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.

OBJECTIVE: To determine whether Bifidobacterium animalis subspecies lactis HN019 (B. lactis HN019) can reduce the sequelae of experimental periodontitis (EP) in rats modulating systemic parameters. BACKGROUND: This study evaluated the effects of probiotic therapy (PROB) in the prevention of local and systemic damage resulting from EP. METHODS: 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). RESULTS: 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. CONCLUSION: Bifidobacterium lactis HN019 can reduce the sequelae of EP in rats modulating intestinal parameters, attenuating expression of lipogenic genes and hepatic steatosis.

Effects of probiotics in rats with experimental metabolic syndrome and periodontitis: An investigation of the intestine-adipose tissue axis.

BACKGROUND: This study evaluated the systemic (intestine and adipose tissue) and local (periodontal tissues) impact of probiotic therapy in rats with metabolic syndrome (MS) associated or not with periodontitis (PE). METHODS: Forty-eight rats received a high-fat diet for induction of MS for 16 weeks. They were subdivided into groups with (+) and without (-) PE, receiving (*) or not (**) receiving probiotics (PROB): MS (-**), MSP (-*), MSPE (+**), and MSPEP (+*). PROB administration (Bifidobacterium animalis subsp. lactis HN019) started on the 8th week of the study and PE was induced on the 14th week by placing ligature on the animals' lower first molars. Euthanasia occurred in the 16th week. Biomolecular, immunoenzymatic assays, and histomorphometric analyses were performed. The data obtained were statistically analyzed (ANOVA, Tukey, p < 0.05). RESULTS: The MSPEP group exhibited reduced alveolar bone loss when compared with the MSPE group, as well as lower levels of hepatic steatosis and proteinuria (p < 0.05). In the intestinal environment, the MSPE group exhibited significantly lower villus height and crypt depth, as well as a greater increase in Bacillota when compared with the MSPEP group (p < 0.05). The MSPEP group showed lower adipokine gene expression (LEPR, NAMPT, and FABP4) in adipose tissue than the MSPE group (p < 0.05). CONCLUSION: The probiotic B. lactis HN019 reduced the severity of experimental periodontitis and modulated the expression of lipogenic genes and intestinal morphological and microbiological parameters in rats with MS.

Adjuvant use of multispecies probiotic in the treatment of peri-implant mucositis: A randomized controlled trial.

AIM: This randomized placebo-controlled clinical trial evaluated the effects of multispecies probiotic containing Lactobacillus rhamnosus HN001™, Lactobacillus paracasei Lpc-37®, and Bifidobacterium animalis subsp lactis HN019™ as an adjunct to mechanical debridement (MD) on changes in bleeding on probing (BOP) in edentulous patients with peri-implant mucositis (PiM). MATERIALS AND METHODS: Patients were randomly assigned to test (probiotic) or control (placebo) groups. All sites with PiM received MD and topical gel application (probiotic or placebo) at baseline and 12 weeks. After initial MD, patients consumed probiotic or placebo capsules twice a day for 12 weeks. Clinical (modified sulcus bleeding index [mSBI]; modified plaque index [mPI]; probing depth [PD]; and BOP) and immunological parameters were collected at baseline and after 12 and 24 weeks. Data were statistically analysed (p < .05). RESULTS: Thirty-six patients with PiM were recruited. The test group presented higher prevalence (p < .05) of cases of restored peri-implant health at 24 weeks than did the control group (72.2% and 33.3%, respectively). No significant difference was observed between test (n = 18) and control (n = 18) groups for mPI and PD. mSBI %-score 0 was higher in the test group than in the control group at 24 weeks (p < .05). When compared with baseline, both groups presented reduced BOP at 12 and 24 weeks (p < .05). BOP was lower in the test group than in the control group at 12 (mean difference = -14.54%; 95% confidence interval [CI] = -28.87 to 0.22; p = .0163) and 24 (mean difference = -12.56%; 95% CI = -26.51 to 1.37; p = .0090) weeks. At 24 weeks, only the test group presented lower levels of interleukin (IL)-1ß, IL-6, IL-8, and tumour necrosis factor (TNF)-α than those at baseline (p < .05). CONCLUSIONS: The multispecies probiotic (administered locally and systemically) containing L. rhamnosus HN001™, L. paracasei Lpc-37®, and B. lactis HN019™ as an adjunct to repeated MD promotes additional clinical and immunological benefits in the treatment of PiM in edentulous patients (ClinicalTrials.gov NCT04187222).

