Periapical bacterial disinfection is critical for dental pulp regenerative cell therapy in apical periodontitis in dogs.

BACKGROUND: Application of pulp regenerative cell therapy for mature teeth with periapical lesions is a critical clinical challenge. The bacterial infection in inaccessible location within the root canal system and in the periapical lesions could cause resistance and impediment, leading to limitations in successful therapy. Thus, the aim of this study was to examine the effect of residual bacteria on the outcome of pulp regeneration in mature teeth with apical periodontitis in dogs. METHODS: Periapical lesions were induced in 32 root canals of 4 dogs in two different models in severities, model A and model B. Model A (moderate infection): the canal exposed to the oral cavity for 2 weeks and then closed for 2 weeks. Model B (severe infection): the canal exposed to the oral cavity for 2 months and then closed for 5 months. All root canals were irrigated with 6% sodium hypochlorite, and 3% EDTA and further with 0.015% levofloxacin-containing nanobubbles, which was also used as an intracanal medicament. The aseptic conditions were examined by bacterial anaerobic culture and/or PCR analyses. The root canal treatment was repeated several times, and allogeneic dental pulp stem cells were transplanted into the root canals. The radiographic evaluation of periapical lesions was performed by cone-beam computed tomography before the first treatment, just after cell transplantation, and after 2 months and 6 months in both model A, model B, respectively. The animals were then sacrificed and the jaw blocks were harvested for histological and histobacteriological evaluations of pulp regeneration and periapical tissue healing. Furthermore, the DiI-labelled DPSCs were transplanted into the root canals after complete disinfection (n = 4) or without root canal treatment (n = 4) in the apical periodontitis model (model A) in one dog, and cell localization was compared 72 h after transplantation. RESULTS: In 8 out of 12 canals from model A, and 10 out of 15 canals from model B, pulp regeneration with good vascularization, innervation, and a significant reduction in the radiolucent area of the periapical lesions were observed. However, in the other 4 canals and 5 canals from model A and model B, respectively, no pulp tissue was regenerated, and inflammation in the periapical tissue, and external resorption or healed external resorption were detected. The presence of residual bacteria in the periapical tissues and severe inflammation were significantly associated with inhibition of regenerated pulp tissue in these 9 unsuccessful canals (P < 0.05, each) (OR = 0.075, each) analyzed by multiple logistic regression analysis. For cellular kinetics, transplanted cells remained in the disinfected root canals, while they were not detected in the infected root canals, suggesting their migration through the apical foramen under the influence of inflammation. CONCLUSIONS: A true pulp-dentin complex was regenerated in the root canal by the pulp regenerative therapy in mature teeth with apical lesions. The successful pulp regeneration was negatively associated both with residual bacteria and inflammation in the periapical tissue.

Curcumin reduces inflammation in rat apical periodontitis.

AIM: The aim of the study was to evaluate the effect of systemic curcumin administration on the severity of apical periodontitis (AP). METHODOLOGY: Forty male Wistar rats weighing 250-280 g each, age 2.5 months, were distributed into four groups (n = 10): control untreated rats (C), control rats treated with curcumin (CUR), rats with pulp exposure-induced apical periodontitis (AP) and rats with pulp exposure-induced apical periodontitis treated with curcumin (AP-CUR). Curcumin treatment was administered orally once daily for 15 days before pulp exposure and continued for 30 days after pulp exposure. The rats were sacrificed at 30 days, and the jaws were collected and reconstructed in a programme specific for micro-CT. The jaws were processed for analysis of the inflammatory process using haematoxylin and eosin staining and immunohistochemical assays for interleukin tumour necrosis factor alpha (TNF-α), interleukin (Il)-6 and Il-1ß. Tartrate-resistant acid phosphatase (TRAP) and osteocalcin (OCN) staining were used to analyse the resorptive process on the bone surface of periapical area. Kruskal-Wallis with Dunn's test was performed for nonparametric data and anova with Tukey's test for parametric data, p < .05. RESULTS: Micro-CT revealed no statistically significant differences in bone resorption between the AP and AP-CUR groups (p > .05). The levels of inflammatory cell infiltration and immunoreactivity for the proinflammatory cytokines TNF-α, Il-6 and Il-1ß were significantly higher in the periapical lesions of the AP group than in the AP-CUR group (p < .05). The number of TRAP-positive multinucleated cells was higher in the AP group than in the AP-CUR group (p < .05). In OCN-positive cells, no differences were observed between the AP and AP-CUR groups (p > .05). CONCLUSIONS: Oral supplementation with curcumin had a significant effect on the AP severity in rats, suggesting an anti-inflammatory effect of curcumin on AP development.

