Inflammatory profile of apical periodontitis exacerbated by cigarette smoke inhalation: Histological and immunohistochemical analysis in rats.

AIM: The aim of this study was to evaluate the effects of cigarette smoke inhalation (CSI) on inflammation, pro-inflammatory mediators and haematological parameters in rats with induced apical periodontitis (AP). METHODOLOGY: Thirty-two 3-month-old male Wistar rats were divided into four experimental groups (n = 8): C-Control; S-rats with CSI; AP-rats with AP; and SAP-rats with CSI + AP. Animals in groups S and SAP inhaled cigarette smoke by remaining inside a smoking chamber for 8 min, three times daily, for 50 days. After 20 days of smoke inhalation, animals in AP and SAP groups had the pulps of the lower right first molar exposed to oral environment for 30 days to induce AP. In these subsequent 30 days, animals in group S and SAP continued with CSI. On Day 50, animals were euthanized and mandibles were histologically processed to assess inflammatory infiltrate, immunohistochemical interleukins (IL-1ß, IL-6 and TNF-α), and blood samples collected for laboratory analysis. The Mann-Whitney test was performed for non-parametric data and the pairwise analyses of Student's t-test for parametric data, with a significance level of p < .050. RESULTS: Inflammatory infiltrate was moderate in AP group and more severe in the SAP (p = .010). The interleukins IL-6, IL-1ß and TNF-α were higher in SAP group (p < .001) when compared to the AP group. A greater number of red blood cells (p = .010), haemoglobin (p = .007) and neutrophils (p = .014) were observed in the SAP group in comparison with the AP group. CONCLUSION: Cigarette smoke inhalation induced a more severe inflammatory infiltrate, with increased levels of pro-inflammatory cytokines and changes in haematological parameters in rats with induced AP. Thus, CSI aggravated AP, exacerbating the inflammatory response.

Insights into the aetiologies of hypercementosis: A systematic review and a scoring system.

OBJECTIVES: This paper aims to better define hypercementosis, investigate its described potential aetiologies, and determine whether there are different patterns of cementum apposition and if they are a function of their supposed aetiology. DESIGN: A literature review was undertaken using the Medline, DOSS, Scopus and Cochrane Library electronic databases. Two co-authors selected the published works independently, extracted the data in accordance with the PRISMA statement. RESULTS: Among 546 articles, 75 articles were finally selected. Eight different supposed aetiologies were identified: (1) intensive masticatory effort, (2) systemic disease, (3) carious lesion and apical periodontitis, (4) impaction, (5) periodontal disease, (6) concrescence, (7) super-eruption, and (8) drugs. Some of these aetiologies can be combined in the same tooth. Hypercementosis manifestations are various in nature and extent with different patterns that may be aetiology-specific. To improve the description and associated consistency in the characterisation of hypercementosis, in this review but also in future studies, we propose a new qualitative scoring system to quickly characterise hypercementosis and determine its most relevant aetiology. CONCLUSIONS: This systematic review demonstrates that hypercementosis is a complex and not yet well-defined condition. Some forms of apposition are specific to a given aetiology. The hypercementosis characterisation may contribute to document the oral condition and/or the individuals masticatory activity.

Present status and future directions: Imaging techniques for the detection of periapical lesions.

Diagnosing and treating apical periodontitis (AP) in an attempt to preserve the natural dentition, and to prevent the direct and indirect systemic effects of this condition, is the major goal in endodontics. Considering that AP is frequently asymptomatic, and is most often associated with a lesion in the periapex of the affected tooth, within the maxillary bones, imaging becomes of paramount importance for the diagnosis of the disease. The aim of this narrative review was to investigate the most relevant classic and current literature to describe which are, to date, the diagnostic imaging systems most reliable and advanced to achieve the early and predictable detection of AP, the best measures of the lesions and the disclosure of the different features of the disease. Dental panoramic tomography (DPT) is a classic exam, considered still useful to provide the basic diagnosis of AP in certain districts of the maxillary bones. Periapical radiographs (PRs) represent a valid routine examination, with few, known limitations. Cone-beam computed tomography (CBCT) is the only system that ensures the early and predictable detection of all periapical lesions in the jaws, with the minor risk of false positives. These techniques can be successfully implemented, with ultrasounds (USI) or magnetic resonance (MRI) imaging, exams that do not use ionising radiations. MRI and USI provide information on specific features of the lesions, like the presence and amount of vascular supply, their content and their relationship with the surrounding soft tissues, leading to differential diagnoses. Further, all the three-dimensional systems (CBCT, USI and MRI) allow the volumetric assessment of AP. Pioneering research on artificial intelligence is slowly progressing in the detection of periapical radiolucencies on DPTs, PRs and CBCTs, however, with promising results. Finally, it is established that all imaging techniques have to be associated with a thorough clinical examination and a good degree of calibration of the operator.