Comparison Between Different Delivery Vehicles for the Probiotic Bifidobacterium animalis subsp. lactis HN019 on Experimental Periodontitis in Rats.

This study aimed to assess the effects of the probiotic (PROB) Bifidobacterium animalis subsp. lactis HN019 in two different delivery vehicles in experimental periodontitis (EP), including the gene expression for IL-10, IFN-γ, and FOXP3. In total, 32 rats were assigned into groups (n=8): C (control), EP, EP-PROB/Water, and EP-PROB/Milk. The probiotic was administered for 4 weeks, from baseline to euthanasia. Periodontitis was induced by ligatures 14 days after baseline. Data were statistically analyzed (p<0.05). Both probiotic groups presented decreased alveolar bone loss and increased interproximal attachment level than group EP. Also, these parameters were significantly improved in the Milk group when compared with the Water group. EP-PROB/Milk showed higher gene expression for IL-10 and lower for FOXP3 in relation to EP-PROB/Water and EP groups. The use of milk was able to potentiate the protective effects of B. lactis HN019 in rats under EP.

The use of probiotics can reduce the severity of experimental periodontitis in rats with metabolic syndrome: An immunoenzymatic and microtomographic study.

BACKGROUND: This study evaluated the effects of Bifidobacterium animalis subsp. lactis HN019 (B. lactis HN019) in the development of periodontitis (PE), associated or not with metabolic syndrome, (MS) in rats. METHODS: Ninety-six rats were grouped according to a food protocol: high-fat diet for induction of MS or standard diet for the control groups (C). They were subdivided into groups with (+) and without (-) PE, receiving (*) or not (**) probiotic (PROB): C-**, CP-*, PE+**, PEP+*, MS- MSP-*, MSPE+**, and MSPEP+*. PROB administration started on the eighth week of the study and PE was induced on the 14th week by placing ligature on the animals' lower first molars. Euthanasia occurred in the 16th week. Biomolecular analyzes, immunoenzymatic assays, and microtomographic analyses were performed. The data obtained were analyzed statistically (P < 0.05). RESULTS: The PEP and MSPEP groups showed lower levels of alveolar bone loss when compared with the PE and MSPE groups, respectively (P < 0.05). The immunoenzymatic analysis showed higher levels of interleukin (IL)-1ß and a higher receptor activator of NF-kappaB ligand (RANKL)/osteoprotegerin (OPG) ratio in the MSPE group when compared with the MSPEP group (P < 0.05). The PEP group showed lower levels of tumor necrosis factor (TNF)-α and IL-6 when compared with the PE group. The use of PROB attenuated dyslipidemia parameters in animals with MS, with or without PE. CONCLUSION: B. lactis HN019 reduced more significantly the severity of PE in rats with MS, modulating both systemic metabolic and immunoinflammatory parameters in periodontal tissues.

Bifidobacterium animalis subsp lactis HN019 presents antimicrobial potential against periodontopathogens and modulates the immunological response of oral mucosa in periodontitis patients.

