Alveolar bone loss with microarchitecture changes in aged rat / Pérdida ósea alveolar con alteraciones en la microarquitectura en ratas envejecidas

SUMMARY: The aim of the present study was to evaluate the changes of alveolar bone in aged rats. The mandibles of the 4- month and 22-month aged rats were scanned by micro-CT. After the reconstruction of the alveolar bone,the distance between the cemento enamel junction (CEJ) and the alveolar bone crest (ABC) was measured. The micro architectures of the inter-radicular alveolar bone were analyzed. The 22-month rats experienced the reduction in alveolar crest height in the buccal side and the lingual side, and significant increase in alveolar bone loss compared with the 4-month rats. The 22-month rats had a porous microarchitecture, the trabecular arrangement was obviously dissociated with the expanded inter-bone spaces of marrow, and the bone histomorphometry analysis showed the decreased bone volume/tissue volume and trabecular thickness in the 22-month rats. These results suggest that alveolar bone loss and alveolar trabecular bone deterioration might contribute to the weakening of molar support in the elderly.

RESUMEN: El objetivo del presente estudio fue evaluar los cambios del hueso alveolar en ratas envejecidas. Las mandíbulas de las ratas de 4 y 22 meses se escanearon mediante micro-TC. Después de la reconstrucción del hueso alveolar, se midió la distancia entre la unión cementoesmalte (CEJ) y la cresta ósea alveolar (ABC). Se analizaron las microarquitecturas del hueso alveolar interradicular. Las ratas de 22 meses experimentaron la reducción de la altura de la cresta alveolar. en el lado bucal y lingual, y un aumento significativo en la pérdida de hueso alveolar en comparación con las ratas de 4 meses. Las ratas de 22 meses tenían una microarquitectura porosa, la disposición trabecular estaba obviamente disociada con los espacios interóseos expandidos de la médula y el análisis de histomorfometría ósea mostró una disminución del volumen óseo / volumen tisular y del grosor trabecular en las ratas de 22 meses. Estos resultados sugieren que la pérdida ósea alveolar y el deterioro del hueso trabecular alveolar podrían contribuir al debilitamiento del soporte molar en los ancianos.

Effect of Obesity and/or Ligature-induced Periodontitis on Aortic Wall Thickness in Wistar Rats / Efeito da obesidade e/ou periodontite induzida por ligadura na espessura da parede da aorta em ratos wistar

ABSTRACT The purpose of this study was to evaluate aortic wall thickness after periodontal disease and/or obesity induction in a Wistar rat model. Sixty male Wistar rats were randomly divided into four groups: control (CT), periodontal disease (PD), obesity (OB), and obesity plus periodontal disease (OB+PD). Groups OB and OB+PD received cafeteria diet for 17 weeks. After they had acquired obesity (week 12), periodontal disease was induced by placing a silk ligature on the maxillary right second molar of groups PD and OB+PD. During the experimental period, body weight and Lee index were assessed. Mean alveolar bone loss (ABL) was evaluated, and aortas were prepared for histometric analysis of the aortic wall by ImageJ software. Body weight and Lee index increased in rats exposed to cafeteria diet. Mean ABL was higher in Groups PD and OB+PD than in control and OB (p<0.05). ABL was 18% higher in Group OB+PD than in Group PD, with statistically significant difference (p<0.001). Aortas were thicker in Groups OB and OB+PD than in control and PD groups, respectively (2.31mm ± 0.28 and 2.33 ± 0.29 vs. 2.18 ± 0.26 and 2.14 ± 0.27). Group OB differed significantly from the control group (p=0.036), and OB+PD and OB differed significantly from PD (p=0.004 and p= 0.001, respectively). Obesity alters aortic wall thickness in Wistar rats. However, the presence of periodontal disease did not affect the aortic wall thickness under the conditions of the present study.

RESUMO O objetivo deste estudo foi avaliar a espessura da parede da aorta após modelos de indução de doença periodontal e/ou obesidade em ratos Wistar. Sessenta ratos Wistar machos foram aleatoria mente divididos em quatro grupos: controle (CT), doença periodontal (DP), obesidade (OB), obesidade mais doença periodontal (OB+DP). Os grupos OB e OB+DP rece beram dieta de cafeteria por 17 semanas. Após de adquirirem obesidade, (semana 12), doença periodontal foi induzido pela colocação de ligaduras de seda no segundo molar superior direito dos grupos DP e OB+DP. Durante o período experi mental, o peso corporal e índice de Lee foram obtidos. Média de perda óssea alveolar (POA) foi avaliada e as aortas preparadas para análise histométrica da parede aórtica (em mm) pelo software ImageJ. Ratos expostos a dieta de cafeteria demonstraram um aumento do peso corporal e do índice de Lee. Uma POA media maior foi observada nos grupos DP e OB+DP comparado aos grupos controle e OB (p<0.05). O grupo OB+DP, quando comparado ao grupo DP, apresentou POA 18% maior e essa diferença foi estatisticamente significativa (p<0.001). Os grupos OB e OB+DP exibiram uma espessura de aorta maior comparado aos grupos DP e controle, respectivamente (2.31 ± 0.28 e 2.33 ± 0.29 vs. 2.18 ± 0.26 e 2.14 ± 0.27). Diferenças significativas foram observadas nas comparações dos grupos OB e controle (p=0,036), e OB+DP e OB comparado ao grupo DP (p=0.004 e p= 0.001, respectivamente). A obesidade parece afetar a espessura da parede da aorta em ratos Wistar. Entretanto, a presença de doença periodontal não afetou a espessura da parede da aorta sob as condições do presente estudo.