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.

Absence of Tumor Necrosis Factor Receptor 1 Inhibits Osteoclast Activity in Apical Dental Resorption Caused by Endodontic Infection in Mice.

INTRODUCTION: The aim of this study was to evaluate osteoclastogenesis and dental resorption resulting from endodontic infection in wild-type (WT) and tumor necrosis factor receptor 1 genetically deficient (TNFR1 KO) mice. METHODS: After approval by the ethics committee on the use of animals, 40 mice were distributed into 2 experimental groups based on time periods: 14 days (n = 10 WT mice and n = 10 TNFR1 KO mice) and 42 days (n = 10 WT mice and n = 10 TNFR1 KO mice). After these periods, morphometric analysis was performed using bright field and fluorescence microscopy and tartrate-resistant acid phosphatase histoenzymology to identify osteoclasts. One-way analysis of variance followed by the Tukey post hoc test was used for the statistical analysis (α = 0.05). RESULTS: WT mice in the 42-day period had a greater apical dental resorption in the distal root of the first molar than TNFR1 KO mice (P < .05). On the other hand, TNFR1 KO mice showed a smaller number of osteoclasts on the dental surface than WT mice (P < .05). CONCLUSIONS: WT mice with apical periodontitis had more extensive apical dental resorptions and a larger number of osteoclasts on the tooth surface than TNFR1 KO mice.

Genetic, Cellular and Molecular Aspects involved in Apical Periodontitis.

The development, establishment and repair of apical periodontitis (AP) is dependent of several factors, which include host susceptibility, microbial infection, immune response, quality of root canal treatment and organism's ability to repair. The understanding of genetic contributions to the risk of developing AP and presenting persistent AP has been extensively explored in modern Endodontics. Thus, this article aims to provide a review of the literature regarding the biochemical mediators involved in immune response signaling, osteoclastogenesis and bone neoformation, as the genetic components involved in the development and repair of AP. A narrative review of the literature was performed through a PUBMED/MEDLINE search and a hand search of the major AP textbooks. The knowledge regarding the cells, receptors and molecules involved in the host's immune-inflammatory response during the progression of AP added to the knowledge of bone biology allows the identification of factors inherent to the host that can interfere both in the progression and in the repair of these lesions. The main outcomes of studies evaluated in the review that investigated the correlation between genetic polymorphisms and AP in the last five years, demonstrate that genetic factors of the individual are involved in the success of root canal treatment. The discussion of this review gives subsides that may help to glimpse the development of new therapies based on the identification of therapeutic targets and the development of materials and techniques aimed at acting at the molecular level for clinical, radiographic and histological success of root canal treatment.

Inhibition of RGS10 Aggravates Periapical Periodontitis via Upregulation of the NF-κB Pathway.