OBJECTIVE: To evaluate the effects of Bifidobacterium animalis subsp. lactis HN019 (HN019) on clinical periodontal parameters (plaque accumulation and gingival bleeding), on immunocompetence of gingival tissues [expression of beta-defensin (BD)-3, toll-like receptor 4 (TLR4), cluster of differentiation(CD)-57 and CD-4], and on immunological properties of saliva (IgA levels) in non-surgical periodontal therapy in generalized chronic periodontitis (GCP) patients. Adhesion to buccal epithelial cells (BEC) and the antimicrobial properties of HN019 were also investigated. MATERIALS AND METHODS: Thirty patients were recruited and monitored clinically at baseline (before scaling and root planing-SRP) and after 30 and 90 days. Patients were randomly assigned to Test (SRP+Probiotic, n = 15) or Control (SRP+Placebo, n = 15) group. Probiotic lozenges were used for 30 days. Gingival tissues and saliva were immunologically analyzed. The adhesion of HN019 with or without Porphyromonas gingivalis in BEC and its antimicrobial properties were investigated in in vitro assays. Data were statistically analyzed (p<0.05). RESULTS: Test group presented lower plaque index (30 days) and lower marginal gingival bleeding (90 days) when compared with Control group. Higher BD-3, TLR4 and CD-4 expressions were observed in gingival tissues in Test group than in Control group. HN019 reduced the adhesion of P. gingivalis to BEC and showed antimicrobial potential against periodontopathogens. CONCLUSION: Immunological and antimicrobial properties of B. lactis HN019 make it a potential probiotic to be used in non-surgical periodontal therapy of patients with GCP. CLINICAL RELEVANCE: B. lactis HN019 may be a potential probiotic to improve the effects of non-surgical periodontal therapy. Name of the registry and registration number (ClinicalTrials.gov): "Effects of probiotic therapy in the treatment of periodontitis"-NCT03408548.

Root coverage using coronally advanced flap with porcine-derived acellular dermal matrix or subepithelial connective tissue graft: a randomized controlled clinical trial.

OBJECTIVE: The aim of this study is to investigate the use of a porcine-derived acellular dermal matrix (MD) in root coverage procedures combined with extended coronally positioned flap (eCAF), in comparison to the subepithelial connective tissue graft (SCTG) associated with the eCAF. MATERIAL AND METHODS: Eighteen adult patients presenting bilateral type 1 gingival recession were randomly assigned to SCTG or MD groups. Clinical and patient-based outcomes were recorded at 3 and 6 months after the surgical procedure. RESULTS: Both groups showed a significant reduction in the mean recession height of 3.33 ± 0.89 mm to 1.24 ± 1.10 mm (MD) and 3.21 ± 0.8 mm to 0.83 ± 0.86 mm (SCTG) without difference between groups. Six patients in the test group and eight in the control group obtained complete root coverage. The keratinized tissue height and thickness (KTT) showed a significant increase after 3 and 6 months in both groups. The average KTT gains were 0.39 ± 0.4 mm (MD) and 0.51 ± 0.5 mm (SCTG) (p < 0.05). Performing multivariate analysis suggests that MD addition to coronally advanced flaps may be similar to SCTG. CONCLUSION: The MD had similar results in comparison to SCTG and in the context of reducing patient morbidity it can be used as an alternative for the treatment of gingival recessions. CLINICAL RELEVANCE: The SCTG is the gold standard therapy for root coverage. The MD has been widely used in mucogingival surgery as a substitute for SCTG and proposed similar results. A substitute is very important for clinicians and patients. It will give a better postoperative and possibilities to treat multiples recession. (Clinicaltrials.gov Identifier: NCT03675334).

The impact of predatory bacteria on experimental periodontitis.

BACKGROUND: This study evaluated the effects of topical administration of Bdellovibrio bacteriovorus HD100 on experimental periodontitis (EP) in rats. METHODS: Thirty-two rats were divided into groups C (control), EP, C-HD100, and EP-HD100. At day 0, animals of groups EP and EP-HD100 received cotton ligatures around mandibular first molars (MFM). In groups C-HD100 and EP-HD100, 1 mL of suspensions containing B. bacteriovorus HD100 was topically administered in the subgingival region of MFMs at days 0, 3, and 7. Animals were euthanized at day 14. Gingival tissue, hemimandibles, and oral biofilm were collected. Data were statistically analyzed. RESULTS: Group EP-HD100 presented greater bone volume and lower connective tissue attachment loss (CTAL) than group EP (P < 0.05). Group EP-HD100 presented greater proportions of Actinomyces and Streptococcus-like species and lower proportions of Prevotella intermedia, Peptostreptococcus micros, Fusobacterium nucleatum, Fusobacterium polymorphum, Eikenella corrodens, Eubacterium nodatum, Campylobacter gracilis, Capnocytophaga sputigena, and Veillonella parvula-like species than group EP. Group EP-HD100 presented greater levels of osteoprotegerin and gene expression of interleukin (IL)-17, IL-10, and forkhead box P3 than group EP (P < 0.05). CONCLUSION: Topical use of B. bacteriovorus HD100 promotes a protective effect against alveolar bone loss and CTAL in rats with EP.

