Effects of Calcium Hydroxide Intracanal Medications on T Helper (Th1, Th2, Th9, Th17, and Tfh) and Regulatory T (Treg) Cell Cytokines in Apical Periodontitis: A CONSORT RCT.

INTRODUCTION: This Consolidated Standards of Reporting Trials randomized clinical trial investigated T helper (Th1, Th2, Th9, Th17, and Tfh) and regulatory T (Treg) cell-type cytokines and their networks in apical periodontitis (AP). We also assessed the effects of calcium hydroxide [Ca(OH)2] intracanal medications (ICMs) on helper T and Treg cell-type cytokines. METHODS: Twenty teeth with primary endodontic infection and apical periodontitis were randomly divided into two groups: Ca(OH)2 + saline solution (n = 10) and Ca (OH)2 + 2% chlorhexidine-gel (n = 10). Samples were collected from the periradicular tissue fluid (PTF) before (PTFs1) and after 14 days of ICMs (PTFs2). The Human High Sensitivity T Cell Panel was used to quantify target T-helper (Th)1: interelukin (IL)-2, IL-12, and interferon-gamma (IFN-γ); Th2: IL-4, IL-5, and IL-13; Th9: IL-9; Th17: IL-17; T follicular helper cells (Tfh): IL-21; and Treg-cell-type cytokine: IL-10. RESULTS: Th1-type cytokines were higher than Th2-type ones, at PTFs1. Positive (+) associations were found among all Th1-type cytokines and all Th2-type cytokines. There were negative (-) correlations between all Th1- and Th2-type cytokines. Size of radiolucent lesions and symptoms (tenderness to percussion and/or pain on palpation) were positively correlated with Th1-type cytokines, IL-17, and IL-21 but negatively correlated with Th2-type cytokines and IL-10 (all, P < .001). Both ICMs increased Th2-type cytokines and IL-10 (P < .05) and decreased Th1-type cytokines, IL-17, and IL-21 (P < .05), with no differences among them (P > .05). CONCLUSIONS: Complex T-cell cytokine networks are involved in AP. Both Ca(OH)2 ICMs effectively increased IL-4, IL-5, IL-10, and IL-13 and lowered IL-2, IL-12, IL-17, IL-21, and IFN-γ.

Clinical influence of calcium hydroxide intracanal medications on matrix metalloproteinases and tissue inhibitors of metalloproteinases in apical periodontitis.

OBJECTIVES: This study investigated the influence of calcium hydroxide intracanal medications on the levels of metalloproteinases (MMPs) and their tissue inhibitors (TIMPs) in apical periodontitis (AP). MATERIALS AND METHODS: Twenty primarily infected root canals with AP were randomly divided into two groups: Ca(OH)2 + sterile saline solution (SSL) group and Ca(OH)2 + 2% chlorhexidine gel (CHX gel) group. We collected samples from the periradicular tissue fluid (PTF) before (s1) and after 14 days of intracanal medication (s2). MMP-1, MMP-2, MMP-9, TIMP-1, and TIMP-2 were measured by ELISA assay. RESULTS: MMP-1, MMP-2, MMP-9, TIMP-1, and TIMP-2 were detected in all PTF samples at s1 and s2 (20/20). At s1, MMP-2 and MMP-9 were detected at higher levels than MMP-1 (p < .05). Higher levels of TIMP-1 than TIMP-2 were found in AP (p < .05). Additionally, we detected higher MMP-1, MMP-2, and MMP-9 over TIMP-1 and TIMP-2 levels in AP (p < .05). At s2, Ca(OH)2 + SSL was as effective as Ca(OH)2 + 2% CHX gel in lowering the levels of MMP-1, MMP-2, and MMP-9 after 14 days of intracanal medication, with no significant difference between them (p > .05). Both Ca(OH) 2 intracanal medications had no significant impact on the levels of TIMP-1 and TIMP-2 (both p > .05). At s2, TIMP-1 levels were higher than TIMP-2 (p < .05). Moreover, there were positive correlations between the levels of MMP-1 and TIMP-1 and MMP-1 and TIMP-2 (p < .05). CONCLUSIONS: Calcium hydroxide medications effectively lowered the levels of MMP-1, MMP-2, and MMP-9 in periapical tissues after 14 days of treatment, with no difference between them. Moreover, the calcium hydroxide intracanal medications tested here had no impact in TIMP-1 and TIMP-2 in periapical tissues. CLINICAL RELEVANCE: MMPs and TIMPs play an essential role in the degradation of the extracellular matrix. The imbalance MMPs and TIMPs can cause periapical tissue destruction. Therefore, the reestablishment of the balance between activated MMPs and TIMPs with root canal therapy is essential to restore tissue homeostasis.

