Periapical bone response to bacterial lipopolysaccharide is shifted upon cyclooxygenase blockage

Abstract Objectives: Infection, inflammation and bone resorption are closely related events in apical periodontitis development. Therefore, we sought to investigate the role of cyclooxygenase (COX) in osteoclastogenesis and bone metabolism signaling in periapical bone tissue after bacterial lipopolysaccharide (LPS) inoculation into root canals. Methodology: Seventy two C57BL/6 mice had the root canals of the first molars inoculated with a solution containing LPS from E. coli (1.0 mg/mL) and received selective (celecoxib) or non-selective (indomethacin) COX-2 inhibitor. After 7, 14, 21 and 28 days the animals were euthanized and the tissues removed for total RNA extraction. Evaluation of gene expression was performed by qRT-PCR. Statistical analysis was performed using analysis of variance (ANOVA) followed by post-tests (α=0.05). Results: LPS induced expression of mRNA for COX-2 (Ptgs2) and PGE2 receptors (Ptger1, Ptger3 and Ptger4), indicating that cyclooxygenase is involved in periapical response to LPS. A signaling that favours bone resorption was observed because Tnfsf11 (RANKL), Vegfa, Ctsk, Mmp9, Cd36, Icam, Vcam1, Nfkb1 and Sox9 were upregulated in response to LPS. Indomethacin and celecoxib differentially modulated expression of osteoclastogenic and other bone metabolism genes: celecoxib downregulated Igf1r, Ctsk, Mmp9, Cd36, Icam1, Nfkb1, Smad3, Sox9, Csf3, Vcam1 and Itga3 whereas indomethacin inhibited Tgfbr1, Igf1r, Ctsk, Mmp9, Sox9, Cd36 and Icam1. Conclusions: We demonstrated that gene expression for COX-2 and PGE2 receptors was upregulated after LPS inoculation into the root canals. Additionally, early administration of indomethacin and celecoxib (NSAIDs) inhibited osteoclastogenic signaling. The relevance of the cyclooxygenase pathway in apical periodontitis was shown by a wide modulation in the expression of genes involved in both bone catabolism and anabolism.

A comparison of the sealing abilities between Biodentine and MTA as root-end filling materials and their effects on bone healing in dogs after periradicular surgery

ABSTRACT Objectives: To compare the sealing ability and biocompatibility of Biodentine with mineral trioxide aggregate (MTA) when used as root-end filling materials. Methodology: The Cell Counting Kit-8 (CCK-8) assay was used to compare the cytotoxicity of MTA and Biodentine. Twenty-one extracted teeth with a single canal were immersed in an acidic silver nitrate solution after root-end filling. Then, the volume and depth of silver nitrate that infiltrated the apical portion of the teeth were analyzed using micro-computed tomography (micro-CT). Seventy-two roots from 3 female beagle dogs were randomly distributed into 3 groups and apical surgery was performed. After six months, the volume of the bone defect surrounding these roots was analyzed using micro-CT. Results: Based on the results of the CCK-8 assay, MTA and Biodentine did not show statistically significant differences in cytotoxicity (P>0.05). The volume and the depth of the infiltrated nitrate solution were greater in the MTA group than in the Biodentine group (P<0.05). The volume of the bone defect was larger in the MTA group than in the Biodentine group. However, the difference was not significant (P>0.05). The volumes of the bone defects in the MTA and Biodentine groups were smaller than the group without any filling materials (P<0.05). Conclusions: MTA and Biodentine exhibited comparable cellular biocompatibility. Biodentine showed a superior sealing ability to MTA in root-end filling. Both Biodentine and MTA promoted periradicular bone healing in beagle dog periradicular surgery models.

Periapical tissue reaction to calcium phosphate root canal sealer in porcine model.

Background: Calcium phosphate cements (CPC) are a group of biomaterials possessing wide scope of use in various branches of medical science. These materials have been proposed to be highly biocompatible and osteoconductive. This study is based on a newly developed CPC formulation (Chitra-CPC) and is aimed at the evaluation of its biocompatibility through an Endodontic Usage Test in a porcine study model. Objective: To evaluate the periapical tissue reaction to Chitra-CPC when used as a root canal sealer/filler material in comparison with a resin sealer, AH Plus (Dentsply). Materials and Methods: The procedure was done on porcine animal model following the ISO 7405 criteria. The material was implanted intentionally into the periapical area of 36 teeth through a root canal procedure carried out in six animals which were divided equally among 1-month and 3-month time periods. Results were based on the histological evaluation of the autopsied specimens after the prescribed time periods. Results: Mild to moderate periapical tissue reaction was found in Chitra-CPC samples belonging to the 1-month time period, whereas majority of the 3-month CPC samples showed an absence of inflammation. Samples of AH Plus in 1-month period showed severe to moderate inflammation, whereas 3-month AH Plus samples had a mild to moderate inflammation. Conclusions: Chitra-CPC is a biocompatible material.

