Efficacy of reciprocating and ultrasonic activation of 6% sodium hypochlorite in the reduction of microbial content and virulence factors in teeth with primary endodontic infection.

AIM: To evaluate in a clinical trial the efficacy of reciprocating and ultrasonic activation of 6% sodium hypochlorite (NaOCl) in the microbial composition and reduction in microbial load as well as in levels of lipopolysaccharide (LPS) and lipoteichoic acid (LTA) in teeth with primary endodontic infections. METHODOLOGY: Samples were collected from 24 root canals with pulp necrosis and periapical lesions, before and after chemo-mechanical canal preparation. The teeth were randomly divided according to the activation protocol as follows: control group without activation (WA, n = 8), reciprocating activation group using Easy Clean tip (EC, n = 8) and ultrasonic activation group using Irrisonic insert (US, n = 8). Microbiological specimens were processed using a culture technique and microbiota composition was analysed using the checkerboard technique. The levels of LPS and LTA were quantified using limulus amebocyte lysate (LAL) and enzyme-linked immunosorbent assay (ELISA), respectively. The Fisher's exact test, Kruskal-Wallis, Dunn's and Wilcoxon's test with a significance level of P < 0.05 were used for statistical analysis. RESULTS: All initial specimens had growth of viable bacteria in fastidious anaerobe agar (FAA), with an average of 105  CFU mL-1 , whereas only one case had such growth after chemo-mechanical canal preparation. LPS and LTA were recovered in 100% of the cases. Chemo-mechanical canal preparation significantly decreased the levels of LPS and LTA (P < 0.05), but no significant differences were found between the groups (P > 0.05). Through the checkerboard technique, bacteria were found in 100% of the initial specimens with concentrations between <105 and 106 . The most frequently identified microorganisms were Prevotella nigrescens and Enterococcus hirae. After chemo-mechanical canal preparation, many species were not detected in any of the three groups tested. A significant reduction occurred in Group US, followed by Groups EC and WA. CONCLUSIONS: Activation of 6% NaOCl reduced the levels of LPS and LTA with no differences between the groups. However, ultrasonic activation was associated with a greater reduction in microbial load within root canals.

Micro-CT assessment of the shaping ability of four root canal instrumentation systems in oval-shaped canals.

AIM: To compare the shaping ability of four root canal instrumentation systems in oval-shaped canals using micro-computed tomographic analysis. METHODOLOGY: Forty anatomically matched mandibular incisors were scanned and assigned to four groups (n = 10), according to the canal preparation protocol: BioRace, Reciproc, Self-Adjusting File (SAF) and TRUShape systems. After canal instrumentation, the specimens were rescanned, and the registered pre- and postoperative datasets were examined to evaluate the percentages of accumulated hard-tissue debris, untouched canal walls and dentine removed. Kruskal-Wallis and Mann-Whitney U-tests with Bonferroni correction were used to compare the variables in the groups (α = 5%). RESULTS: The preparation techniques did not affect the percentage of accumulated hard-tissue debris (P = 0.126). The percentage of untouched canal areas was significantly higher for BioRace (32.38%)compared to Reciproc (18.95%) and SAF (16.08%) systems (P < 0.05). Reciproc removed significantly more dentine (4.18%) than BioRace (2.21%) and SAF (2.56%) (P < 0.05). The TRUShape system had intermediate results for both untouched canal walls (19.20%) and dentine removed (3.77%), with no significant difference compared to BioRace, Reciproc and SAF systems. CONCLUSIONS: The preparation techniques resulted in the same level of accumulated hard-tissue debris. Compared to the other tested systems, BioRace was associated with more untouched canal walls and Reciproc produced the greatest amount of removed dentine. Although it touched more of the root canal walls, the SAF system removed less dentine, whereas TRUShape had intermediate results for these same parameters. None of the systems tested were able to provide optimal shaping ability in oval-shaped canals.