Pam2CSK4 (TLR2 agonist) induces periodontal destruction in mice

Abstract Lipoproteins are important bacterial immunostimulating molecules capable of inducing receptor activator of nuclear factor-κB (RANKL) and osteoclast formation in vitro and in vivo . Although these molecules are present in periodontopathogenic bacteria, their role in periodontitis is not known. In this study, we used Pam2CSK4 (PAM2), a synthetic molecule that mimics bacterial lipoprotein, to investigate the effects of lipoproteins on periodontitis in mice. C57BL/6 male mice were randomly divided into three experimental groups: 1) Negative control group: animals received vehicle injection; 2) Positive control group: animals received injection of Escherichia coli lipopolysaccharide (LPS); 3) PAM2 group: animals received PAM2 injection. All the injections were performed bilaterally every other day into the palatal mucosa between first and second molars. After twenty-four days, the animals were euthanized to assess alveolar bone volume (micro-CT), cellular and extracellular composition in the gingiva (stereometric analysis), and osteoclast numbers (TRAP staining). Treatment with either PAM2 or LPS induced gingival inflammation, as demonstrated by increased infiltration of inflammatory cells and enhanced angiogenesis, associated with a smaller number of fibroblasts and decreased extracellular matrix. Importantly, treatment not only with LPS but also with PAM2 resulted in a larger number of TRAP+ multinucleated osteoclasts and significant loss of alveolar bone. Collectively, our data demonstrate that PAM2 can induce gingival inflammation and bone loss in mice, broadening the avenues of investigation into the role of lipoproteins in the pathogenesis of periodontal disease.

Prevalence of horizontal alveolar changes in edentulous patients: a retrospective tomographic study

Abstract Horizontal bone loss after tooth extraction is a common finding that demands bone reconstruction in various cases. The aim of this study was to assess the horizontal alveolar status in partially and completely edentulous patients using cone-beam computed tomography (CBCT). In total, 1516 CBCT scans of 1404 adult patients were analyzed. Assessment of the images was performed in accordance with the previously published horizontal alveolar change (HAC) classification, which categorizes horizontal bone defects into four classes: HAC 1, HAC 2, HAC 3 and HAC 4 (from the least severe to the most severe condition). Analysis of 1048 scans from partially edentulous patients presented a distribution of 63.55%, 22.14%, 13.36% and 0.95% in HAC 1, HAC 2, HAC 3 and HAC 4, respectively. Analysis of 468 scans from completely edentulous patient images presented a distribution of 19.87%, 28.63%, 41.67% and 9.83% in HAC 1, HAC 2, HAC 3 and HAC 4, respectively. Based on these results, as in HAC 4, no cancellous bone was found between the cortical buccal and lingual/palatal bone plates, it seems reasonable to state that the absence of cancellous bone is higher in completely edentulous patients than in partially edentulous patients. Therefore, the absence of cancellous bone seems to be higher in completely edentulous than in partially edentulous patients.

Evaluation of the effect of topical and systemic ozone application in periodontitis: an experimental study in rats

Abstract Objective: The goal of the present study was to determine the effect of systemic and topical ozone application on alveolar bone loss (ABL) by evaluating the effect of Hypoxia-inducible factor −1 alpha (HIF-1-α) and receptor activator of NF-kB ligand (RANKL)-positive cells on histopathological and immunohistochemical changes in a rat periodontitis model. Methodology: Thirty male Wistar rats were divided into three groups: 1) Group C (control group); 2) Group SO (systemic ozone group) and 3) Group TO (topical ozone group). Experimental periodontitis was induced with a 3/0 silk suture placed at the mandibular left first molars of rats, and the suture was removed 14 days later. Ozone gas was injected intraperitoneally (0.7 mg/kg) in SO group. Topical ozone application protocol was performed using an ozone generator at 80% concentration (4th grade) 90- degree probe for the duration of 30 s. Both ozone applications were carried out for two weeks at intervals of two days. Histomorphometric and immunohistochemical analysis were performed. Results: ABL was significantly lower in Group SO compared to Group C (p: 0.0052). HIF-1α- positive cells were significantly lower in Group TO than in Group C (p: 0.0043). RANKL-positive cells were significantly lower in Group SO and in Group TO compared to the control group (p: 0.0033, p: 0.0075, respectively). Conclusion: Both ozone applications decreased RANKL-positive cell counts, TO application decreased HIF-1-α positive cells counts, and SO application was found to be more effective in reducing ABL compared to control group.