Effect of MTA-based sealer on the healing of periapical lesions

Some manufacturers have recently added specific components to improve the ease of handling and insertion material properties of MTA in order to create MTA-based sealers. Objective The aim of this study was to evaluate the healing of periapical lesions in canine teeth after a single session of endodontic treatment with MTA Fillapex® compared with Sealapex® or Endo-CPM-Sealer®. Material and Methods Sixty-two root canals were performed on two 1-year-old male dogs. After coronal access and pulp extirpation, the canals were exposed to the oral cavity for 6 months in order to induce periapical lesions. The root canals were prepared, irrigated with a solution of 2.5% sodium hypochlorite and filled with gutta-percha and different sealers, according to the following groups: 1) Sealapex®; 2) Endo-CPM-Sealer®; and 3) MTA Fillapex®. Some teeth with periapical lesions were left untreated for use as positive controls. Healthy teeth were used as negative controls. After 6 months, the animals were sacrificed and serial sections from the roots were prepared for histomorphologic analysis and stained with hematoxylin and eosin and the Brown and Brenn technique. The lesions were scored according to pre-established histomorphologic parameters and the scores statistically analyzed using the Kruskal-Wallis test. Results All 3 materials produced similar patterns of healing (p>0.05); in particular, persistent inflammation and absence of complete periapical tissue healing were consistently noted. Conclusions Preparation of the infected root canals followed by filling with the materials studied was insufficient to provide complete healing of the periapical tissues. .

The effect of calcium hydroxide placement on pH and calcium concentration in periapical environment: An in vitro study.

Aims: Comparing the calcium concentration and pH levels of Ca(OH) 2 medicament placing in pulp chamber and root canal. Materials and Methods: Ninety-nine extracted human mandibular second premolars were instrumented to size #40 k file. Nine teeth served as the control group and the remaining teeth were assigned into two groups. Group 1-Ca(OH) 2 was placed in the dried pulp chamber, while root canals remained wet with normal saline; group 2-Ca(OH) 2 was placed in dried root canals. In control group, canals remained wet without medication. Each group was divided into 3 sub-groups of 15 teeth in which pH and calcium concentration were measured in three intervals of 2 days, 1 week, and 2 weeks by pH meter and atomic absorption spectrometer system, respectively. Findings were assessed using Kruskal-Wallis and t-test. Results: At 1 and 2 weeks, the calcium concentration had increased without being significantly different from Ca(OH) 2 placed either in the root canal or in the pulp chamber. Ca(OH) 2 placed in the pulp chamber or root canal provided similar pH values (P=0.362). Conclusions: Placing Ca(OH) 2 in pulp chamber is as effective as placing it in the root canal.

Histopathological analysis of corticosteroid-antibiotic preparation and propolis paste formulation as intracanal medication after pulpectomy: an in vivo study

Intracanal medication in pulpectomy therapy is used between appointments with the objective of reducing pain and inflammatory processes in pulp and periapical tissues. Propolis has been known as a natural antibiotic and has been subject of medical and dental research due to its therapeutic properties such as antibiotic, analgesic and anti-inflammatory effects. OBJECTIVE: The aim was to carry out an in vivo evaluation of the periapical tissue response to propolis paste when used as an intracanal medication in the teeth of dogs after pulpectomy. MATERIAL AND METHODS: 72 dog's incisors were selected for the experiment. After biomechanical preparation the root canal was filled with a corticosteroid-antibiotic preparation, experimental propolis paste, non-medicament (negative control) or non-pulpectomy at all (positive control). The medications were left inside the root canal for 7, 14 or 28 days. At the end of the experimental period histological sections were prepared and all laboratories processes for Harris hematoxylin and eosin staining was proceeded followed by the analysis using an optical microscope. Sections were classified according to a score representing the inflammatory events observed: the presence of polymorphonuclear neutrophils, polymorphonuclear eosinophils, lymphocytes and plasma cells, macrophages and/or giant cells, fibrous condensation and abscesses. RESULTS: There were statistically significant differences between the tissue reactions caused by the two substances being tested, after different experimental periods, with the periapical tissue that was in contact with propolis paste exhibiting fewer inflammatory reactions in comparison to corticosteroid-antibiotic preparation. CONCLUSIONS: The low tissue responses from propolis paste suggest that this material could be considered as an option for root canal medication after pulpectomy.