Clinical efficacy of EDTA ultrasonic activation in the reduction of endotoxins and cultivable bacteria.

AIM: This clinical study was conducted to investigate the influence of 17% ethylenediaminetetraacetic acid (EDTA) ultrasonic activation after chemomechanical preparation (CMP) on eliminating/reducing oral bacterial lipopolysaccharides (known as endotoxins) and cultivable bacteria in teeth with pulp necrosis and apical periodontitis. METHODOLOGY: Samples were taken from 24 root canals at several clinical periods: S1 - before CMP; S2 - after CMP; S3 - after EDTA: G1 - with ultrasonic activation (n = 12) and G2 - without ultrasonic activation (n = 12). Root canals were instrumented using Mtwo rotary files. Culture techniques were used to determine the number of colony-forming units (CFU). Limulus amebocyte lysate (LAL) was used to measure endotoxin levels. Friedman's and Wilcoxon signed-rank tests were used to compare the amount of bacteria and endotoxin levels in each period (P < 0.05). RESULTS: Endotoxins and cultivable bacteria were recovered in 100% of the initial samples (S1). CMP was effective in reducing endotoxins and bacterial load (all with P < 0.05). Higher values of endotoxin reduction were achieved with EDTA ultrasonic activation [G1, 0.02 EU mL-1 (range 0.01-0.75)] compared with the no activation group [G2, 1.13 EU mL-1 (range 0.01-8.34)] (P < 0.05). Regarding bacterial reduction, no statistically significant difference was found in S3, regardless of the group (G1, G2, P > 0.05). CONCLUSIONS: Chemomechanical preparation was effective in reducing bacteria and endotoxins, but could not completely eliminate them. The ultrasonic activation of EDTA was effective in further reducing endotoxin levels in the root canals of teeth with pulp necrosis and apical periodontitis.

Cytocompatibility of Biodentine using a three-dimensional cell culture model.

AIM: To evaluate the cytotoxic effects of Biodentine, using a three-dimensional (3D) cell culture associated with an in situ root-end filling experimental model. White mineral trioxide aggregate (MTA) and zinc oxide cement were used as reference for comparison. IL-1α and TNF-α cytokine production were also evaluated. METHODOLOGY: The root canals of 24 human maxillary incisor teeth were prepared using a single-file reciprocating technique. After root filling, a 3-mm root-end resection was performed and 3 mm of gutta-percha was removed from the canal. The teeth were randomly distributed to receive one of the following root-end filling materials: Biodentine, white MTA or zinc oxide cement (positive control group). In the negative control group, the root canal was not retro-filled. The cytocompatibility of the materials was evaluated using the methyl-thiazol-diphenyl-tetrazolium (MTT) assay in an in situ root-end filling experimental model. Balb/c 3T3 fibroblasts, cultured in rat tail collagen type I 3D scaffold, were exposed to the root apex for 24 h, and cell viability was measured by means of reduction MTT salt. IL-1α and TNF-α production were analysed using enzyme-linked immunosorbent assay. One-way analysis of variance was performed and, when the F-ratios were significant, data were compared by Duncan's multiple-range test. The alpha-type error was set at 0.05. RESULTS: Biodentine and MTA groups had similar cell activity to the negative control group (P > 0.05), indicating low cytotoxicity for both materials. The stronger cytotoxicity effect was identified on the zinc oxide cement (P < 0.05). Zinc oxide cement caused a significant up-regulation in IL-1α and TNF-α (P < 0.05). No significant differences amongst MTA, Biodentine and the negative control group were observed for TNF-α (P > 0.05); however, both MTA and Biodentine were associated with overproduction of IL-1α when compared to the control group (P < 0.05). CONCLUSIONS: Biodentine and MTA had similar cytocompatibility in a 3D cell culture model associated with an in situ root-end filling model. The methodology could be used as an alternative to assess the cytocompatibility of endodontic cements because it is more closely related to the in vivo situation.