Sub-antimicrobial doses of doxycycline decreased bone loss related to ligature-induced periodontitis in hypertensive rats.

BACKGROUND/OBJECTIVE: The beneficial effects of sub-antimicrobial dose doxycycline (SDD) associated with nonsurgical periodontal therapy are well documented. Recently, the effects of SDD on metalloproteinases have been investigated in the treatment of hypertension. The aim of this study was to evaluate the effects of SDD on ligature-induced periodontitis in spontaneously hypertensive rats (SHR). METHODS: Fifty-four adult male rats were divided into three groups: SHR-C, SHR-L and SHR-L-DOX (C - Control; L - Ligature). In group SHR-L-DOX, animals were treated with daily 5 mg/kg SDD administration. In L groups, a ligature remained around mandibular first molars for the first 10 days. Each group was divided for euthanasia at 10 or 21 days. Microtomographic and histometric analyses were performed. Osteoclastogenesis was evaluated by tartrate-resistant acid phosphatase (TRAP) assay and gene expression of 84 inflammatory mediators by polymerase chain reaction (PCR) array. RESULTS: Group SHR-L-DOX presented reduced systolic blood pressure when compared with group SHR-L at both 10 and 21 days (p < 0.05). Group SHR-L-DOX showed decreased bone and attachment loss in comparison with group SHR-L at both 10 and 21 days (p < 0.05). SDD treatment reduced the amount of TRAP-positive cells at 10 days (p < 0.05). Group SHR-L-DOX showed a downregulated inflammatory genes profile in comparison with SHR-L at 10 and 21 days. CONCLUSION: SDD therapy exerted systemic modulatory effect on inflammation with reduced periodontal tissue destruction in hypertensive rats.

Effects of Bifidobacterium probiotic on the treatment of chronic periodontitis: A randomized clinical trial.

AIM: This randomized placebo-controlled clinical trial evaluated the effect of Bifidobacterium animalis subsp. lactis (B. lactis) HN019-containing probiotic lozenges as adjuvant to scaling and root planing (SRP) in patients with generalized chronic periodontitis. MATERIALS AND METHODS: Forty-one chronic periodontitis patients were recruited and monitored clinically, immunologically, and microbiologically at baseline (before SRP) and 30 and 90 days after SRP. All patients were randomly assigned to a Test (SRP + Probiotic, n = 20) or Control (SRP + Placebo, n = 21) group. The probiotic lozenges were used twice a day for 30 days. The data were statistically analysed. RESULTS: The Test group presented a decrease in probing pocket depth and a clinical attachment gain significantly higher than those of the Control group at 90 days. The Test group also demonstrated significantly fewer periodontal pathogens of red and orange complexes, as well as lower proinflammatory cytokine levels when compared to the Control group. Only the Test group showed an increase in the number of B. lactis HN019 DNA copies on subgingival biofilm at 30 and 90 days. CONCLUSION: The use of B. lactis HN019 as an adjunct to SRP promotes additional clinical, microbiological, and immunological benefits in the treatment of chronic periodontitis (NCT03408548).

Effectiveness of Multistrain Versus Single-strain Probiotics: Current Status and Recommendations for the Future.

