity, and partially repaired inflammation-induced alveolar bone loss. These results suggest that Mg2+-containing thermo-sensitive porous hydrogels might be promising candidates for treating periodontitis.

Periodontitis therapy with iron alginate and glucose oxidase based on the Fenton reaction

Periodontitis is a frequent chronic inflammatory disease brought on by bacterial infection in the biofilm that forms on the teeth. It can cause the gingival recession and tooth loss, gravely jeopardizing the oral health of people. Current therapies still suffer from issues like bleeding, pain, injured gums, limited treatment effectiveness, a lengthy treatment cycle, and bacterial resistance when using a combination of surgical intervention and antibiotic therapy. Herein, a simple one-step cross-linking process was used to create glucose oxidase (GOD) loaded upon iron alginate (FeAlg/GOD), which has a strong anti-biofilm activity, good biocompatibility, and was successfully used to treat periodontitis caused by biofilms. The results demonstrated that in the presence of a high glucose concentration in the gingival crevicular fluid, FeAlg/GOD could catalyze the conversion of glucose into gluconic acid and hydrogen peroxide. The produced hydrogen peroxide could convert Fe3+ to Fe2+ when it got to the deep periodontal pockets. Following that, the Fenton reaction was activated, producing large amounts of hydroxyl radical (OH). In conclusion, oral plaque biofilms were removed by the FeAlg/GOD. Additionally, successful in vivo treatment was accomplished in rat's experimental periodontitis models, as evidenced by a markedly reduced inflammatory response, decreased bone loss, and made healthier gum tissue. Overall, our biocompatible and antibacterial FeAlg/GOD study presents a novel strategy for effectively speeding periodontal tissue repair and eliminating oral pathogens to tackle the grave threat of biofilm-associated illnesses.

Desmarestia anceps Montagne modulates inflammatory response in experimental periodontitis in male Wistar rats

ObjectiveDesmarestia anceps Montagne (D. anceps) seaweed has been shown to play a range of biologic activities, such as anti-inflammatory and antioxidant properties. The purpose of this study was to investigate the effects of D. anceps on pathogenesis of ligature-induced experimental periodontitis in rats. DesignFifty-four male Wistar rats were randomly divided into six groups. Doses of D. anceps (100 and 200 mg/kg) were administered for 14 days to animals with or without ligature-induced periodontitis, in order to compare to control groups (vehicle solution). Morphometric registration of alveolar bone loss and histological analysis were conducted. Inflammatory cells were counted on hematoxylin-eosin-stained slides. Immunohistochemical expressions for interleukin-1 beta (IL-1β), tumor necrosis factor (TNF-α), receptor activator of nuclear factor kappa-Β ligand (RANKL) and bone morphogenetic protein 2 (BMP-2) were evaluated. ResultsAlveolar bone loss was higher in groups exposed to ligature-induced periodontitis, but no significant differences were observed comparing different treatment and control groups. Animals that received D. anceps extracts showed significant lower counts of inflammatory cells. D. anceps promoted increased expression of BMP-2 and inhibited IL-1β, TNF-α and RANKL. ConclusionD. anceps modulates inflammatory response in experimental periodontitis in rats. Further pharmacological and immunological analyses are warranted.

Local application of gingiva-derived mesenchymal stem cells on experimental periodontitis in rats.