Pam2CSK4 (TLR2 agonist) induces periodontal destruction in mice

Abstract Lipoproteins are important bacterial immunostimulating molecules capable of inducing receptor activator of nuclear factor-κB (RANKL) and osteoclast formation in vitro and in vivo . Although these molecules are present in periodontopathogenic bacteria, their role in periodontitis is not known. In this study, we used Pam2CSK4 (PAM2), a synthetic molecule that mimics bacterial lipoprotein, to investigate the effects of lipoproteins on periodontitis in mice. C57BL/6 male mice were randomly divided into three experimental groups: 1) Negative control group: animals received vehicle injection; 2) Positive control group: animals received injection of Escherichia coli lipopolysaccharide (LPS); 3) PAM2 group: animals received PAM2 injection. All the injections were performed bilaterally every other day into the palatal mucosa between first and second molars. After twenty-four days, the animals were euthanized to assess alveolar bone volume (micro-CT), cellular and extracellular composition in the gingiva (stereometric analysis), and osteoclast numbers (TRAP staining). Treatment with either PAM2 or LPS induced gingival inflammation, as demonstrated by increased infiltration of inflammatory cells and enhanced angiogenesis, associated with a smaller number of fibroblasts and decreased extracellular matrix. Importantly, treatment not only with LPS but also with PAM2 resulted in a larger number of TRAP+ multinucleated osteoclasts and significant loss of alveolar bone. Collectively, our data demonstrate that PAM2 can induce gingival inflammation and bone loss in mice, broadening the avenues of investigation into the role of lipoproteins in the pathogenesis of periodontal disease.

Treatment of peri-implant soft tissue defects: a narrative review

Abstract Soft tissue defects around dental implants, such as papilla or volume loss, peri-implant recession and alterations of the ridge color and/or texture, lead to esthetic and functional complaints. Treatments of these defects in implants are more demanding than in teeth because peri-implant tissue exhibits different anatomical and histological characteristics. This narrative review discusses the proposed treatments for soft tissue defects around implants in the current literature. Several clinical and pre-clinical studies addressed methods to augment the quantity of the peri-implant keratinized mucosa. Autogenous grafts performed better than soft tissue substitutes in the treatment of soft tissue defects, but there is no clinical consensus on the more appropriate donor area for connective tissue grafts. Treatment for facial volume loss, alterations on the mucosa color or texture and shallow peri-implant recessions are more predictable than deep recessions and sites that present loss of papilla. Correction of peri-implant soft tissue defects may be challenging, especially in areas that exhibit larger defects and interproximal loss. Therefore, the regeneration of soft and hard tissues during implant treatment is important to prevent the occurrence of these alterations.

The effect of colchicine on alveolar bone loss in ligature-induced periodontitis

Abstract Colchicine is widely used in the treatment of several inflammatory diseases due to its anti-inflammatory effect, but effects on bone metabolism are unclear. The aim of this study was to evaluate the effects of systemically-administered colchicine on healthy periodontium and experimentally-induced periodontitis. In total, 42 male Wistar rats were included in this study. A non-ligated group constituting the negative control group (Control, C, n = 6) and a ligature-only group forming the positive control group (LO, n = 12) were created separately. Twelve rats were treated with 0.4 mg/kg colchicine and another 12 with 1 mg/kg colchicine. In the colchicine-administered groups, right mandibles constituted the ligated groups (1 mgC-L or 0.4 mgC-L) and left mandibles formed the corresponding non-ligated controls (1mgC or 0.4mgC). Silk ligatures were placed at the gingival margin of the lower first molars. The animals were euthanized at different time-points of healing (11 or 30 days). Alveolar bone loss was clinically measured and TRAP+ osteoclasts, osteoblastic activity, and MMP-1 expression were examined histologically. There was no increase in alveolar bone loss with either colchicine dose in healthy periodontium (p > 0.05) and the highest level of alveolar bone loss, TRAP+ osteoclast number, and MMP-1 expression were measured in the LO group (p < 0.05). The 0.4 mgC-L group showed less alveolar bone loss at 11 days (p < 0.05), but greater loss at 30 days. The 1 mgC-L group showed higher osteoblast number than the other ligated groups (p < 0.05) at both time-points. In summary, colchicine did not increase alveolar bone loss in healthy periodontium and also may tend to reduce periodontitis progression. However, further extensive study is necessary to understand the mechanism of colchicine action on alveolar bone loss in periodontitis.

Gliclazide reduced oxidative stress, inflammation, and bone loss in an experimental periodontal disease model