Probiotics are investigated as single-strain and multistrain products. In the market, however, there is an increasing tendency to work with multistrain probiotics, in particular, products with a high number of different strains. There are some thoughts behind this: more strains imply more chances of success; it can mean a broader spectrum of efficacy, and there is often the hope that there are at least additive and, potentially, even synergistic effects. The present review did not find convincing evidence that these assumptions are valid. There is, however, also no strong evidence that the assumptions are incorrect and/or that there is antagonistic activity between strains in a combination. We suggest that, to answer these questions, structured research is conducted. Starting with a systematic review of meta-analyses that have compared single-strain and multistrain probiotic efficacy, dedicated human studies need to be performed, comparing single-strain and multistrain probiotics to each other and placebo. In vitro and animal studies can provide indications and may help understand mechanisms. For human, animal, and in vitro studies, it is recommended to work with the simple setup of 2 single strains, a 2-strain combination, and placebo. It is also important in such research to take into consideration the doses, as a combination product will have a higher total dose.

Aggressive periodontitis presents a higher degree of bilateral symmetry in comparison with chronic periodontitis.

The purpose of this cross-sectional study was to evaluate the occurrence of bilateral symmetry in the distribution of clinical parameters in subjects with generalized aggressive periodontitis (GAP) and severe chronic periodontitis (SCP). The sample comprised 53 subjects with GAP and 33 with SCP. Probing depth (PD) and clinical attachment loss (CAL) were recorded from both buccal and lingual interproximal sites of incisors and molars. The symmetry of periodontal destruction was analyzed in terms of intraclass coefficient correlations (ICC) for pairs of contralateral sites at which PD and/or CAL was ≥5 mm at one of the sites of the subjects in each group. GAP patients had a higher proportion of both PD and CAL ≥ 5 mm and also a higher mean proportion of subjects having PD and/or CAL ≥ 5 mm at one or both sites. The GAP group had 20 pairs of contralateral sites with PD (ICC = 0.22-0.63) and 26 pairs with CAL (ICC = 0.20-0.63), the correlation being statistically significant, while the SCP group had only 2 pairs (ICC = 0.36-0.48) with PD and 5 pairs with CAL (ICC = 0.33-0.58) showing a significant correlation. It can be concluded that GAP shows more symmetric periodontal destruction than SCP.

Local administration of Tiludronic Acid downregulates important mediators involved in periodontal tissue destruction in experimental periodontitis in rats.

OBJECTIVE: The purpose of this study was to evaluate whether local administration of TIL could influence the expression of the inflammatory mediators IL-1ß, TNF-α, MMP-8 and COX-2 in rats with experimental periodontitis (EP). METHODS: Twenty-four adult male rats (Rattus norvegicus, albinus, Wistar) were assigned to groups C, EP, EP-TIL (CControl group, EP-Periodontitis groups). On EP groups, a ligature was placed around maxillary 2nd molars on day 1. On group EP-TIL, 20 µL of TIL solution (1 mg/kg body weight) was injected into the subperiosteal palatal area adjacent to the maxillary 2nd molar every other day until euthanasia (day 11). Alveolar bone loss was morphometrically analyzed. mRNA expressions of IL-1ß, TNF-α, MMP-8 and COX-2 were assessed by qPCR. IL-1ß, TNF-α, MMP-8 and COX-2 were immunohistochemically analyzed. Data were analyzed statistically. RESULTS: Group EP-TIL presented reduced alveolar bone loss when compared with group EP (p < 0.05). Group EP-TIL presented decreased mRNA expressions of IL-1ß, TNF-α, MMP-8 and COX-2 and reduced immunolabeling of IL-1ß, TNF-α and MMP-8 when compared with group EP (p < 0.05). No differences regarding the immunolabeling of COX-2 were found when group EP-TIL was compared with the other groups (p > 0.05). CONCLUSION: Within the limits of this study, it can be concluded that local administration of TIL downregulates important mediators involved in periodontal tissue destruction in ligature-induced periodontitis in rats.

Effects of local administration of tiludronic acid on experimental periodontitis in diabetic rats.