Resolution of Nasal Sinus Tract after Endodontic Therapy: A Case Report with Microbial Analysis.

This case report describes the resolution of a 20-year misdiagnosed nasal sinus tract after root canal therapy with multiple sessions of calcium hydroxide (Ca[OH]2) intracanal medication. Clinical evaluation, including diagnostic testing and sinus tract tracing, was performed followed by a cone-beam computed tomographic scan and 3-dimensional reconstruction of the apical lesion. Bacteria and endotoxin analyses were performed from the nasal sinus tract and paired root canal infection before (s1) and after instrumentation (s2) and after 7 (s3), 14 (s4), and 21 (s5) days of Ca(OH)2 medication. The bacteria analysis was performed using the checkerboard DNA-DNA hybridization method and endotoxin quantified by the limulus amebocyte lysate method. A similar microbiota profile was found in the sinus tract and paired root canal infection. No target bacterial species were detected in the root canal at s2, s3, and s5. In contrast, Actinomyces israellii and Eubacterium nodatum were detected at s4. Differences in bacterial detection were found between s1 × s2, s3 × s4, and s4 × s5 (all P < .05). Endotoxin was detected in the root canal at all sampling times. Differences in the levels of endotoxin were found between s1 × s2, s2 × s3, and s3 × s4 (all P < .05).The bacterial analysis revealed similar microbiota profiles present in the nasal sinus tract and paired root canal infection with the participation of a wide variety of gram-positive and -negative species. Additionally, root canal therapy with multiple sessions of Ca(OH)2 intracanal medication for 21 days was effective in disinfecting the root canal system and resolving the nasal sinus tract.

Clinical Investigation of Matrix Metalloproteinases, Tissue Inhibitors of Matrix Metalloproteinases, and Matrix Metalloproteinase/Tissue Inhibitors of Matrix Metalloproteinase Complexes and Their Networks in Apical Periodontitis.

INTRODUCTION: This clinical study investigated the levels of metalloproteinases (MMPs) and their tissue inhibitors (TIMPs) and respective forms (MMP/TIMP complexes) in apical periodontitis to determine their networks in the development of clinical/radiographic features, thus quantifying the levels of endotoxins (lipopolysaccharides) present in primarily infected root canals with apical periodontitis. METHODS: Twenty primarily infected root canals with apical periodontitis were selected. The presence of pain on palpation, tenderness to percussion, and the size of the radiographic lesion were recorded. The levels of MMPs (MMP-1, -2, and -9), TIMPs (TIMP-1 and -2), and their MMP/TIMP complexes (MMP-1/TIMP-1, MMP-1/TIMP-2, MMP-2/TIMP-1, MMP-2/TIMP-2, MMP9/TIMP-1, and MMP-9/TIMP-2) present in the periapical interstitial fluid were measured using the enzyme-linked immunosorbent assay. The kinetic chromogenic LAL test was used to quantify endotoxins. RESULTS: A higher mean level of MMP-9 (968.35 ± 342.00 pg/mL) was followed by MMP-2 (894.00 ± 591.62 pg/mL) and MMP-1 (789.43 ± 342.83 pg/mL). The linear regression analysis revealed a positive association of MMP-1 with both MMP-2 and MMP-9 (all P < .001). TIMP-1 (481.79 ± 86.09 pg/mL) (24/24) was found in higher levels than TIMP-2 (206.45 ± 86.09 pg/mL) (P < .05), including a positive correlation of MMP-1 with both TIMP-1 and TIMP-2 (all P < .05). Higher mean levels of MMP1, -2, and -9 were found in teeth with larger-size radiolucent lesions (>7 mm) compared with smaller ones (≤7 mm) (all P < .01). Higher levels of MMP-1 decreased the chance of TTP, whereas MMP-9 (odds ratio = 0.97) increased the chance of pain on percussion (odds ratio = 1.01). Higher levels of endotoxins present in root canals were positively correlated with larger amounts of MMP -9 (P < .05). CONCLUSIONS: MMPs, TIMPs, and their complexes (MMP/TIMP) are involved in apical periodontitis by interacting with complex networks in the development of clinical features and the severity of bone destruction.