INTRODUCTION: Periapical periodontitis develops due to the interplay between root canal microorganisms and host defenses. The mechanism underlying the pathogenesis of periapical periodontitis remains unclear. Regulator of G protein signaling protein 10 (RGS10) has been suggested to play a role in regulating inflammation. This study explored the potential regulatory effects of RGS10 on periapical periodontitis and the proinflammatory pathway of nuclear factor (NF)-κB. METHODS: Disease models of periapical inflammation in mice were established, and adenovirus-associated virus (AAV) was used to inhibit RGS10 expression. Periapical lesions were detected using micro-computed tomography. Quantitative reverse transcriptase PCR (qRT-PCR), western blotting (WB), enzyme-linked immunosorbent assay (ELISA), enzyme activity staining of tartrate-resistant acid phosphatase, and immunohistochemistry were conducted to assess the role of RGS10 expression on NF-κB proinflammatory signaling, OPG, RANKL, and osteoclasts in the periapical regions of each group. TNFα was used to stimulate L929 cells alone or with small interfering RNA (siRNA). To assess the expression of associated molecules, WB, immunofluorescence, qRT-PCR, and ELISA were performed. RESULTS: RGS10 inhibition increased alveolar bone destruction in periapical periodontitis lesions and substantially enhanced the NF-κB proinflammatory signaling pathway activation level. Furthermore, RGS10 inhibition upregulated the ratio of OPG/RANKL and the maturation of osteoclasts during alveolar bone resorption. L929 cell TNFα stimulation and siRNA transfection confirmed these in vivo results. CONCLUSION: RGS10 negatively regulates NF-κB proinflammatory signaling in periapical periodontitis and participates in bone remodeling. Therefore, RGS10 is a promising treatment option for long-term chronic periapical inflammation and may be a new target for the artificial regulation of inflammation.

Interferon-gamma inducible protein 16 and type I interferon receptors expression in experimental apical periodontitis induced in wild-type mice.

AIM: The aim of this study was to evaluate the IFI16 and IFN-α/ß receptors expression during the genesis and development of experimental apical periodontitis (AP) in mice teeth. METHODOLOGY: Apical periodontitis was induced in the lower first molars of 40 C57BL/6 mice. They were divided according to the experimental periods 2, 7, 14, 21 and 42 days (n = 8 per group). Five animals were used as a control group (without AP). Specimens were submitted to histological processing for description of the inflammatory process, immunostaining for the presence/absence and localization of IFI16 and IFN-α/ß receptors (qualitative and semi-quantitative analysis) and tartrate-resistant acid phosphatase (TRAP) histoenzimology. RESULTS: The results showed a gradual development of AP over the experimental times. The expression of IFI16 was noticeably more exacerbated in the experimental early period (day 2) whilst the lowest expression was observed in the control group (p = .02). For IFN-α/ß receptors, a higher intensity staining was observed 42 days after AP induction, that was statistically different from the control group (p = .02). In addition, the number of TRAP-positive cells was higher on the later periods (days 21 and 42; p < .001). CONCLUSION: IFI16 protein expression was highest during the early periods after AP induction in mice teeth, whilst IFN-α/ß receptor expression was highest after AP became established.

Role of NOD2 and hepcidin in inflammatory periapical periodontitis.

The immunological response occurring during periapical inflammation includes expression of nucleotide binding oligomerization domain containing 2 and hepcidin. Nucleotide binding oligomerization domain containing 2 deficiency increases infiltration of inflammatory cells close to alveolar bone. Hepcidin has an important role in iron metabolism affecting bone metabolism.We investigated the role of nucleotide binding oligomerization domain containing 2 and hepcidin in inflammatory periapical periodontitis. Periapical periodontitis was induced in rats and confirmed by micro-computed tomography. Nucleotide binding oligomerization domain 2 and hepcidin were evaluated through immunohistochemistry. Bioinformatics analysis was undertaken usingthe Kyoto Encyclopedia of Genes and Genomes and Gene Ontology databases. Micro-computer tomography revealed alveolar bone resorption in the periapical region and furcation area of mandibular molars in rats of the periapical periodontitis group. Immunohistochemistry showed increased expressionof nucleotide binding oligomerization domain containing 2 and hepcidin around root apices in rats of the periapical periodontitis group. Bioinformatics analysis of differentially expressed genes in inflamed and non-inflamed tissues revealed enrichment in the NOD-like receptor signaling pathway. Our data suggest that nucleotide binding oligomization domain contain2 and hepcidin have important roles in periapical periodontitis severity because they can reduce alveolar bone loss.They could elicit new perspectives for development of novel strategies for periapical periodontitis treatment.

Correlation between Transforming Growth Factor-ß and Periapical Lesions in Patients with Chronic Apical Periodontitis.