Tissue reaction to Endométhasone sealer in root canal fillings short of or beyond the apical foramen

OBJECTIVE: This study evaluated the response of periapical tissues to the endodontic sealer Endométhasone in root canal fillings short of or beyond the apical foramen. MATERIAL AND METHODS: Twenty root canals of premolars and incisors of 2 mongrel dogs were used. After coronal access and pulp extirpation, the canals were instrumented up to a size 55 K-file and the apical cemental barrier was penetrated with a size 15 K-file to obtain a main apical foramen, which was widened to a size 25 K-file. The canals were irrigated with saline at each change of file. The root canals were obturated either short of or beyond the apical foramen by the lateral condensation of gutta-percha and Endométhasone, originating 2 experimental groups: G1: Endométhasone/short of the apical foramen; G2: Endométhasone/beyond the apical foramen. The animals were killed by anesthetic overdose 90 days after endodontic treatment. The individual roots were obtained and serial histological sections were prepared for histomorphological analysis (H&E and Brown & Brenn techniques) under light microscopy. The following parameters were examined: closure of the apical foramen of the main root canal and apical opening of accessory canals, apical cementum resorption, intensity of the inflammatory infiltrate, presence of giant cells and thickness and organization of the apical periodontal ligament. Each parameter was scored 1 to 4, 1 being the best result and 4 the worst. Data were analyzed statistically by the Wilcoxon nonparametric tests (p=0.05). RESULTS: Comparing the 2 groups, the best result (p<0.05) was obtained with root canal filling with Endométhasone short of the apical foramen but a chronic inflammatory infiltrate was present in all specimens. CONCLUSIONS: Limiting the filling material to the root canal space apically is important to determine the best treatment outcome when Endométhasone is used as sealer.

Histopathological evaluation of root canal filling materials for primary teeth

This study aimed to assess the response of apical and periapical tissues of dogs' teeth after root canal filling with different materials. Forty roots from dogs' premolars were prepared biomechanically and assigned to 4 groups filled with: Group I: commercial calcium hydroxide and polyethylene glycol-based paste (Calen®) thickened with zinc oxide; Group II: paste composed of iodoform, Rifocort® and camphorated paramonochlorophenol; Group III: zinc oxide-eugenol cement; Group IV: sterile saline. After 30 days, the samples were subjected to histological processing. The histopathological findings revealed that in Groups I and IV the apical and periapical regions exhibited normal appearance, with large number of fibers and cells and no resorption of mineralized tissues. In Group II, mild inflammatory infiltrate and mild edema were observed, with discrete fibrogenesis and bone resorption. Group III showed altered periapical region and thickened periodontal ligament with presence of inflammatory cells and edema. It may be concluded that the Calen paste thickened with zinc oxide yielded the best tissue response, being the most indicated material for root canal filling of primary teeth with pulp vitality.

O objetivo deste estudo foi avaliar a resposta dos tecidos apicais e periapicais de dentes de cães, após obturação dos canais radiculares com diferentes materiais indicados para dentes decíduos. Foram utilizados pré-molares de cães, totalizando 40 raízes que, após pulpectomia e preparo biomecânico, foram divididas em 4 grupos, nos quais os canais radiculares foram obturados com os seguintes materiais: Grupo I - pasta comercial composta de hidróxido de cálcio e polietileno glicol 400 (Calen®) espessada com óxido de zinco; Grupo II - pasta composta de iodofórmio, Rifocort® e paramonoclorofenol canforado; Grupo III - cimento de óxido de zinco e eugenol; e Grupo IV - solução salina. Decorridos 30 dias, as peças foram submetidas ao processamento histológico. De acordo com os resultados da análise histopatológica observou-se que nos Grupos I e IV as regiões apical e periapical apresentaram aspecto de normalidade, com grande número de fibras e células e ausência de reabsorção dos tecidos mineralizados. No Grupo II observou-se infiltrado inflamatório e edema leves, com discreta fibrogênese e reabsorção óssea. O Grupo III apresentou alteração na região periapical e ligamento periodontal ampliado, com presença de células inflamatórias e edema. Os resultados obtidos permitiram concluir que a pasta Calen espessada com óxido de zinco apresentou a melhor resposta tecidual, sendo a mais indicada para a obturação de canais radiculares de dentes decíduos com vitalidade pulpar.