Stem cell-based approaches in regenerative periodontal therapy have been used in different experimental models. In this study, the effect of local application of gingival mesenchymal stem cells (GMSC) in fibroin/chitosan oligosaccharide lactate hydrogel (F/COS) on periodontal regeneration was evaluated using experimental periodontitis model in rats. Mesenchymal stem cells were isolated from the gingiva of rats and characterized. Viability tests and confocal imaging of GMSC in hydrogels were performed. Healthy control without periodontitis (Health; H; n=10), control with periodontitis but no application (Periodontitis; P; n=10), only hydrogel application (F/COS; n=10), and GMSC+F/COS (n=10) four groups were formed for in vivo studies. Experimental periodontitis was created with silk sutures around the maxillary second molars. GMSC labeled with green fluorescent protein (GFP) (250,000 cells/50μL) in F/COS were applied to the defect. Animals were sacrificed at 2nd and 8th weeks and maxillae of the animals were evaluated by micro-computed tomography (micro-CT) and histologically. The presence of GFP-labeled GMSC was confirmed at the end of 8weeks. Micro-CT analysis showed statistically significant new bone formation in the F/COS+GMSC treated group compared with the P group at the end of 8weeks (p<0.05). New bone formation was also observed in the F/COS group, but the statistical analysis revealed that this difference was not significant when compared with the P group (p>0.05). Long junctional epithelium formation was less in the F/COS+GMSC group compared with the P group. Periodontal ligament and connective tissue were well-organized in F/COS+GMSC group. The results showed that local GMSC application in hydrogel contributed to the formation of new periodontal ligament and alveolar bone in rats with experimental periodontitis. Since gingiva is easly accessible tissue, it is promising for autologous cell-based treatments in clinical applications.

The Development of Thermoresponsive Polymeric Simvastatin Prodrug for the Treatment of Experimental Periodontitis in Rats.

Periodontitis (PD) is a severe inflammatory gum pathology that damages the periodontal soft tissue and bone. It is highly prevalent in the US, affecting more than 47% of adults. Besides routine scaling and root planing, there are few effective treatments for PD. Developed as an effective treatment for hyperlipidemia, simvastatin (SIM) is also known for its well-established anti-inflammatory and osteogenic properties, suggesting its potential utility in treating PD. Its clinical translation, however, has been impeded by its poor water-solubility, lack of osteotropicity, and side effects (e.g., hepatoxicity) associated with systemic exposure. To address these challenges, an N-(2-hydroxypropyl) methacrylamide (HPMA) copolymer-based thermoresponsive polymeric prodrug of SIM (ProGel-SIM) was developed as a local therapy for PD. Its aqueous solution is free-flowing at 4 °C and transitions into a hydrogel at ∼30 °C, allowing for easy local application and retention. After a thorough characterization of its physicochemical properties, ProGel-SIM was administered weekly into the periodontal pocket of an experimental rat model of PD. At 3 weeks post initiation of the treatment, the animals were euthanized with palate isolated for μ-CT and histological analyses. When compared to dose equivalent simvastatin acid (SMA, active form of SIM) treatment, the rats in the ProGel-SIM treated group showed significantly higher periodontal bone volume (0.34 mm3 vs 0.20 mm3, P = 0.0161) and less neutrophil (PMN) infiltration (P < 0.0001) and IL-1β secretion (P = 0.0036). No measurable side effect was observed. Collectively, these results suggest that ProGel-SIM may be developed as a promising drug candidate for the effective clinical treatment of PD.

Preventive effects of Capparis Spinose extract on experimental periodontitis in rats: a histopathological and biochemical study.

This study aimed to assess the effectiveness of Capparis Spinose (CS) in preventing the initiation and progression of experimental periodontitis and to evaluate the effect of its on systemic oxidative stress in rats by experimental periodontitis model. Twenty-four male rats were equally divided into; Ligatured (L), non-ligatured (NL), and Ligatured with CS (11 days/day per 20 mg/kg) (LC) groups. Experimental periodontitis was induced with the silk suture technic. Alveolar bone loss was examined, and total antioxidant capacity(TAOC), total oxidant status(TOS), and oxidative stress index(OSI) were analyzed in rat serum. Although; alveolar bone loss showed statistically significant lower values in the LC group compared to L (p < 0.05), not NL. In the LC group, osteoclast and osteoblast numbers were statistically significant compared to L, but there were no statistical differences between LC and NL. Serum TAOC levels were significantly lower in group L compared to others and also LC group showed significant differences from NL. TOS and OSI levels were significantly higher in group L than in other groups. Within the limitation of the present study, it can be said that the destruction via local inflammation that may occur after the experimental periodontitis can be prevented by using CS.

Dental Pulp Stem Cell-Derived Exosomes Regulate Anti-Inflammatory and Osteogenesis in Periodontal Ligament Stem Cells and Promote the Repair of Experimental Periodontitis in Rats.