Objective: To examine the expression of transforming growth factor-ß (TGF-ß) in the periapical granulation tissue and serum of patients with chronic apical periodontitis and to conduct immunohistochemical analysis so as to explore the relationship between TGF-ß and the degree of periapical lesions. Methods: Periapical granulation tissues of 20 cases of chronic apical periodontitis were collected as the experimental group. Healthy gingival tissues without eruption of third molars of 5 cases were collected as the control group. Immunohistochemistry, enzyme-linked immunosorbent assay, and real-time PCR (RT-PCR) were utilized to determine the expression of TGF-ß mRNA and protein, and the difference in the expression of TGF-ß was compared between groups. In the experimental group, oral CBCT was taken to measure the periapical bone resorption area. Spearman's correlation method was applied to analyze the correlation between TGF-ß protein and gene expression levels and periapical bone resorption area. Results: Immunohistochemistry and enzyme-linked immunosorbent assay demonstrated that the expression of TGF-ß protein in chronic apical periodontitis tissue and serum was higher than that in the controls (P < 0.05). RT-PCR revealed that the expression of TGF-ß mRNA was higher in chronic apical periodontitis tissue than that of the controls (P < 0.05). Spearman's correlation analysis showed that in the experimental group, the mRNA expression of TGF-ß was positively correlated with the periapical bone resorption area (P < 0.01), and the protein expression level was not correlated with the periapical bone resorption area (P < 0.05). Conclusion: The increased expression of TGF-ß in the periapical granulation tissue and serum of patients with chronic apical periodontitis has a certain correlation with the progression of periapical periodontitis. The correlation between TGF-ß at the mRNA level and the degree of early stage disease as well as the high expression of TGF-ß in inflammatory cells in immunohistochemistry have confirmed that TGF-ß promotes bone resorption in early periapical periodontitis, and its mechanism of action deserves further investigation.

Chronic systemic corticosteroid therapy influences the development of pulp necrosis and experimental apical periodontitis, exacerbating the inflammatory process and bone resorption in rats.

AIM: The main aim of the study was to evaluate the inflammatory response and development of apical periodontitis in rats chronically treated with glucocorticoids. METHODOLOGY: Male Wistar rats (Rattus novergicus) were randomly divided into two groups: the experimental group, which was treated with prednisone (5 mg/kg/day) and a control group, which was administered saline solution for 30 days before induction of apical periodontitis, continuing until the day of euthanasia days 0, 7, 14 and 28 after injury induction. The mandibles were subjected to histological evaluation to determine the size of the lesion, was also performed for the presence and absence of pulp necrosis, bone resorption and micro abscesses, and histomorphometric analyses were performed based on the number of polymorphonuclear cells and mononuclear cells. Histochemical analysis was also performed to assess the percentage of collagen fibres and their typification, in addition to immunohistochemistry for the inflammatory markers interleukin IL-1ß, IL-6, TNF-α and TRAP. RESULTS: Despite after 7 days, there was no differences between groups, a significant increase in the root pulp necrosis (p = .001), micro-abscesses (p = .026) and the size of the apical lesion on the 14th day of treatment with prednisone (p = .008). On the same day, there was also an increase in the number of polymorphonuclear cells (p = .042) and cells immunostained for IL-1ß (p = .006), IL-6 (p < .001) and TRAP (p = .002) in animals treated with prednisone. The numbers of mononuclear cells also increase in 28 days (p = .025) and TNF-α ± increases in the prednisone group on the 7th day (p = .041). The prednisone group also showed a decrease in collagen after 14 (both type I [p = .041] and type III [p = .046]) and 28 type III (p = .002) days after the coronary opening. CONCLUSIONS: The glucocorticoids modified the development of experimental apical periodontitis induced in rats, causing an early increase in periapical bone resorption and pulp necrosis. These effects are associated with alterations in cytokine levels, in the inflammatory response and in collagen deposition, in the 14th day after injury induction.

Pcsk9 Knockout Aggravated Experimental Apical Periodontitis via LDLR.