Abstract Objective The aim of this study was to evaluate the effects of gliclazide on oxidative stress, inflammation, and bone loss in an experimental periodontal disease model. Material and Methods Male albino Wistar rats were divided into no ligature, ligature, and ligature with 1, 5, and 10 mg/kg gliclazide groups. Maxillae were fixed and scanned using micro-computed tomography to quantify linear and bone volume/tissue volume (BV/TV) and volumetric bone loss. Histopathological, immunohistochemical and immunofluorescence analyses were conducted to examine matrix metalloproteinase-2 (MMP-2), cyclooxygenase 2 (COX-2), cathepsin K, members of the receptor activator of the nuclear factor kappa-Β ligand (RANKL), receptor activator of nuclear factor kappa-Β (RANK), osteoprotegerin (OPG) pathway, macrophage migration inhibitory factor (MIF), superoxide dismutase-1 (SOD-1), glutathione peroxidase-1 (GPx-1), NFKB p 50 (Cytoplasm), NFKB p50 NLS (nuclear localization signal), PI3 kinase and AKT staining. Myeloperoxidase activity, malondialdehyde and glutathione levels, while interleukin-1 beta (IL-1β) and tumor necrosis factor-alpha (TNF-α) levels were evaluated by spectroscopic ultraviolet-visible analysis. A quantitative reverse transcription polymerase chain reaction was used to quantify the gene expression of the nuclear factor kappa B p50 subunit (NF-κB p50), phosphoinositide 3-kinase (PI3k), protein kinase B (AKT), and F4/80. Results Micro-computed tomography showed that the 1 mg/kg gliclazide treatment reduced linear bone loss compared to the ligature, 5 mg/kg gliclazide, and 10 mg/kg gliclazide treatments. All concentrations of gliclazide increased bone volume/tissue volume (BV/TV) compared to the ligature group. Treatment with 1 mg/kg gliclazide reduced myeloperoxidase activity, malondialdehyde, IL-1β, and TNF-α levels (p≤0.05), and resulted in weak staining for COX-2, cathepsin k, MMP-2, RANK, RANKL, SOD-1, GPx-1,MIF and PI3k. In addition, down-regulation of NF-κB p50, PI3k, AKT, and F4/80 were observed, and OPG staining was strong after the 1 mg/kg gliclazide treatment. Conclusions This treatment decreased neutrophil and macrophage migration, decreased the inflammatory response, and decreased bone loss in rats with ligature-induced periodontitis.

Maxillary sinus floor pneumatization and alveolar ridge resorption after tooth loss: a cross-sectional study

Abstract This is a cross-sectional study that aimed to estimate maxillary sinus floor (MSF) pneumatization in single missing tooth of posterior maxilla, by using cone-beam computed tomography (CBCT). CBCT images were analyzed bilaterally and divided into 2 groups: edentulous site (EdS) - edentulous single region of upper second premolar, first or second molars; Tooth site (TS) - contralateral region homologous to the EdS region, with tooth present. Variables evaluated were: sinus height (SH), estimated sinus pneumatization (eSP: ∆ EdS - TS), healed ridge height (HR) and presence of localized sinus pneumatization (LSP) in molars teeth at TS. HR were categorized according to therapeutic option for posterior maxilla. 183 CBCT scans were included and it was observed that EdS presented a higher SH than the TS (p < 0.001) showing an eSP of 0.9 ± 2.93 mm. First molars presented the highest SH for both sides, although significant differences were detected when compared to second molars. First molars were mostly affected by LSP at TS (36 out of 43). Individuals with LSP at TS presented lower HR than the ones without LSP (p < 0.05). 54% of the cases presenting LSP obtained HR < 5 mm, which indicates sinus lift surgery. The present study showed that tooth loss in posterior maxilla favors sinus pneumatization and the identification of LSP at molar roots seems to indicate a greater necessity for sinus lift surgeries.

Effects of the photobiomodulation using different energy densities on the periodontal tissues under orthodontic force in rats with type 2 diabetes mellitus

Abstract To evaluate the impact of the GaAlAs diode laser with energy densities of 160 J/cm2, 320 J/cm2, and 640 J/cm2 on the periodontal tissues under continuous orthodontic force application and on the rate of orthodontic tooth movement in rats with type-2 diabetes mellitus. The intensity of primary alveolar bone formation was also investigated through the immune-positive osteocytes for OPN antibody. Forty adult male Wistar rats were divided into eight groups of 5 rats: normoglycemic (N), 160 J-laser-normoglycemic (160 J-LN), 320 J-laser-normoglycemic (320 J-LN), 640 J-laser-normoglycemic (640 J-LN), diabetic (D), 160 J-laser-diabetic (160 J-LD), 320 J-laser-diabetic (320 J-LD), and 640 J-laser-diabetic (640 J-LD) rats. Diabetes mellitus was induced by a single intravenous injection of 40 mg/kg monohydrated-alloxan. An orthodontic force magnitude of 20cN was applied. The laser parameters were continuous emission of 780-nm wavelength, output power of 20mW, and fiber probe with a spot size of 0.04 cm in diameter. Radiographic, histomorphological, and immunohistochemical analysis were performed after a period of 21 days. The photobiomodulation using the energy density of 640 J/cm2 strongly stimulated the alveolar bone formation and contributed the reorganization of the soft periodontal tissues, followed by the 320 J/cm2. Extensive alveolar bone loss, intense infiltration of inflammatory cells, and degradation of the PDJ tissue were mainly found in the D and 160 J-LD groups. The rate of orthodontic tooth movement was represented by the interdental distance between the cementoenamel junctions of the right mandibular first and second molars . This distance was larger in the diabetic groups (D: 39.98±1.97, 160 J-LD: 34.84±6.01, 320 J-LD: 29.82±1.73, and 640 J-LD: 35.47±4.56) than in the normoglycemic groups (N: 21.13±1.19; 160 J-LN: 22.69±0.72, 320 J-LN: 22.28±0.78, and 640 J-LN: 24.56±2.11). The number of osteopontin-positive osteocytes was significantly greater in the 640 J-LD (14.72 ± 0.82; p < 0.01) and 640 J-LN (13.62 ± 1.33; p < 0.05) groups than with D (9.82 ± 1.17) and 160 J-LD (9.77 ± 1.10) groups. Therefore, the energy density of 640 J/cm2 provided the best maintenance and integrity of the periodontal tissue microarchitecture under continuous orthodontic force when compared with the other dosages, mainly in the uncontrolled diabetic rats. The interdental distance was greater in the D and 160 J-LD groups due to presence of severe periodontitis caused by diabetes plus the mechanical stress generated by continuous orthodontic forces, implying, thus, an insufficient biostimulatory effect for the dosage of 160 J/cm2.