Dental pulp stem cell-derived exosomes (DPSC-EXO), which have biological characteristics similar to those of metrocytes, have been found to be closely associated with tissue regeneration. Periodontitis is an immune inflammation and tissue destructive disease caused by plaque, resulting in alveolar bone loss and periodontal epithelial destruction. It is not clear whether DPSC-EXO can be used as an effective therapy for periodontal regeneration. The purpose of this study was not only to verify the effect of DPSC-EXO on reducing periodontitis and promoting periodontal tissue regeneration, but also to reveal the possible mechanism. DPSC-EXO was isolated by ultracentrifugation. Then it characterized by transmission electron microscope (TEM), nanoparticle tracking analysis (NTA) and Western Blot. In vitro, periodontal ligament stem cells (PDLSCs) were treated with DPSC-EXO, the abilities of cell proliferation, migration and osteogenic potential were evaluated. Furthermore, we detected the expression of IL-1β, TNF-αand key proteins in the IL-6/JAK2/STAT3 signaling pathway after simulating the inflammatory environment by LPS. In addition, the effect of DPSC-EXO on the polarization phenotype of macrophages was detected. In vivo, the experimental periodontitis in rats was established and treated with DPSC-EXO or PBS. After 4 weeks, the maxillae were collected and detected by micro-CT and histological staining. DPSC-EXO promoted the proliferation, migration and osteogenesis of PDLSCs in vitro. DPSC-EXO also regulated inflammation by inhibiting the IL-6/JAK2/STAT3 signaling pathway during acute inflammatory stress. In addition, the results showed that DPSC-EXO could polarize macrophages from the M1 phenotype to the M2 phenotype. In vivo, we found that DPSC-EXO could effectively reduce alveolar bone loss and promote the healing of the periodontal epithelium in rats with experimental periodontitis. DPSC-EXO plays an important role in inhibiting periodontitis and promoting tissue regeneration. This study provides a promising acellular therapy for periodontitis.

Bifidobacterium animalis subspecies lactis HN019 can reduce the sequelae of experimental periodontitis in rats modulating intestinal parameters, expression of lipogenic genes, and levels of hepatic steatosis.

To determine whether Bifidobacterium animalis subspecies lactis HN019 (B. lactis HN019) can reduce the sequelae of experimental periodontitis (EP) in rats modulating systemic parameters. This study evaluated the effects of probiotic therapy (PROB) in the prevention of local and systemic damage resulting from EP. Forty-eight rats were allocated into four groups: C (control), PROB, EP, and EP-PROB. PROB (1 × 1010  CFU/mL) administration lasted 8 weeks and PE was induced on the 7th week by placing ligature on the animals' lower first molars. All animals were euthanized in the 9th week of the experiment. Biomolecular analyses, RT-PCR, and histomorphometric analyses were performed. The data obtained were analyzed statistically (ANOVA, Tukey, p < .05). The EP group had higher dyslipidemia when compared to the C group, as well as higher levels of insulin resistance, proteinuria levels, percentages of systolic blood pressure, percentage of fatty hepatocytes in the liver, and expression of adipokines was up-regulated (LEPR, NAMPT, and FABP4). All these parameters (except insulin resistance, systolic blood pressure, LEPR and FABP4 gene expression) were reduced in the EP-PROB group when compared to the EP group. The EP group had lower villus height and crypt depth, as well as a greater reduction in Bacteroidetes and a greater increase in Firmicutes when compared to the EP-PROB group. Greater alveolar bone loss was observed in the EP group when compared to the EP-PROB group. Bifidobacterium lactis HN019 can reduce the sequelae of EP in rats modulating intestinal parameters, attenuating expression of lipogenic genes and hepatic steatosis.

Cannabidiol attenuates periodontal inflammation through inhibiting TLR4/NF-κB pathway.