Relationship between the size of patency file and apical extrusion of sodium hypochlorite.

Background: Sodium hypochlorite (NaOCl) is the most widely used endodontic irrigant because of its excellent antimicrobial, organic tissue dissolving, and lubricating properties. However, it is highly cytotoxic to the periapical tissues. Aim: This study evaluated in vitro the extrusion of 5.25% NaOCl through the apical foramina of mesiobuccal (MB) root canals of maxillary first molars in two experimental conditions: Before apical debridement and after apical debridement with different instrument sizes to ensure direct access to the apical foramen (apical patency). Materials and Methods: Coronal accesses were prepared in 17 teeth and the apical foramina of the distobuccal and palatal root canals were sealed. The teeth were held in acrylic receptacles with the roots turned upwards to reproduce their position in the maxillary dental arch. The receptacles were filled with a starch/KI solution (a reagent that changes its color to blue after contacting NaOCl) covering the roots. The experiment had two phases: P1: Irrigation of the MB canals with 5.25% NaOCl without previous establishment of apical patency; P2: Canal irrigation after use of size 10 K-file and size 15 Flexofile as patency files. Only specimens with no NaOCl extrusion in P1 were assigned to P2. NaOCl was delivered pressureless at the canal entrance. The moment that the starch/KI solution contacted NaOCl was captured on digital photographs. Results and Conclusions: There was no NaOCl extrusion in nine specimens in P1, but all of these teeth had irrigant extrusion in P2. The 5.25% NaOCl used as an endodontic irrigant showed great capacity to extrude beyond both intact and small-sized apical foramina of MB root canals of maxillary first molars.

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.

Emprego do iodofórmio em Endodontia: revisäo da literatura / Iodoform application in Endodontics: a review of the literature

O objetivo deste estudo foi realizar uma revisäo da literatura sobre o iodofórmio, referente às suas propriedades anti-sépticas e de estimulaçäo biológica, e também ao seu emprego clínico em Endodontia. Apesar dos excelentes resultados clínicos obtidos com o uso dessa droga, ainda existem divergências entre os autores sobre o seu mecanismo de açäo. Portanto, novos estudos säo necessários para que tal mecanismo seja esclarecido, de forma a justificar o seu uso como medicaçäo intracanal no tratamento de lesöes periapicais refratárias à terapia endodôntica convencional

Contribuição para o estudo da reação inflamatória do tecido periapical em dentes de cães com canais radiculares obturados segundo as técnicas convencional e Sistema Ultrafil / Contribution to the study of the inflammatory reaction of periapical tissue to conventional and Ultrafil technics of canals filling, in dogs

Foi avaliada, por meio de microscopia óptica, a reação inflamatória periapical em dentes de cães frente a 3 técnicas diferentes de obturação de canal. Dentes de cães foram devidamente instrumentados, e os canais obturados pelas técnicas de injeção de guta-percha termoplastificada a baixa temperatura (Sistema Ultrafil), com e sem o uso de cones de guta-percha seccionados na região apical, e técnica de obturação convencional, acorde PAIVA e ANTONIAZZI (1991), sem preparo apical. As observações histopatológicas do tecido periapical, aos 90 dias, permitiu afirmar que a técnica de obturação que utilizou o cone de guta-percha seccionado e preenchimento do canal com Sistema Ultrafil, apresentou consolidação do processo de reparação , enquanto que, nas outras duas técnicas, a reparação foi incompleta

Resinifying therapy in endodontics. II--Histological and bacteriological evaluation.

This study was conducted on six rabbits, six rats and six human permanent teeth to evaluate histologically the effect of resin resorcinol in the periapical areas, the bacteriological aspect of the resin and the penetration of resin in the dentinal tubules. After 12 days period of observation plenty of lymphocytes and macrophages were evident along with granulomas which are suggestive of the tendency of the resin to stay longer. It was observed that the resin penetrate approximately three fourth distance inside the dentinal tubules. In the last phase the resin was found to be effective against the eight commonly found micro organisms. Though the histological results indicated that the periapical tissue would take longer time for complete repair, yet clinically the tooth functions well. Resinifying therapy is effective and economical method in root canal treatment.