Omega 3 Fatty Acids Reduce the Triglyceride Levels in Rats with Apical Periodontitis.

The aim of this study was to evaluate the influence of the prophylactic and therapeutic supplementation with omega 3 polyunsaturated fatty acids (w-3 PUFAs) on the lipid profile and periapical bone resorption in rats with apical periodontitis. Forty male rats were divided into groups: control rats (C), rats treated with w-3 PUFAs (C+O), rats with pulp exposure-induced apical periodontitis (AP), and rats with AP treated with w-3 PUFAs (AP+O). The administration of w-3 PUFAs was carried out orally once a day for 15 days before pulp exposure and, subsequently, for an additional 30 days after pulp exposure. AP was induced by exposing pulpal tissues to the oral environment. The samples were collected after 30 days. Triglycerides and cholesterol levels were enzymatically measured using the Trinder method. The jaws were collected and submitted for histological analysis. Two-way analysis of variance and Kruskal-Wallis tests were used for statistical analysis, and the significance was set at p<0.05. The triglyceride levels of the AP group were significantly higher than those of the C, C+O and AP+O groups (p<0.05). However, the difference in the cholesterol levels among the groups was not significant (p>0.05). Rats with AP showed larger areas of bone resorption as well as higher inflammatory intensity compared with rats with AP supplemented with w-3 PUFAs. It may be concluded that the presence of multiple AP foci increased the triglyceride levels. In addition, omega 3 supplementation might reduce these levels in rats with AP, as well as the bone resorption areas of periapical tissues.

Omega 3 fatty acids reduce the triglyceride levels in rats with apical periodontitis

Abstract The aim of this study was to evaluate the influence of the prophylactic and therapeutic supplementation with omega 3 polyunsaturated fatty acids (w-3 PUFAs) on the lipid profile and periapical bone resorption in rats with apical periodontitis. Forty male rats were divided into groups: control rats (C), rats treated with w-3 PUFAs (C+O), rats with pulp exposure-induced apical periodontitis (AP), and rats with AP treated with w-3 PUFAs (AP+O). The administration of w-3 PUFAs was carried out orally once a day for 15 days before pulp exposure and, subsequently, for an additional 30 days after pulp exposure. AP was induced by exposing pulpal tissues to the oral environment. The samples were collected after 30 days. Triglycerides and cholesterol levels were enzymatically measured using the Trinder method. The jaws were collected and submitted for histological analysis. Two-way analysis of variance and Kruskal-Wallis tests were used for statistical analysis, and the significance was set at p<0.05. The triglyceride levels of the AP group were significantly higher than those of the C, C+O and AP+O groups (p<0.05). However, the difference in the cholesterol levels among the groups was not significant (p>0.05). Rats with AP showed larger areas of bone resorption as well as higher inflammatory intensity compared with rats with AP supplemented with w-3 PUFAs. It may be concluded that the presence of multiple AP foci increased the triglyceride levels. In addition, omega 3 supplementation might reduce these levels in rats with AP, as well as the bone resorption areas of periapical tissues.