Periodontitis is a chronic inflammatory disease involving soft and hard tissue destruction in the periodontal region. Cannabidiol (CBD) is a natural compound isolated from cannabis, which has the effect of inhibiting inflammation. However, the role of CBD in periodontitis remains unclear. The aim of this study was to investigate the anti-inflammatory effects and osteoprotective actions of CBD in periodontitis and its molecular mechanisms. After establishing the rat periodontitis model by ligatures, the specimens were processed for morphometric analysis by Micro-CT. The gingival tissues were collected, and the levels of TNF-α, IL-1β, and TLR4 were measured by enzyme-linked immunosorbent assay. LPS was used to induce the inflammatory response of human periodontal ligament cells (hPDLCs) in vitro. QPCR and western blot were carried out to detect the expression of related inflammatory cytokines and signaling pathways. Cannabidiol significantly inhibits bone loss in experimental rat periodontitis models. CBD downregulated the pro-inflammatory mediator TNF-α, related to the decrease of TLR4 protein expression. Overexpression of TNF-α and TLR4 caused by LPS in hPDLCs. CBD inactivated the TLR4/NF-κB signaling pathway by inhibiting TLR-4 expression and p65 NF-κB phosphorylation. CBD can be considered as a therapeutic agent for periodontitis. Our study demonstrated that CBD attenuates ligature-induced periodontitis in rats and LPS-induced inflammation in hPDLCs by inhibiting TLR4/NF-κB pathway activation. It indicates that topical CBD application is effective in treating periodontitis.

Pathomorphological analysis of the gum tissues in the application of the combination of Soderm®-Forte gel with Cytoflavin® in experimental periodontitis in rats

Introduction: Periodontitis is one of the most urgent problems of modern dentistry. The development of new paradigms and regimens of combination therapy of patients with periodontitis is a strategic task for pharmacologists and dentists. In view of this, pathomorphological examination is of high importance, since it allow us to conclude about the therapeutic effectiveness of the administered drugs with high objectivity. Aim of the study: to evaluate the effect of the composition of Soderm®-Forte and Cytoflavin® on the pathomorphological pattern of gum tissues of rats with experimental periodontitis (EP). Materials and Methods: EP was simulated in rats by ligature method. Study design: animals with intact periodontium; animals with untreated EP; animals with EP treated with traditional drug therapy (TDT); animals with EP treated with the combination of TDT and Soderm®-Forte gel; and animals with EP treated with the combination of TDT, Soderm®-Forte and Cytoflavin®. For pathomorphological examination, biopsy specimen was taken from the gingival margin of the lower incisors. Slides were stained with hematoxylin and eosin, as well as by Masson. Computer morphometry was performed using the ImageJ software. Results: In EP, TDT has a moderate positive effect on pathomorphological changes in the gum. The combination of TDT and Soderm®-Forte and, to a greater degree, the combination of TDT, Soderm®-Forte and Cytoflavin® have high therapeutic efficacy, characterized by rapid regeneration of the gum tissues.  Conclusion: The combination of TDT, Soderm®-Forte and Cytoflavin® in EP has a more pronounced therapeutic effect, manifested by early regression of pathological changes and acceleration of tissue regeneration in the gum. Graphical Abstract

Metal-organic framework-based nanoplatform enhance fibroblast activity to treat periodontitis.

After periodontal tissue injury, reconstruct soft tissue sealing around the tooth surface is of fundamental importance to treat periodontitis. Among multiple cell types, fibroblast plays a central role in reestablishing functional periodontium. To enhance fibroblast activity, a novel metal-organic framework-based nanoplatform is fabricated using mesoporous Prussian blue (MPB) nanoparticles to load baicalein (BA), named MPB-BA. Drug release test displayed sustained BA release of MPB-BA. Cell proliferation, transwell migration and wound healing tests revealed accelerated fibroblast proliferation and migration for the established MPB-BA nanoplatform. Moreover, vinculin immunofluorescence staining, western blot and quantitative real-time PCR analysis showed up-regulated vinculin protein and integrin α5 and integrin β1 gene expressions for MPB-BA, suggesting improved cell adhesion. In addition, hematoxylin and eosin (H&E) and Masson trichromatic staining suggested superior anti-inflammatory and collagen fiber reconstruction effects for MPB-BA in a rat experimental periodontitis model in vivo. Our study may provide a promising strategy for the treatment of periodontitis.