Bioactive glass added to autogenous bone graft in maxillary sinus augmentation: a prospective histomorphometric, immunohistochemical, and bone graft resorption assessment

Abstract Objective The aim of this study was to compare the bone resorption rate, histomorphometry and immunohistochemical findings of bioactive glass (Biogran; Biomet, Warsaw, IN, USA) mixed with autogenous bone grafts (1:1) and autogenous bone graft isolate in maxillary sinus elevation surgery. Material and Methods A total of 9 maxillary sinuses were grafted with Biogran with autogenous bone graft (group 1) and 12 were mixed with autogenous bone graft (group 2). Postoperative cone beam computed tomography (CBCT) was used to measure the initial graft volume after 15 days (T1), and 6 months later, another CBCT scan was performed to evaluate the final graft volume (T2) and determine the graft resorption rate. The resorption outcomes were 37.9%±18.9% in group 1 and 45.7%±18.5% in group 2 (P=0.82). After 6 months, biopsies were obtained concurrent with the placement of dental implants; these implants were subjected to histomorphometric analysis and immunohistochemical analysis for tartrate-resistant acid phosphatase (TRAP). Results The average bone formation in group 1 was 36.6%±12.9 in the pristine bone region, 33.2%±13.3 in the intermediate region, and 45.8%±13.8 in the apical region; in group 2, the values were 34.4%±14.4, 35.0%±13.9, and 42.0%±16.6 of new bone formation in the pristine bone, intermediate, and apical regions, respectively. Immunostaining for TRAP showed poor clastic activity in both groups, which can indicate that those were in the remodeling phase. Conclusions The similarity between the groups in the formation and maintenance of the graft volume after 6 months suggests that the bioactive glass mixed with autogenous bone (1:1) can be used safely as a bone substitute for the maxillary sinus lift.

Dehiscences and fenestrations: methodological care necessary to avoid errors in diagnosis and measurement

ABSTRACT The low prevalence of gingival recessions observed in orthodontic clinical practice may be assigned to the fact that in studies in which dehiscences and bone fenestrations are described as frequent, they were diagnosed based on: 1) dry skull studies; 2) areas with periosteal reflection together with flap; and 3) imaging techniques with low sensitivity to detect these defects, which have a delicate structure and function. In areas of pseudo-dehiscences and fenestrations, the periosteum and the alveolar cortical bone are very thin; also, they either have been removed during preparation of the dry specimens in the areas for analysis, or, alternatively, have not been investigated using an ideal imaging method.

RESUMO A ausência de prevalência elevada das recessões gengivais relacionadas à prática clínica ortodôntica se explica, provavelmente, porque, nos trabalhos em que as deiscências e fenestrações ósseas são descritas como frequentes, essas foram diagnosticadas a partir de: 1) estudos em crânios secos; 2) áreas com rebatimento periosteal junto com o retalho; e 3) uso de métodos imagiológicos que falham em sensibilidade para captá-las, pela sua delicadeza estrutural e funcional. Nessas áreas de pseudodeiscências e fenestrações, existe periósteo e cortical óssea alveolar muito fina, que foram eliminados nos procedimentos de preparação dos espécimes secos, nas áreas para a análise ou, então, não se aplicou um método imagiológico ideal.

Agreement, correlation, and kinetics of the alveolar bone-loss measurement methodologies in a ligature-induced periodontitis animal model

Abstract Periodontal research involves the use of animal models to better understand the biological processes of periodontal diseases and the potential of new or existing therapies. Currently, ligature-induced periodontitis in rats is the main model used in periodontal research, in this model, alveolar bone loss (ABL) is the main parameter evaluated by radiographic, morphometric, and histological techniques. Interestingly, although these methodologies are widely used, it is not totally clarified neither the kinetics of ABL over the induction time nor the agreement degree (repeatability and reproducibility) of these techniques. Objective: To characterize ABL kinetics at 0, 3, 7, 15, 30, and 60 days after ABL induction by ligature and to evaluate the intra- (repeatability) and inter-examiner (reproducibility) agreement and the correlation among the radiographic, morphometric, and histological methodologies. Material and Methods: 60 male Wistar rats with induced ABL were randomly divided into 6 experimental groups (n = 10 animals/group). After 0, 3, 7, 15, 30, and 60 days, the animals were euthanized and their hemimandibles were removed for ABL determination using radiographic, morphometric and histological techniques. Results: Radiographic and morphometric/linear techniques allowed the detection of statistically significant ABL on the third day, while histological and morphometric/area techniques could only detect ABL after the seventh day (ANOVA/Tukey, p<0.05). After the fifteenth day, except for histological analysis, the ABL was stabilized. Concerning the agreement of the methodologies, Bland Altman's test (intra and inter-examiner evaluations) showed no difference among the measurements (p>0.05). In addition, high correlations (Pearson's test, r2>0.9, p<0.05) were observed. Conclusion: The results indicated that the minimum time for ABL induction could vary from 3 to 7 days, according to the chosen analysis methodology. Agreement and correlation data support the comparison of results between studies with same induction time.