Resumo O objetivo deste estudo foi avaliar a influência da suplementação profilática e terapêutica com os ácidos graxos ômega-3 no perfil lipídico e na reabsorção óssea, em ratos com periodontite apical. Quarenta ratos machos foram divididos em grupos: ratos controle (C), ratos tratados com ácidos graxos ômega-3 (C+O), ratos com periodontite apical induzida por meio de exposição pulpar (PA), ratos com PA tratados com ácidos graxos ômega-3 (PA+O). A administração do ômega-3 foi realizada oralmente, uma vez ao dia durante 15 antes da exposição pulpar e, subsequentemente, por mais 30 dias depois da exposição pulpar. A PA foi induzida por meio da exposição do tecido pulpar ao ambiente oral. Após 30 dias, os ratos foram mortos e os níveis de triglicérides e colesterol foram mensurados pelo método enzimático de Trinder. As mandíbulas foram coletadas e submetidas à análise histológica. Análise de variância de dois fatores e teste de Kruskal-Wallis foram utilizados para análise estatística, e o nível de significância foi de p < 0,05. Os níveis de triglicérides do grupo PA foram significativamente maiores que dos grupos C, C+O e PA+O (p<0,05). Entretanto, não houve diferença significativa nos níveis de colesterol entre os grupos (p>0,05). Ratos com PA apresentaram maior área de reabsorção óssea bem como maior intensidade no infiltrado inflamatório comparados aos ratos com PA suplementados com ômega-3. Pode-se concluir que a presença de múltiplos focos de PA aumentou os níveis de triglicérides. Além disso, a suplementação com ômega-3 pode reduzir estes níveis em ratos com PA, bem como a área de reabsorção óssea dos tecidos periapicais.

Omega-3 Fatty Acids Reduce Inflammation in Rat Apical Periodontitis.

INTRODUCTION: The effects of omega-3 polyunsaturated fatty acids (ω-3 PUFAs) on pro- and anti-inflammatory mediators were evaluated in a rat model of pulp exposure-induced apical periodontitis (AP). METHODS: Twenty-eight male Wistar rats were divided into 4 groups: control, untreated rats (group C); control rats treated with ω-3 PUFAs (group C-O); rats with pulp exposure-induced AP (group AP); and rats with pulp exposure-induced AP treated with ω-3 PUFAs (group AP-O). Omega-3 PUFAs were administered orally once a day for 15 days before pulp exposure; this treatment was continued for 30 days after pulp exposure. The rats were sacrificed 30 days after pulp exposure, and their dissected jaws were subjected to immunohistochemical analysis to detect immunoreactivity for tumor necrosis factor alpha (TNF-α), interleukin (IL)-6, IL-1ß, IL-17, and IL-10 on the periapical bone surface. The results were statistically evaluated using analysis of variance and the Tukey post-test. The significance level was set at 5%. RESULTS: Immunoreactivity for the proinflammatory cytokines TNF-α, IL-6, IL-1ß, and IL-17 was higher in the AP group than in the AP-O, C, and C-O groups (P < .05). Immunoreactivity for the anti-inflammatory cytokine IL-10 was lower in the AP group than in the AP-O group (P < .05). CONCLUSIONS: Supplementation with ω-3 PUFAs can modulate the inflammatory response in rat AP, decreasing levels of TNF-α, IL-6, IL-1ß, and IL-17 but increasing levels of IL-10.

Omega 3 Fatty Acids Reduce Bone Resorption While Promoting Bone Generation in Rat Apical Periodontitis.