BACKGROUND: Tiludronic acid (TIL) presents antiresorptive and anti-inflammatory properties and has not been evaluated in the periodontitis-diabetes mellitus (DM) association to date, to the best knowledge of the authors. This study evaluates effects of local administration of TIL on experimental periodontitis (EP) in rats with streptozotocin-induced DM. METHODS: Thirty-two animals (Rattus norvegicus albinus, Wistar) were divided into groups DM/C (Control), DM/EP, DM/EP/TIL1, and DM/EP/TIL3. In EP groups, a ligature was placed around mandibular first molars. In groups DM/EP/TIL1 and DM/EP/TIL3, TIL solutions (1 and 3 mg/kg, respectively) were injected into the gingival tissue of mandibular molars every other day for 10 days, until euthanasia. Periodontal tissues were analyzed by microcomputed tomography (micro-CT), histomorphometry, immunohistochemistry (tartrate-resistant acid phosphatase [TRAP], receptor activator of nuclear factor-κB ligand [RANKL], osteoprotegerin, cleaved caspase 3), and quantitative reverse transcription-polymerase chain reaction (interleukin [IL]-1ß, vascular endothelial growth factor [VEGF]). RESULTS: In micro-CT analyses, groups DM/EP/TIL1 and DM/EP/TIL3 presented reduced alveolar bone resorption (P < 0.05). Group DM/EP/TIL3 presented decreased attachment loss (P < 0.05). The amount of TRAP-positive multinucleated cells was decreased in TIL groups (P < 0.05). Group DM/EP/TIL3 presented a lower immunolabeling pattern for RANKL (P < 0.05). TIL treatment decreased genic expression of IL-1ß, and in group DM/EP/TIL3, expression of VEGF was increased (P < 0.05). CONCLUSION: Local administration of TIL promoted a protective effect against tissue destruction in EP in diabetic rats, and the dosage of 3 mg/kg of TIL promoted the best results regarding its antiresorptive and anti-inflammatory effects.

Anti-inflammatory and antiresorptive effects of Calendula officinalis on inflammatory bone loss in rats.

OBJECTIVE: The aim of this work was to evaluate the anti-inflammatory and antiresorptive effects of Calendula officinalis (CLO) on alveolar bone loss (ABL) in rats. MATERIAL AND METHODS: Male Wistar rats were subjected to ABL by ligature with nylon thread around the second upper left molar. The contralateral hemimaxillae were used as control. Rats received saline solution (SAL) or CLO (10, 30, or 90 mg/kg) 30 min before ligature and daily until the 11th day. The maxillae were removed and prepared for macroscopic, radiographic, micro-tomographic, histopathologic, histometric analysis, and immunohistochemical localization of receptor activator of nuclear factor kappa-B ligand (RANKL) and osteoprotegerin (OPG). The gingival tissues were used to quantify the myeloperoxidase (MPO) activity, tumor necrosis factor-alpha (TNF-α), and interleukin-1ß (IL-1ß) concentrations by ELISA. Blood samples were collected for leukogram and to evaluate the bone-specific alkaline phosphatase (BALP) activity and serum levels of aspartate and alanine transaminases (AST/ALT). RESULTS: The bone loss induced by 11 days of ligature induced bone loss, reduced levels of BALP, leukocyte infiltration, increased MPO activity, gingival concentrations of TNF-α and IL-1ß, and RANKL while reduced OPG immunoexpressions in the periodontal tissue and leukocytosis. Of the CLO, 90 mg/kg reduced bone loss, neutrophilia, the levels of pro-inflammatory mediators, and RANKL expression, while it increased OPG immunopositive cells and BALP serum levels, when compared to SAL. CLO did not affect either kidney or liver function, indicated by serum AST/ALT levels. CONCLUSION: The present data suggests that CLO reduced inflammatory bone resorption in experimental periodontitis, which may be mediated by its anti-inflammatory properties and its effects on bone metabolism. CLINICAL RELEVANCE: CLO can be a potential therapeutical adjuvant in the treatment of periodontitis.

Long-term therapy with intravenous zoledronate increases the number of nonattached osteoclasts.