Suppressor of cytokine signaling 1 expression during LPS-induced inflammation and bone loss in rats

Abstract This study aimed to characterize the dynamics of suppressor of cytokine signaling (SOCS1) expression in a rat model of lipopolysaccharide-induced periodontitis. Wistar rats in the experimental groups were injected three times/week with LPS from Escherichia coli on the palatal aspect of the first molars, and control animals were injected with vehicle (phosphate-buffered saline). Animals were sacrificed 7, 15, and 30 days after the first injection to analyze inflammation (stereometric analysis), bone loss (macroscopic analysis), gene expression (qRT-PCR), and protein expression/activation (Western blotting). The severity of inflammation and bone loss associated with LPS-induced periodontitis increased from day 7 to day 15, and it was sustained through day 30. Significant (p < 0.05) increases in SOCS1, RANKL, OPG, and IFN-γ gene expression were observed in the experimental group versus the control group at day 15. SOCS1 protein expression and STAT1 and NF-κB activation were increased throughout the 30-day experimental period. Gingival tissues affected by experimental periodontitis express SOCS1, indicating that this protein may potentially downregulate signaling events involved in inflammatory reactions and bone loss and thus may play a relevant role in the development and progression of periodontal disease.

Effect of sumac extract on serum oxidative status, RANKL/OPG system and alveolar bone loss in experimental periodontitis in rats

Objectives Sumac (Rhus coriaria L.) is widely used spice which has several properties such as antioxidant, anti-inflammatory and antimicrobial. The purpose of this animal study was to evaluate the effects of sumac extract on levels of receptor activator of nuclear factor-kappa B ligand (RANKL), osteoprotegerin (OPG) expression, serum oxidative status, and alveolar bone loss in experimental periodontitis. Material and Methods Twenty-four Wistar rats were separated into three groups: non-ligated (NL, n=8), ligature only (LO, n=8), and ligature and treated with sumac extract (S, n=8) (20 mg/kg per day for 11 days). A 4/0 silk suture was placed around the mandibular right first molars subgingivally; after 11 days, the rats were sacrificed, and alveolar bone loss was histometrically measured. The detection of RANKL and OPG were immunohistochemically performed. Levels of serum total antioxidant status (TAS)/total oxidant status (TOS), and oxidative stress index (OSI) were also analyzed. Results Alveolar bone loss was significantly greater in the LO group compared to the S and NL groups (p<0.05). The number of inflammatory cell infiltrate (ICI) and osteoclasts in the LO group was significantly higher than that of the NL and S groups (p<0.05). The number of osteoblasts in the LO and S groups was significantly higher than that of the NL group (p<0.05). There were significantly more RANKL-positive cells in the LO group than in the S and NL groups (p<0.05). OPG-positive cells were higher in S group than in LO and NL groups (p<0.05). TOS and OSI levels were significantly reduced in S group compared to LO group (P<0.05) and TAS levels were similar in S and NL group (p>0.05). Conclusions The present study showed that systemic administration of sumac extract may reduce alveolar bone loss by affecting RANKL/OPG balance, TOS and OSI levels in periodontal disease in rats. .

Effects of Alcohol and/or Tobacco Exposure on Spontaneous Alveolar Bone Loss in Rat

The aim of the present study was to evaluate the effect of alcohol and/or tobacco exposure on spontaneous alveolar bone loss in Wistar rats. Twenty-four, male, 60 day-old, Wistar rats were assigned to 4 groups: Group 1 received 10 mL/kg of glucose solution (5%). Group 2 received 2 g/kg alcohol (20%). Group 3 was exposed to tobacco smoke (6 cigarettes/60 min). Group 4 received both interventions of groups 2 and 3. Alcohol was given by gastric gavage and cigarette exposure was performed using a forced ventilation chamber. After 30 days, animals were sacrificed and the upper maxillae removed and defleshed. Morphometric analysis of alveolar bone loss (ABL) around the second molar was performed in standardized digital photographs. Statistical analysis was conducted using paired t-test, one-way ANOVA and occurrence of spontaneous periodontal disease (ABL ≥ 0.39 mm) was analyzed by Fisher's exact test. Significant differences in body weight were observed between all groups. Group 2 presented higher body weight as compared to the 3 other groups at 4 weeks (p≤0.05). Mean ABL values were 0.31 mm (±0.08), 0.29 mm (±0.07), 0.33 mm (±0.10), and 0.33 mm (±0.08) for groups 1, 2, 3, and 4, respectively. No significant differences were found among groups. In the analysis of occurrence of periodontal breakdown, alcohol exposure decreased the occurrence of ABL and cigarette exposure increased ABL. The combination of alcohol and cigarette exposure did not differ from the control group. Alcohol consumption decreased the occurrence of periodontal breakdown, while tobacco increased this rate.