INTRODUCTION: This study evaluated the effects of the dietary supplement omega 3 polyunsaturated fatty acids (ω-3 PUFAs) on pulp exposure-induced apical periodontitis (AP) in rats. METHODS: Twenty-eight male rats were divided into groups: control untreated rats (C), control rats treated with ω-3 PUFAs alone (C-O), rats with pulp exposure-induced AP, and rats with pulp exposure-induced AP treated with ω-3 PUFAs (AP-O). The ω-3 PUFAs were administered orally, once a day, for 15 days before pulp exposure and, subsequently, 30 days after pulp exposure. Rats were killed 30 days after pulp exposure, and jaws were subjected to histologic and immunohistochemical analyses. Immunohistochemical analyses were performed to detect tartrate-resistant acid phosphatase-positive osteoclasts and osteocalcin-positive osteoblasts on the bone surface of periapical area. Results were statistically evaluated by using analysis of variance and Tukey honestly significant difference, and P < .05 was considered statistically significant. RESULTS: The bone resorption lesion was significantly larger in the AP group compared with AP-O, C, and C-O groups (P < .05). The level of inflammatory cell infiltration was significantly elevated, and the number of tartrate-resistant acid phosphatase-positive osteoclasts was significantly higher in the periapical lesions of the AP group compared with AP-O, C, and C-O groups (P < .05). The number of osteocalcin-positive osteoblasts was significantly increased in the AP-O group compared with the AP group (P > .05). CONCLUSIONS: Supplementation with ω-3 PUFAs not only suppresses bone resorption but also promotes new bone formation in the periapical area of rats with AP in conjunction with downregulation of inflammatory cell infiltration into the lesion.

Raloxifene modulates regulators of osteoclastogenesis and angiogenesis in an oestrogen deficiency periapical lesion model.

AIM: To analyse the local regulatory mechanisms of osteoclastogenesis and angiogenesis during the progression of periapical lesions in female rats with oestrogen deficiency and treatment with raloxifene (RLX). METHODOLOGY: Female Wistar rats were distributed into groups: SHAM-veh, subjected to sham surgery and treated with a vehicle; OVX-veh, subjected to ovary removal and treated with a vehicle; and OVX-RLX, subjected to ovary removal and treated with RLX. Vehicle or RLX was administered orally for 90 days. During treatment, the dental pulp of mandibular first molars was exposed to the oral environment for induction of periapical lesions, which were analysed after 7 and 30 days. After the experimental periods, blood samples were collected for measurement of oestradiol, calcium, phosphorus and alkaline phosphatase. The rats were euthanized and the mandibles removed and processed for immunohistochemical detection of receptor activator of nuclear factor kappa-B ligand (RANKL), osteoprotegerin (OPG), hypoxia-inducible factor-1 alpha (HIF-1α) and bone-specific alkaline phosphatase (BALP). Data were compared using Kruskal-Wallis followed by Dunn test (nonparametric values) and anova followed by the Tukey's test (parametric values). RESULTS: The plasma concentration of oestradiol showed hypo-oestrogenism in the rats subjected to ovary removal. On day 7, alkaline phosphatase activity, calcium and phosphorus were higher in the OVX-RLX group than in the OVX-veh group (P < 0.001), but immunolabelling for RANKL and HIF-1α was lower in OVX-RLX group (P < 0.001). On day 30, the OVX-veh group had higher immunolabelling for RANKL than the OVX-RLX group (P < 0.05). There were no significant differences in the immunoreactivity of OPG and BALP between any groups at either time-point (P > 0.05). CONCLUSION: RLX therapy reversed the increased levels of the local regulators of both osteoclastogenesis and angiogenesis induced by oestrogen deficiency.

Tratamento endodôntico de incisivo lateral superior após falha da cirurgia parendodôntica / Endodontic treatment of maxillary lateral incisor after failed surgery paraendodontic

O propósito deste artigo é relatar um caso clínico no qual foi realizada uma intervenção cirúrgica parendodôntica no incisivo lateral superior esquerdo em um paciente para remoção de uma lesão no periodonto apical, sem um prévio tratamento endodôntico, tendo a incisão, a enucleação da lesão e a sutura sido feitas de maneira equivocada, provocando uma fenestração tecidual no tecido gengival, expondo o tecido ósseo ao meio bucal, com consequente infecção e grande quantidade de exsudato purulento. Foi realizado um tratamento endodôntico no dente em questão, utilizando-se a limpeza e modelagem do canal com instrumentação rotatória e hipoclorito de sódio a 5,25%, cavitação ultrassônica com EDTA a 17%, curativo intracanal com hidróxido de cálcio e, em uma sessão posterior,obturação com guta-percha termoplastificada e cimento resinoso. Durante o exame clínico radiográfico de proservação realizado após quatro meses de concluído o tratamento endodôntico, foi observado a recuperação do tecido gengival e a radiografia periapical mostrou uma regeneração do tecido ósseo, onde antes existia a lesão.