Root canal content from primary endodontic infection and upregulation of gelatinases in fibroblast cells.

AIM: To investigate endotoxin levels from primary endodontic infections before and after chemomechanical preparation (CMP) and to determine their antigenicity against 3T3 fibroblasts through gelatinolytic activity of matrix metalloproteinases (MMPs). METHODOLOGY: Twenty-four root canals with primary endodontic infection and apical periodontitis were selected. Samples were collected using paper points before (S1) and after chemomechanical preparation (CMP) (S2). The limulus amebocyte lysate assay was used for endotoxin measurement. Fibroblasts were stimulated with root canal contents for 24 h. Supernatants of cell cultures stimulated with root canal contents were collected after 24 h to determine the levels of MMP-2 and MMP-9 gelatinolytic activity using the zymography technique. Friedman and Wilcoxon tests were used to compare the amount of endotoxin before (S1) and after CMP (S2) (P < 0.05). Data obtained from gelatinolytic activity were analysed using anova and Tukey's tests (P < 0.05). RESULTS: Endotoxin was recovered in 100% of the samples. There was a significant reduction in endotoxin levels after CMP (P < 0.05). A correlation was found between the levels of endotoxins and MMP-2 expression (P < 0.05). Root canal contents of initial samples (S1) induced significantly greater MMP-2 expression by fibroblasts when compared to S2 and the nonstimulated group (P < 0.05). No gelatinolytic activity of MMP-9 was observed in S1, S2 and control group. CONCLUSIONS: Root canal contents from primary endodontic infections had gelatinolytic activity for MMP-2. Moreover, CMP was effective in reducing endotoxin levels and their antigenicity against fibroblasts on gelatinolytic activity.

Microbiological profile and antimicrobial susceptibility pattern of infected root canals associated with periapical abscesses.

The aim of this investigation was to identify microorganisms from root canals with periapical abscesses and assess the susceptibility of specific anaerobic bacteria to selected antimicrobials and their ß-lactamase production. Sixty root canals were microbiologically investigated. The susceptibility of Anaerococcus prevotii, Fusobacterium necrophorum, F. nucleatum, Parvimonas micra, and Prevotella intermedia/nigrescens to antimicrobials was evaluated with the Etest, whereas ß-lactamase production was assessed with nitrocefin. A total of 287 different bacterial strains were recovered, including 201 strict anaerobes. The most frequently strict isolated anaerobes were A. prevotii, P. micra, and F. necrophorum. The selected bacteria were susceptible to all the tested antibiotics, except A. prevotii and Fusobacterium species to azithromycin and erythromycin, as well as A. prevotii and F. necrophorum to metronidazole. None of the microorganisms produced ß-lactamase. Gram-positive anaerobic bacteria predominated in the root canals with periapical abscesses. All microorganisms tested were susceptible to benzylpenicillin, amoxicillin, amoxicillin + clavulanate, cefaclor, and clindamycin, producing no ß-lactamase.

Quantification of cultivable bacteria and endotoxin in post-treatment apical periodontitis before and after chemo-mechanical preparation.

This clinical study was conducted to quantify cultivable bacteria and endotoxin in root canals with post-treatment apical periodontitis by correlating their levels with clinical features and to evaluate the effect of chemo-mechanical preparation (CMP) with 2 % chlorhexidine gel + 17 % EDTA on bacterial and endotoxin removal/elimination. Moreover, target strict Gram-negative anaerobic bacteria were investigated by polymerase chain reaction (PCR). Fifteen teeth with post-treatment apical periodontitis were sampled before (s1) and after (s2) CMP. Culture techniques determined the number of colony-forming units (CFU). PCR (16S rDNA) and limulus amebocyte lysate (LAL) assay were used for bacterial and endotoxin detection, respectively. Prevotella nigrescens (4/15), Prevotella intermedia (2/15), and Tannerella forsythia (2/15) were the most frequently detected species. Endotoxin was recovered in 100 % of the samples. At s1, bacteria and endotoxin were detected at a median value of 5.14 × 10(3) CFU/mL and 3.96 EU/mL, respectively. Higher levels of endotoxin were related to a larger size of radiolucent area (>5 mm) (p < 0.05). CMP was more effective in reducing bacteria (99.61 %) than endotoxin (60.6 %) (both p < 0.05). Our findings indicated that the levels of endotoxin found in infected root canals were related to a larger size of radiolucent area in the periapical region. Moreover, CMP was effective in reducing both bacterial and endotoxin contents in post-treatment apical periodontitis.