Apical periodontitis (AP), an inflammatory lesion around the apex of tooth roots, is mostly caused by dental pulp infection. Proprotein convertase subtilisin/kexin type 9 (PCSK9) plays a vital role in regulating cholesterol homeostasis by targeting low-density lipoprotein receptor (LDLR) and participates in bacterium-induced chronic periodontitis. However, the roles of PCSK9 in AP are unknown. Here, we investigated its role in AP by using Pcsk9-/- mice. Micro-computed tomography scanning and histological staining revealed that the periapical bone loss of Pcsk9-/- mice was greater than that of wild-type (WT) mice, and increased expression of inflammation-related factors tumor necrosis factor α (TNF-α) and interleukin (IL)-6 was also observed. Immunofluorescence staining and quantitative real-time polymerase chain reaction showed PCSK9 expression in bone marrow macrophages (BMMs) was increased after treatment with lipopolysaccharide (LPS). This finding was consistent with the in vivo results that the expression level of PCSK9 in exposed WT mice increased compared with that in unexposed WT mice. After LPS challenge, the expression levels of TNF-α, IL-1ß, and IL-6 in BMMs were increased, and Pcsk9 knockout aggravated the expression of these inflammatory factors. The number of osteoclasts positive for tartrate-resistant acid phosphatase staining around the apical lesion in Pcsk9-/- mice was higher than that in WT mice. Then BMMs underwent the osteoclast differentiation. Pcsk9 knockout BMMs induced increased and larger osteoclasts. While this effect of Pcsk9 knockout was abolished by the addition of Ldlr small interfering RNA, revealing that Pcsk9 knockout increased osteoclastogenesis was dependent on the LDLR. Immunohistochemistry staining showed increased expression level of LDLR in exposed Pcsk9-/- periapical areas. In vitro experiments showed that LPS promoted the expression level of LDLR in Pcsk9-/- BMMs and increased osteoclast formation ability, indicating that LPS promoted the elevation of osteoclasteogenesis caused by the Pcsk9 knockout. In conclusion, Pcsk9 deficiency aggravated the inflammatory response and promoted the osteoclastogenesis in an LDLR-dependent manner in AP experimental mice.

Wnt signalling in oral and maxillofacial diseases.

Wnts include more than 19 types of secreted glycoproteins that are involved in a wide range of pathological processes in oral and maxillofacial diseases. The transmission of Wnt signalling from the extracellular matrix into the nucleus includes canonical pathways and noncanonical pathways, which play an important role in tooth development, alveolar bone regeneration, and related diseases. In recent years, with the in-depth study of Wnt signalling in oral and maxillofacial-related diseases, many new conclusions and perspectives have been reached, and there are also some controversies. This article aims to summarise the roles of Wnt signalling in various oral diseases, including periodontitis, dental pulp disease, jaw disease, cleft palate, and abnormal tooth development, to provide researchers with a better and more comprehensive understanding of Wnts in oral and maxillofacial diseases.

An evaluation of an array of viruses and fungi in adult Lebanese patients presenting with various dental infections: A cross-sectional study.

INTRODUCTION: The role of bacteria in the pathogenesis of periodontitis, pericoronitis, and periapical infections has been well-established. However, the variation in the severity and prognosis of these lesions could suggest a potential role of other microorganisms, such as viruses and fungi. This study aims to evaluate the presence of adenovirus, human papillomavirus-16, Epstein-Barr virus, Candida, and non-Candida fungi in these infections. METHODOLOGY: A cross-sectional study including 120 healthy adult patients presenting with dental infections requiring dental extractions were conducted to assess the prevalence and the relative quantity of viruses and fungi in saliva, infected, and healthy tissues using quantitative polymerase chain reaction tests. Samples were collected, and a categorical scale was used for the prevalence and a continuous scale for the relative quantification. Statistical analyses were performed using Chi-square for the prevalence and Wilcoxon rank test for the relative quantification. RESULTS: Except for the Epstein-Barr virus and Candida, the presence of viruses and fungi was significantly associated with dental infections. Adenovirus showed an association with pericoronitis, while human papilloma virus-16 exhibited an association with periapical infections. Non-Candida fungi, on the other hand, showed a positive association with all infected tissues and saliva as compared to healthy control lesions except for periapical infections. CONCLUSIONS: According to this study, viruses and fungi were found to be prevalent in dental infections. However, their associations with those infections vary depending on the types of viruses or fungi involved and the category of dental infections.

VISTA Blockade Aggravates Bone Loss in Experimental Murine Apical Periodontitis.