The intention of this article is to tell a clinical case in which was made a endodontic surgery in a patient to remove a lesion in the apical periodonto and was carried throughan endodontic surgical intervention in the maxillary lateral incisor without a previous endodontic treatment, having the incision, the enucleation of the injury and the been suture made in a mistake way, provoking a tecidual fenestration in the gum of the patient,displaying the osseous tissue to the buccal way, with consequent infection and greatamount of purulent exudates. An endodontic treatment in the tooth in question was carried through, using itself cleaning and shaping of the canal with rotatory instrumentation and 5,25% sodium hypochlorite, ultrasonic cavitation with 17% EDTA, dressing intra canalwith of calcium hydroxide and, in a posterior session, obturation with termoplastificad gutta-percha and resinous sealer. During the clinical examination of flare-up conducted after four months of carried through the endodontic treatment was observed a recovery of the gum tissue and the periapical x-ray showed a regeneration of the osseous tissue,where before the injury existed.

Response of the periapical tissue of dogs' teeth to the action of citric acid and EDTA.

The purpose of this study was to analyze the inflammatory response of dog's periapical tissues to 17% trisodium EDTA salt (pH 8.0) and 1% citric acid (pH 2.0). Saline was used as a control. Six adult dogs were used as the biological model of the study. The experimental units comprised 56 roots of mandibular molars (first and second) and premolars (first, second and third). After coronal opening, pulpectomy and root canal instrumentation were performed using the above-mentioned irrigating solutions. After 24 and 48 hours, the animals were euthanized and the teeth and their supporting tissues were removed and histologically processed. The sections were stained with hematoxylin and eosin and analyzed histopathologically with a light microscope at x100 magnification. The histological analysis focused on the occurrence of acute inflammatory response. The presence of swelling, vasodilatation and inflammatory cells were evaluated and the degree of inflammation was determined for each case. Data were analyzed by Fisher's exact test using the SPSS software with a confidence interval of 95% (p<0.05). 17% EDTA and 1% citric acid caused inflammatory responses in dog's periapical tissues with no significant differences to each other or to saline (control) at either the 24-hour (p=0.482) or 48-hour (p=0.377) periods. It may be concluded that the inflammatory response was of mild intensity for the tested substances.

Response of the periapical tissue of dogs' teeth to the action of citric acid and EDTA

The purpose of this study was to analyze the inflammatory response of dog's periapical tissues to 17 percent trisodium EDTA salt (pH 8.0) and 1 percent citric acid (pH 2.0). Saline was used as a control. Six adult dogs were used as the biological model of the study. The experimental units comprised 56 roots of mandibular molars (first and second) and premolars (first, second and third). After coronal opening, pulpectomy and root canal instrumentation were performed using the above-mentioned irrigating solutions. After 24 and 48 hours, the animals were euthanized and the teeth and their supporting tissues were removed and histologically processed. The sections were stained with hematoxylin and eosin and analyzed histopathologically with a light microscope at x100 magnification. The histological analysis focused on the occurrence of acute inflammatory response. The presence of swelling, vasodilatation and inflammatory cells were evaluated and the degree of inflammation was determined for each case. Data were analyzed by Fisher's exact test using the SPSS software with a confidence interval of 95 percent (p<0.05). 17 percent EDTA and 1 percent citric acid caused inflammatory responses in dog's periapical tissues with no significant differences to each other or to saline (control) at either the 24-hour (p=0.482) or 48-hour (p=0.377) periods. It may be concluded that the inflammatory response was of mild intensity for the tested substances.