A feasibility study for the analysis of reparative dentinogenesis in pOBCol3.6GFPtpz transgenic mice.

AIM: To examine the feasibility of using the pOBCol3.6GFPtpz [3.6-green fluorescent protein (GFP)] transgenic mice as an in vivo model for studying the biological sequence of events during pulp healing and reparative dentinogenesis. METHODOLOGY: Pulp exposures were created in the first maxillary molar of 12-16-week-old 3.6-GFP transgenic mice with CD1 and C57/Bl6 genetic background. Direct pulp capping on exposed teeth was performed using mineral trioxide aggregate followed by restoration with a light-cured adhesive system (AS) and composite resin. In control teeth, the AS was placed in direct contact with the pulp. Animals were euthanized at various time points after pulp exposure and capping. The maxillary arch was isolated, fixed and processed for histological and epifluorescence analysis to examine reparative dentinogenesis. RESULTS: Analysis of teeth immediately after pulp exposure revealed absence of odontoblasts expressing 3.6-GFP at the injury site. Evidence of reparative dentinogenesis was apparent at 4 weeks in 3.6-GFP mice in CD1 background and at 8 weeks in 3.6-GFP mice with C57/Bl6 background. The reparative dentine with both groups contained newly formed atubular-mineralized tissue resembling a dentine bridge and/or osteodentine that was lined by cells expressing 3.6-GFP as well as 3.6-GFP expressing cells embedded within the atubular matrix. CONCLUSION: This study was conducted in a few animals and did not allow statistical analysis. The results revealed that the 3.6-GFP transgenic animals provide a unique model for direct analysis of cellular and molecular mechanisms of pulp repair and tertiary dentinogenesis in vivo. The study also shows the effects of the capping material and the genetic background of the mice in the sequence and timing of reparative dentinogenesis.

Evaluation of cytotoxicity and up-regulation of gelatinases in fibroblast cells by three root repair materials.

AIM: To investigate the effects of root repair materials on the cytotoxicity and gelatinolytic activity of matrix metalloproteinases (MMPs) in 3T3 fibroblasts. METHODOLOGY: Fibroblasts (3T3, 3 × 10(5) cells per well) were incubated with elutes of calcium hydroxide (Biodinâmica, Ibiporã, PR, Brazil), EndoBinder (Binderware, São Carlos, SP, Brazil) and mineral trioxide aggregate (MTA) (Angelus, Londrina, PR, Brazil) for 24 h. The cytotoxicity of all root repair materials was determined using the MTT assay. Supernatants of cell cultures incubated with materials were collected after 24 h to determine the levels of MMP-2 and MMP-9 gelatinolytic activity by gelatin zymography. Data were analysed using anova and Tukey's test. RESULTS: Cells secreted MMP-2 after 24 h with calcium hydroxide inducing significantly greater MMP-2 expression in relation to the control and the other root repair materials (P < 0.05). The cytotoxicity results revealed that there was no significant difference in the cell viability of MTA, EndoBinder and the control group. However, there was a significantly reduced cell viability of 3T3 fibroblasts in association with calcium hydroxide (P < 0.05). CONCLUSIONS: Calcium hydroxide was associated with significantly less cell viability when compared with EndoBinder and MTA. All materials had gelatinolytic activity for MMP-2 with calcium hydroxide being associated with the greatest activity.