V-domain Ig suppressor of T cell activation (VISTA) is a novel coinhibitory immune checkpoint molecule that maintains immune homeostasis. The present study explored the role of VISTA in human and murine inflammatory tissues of apical periodontitis (AP). VISTA was upregulated in inflammatory tissues of human AP. In mice, the expression of VISTA gradually increased with the development of mouse experimental apical periodontitis (MAP), the CD3+ T cells, CD11b+ myeloid cells, and FOXP3+ regulatory T cells also gradually accumulated. Moreover, a blockade of VISTA using a mouse in vivo anti-VISTA antibody aggravated periapical bone loss and enhanced the infiltration of immune cells in an experimental mouse periapical periodontitis model. The collective results suggest that VISTA serves as a negative regulator of the development and bone loss of apical periodontitis.

A Mouse Model for Studying the Development of Apical Periodontitis with Age.

Older age is associated with reduced immune function. Our aim was to study how age affects the development of apical periodontitis (AP). AP was induced in two age groups of mice (young vs. adult). Histological samples were stained by Hematoxylin Eosin, Brown and Brenn, and Tartrate-Resistant Acid Phosphatase. In addition, the samples were scanned by Micro-Computerized-Tomography (micro-CT) to evaluate apical constriction and periapical lesion size. Cell density in the periapical region was computationally assessed. Moreover, lesion immune cell populations were characterized by flow cytometry and immunofluorescence. The young group presented more canals with necrotic radicular pulp compared to the adults. There was no difference in bacteria location in the canals between the groups. Apical constriction size was larger in the young mice compared to the adults. The periapical cell density was higher in the young group, while the dominant immune cells in the lesions were neutrophils, which also exhibited the highest young/adult ratio. Immunofluorescence demonstrated neutrophils in the lesion. More osteoclasts were present in the lesions of the young mice, in correlation to the higher volume of bone resorption in this group. Overall, we conclude that the immune reaction to AP stimuli was attenuated in the adult mice compared to the young.

Inflammatory Response Mechanisms of the Dentine-Pulp Complex and the Periapical Tissues.

The macroscopic and microscopic anatomy of the oral cavity is complex and unique in the human body. Soft-tissue structures are in close interaction with mineralized bone, but also dentine, cementum and enamel of our teeth. These are exposed to intense mechanical and chemical stress as well as to dense microbiologic colonization. Teeth are susceptible to damage, most commonly to caries, where microorganisms from the oral cavity degrade the mineralized tissues of enamel and dentine and invade the soft connective tissue at the core, the dental pulp. However, the pulp is well-equipped to sense and fend off bacteria and their products and mounts various and intricate defense mechanisms. The front rank is formed by a layer of odontoblasts, which line the pulp chamber towards the dentine. These highly specialized cells not only form mineralized tissue but exert important functions as barrier cells. They recognize pathogens early in the process, secrete antibacterial compounds and neutralize bacterial toxins, initiate the immune response and alert other key players of the host defense. As bacteria get closer to the pulp, additional cell types of the pulp, including fibroblasts, stem and immune cells, but also vascular and neuronal networks, contribute with a variety of distinct defense mechanisms, and inflammatory response mechanisms are critical for tissue homeostasis. Still, without therapeutic intervention, a deep carious lesion may lead to tissue necrosis, which allows bacteria to populate the root canal system and invade the periradicular bone via the apical foramen at the root tip. The periodontal tissues and alveolar bone react to the insult with an inflammatory response, most commonly by the formation of an apical granuloma. Healing can occur after pathogen removal, which is achieved by disinfection and obturation of the pulp space by root canal treatment. This review highlights the various mechanisms of pathogen recognition and defense of dental pulp cells and periradicular tissues, explains the different cell types involved in the immune response and discusses the mechanisms of healing and repair, pointing out the close links between inflammation and regeneration as well as between inflammation and potential malignant transformation.

Inhibition of the CXCL9-CXCR3 axis suppresses the progression of experimental apical periodontitis by blocking macrophage migration and activation.