The purpose of this study was to investigate the influence of long-term therapy with intravenous zoledronate (ZA) on the healing of extraction sockets in rats. Forty rats, divided into groups C (Control) and Z (Zoledronate), received intravenous injections of either saline solution or ZA for 24 weeks. Their right maxillary incisor was extracted. Euthanasia was performed at 7 or 28 days postoperative. Histomorphometric (Newly Formed Bone Area) and immunohistochemical (RANKL, OPG and TRAP) analyses were performed. Data were statistically analyzed (ANOVA, Tukey's test and Kruskal-Wallis, Dunn's Multiple Comparison test).Groups C and Z showed similar new bone area, RANKL and OPG immunolabeling. The number of TRAP-positive multinucleated cells was significantly higher in Group Z than in Group C at 28 days. A significantly higher proportion of nonattached osteoclasts were seen in Group Z than in Group C at both periods of analysis. Long-term therapy with intravenous ZA stimulated nonattached osteoclast formation in extraction sockets in rats, thus decreasing local bone resorption. However, it did not influence bone formation by osteoblasts.

Effects of the probiotic Bifidobacterium animalis subsp. lactis on the non-surgical treatment of periodontitis. A histomorphometric, microtomographic and immunohistochemical study in rats.

Lactobacillus probiotics have been investigated in periodontitis. However, the effects of the genus Bifidobacterium on periodontitis are hardly known. This study evaluated the effects of the probiotic (PROB) Bifidobacterium animalis subsp. lactis (B. lactis) HN019 as an adjunct to scaling and root planing (SRP) in rats with experimental periodontitis (EP). At baseline, 32 rats were assigned to 4 groups: C (control), PROB, EP-SRP and EP-SRP-PROB. In groups EP-SRP and EP-SRP-PROB, the mandibular first molars of the animals received a ligature. At day 14, the ligatures were removed and SRP was performed. Animals of groups PROB and EP-SRP-PROB were orally administered with 10 mL/day of 109 colony forming units of B. lactis HN019 for 15 days, starting at day 14. Animals were euthanized at day 29. Histomorphometric, microtomographic and immunohistochemical analyses were performed. Microbiological effects of B. lactis on biofilm were also evaluated. Data were statistically analyzed (ANOVA, Tukey; Kruskal-Wallis, Dunn's; Two-tailed t-test; p<0.05). Group EP-SRP-PROB presented reduced alveolar bone resorption and attachment loss when compared with Group EP-SRP (p<0.05). Group EP-SRP-PROB showed significantly fewer osteoclasts, increased expression of anti-inflammatory cytokines and reduced expression of proinflammatory cytokines compared with Group EP-SRP (p<0.05). B. lactis promoted a higher ratio between aerobic and anaerobic bacteria in biofilm samples (p<0.05). B. lactis HN019 may have a role in the treatment of EP in rats, as an adjunct to SRP.

Benefits of Bifidobacterium animalis subsp. lactis Probiotic in Experimental Periodontitis.

BACKGROUND: This study evaluates effects of topical administration of probiotic bacteria of the genus Bifidobacterium on experimental periodontitis (EP) in rats. METHODS: Thirty-two rats were divided into groups C (control; without EP), EP (EP only), C-HN019 (control+probiotic), and EP-HN019 (EP+probiotic). On day 0 of the experiment, animals of groups EP and EP-HN019 received cotton ligatures around mandibular first molars (MFMs). In groups C-HN019 and EP-HN019, 1 mL of suspensions containing Bifidobacterium animalis subsp. lactis (B. lactis) HN019 was topically administered in the subgingival region of MFMs on days 0, 3, and 7. In groups C and EP, topical administrations were performed using a sham suspension (without probiotic). All animals were euthanized at day 14. Gingival tissue, hemimandibles, and oral biofilm were collected. Data were statistically analyzed (P <0.05). RESULTS: Group EP presented greater bone porosity, trabecular separation, and connective tissue attachment loss (CTAL) as well as reduced bone volume than all other groups (P <0.05). In group EP-HN019, there were greater proportions of Actinomyces and Streptococcus-like species and lower proportions of Veillonella parvula, Capnocytophaga sputigena, Eikenella corrodens, and Prevotella intermedia-like species than group EP. Group EP-HN019 presented greater expressions of osteoprotegerin and ß-defensins than group EP (P <0.05). Group EP presented greater levels of interleukin-1ß and receptor activator of nuclear factor-kappa B ligand than group EP-HN019 (P <0.05). CONCLUSION: Topical use of B. lactis HN019 promotes a protective effect against alveolar bone loss and CTALs attributable to EP in rats, modifying immunoinflammatory and microbiologic parameters.