Evaluation of cytotoxicity and up-regulation of gelatinases in human fibroblast cells by four root canal sealers.

AIM: To investigate the effects of root canal sealers on the cytotoxicity and gelatinolytic activity of matrix metalloproteinases (MMPs) in human fibroblasts. METHODOLOGY: Human fibroblasts (MRC5, 3×10(5) cells per well) were incubated directly or indirectly with AH Plus, Endomethasone N, Pulp Canal Sealer EWT or Sealapex for 30 min, 1, 4 or 24 h (time-points). The cytotoxicity of all root canal sealers was determined by counting viable cells using the trypan blue exclusion assay. Supernatants of cell cultures incubated with root sealers directly or indirectly were collected after each time-point to determine the levels of MMP-2 and MMP-9 gelatinolytic activity by gelatin zymography. Data were analysed using anova and the Tukey's tests. RESULTS: Cells secreted MMP-2 after periods of 4 and 24 h; however, there were no significant differences between the sealers. Secretion of gelatinases was elevated by root canal sealers in direct contact with the cell monolayer when compared to indirect contact (P < 0.05). At the time-points tested, no gelatinolytic activity could be detected in the control group without the sealers. The cytotoxicity results revealed that all sealers were cytotoxic in both contact forms. Sealapex had the lowest cytotoxicity and AH Plus the most cytotoxicity. CONCLUSIONS: All root canal sealers induced the expression of MMP-2 in MRC5 fibroblasts. AH Plus had the highest cytotoxicity amongst the tested sealers, but all were associated with cytotoxic effects.

Thickness of dentine in mesial roots of mandibular molars with different lengths.

AIM: To measure the minimum thickness of the distal (furcal) root dentine associated with the buccal and lingual canals of the mesial roots of mandibular first molars with different lengths. METHODOLOGY: The mesial roots of 285 mandibular first molars were allocated into three groups according to their length: group I - long (24.14 mm +/- 0.85), group II - medium (22.10 mm +/- 0.65) and group III - short (19.97 mm +/- 0.75). The minimum thickness of the distal (furcal) root dentine associated with the buccal and lingual canals of the mesial roots 2 mm below the furcation was measured. The distance between the buccal and lingual canals, and the depth of concavity in the distal surface of the mesial roots were also measured. anova and Tukey-Kramer were used to test for significant differences among the groups. RESULTS: The minimum thickness of the distal wall of the mesiobuccal canal was significantly different (P < 0.05) between group I (long) and III (short), with long teeth having the smallest mean values. No significant difference was found in the thickness of the distal wall of the mesiolingual canal among the groups studied (P > 0.05). The shortest distance between the mesiobuccal and the mesiolingual canals was observed in group III (P < 0.05). The distal (furcal) concavity was deeper in group I (P < 0.05) when compared with the other groups. CONCLUSION: There was a significant difference in the minimum thickness of the distal (furcal) root wall of the mesiobuccal canal of mandibular first molars 2 mm below the furcation between group I (long) and group III (short) teeth. The thinnest walls were found in the longest teeth. The deepest concavities in the distal (furcal) walls of the mesial roots were found in the longest roots.

Ex vivo antimicrobial activity of several bleaching agents used during the walking bleach technique.