O objetivo do presente estudo foi avaliar o efeito da exposição do álcool e/ou tabaco sobre a perda óssea alveolar (POA) espontânea em ratos Wistar. Vinte e quatro ratos, machos, com 60 dias de vida foram divididos em 4 grupos: Grupo 1 recebeu 10 mL/kg de solução de glicose (5%). Grupo 2 recebeu 2 g/kg de álcool (20%). Grupo 3 foi exposto a fumaça do tabaco (6 cigarros/60 min). Grupo 4 recebeu a mesma intervenção dos grupos 2 e 3. A solução de álcool foi dada por meio de gavagem e a exposição ao tabaco foi realizada por meio de câmara de ventilação forçada. Após 30 dias de experimento, os animais foram sacrificados e as maxilas removidas. Análise morfométrica da POA ao redor do segundo molar superior foi realizada de modo padronizada. A análise estatística dos dados foi realizada por meio de teste t pareado e ANOVA. Ocorrência de doença periodontal espontânea (POA ≥ 0,39 mm) foi realizada pelo teste exato de Fisher. Diferenças significativas no peso corporal médio foram observadas em todos os grupos. Grupo 2 apresentou maior peso corporal médio quando comparado aos outros 3 grupos ao fim do experimento (p≤0,05). A média de POA foi 0,31 mm (±0,08); 0,29 mm (±0,07); 0,33 mm (±0,10) e 0,33 mm (±0,08) para os grupos 1, 2, 3 e 4, respectivamente. Não houve diferenças significativas entre os grupos. Na análise de ocorrência de destruição periodontal, a exposição de álcool diminuiu sua ocorrência, enquanto que exposição ao tabaco aumentou a POA espontânea. A combinação de álcool e tabaco não diferiu do grupo controle.

Microscopic evaluation of induced tooth movement after subluxation trauma: an experimental study in rats

OBJECTIVE: The objective of this study was to assess the histological alterations that occurred in the periodontal area of rat molars submitted to induced tooth movement (ITM) right after an intentional trauma (subluxation). METHODS: Forty adult male Wistar rats (Rattus norvegicus albinus) were selected. The animals were divided into eight groups (n = 5), according to the combination of variables: Group 1 - control (neither trauma nor ITM); Group 2 - ITM; Groups 3, 4, 5 and 6 - dentoalveolar trauma groups corresponding, respectively, to 1, 3, 8 and 10 days after trauma; Groups 7 and 8 - the animals' molars were subjected to a 900 cN impact and, one and three days after the trauma event, tooth movement was induced. The rats' maxillary first molars were mesially moved during seven days with a closed coil (50 cN). After the experimental period of each group, the animals were sacrificed by anesthetic overdose and the right maxillas were removed and processed for histological analysis under light microscopy. RESULTS: In the animals of group 3, 4, 5 and 6, the histological alterations were not very significant. Consequently, the effect of induced tooth movement right after a subluxation event (groups 7 and 8) was very similar to those described for Group 2. CONCLUSION: There was no difference in the quality of periodontal repair when ITM was applied to teeth that had suffered a subluxation trauma. .

OBJETIVO: avaliar as alterações histológicas ocorridas na área periodontal de molares de ratos submetidos à movimentação dentária induzida (MDI), logo após um trauma intencional (subluxação). MÉTODOS: quarenta ratos Wistar machos adultos (Rattus norvegicus albinus) foram selecionados. Os animais foram divididos em oito grupos (n = 5), de acordo com a combinação das variáveis: Grupo 1 - controle (sem trauma e sem MDI); Grupo 2 - MDI; Grupos 3, 4, 5 e 6 - grupos de trauma dentoalveolar correspondendo, respectivamente, para 1, 3, 8 e 10 dias após o trauma; Grupos 7 e 8 - os molares murinos foram submetidos a um impacto de 900cN e, de um e três dias após o evento trauma, o movimento do dente foi induzido. Os primeiros molares superiores dos animais foram movidos mesialmente durante sete dias, com uma mola fechada (50cN). Após período experimental de cada grupo, os animais foram sacrificados por overdose anestésica e as maxilas direitas foram removidas e processadas para análise histológica qualitativa. RESULTADOS: nos animais dos grupos 3, 4, 5 e 6, as alterações histológicas não foram muito significativas. Consequentemente, o efeito do movimento dentário induzido logo após um evento de subluxação (grupos 7 e 8) foi muito semelhante ao descrito para o grupo 2. CONCLUSÃO: não houve diferença na qualidade do reparo periodontal quando a MDI foi aplicada aos dentes que sofreram um trauma de subluxação. .