Favourable effects of Bacillus subtilis and Bacillus licheniformis on experimental periodontitis in rats.

OBJECTIVES: The purposes of this study were to evaluate, in rats: i) the effects of Bacillus species on the development of experimental periodontitis (EP) via microtomographic, immunological and hematological assays (Experiment 1-E1); ii) the effects of Bacillus species as adjuncts to scaling and root planing (SRP) for the treatment of EP via histomorphometric and immunohistochemical analyses (Experiment 2-E2). METHODS: In E1, 24 rats were divided into groups C1 (control), PROB1, EP1 and EP-PROB1. In groups with EP, the mandibular first molar of each animal received a ligature for 14 days. In groups PROB1, animals received Bacillus species for 44 days, starting 30 days before EP induction in Group EP-PROB1. In E2, 24 rats were assigned to groups C2 (control), PROB2, EP-SRP2 and EP-SRP-PROB2. In groups with SRP, EP was induced as described in E1. The ligatures were removed after 14 days and SRP was performed. In groups PROB2, animals received Bacillus species for 15 days, starting after SRP in Group EP-SRP-PROB2. RESULTS: In E1, Group EP1 presented bone loss (BL) and eosinophil numbers greater than Group EP-PROB1 (P<0.05). In Group EP-PROB1, the receptor activator of nuclear factor-kB ligand (RANKL)/osteoprotegerin (OPG) ratio was similar to that of groups without EP. In E2, Group EP-SRP-PROB2 presented fewer TRAP-positive osteoclasts, lower immunolabeling pattern for a proinflammatory cytokine and decreased BL and attachment loss than Group EP-SRP2 (P<0.05). CONCLUSIONS: Bacillus species supplementation provided a protective effect against BL and enhanced the effects of SRP in the treatment of EP in rats.

Effects of electroacupuncture on experimental periodontitis in rats.

BACKGROUND: Acupuncture has shown the capability of modulating the immuno-inflammatory response of the host. This study aims to evaluate the effects of electroacupuncture (EA) on ligature-induced periodontitis in rats. METHODS: Thirty-two animals were divided into four groups: 1) control; 2) experimental periodontitis (EP); 3) sham-treated (EP/EA-sham); and 4) treated with EA (EP/EA). For the EP groups, a ligature was placed around the right mandibular first molars at day 1. Sessions of EA or EA-sham were assigned every other day. For EA treatment, large intestine meridian points LI4 and LI11 and stomach meridian points ST36 and ST44 were used. EA-sham was performed in off-meridian points. Animals were euthanized at day 11. Histomorphometric and microtomographic analyses were performed. Immunolabeling patterns for the receptor activator of nuclear factor κB ligand (RANKL), osteoprotegerin (OPG), and tartrate-resistant acid phosphatase (TRAP) were assessed. Expressions of interleukin (IL)-1ß, matrix metalloproteinase (MMP)-8, IL-6, and cyclooxygenase (COX)-2 messenger RNAs (mRNAs) were evaluated by quantitative reverse transcription-polymerase chain reaction. Data were analyzed statistically (P <0.05, analysis of variance). RESULTS: Histomorphometric and microtomographic analyses demonstrated that group EP/EA presented reduced alveolar bone loss when compared to group EP (P <0.05). Reduced RANKL immunolabeling and fewer TRAP-positive multinucleated cells were observed in the EA-treated group in relation to group EP. No differences were observed in OPG expression among groups. EA treatment decreased the genic expression of IL-1ß and MMP-8 (P <0.05), increased the mRNA expression of IL-6 (P <0.05), and did not modify the genic expression of COX-2 in animals with EP (P >0.05). CONCLUSION: It can be concluded that EA reduced periodontal tissue breakdown and the expression of some proinflammatory mediators and a proresorptive factor in EP in rats.