A modified method for constructing experimental rat periodontitis model.

Background: Periodontitis is a prevalent disease caused teeth lost. The present rat models inducing periodontitis with thread ligature and metal steel ligature have some disadvantages. Methods: We modified the existing rat ligature periodontitis model by fixing the thread ligature on the metal steel ligature passed through the gap between the first and secondmolars of rats with detailed modeling steps and illustrations. We research the pathological process of the periodontitis induced by the modified model, and briefly compared the modified model with the thread ligature model and the metal steel ligature model. Result: Our experimental results showed that there was an aggravation in inflammatory infiltration and alveolar bone resorption in modeling area within 14days of initial induction. After that, the inflammatory infiltration was reduced. And no significant increase in alveolar bone destruction appeared. The modified model was more reliable compared to the thread ligature model, and had greater ability of bacterial aggregation compared to the metal steel ligature model. Conclusion: The modified method covered pathological process of the periodontitis, and showed sufficient efficiency and reliability in inducing rat periodontitis.

Evaluation of the behaviour of periodontal collagen fibers during the progression of experimental periodontitis in rats

Resumo Introdução A periodontite é uma doença infecto-inflamatória, resultante da disbiose microbiana e da resposta do hospedeiro, que leva à destruição dos tecidos de suporte dentário, inclusive das fibras colágenas periodontais, podendo culminar na perda do elemento dental. Objetivo Avaliar o comportamento das fibras colágenas periodontais durante a progressão da periodontite experimental induzida em ratos. Material e método Doze ratos Wistar foram distribuídos nos grupos: Controle (C), Periodontite Experimental 14-dias (PE-14d), Periodontite Experimental 21-dias (PE-21d) e Periodontite Experimental 42-dias (PE-42d). No dia 0, os animais do grupo C foram eutanasiados. Neste mesmo dia, os animais remanescentes foram submetidos à instalação de uma ligadura de algodão ao redor do primeiro molar inferior esquerdo para indução da periodontite experimental. Tais animais foram eutanasiados aos 14 (PE-14d), 21 (PE-21d) e 42 (PE-42d) dias após a instalação da ligadura. Executou-se o processamento histológico das hemimandíbulas e as secções foram submetidas à reação histoquímica pelo vermelho picro-sirius. A análise qualitativa descritiva foi realizada sob microscopia de luz polarizada, na região de furca dental, evidenciando as fibras do ligamento periodontal. Resultado O grupo C exibiu feixes espessos e orientados de fibras colágenas maduras, condizentes com aspecto de normalidade. Os grupos com periodontite experimental exibiram desestruturação tecidual severa, com fibras colágenas imaturas e de menor espessura, sendo tais condições mais exacerbadas nos grupos PE-14d e PE-21d. Conclusão As fases iniciais da periodontite apresentam caráter agudo e, portanto, resultam na rápida destruição dos tecidos periodontais de suporte, prejudicando potencialmente a fibrilogênese e a reestruturação do colágeno no ligamento periodontal.

Evaluation of heat-inactivated Limosilactobacillus fermentum CCFM1139 and its supernatant for the relief of experimental periodontitis in rats.

Periodontitis is a chronic inflammatory disease induced by subgingival microbial dysbiosis, characterised by inflammation of the soft tissues of the periodontium and progressive loss of alveolar bone. Limosilactobacillus fermentum CCFM1139 is a probiotic with the potential to relieve periodontitis in vitro and in vivo. Due to the cost of active strain in production applications, we considered the effectiveness of bacterial components and metabolites in alleviating experimental periodontitis. Therefore, this study investigated the effect of heat-inactivated Limosilactobacillus fermentum CCFM1139 and its supernatant in the development of experimental periodontitis through animal experiments. The results showed that active, heat-inactivated Limosilactobacillus fermentum CCFM1139 and its supernatant all significantly reduced IL-1β levels in gingival tissue and serum (p < 0.05). Micro-computed tomography (micro CT) analysis showed that the active and heat-inactivated Limosilactobacillus fermentum CCFM1139 reduced alveolar bone loss in rats with periodontitis by 25.6% and 15.9% respectively (p < 0.05), with no change in percentage of bone volume (p > 0.05). In histomorphometric analysis, active Limosilactobacillus fermentum CCFM1139 showed better results in reducing alveolar bone loss and reducing inflammatory cell recruitment at the second molar. In addition, there was no significant difference in the number of tartrate-resistant acid phosphatase (TRAP) positive cells after in all experimental groups (p > 0.05). Therefore, heat-inactivated Limosilactobacillus fermentum CCFM1139 or its supernatant also have the ability to relieve periodontitis, and their alleviating effect may focus on the regulation of inflammatory response.