Correlation between Volume of Apical Periodontitis Determined by Cone-beam Computed Tomography Analysis and Endotoxin Levels Found in Primary Root Canal Infection.

INTRODUCTION: This clinical study was conducted to correlate the levels of endotoxins and bacterial counts found in primary endodontic infection with the volume of periapical bone destruction determined by cone-beam computed tomography (CBCT) analysis. Moreover, the levels of bacteria and endotoxins were correlated with the development of clinical features. METHODS: Twenty-four root canals with primary endodontic disease and apical periodontitis were selected. Clinical features such as pain on palpation, pain on percussion, and previous episode of pain were recorded. The volume (cubic millimeters) of periapical bone destruction was determined by CBCT analysis. Endotoxins and bacterial samplings were collected by using sterile/apyrogenic paper points. Endotoxins were quantified by using limulus amebocyte lysate assay (KQCL test), and bacterial count (colony-forming units [CFU]/mL) was determined by using anaerobic culture techniques. Data were analyzed by Pearson correlation and multiple logistic regression (P < .05). RESULTS: Endotoxins and bacteria were detected in 100% of the root canal samples (24 of 24), with median values of 10.92 endotoxin units (EU)/mL (1.75-128 EU/mL) and 7.5 × 10(5) CFU/mL (3.20 × 10(5)-8.16 × 10(6) CFU/mL), respectively. The median volume of bone destruction determined by CBCT analysis was 100 mm(3) (10-450 mm(3)). The multiple regression analysis revealed a positive correlation between higher levels of endotoxins present in root canal infection and larger volume of bone destruction (P < .05). Moreover, higher levels of endotoxins were also correlated with the presence of previous pain (P < .05). CONCLUSIONS: Our findings revealed that the levels of endotoxins found in root canal infection are related to the volume of periapical bone destruction determined by CBCT analysis. Moreover, the levels of endotoxin are related to the presence of previous pain.

Microbiological profile resistant to different intracanal medications in primary endodontic infections.

INTRODUCTION: This clinical study aimed to determine the microbiological profile resistant to different intracanal medications in primary endodontic infections by using both microbiological culture and the checkerboard DNA-DNA hybridization technique. METHODS: Twenty primarily infected root canals were selected and then instrumented before being randomly divided into 2 groups according to the intracanal medications: calcium hydroxide (Ca[OH]2) or Ca(OH)2 + chlorhexidine (CHX). Samples were collected before and after root canal procedures, which consisted in submitting them to microbiological culture and processing them for checkerboard DNA-DNA hybridization. RESULTS: No differences were found between the Ca(OH)2 (99.98%) and Ca(OH)2 + CHX groups (99.76%) regarding the median percentage values for the reduction of cultivable bacteria. The most frequently detected species were Capnocytophaga ochracea (70%) and Fusobacterium nucleatum ssp. vincentii (70%) in the initial samples. After instrumentation, the most frequently detected species were E. faecium (60%). After root canal treatments using either Ca(OH)2 or Ca(OH)2 + CHX as intracanal medications, the most frequently detected species were F. nucleatum ssp. vincentii (90%) and Enterococcus faecium (40%), respectively. Both treatments significantly decreased the number of bacterial species compared with the initial sample. However, this reduction was significantly greater in the Ca(OH)2 + CHX group (P < .05). This difference was also observed when evaluating the total bacterial load (P < .05). CONCLUSIONS: The use of Ca(OH)2 associated with CHX as an intracanal medication showed better results by acting on gram-positive and gram-negative microorganisms although such an action to eradicate enterococci should also be sought.