Effect of andiroba oil on periodontitis in Wistar rats

PURPOSE: To evaluate the effects of andiroba oil on the periodontitis in rats. METHODS: The periodontitis was induced by the placement of cotton ligatures around the cervix of the second upper molars on fifteen rats, and waiting fifty days. The animals were randomly distributed into three groups: saline group, andiroba oil group and meloxican group, differentiated by substance used in the treatment of periodontitis. The groups received the respective substance by gavage for seven days, after the periodontitis induced. It was analyzed the score of inflammatory cells and the measurement from the cemento-enamel junction to the bone crest. RESULTS: The andiroba oil group (p=0.008) and meloxican group (p=0.0347) show a less score of inflammatory cells than saline group, however there weren't difference between them (p=0.2754). Regarding the analysis of measurement from the cemento-enamel junction to the bone crest, there was no difference between groups studied (p=0.3451). CONCLUSION: Andiroba oil decreased the quantity of inflammatory cells, however, it didn't have an effect on the measurement of alveolar bone loss, like the treatment with Meloxican®.

Caracterização morfométrica e molecular do papel de citocinas pró e anti-inflamatórias no processo de reparo ósseo alveolar em condições homeostáticas e infecciosas / Morphometric and molecular characterization of the role of pro- and anti-inflammatory cytokines in alveolar bone repair process under homeostatic and infectious conditions

O metabolismo ósseo é influenciado por fatores endócrinos, genéticos, de crescimento, sistema RANK/RANKL/OPG, além de uma variedade de moléculas regulatórias, como as citocinas. Citocinas têm sido implicadas na patogênese de doenças ósseas, no entanto, ainda pouco se sabe sobre os mecanismos envolvidos na interação entre o sistema ósseo e imunológico no processo de reparo ósseo. O objetivo deste estudo foi caracterizar o papel de TNF-α e IL-10 no reparo ósseo alveolar em condições homeostáticas (controle [C]) e infecciosas (alveolite experimental [A]) pós exodontia em camundongos C57Bl/6 (WT), TNFp55KO e IL-10KO. Após a cirurgia nos grupos infectados foi induzida a alveolite por meio de isquemia do alvéolo e uma suspensão de secreção purulenta. As maxilas foram coletadas em 0h, 7, 14 e 21 dias após a extração do incisivo superior para análises histológica, histomorfométrica e molecular (RealTimePCR). Na análise histomorfométrica foram quantificados os parâmetros coágulo, células inflamatórias, fibras, fibroblastos, vasos sanguíneos, matriz óssea, osteoblastos, osteoclastos, e outros espaço do líquido intersticial e medula óssea. Na análise molecular (RealTimePCR) foram quantificados a expressão de fatores de crescimento, marcadores ósseos e de matriz extracelular, citocinas e quimiocinas envolvidos no processo. Os dados obtidos foram submetidos ao teste OneWay ANOVA seguido do teste de comparação múltipla de Tukey. Os resultados demonstraram que nos camundongos WT-C houve a formação inicial de coágulo (0 hora) com início da expressão de BMP2, BMP4, BMP7, TGFb1 and VEGFa que tiveram aumento gradativo com pico em 7 dias. A expressão de TNF-α e IL10 também tiveram seus picos aos 7 dias em paralelo com contagem de leucócitos, associado com a expressão de CCL2, CCL5 e CXCL1. Nos períodos seguintes houve uma diminuição inflamatória e o aumento de marcadores osteoblásticos/osteogênicos. A indução da alveolite experimental em WT-A resultou no aumento marcante...

Bone metabolism is influenced by endocrine, genetic and growth factors, RANK/RANKL/OPG system, besides a variety of regulatory molecules, such as cytokines. Cytokines have been implicated in pathogenesis of bone diseases, however, little is known about the mechanisms involved in the interaction between skeletal and immune system in the bone repair process. The objective of this study was characterized the role of TNF-α and IL-10 in alveolar bone repair under homeostatic (control [C]) and infectious (experimental alveolitis [A]) conditions in C57Bl/6 (WT), TNFp55KO and IL-10KO mice. After surgery, in infectious groups was induced by ischemia alveolitis the well and a suspension of pus. The maxillas were collected at 0h, 7, 14 and 21 days after extraction of the maxillary incisor for histologic, histomorphometric and molecular (RealTimePCR). In histomorphometric analysis parameters were measured clot, inflammatory cells, fibers, fibroblasts, blood vessels, bone matrix, osteoblast, osteoclast, and other space - the interstitial fluid and bone mar row. Molecular analysis (RealTimePCR) were quantified the expression of growth factors, bone markers and extracellular matrix, cytokines and chemokines involved in the process. The data were submitted to the OneWay ANOVA test followed by Tukey's multiple comparison test. The results showed that in WT-C initial clot formation (0 hours) with early expression of BMP2, BMP4, BMP7, and TGFb1 VEGFa who had gradual increase peaking in 7 days. The expression of TNF-α and IL10 also peaked at 7 days in parallel with leukocyte count, associated with CCL2, CCL5 and CXCL1. In late periods there were decrease of inflammation and markers osteoblastic / osteogenic increased. Induction of experimental alveolitis in WT resulted in a marked increase in expression of TNF-α accompanied by increased expression of CXCL1 and CCL5, increased leukocyte count and decreased of IL10 expression that peaked at 14d, besides prominent...