Adjuvant effects of Saccharomyces cerevisiae in the treatment of experimental periodontitis in rats undergoing chemotherapy.

Surgical procedures, radiotherapy, and chemotherapy, individually or in association, are current oncological treatments. Among the most used chemotherapy drugs, 5-fluorouracil (5FU) is an antimetabolite with a broad spectrum of action. This study evaluated the effects of probiotics (PRO) as an adjuvant to the treatment of experimental periodontitis (EP) in rats immunosuppressed with 5FU.108 rats were randomly allocated to six different groups: EP; SS - systemic treatment with saline solution (SS); 5FU - systemic treatment with 5FU; 5FU+PRO - systemic treatment with 5FU, followed by the local administration of Saccharomyces cerevisiae ; 5FU+SRP - systemic treatment with 5-FU, followed by scaling and root planing (SRP); and 5FU+SRP+PRO - systemic treatment with 5FU followed by local treatments with SRP and PRO. Immunosuppression was obtained at two points: at the time of ligature installation and after 48 h. Six animals from each group were euthanized at seven, 15, and 30 d and hemimandibles were collected and processed for histopathological, histometric, and immunohistochemical analysis. Data were subjected to statistical analysis (α=5%). At 7 d, the 5FU+PRO group showed less bone resorption and better structured connective tissue compared with the EP, SS, 5FU+SRP, and 5FU+SRP+PRO groups. At 15 d, the 5FU+SRP group showed a greater intensity of the inflammatory response (p<0.05). At 30 d, the 5FU+SRP+PRO group showed better structured bone tissue and a higher percentage of bone tissue (PBT) than the EP, SS, 5FU, and 5FU+PRO groups (p<0.05). The use of Saccharomyces cerevisiae as monotherapy or as an adjuvant to periodontal therapy may have a positive effect on bone repair in immunosuppressed conditions.

Effects of Lactobacillus reuteri MG5346 on Receptor Activator of Nuclear Factor-Kappa B Ligand (RANKL)-Induced Osteoclastogenesis and Ligature-Induced Experimental Periodontitis Rats.

Effects of culture supernatants of Lactobacillus reuteri MG5346 (CS-MG5346) on receptor activator of nuclear factor-kappa B ligand (RANKL)-induced osteoclastogenesis were examined. CS-MG5346 treatment up to 400 μg/mL significantly reduced tartrate-resistant acid-phosphatase (TRAP) activity, the phenotype biomarker of osteoclast, without affecting cell viability. CS-MG5346 inhibited the expression of osteoclast specific transcriptional factors (c-fos and nuclear factor-activated T cells c1) and their target genes (TRAP, cathepsin, and matrix metallo-proteinase-9) in a dose-dependent manner (p<0.05). The administration of L. reuteri MG5346 (2×108 CFU/day) for 8 wks significantly improved furcation involvement, but no difference was observed in alveolar bone loss in ligature-induced experimental periodontitis rats. The elevated RANKL/ osteoprotegerin ratio, the biomarker of periodontitis, was significantly lowered in the gingival tissue by administration of L. reuteri MG5346 (p<0.05). L. reuteri MG5346 showed excellent stability in simulated stomach and intestinal fluids and did not have antibiotic resistance. Based on the results, L. reuteri MG5346 has the potential to be a promising probiotic strain for oral health.