Apical periodontitis (AP) is an acute or chronic inflammatory disease caused by complex interactions between infected root canal and host immune system. It results in the induction of inflammatory mediators such as chemokines and cytokines leading to periapical tissue destruction. To understand the molecular pathogenesis of AP, we have investigated inflammatory-related genes that regulate AP development. We found here that macrophage-derived CXCL9, which acts through CXCR3, is recruited by progressed AP. The inhibition of CXCL9 by a CXCR3 antagonist reduced the lesion size in a mouse AP model with decreasing IL-1ß, IL-6 and TNFα expression. The treatment of peritoneal macrophages with CXCL9 and LPS induced the transmigration and upregulation of osteoclastogenic cytokines such as IL-1ß, IL-6 and matrix metalloprotease 2, a marker of activated macrophages. This suggests that the CXCL9-CXCR3 axis plays a crucial role in the development of AP, mediated by the migration and activation of macrophages for periapical tissue destruction. Our data thus show that CXCL9 regulates the functions of macrophages which contribute to AP pathogenesis, and that blocking CXCL9 suppresses AP progression. Knowledge of the principal factors involved in the progression of AP, and the identification of related inflammatory markers, may help to establish new therapeutic strategies.

Microbial Communities in the Extraradicular and Intraradicular Infections Associated With Persistent Apical Periodontitis.

Microorganisms in the complex root canal system and the extraradicular regions, including the periapical lesions and extraradicular biofilm may cause root canal treatment failures. However, few studies described the difference between the intraradicular and extraradicular infections from the same tooth associated with persistent apical periodontitis. This study aimed to characterize the microbiome present in the root canal, extraradicular biofilm, and periapical lesions associated with persistent apical periodontitis. The microbial communities in the root canal, extraradicular biofilm, and periapical lesions were investigated by Illumina high-throughput sequencing using Illumina Hiseq 2500 platform. The dominant phyla in the extraradicular and intraradicular infections associated with persistent apical periodontitis were Proteobacteria, Firmicutes, Bacteroidetes, and Actinobacteria, and the genera Fusobacterium, Morganella, Porphyromonas, Streptococcus, and Bifidobacterium dominated across all samples. Although extraradicular infection sites showed higher OTU richness and ß-diversity compared to intraradicular samples, the occurrence of sinus tract rather than the sampling sites demarcated the microbial communities in the infections associated with persistent apical periodontitis. PERMANOVA analysis confirmed that the samples with or without sinus tracts contained significantly different microbial communities. Porphyromonas, Eubacterium, Treponema, and Phocaeicola were found in significantly higher levels with sinus tracts, whilst Microbacterium and Enterococcus were more abundant in samples without sinus tracts. In conclusion, diverse bacteria were detected in both intraradicular and extraradicular infections associated with persistent apical periodontitis, which might be influenced by the occurrence of the sinus tract. The results may provide new insight into the pathogenesis of persistent apical periodontitis.

Immunohistochemical expression of PCNA, STRO-1 and CD 44 in the healing of experimentally induced periapical lesions in rats.

OBJECTIVE: The aim of the study was to determine the expression of cell proliferating marker, anti-proliferating cell nuclear antigen (anti-PCNA) and mesenchymal stem cell (MSC) markers (anti-STRO-1 and anti-CD44) in periapical periodontitis and their role in the healing of periapical lesion in periapical periodontitis. MATERIALS AND METHODS: Ninety Sprague-Dawley male rats (100 g) were divided into 3 groups: Experimental group I (EG I: n = 30), experimental group II (EG II: n=30) and control group (CG: n = 30). Periapical lesions were experimentally developed by leaving the dental pulp of maxillary first molars mesial root open to oral environment for 4 weeks. Conventional root canal treatment was performed in EG II. Maxillary first molars along with alveolar bone were resected and fixed. The processed samples were stained with routine hematoxylin and eosin (H&E), and evaluated immunohistochemically using antibodies against anti-PCNA, anti-STRO-1, and anti-CD44 polyclonal antibodies. Data were analyzed using Chi-square test and a p-value of <0.05 was considered significant. RESULTS: Immunostaining of anti-PCNA showed 30%, 70% and 53.3% positive staining in CG, EG I, and EG II, respectively (p<0.001). Moreover, the CD44 staining was 20% in CG in contrast to 63.6% in EG I and 43.3 in EG II. STRO-1 staining in CG was 10%, 50% in the EG I and 36.6% in the EG II (p<0.001). CONCLUSIONS: Periapical inflammatory tissues expressed significant proliferative cell marker PCNA and mesenchymal stem cell markers STRO-1, and CD44. These findings further reaffirm the promising role of mesenchymal stem cells in the healing of periapical periodontitis.