AIM: To investigate ex vivo the antimicrobial activity of a paste of sodium perborate associated with various vehicles comparing it with 37% carbamide peroxide and 35% hydrogen peroxide. METHODOLOGY: The antimicrobial activity of these agents was evaluated against three microorganisms: Enterococcus faecalis, Streptococcus mutans and Candida albicans. One millilitre of each tested substance was placed on the bottom of wells of 24-well cell culture plates. Six wells were used for each time period and group. Two millilitres of the microbial suspension was ultrasonically mixed for 10 s with the bleaching pastes and placed in contact with them for 10, 30, 45 s; 1, 3, 5, 10, 20, 30 min; and 1 and 2 h. After each period of time, 1 mL from each well was transferred to tubes containing 2 mL of freshly prepared brain heart infusion agar + neutralizers. Agar plates were inoculated in appropriate gaseous conditions. Data were analysed statistically by the Kruskal-Wallis test with the level of significance set at P < 0.05. RESULTS: In all groups containing chlorhexidine (groups 3, 5 and 7), the antimicrobial activity of the bleaching paste was significantly increased when compared with groups with other kinds of vehicle (groups 1, 2, 4, 6 and 8). For all tested groups, the most resistant microorganism was E. faecalis. CONCLUSIONS: Chlorhexidine when used as a vehicle for sodium perborate enhanced its antimicrobial activity.

Evaluation of cone beam computed tomography and periapical radiography in the diagnosis of root resorption.

BACKGROUND: This study aimed to evaluate the accuracy of cone beam computed tomography (CBCT) and periapical radiography in diagnosing root resorption and verify the influence of filling material in detecting these lesions. METHODS: Digital periapical radiographs and CBCT images of patients with root resorption and a history of dental trauma from a radiology clinic were reviewed retrospectively. The sample comprised 40 teeth with root resorption and 20 normal teeth as controls. Images were analysed by two radiologists and two endodontists. The sensitivity, specificity and accuracy were determined. The kappa coefficient assessed interobserver agreement and the t test determined significant differences between the imaging methods. RESULTS: The accuracy of CBCT in diagnosing external (P = 0.0144) and internal (P = 0.0038) inflammatory resorption was significantly higher than for periapical radiography. For replacement resorption, no statistical difference was noted (P > 0.05). In endodontically treated teeth, CBCT was statistically superior in diagnosing root resorption (P = 0.0138). CONCLUSIONS: CBCT was superior to digital periapical radiography in diagnosing external and internal inflammatory root resorption after dental trauma and can be considered in the differential diagnosis of resorptive lesions in teeth with endodontic treatment.

Pulp revascularization for immature replanted teeth: a case report.

Immature avulsed teeth are not usually treated with pulp revascularization because of the possibility of complications. However, this therapy has shown success in the treatment of immature teeth with periapical lesions. This report describes the case of an immature replanted tooth that was successfully treated by pulp revascularization. An 8-year-old boy suffered avulsion on his maxillary left lateral incisor. The tooth showed incomplete root development and was replanted after 30 minutes. After diagnosis, revascularization therapy was performed by irrigating the root canal and applying a calcium hydroxide paste and 2% chlorhexidine gel for 21 days. In the second session, the intracanal dressing was removed and a blood clot was stimulated up to the cervical third of the root canal. Mineral trioxide aggregate was placed as a cervical barrier at the entrance of the root canal and the crown was restored. During the follow-up period, periapical repair, apical closure and calcification in the apical 4 mm of the root canal was observed. An avulsed immature tooth replanted after a brief extra-alveolar period and maintained in a viable storage medium may be treated with revascularization.

A simple method for the analysis of root canal preparation.

The purpose of this article is to present a simple and easy method for in vitro analysis of root canal instrumentation that permits one to observe and measure the diameter of a root before and after instrumentation, using the teeth themselves as control.

In vitro assessment of the antimicrobial action and the mechanical ability of chlorhexidine gel as an endodontic irrigant.

The objective of this study was to assess the chlorhexidine gluconate gel as an endodontic irrigant. First the ability of chlorhexidine gel to disinfect root canals contaminated in vitro with Enterococcus faecalis was investigated. A scanning electron microscope was also used to evaluate its cleansing ability compared with endodontic irrigants commonly used, such as sodium hypochlorite and chlorhexidine gluconate liquid. The results indicated that the chlorhexidine gel produced a cleaner root canal surface and had an antimicrobial ability comparable with that obtained with the other solutions tested. It was concluded that chlorhexidine gluconate in gel form has potential for use as an endodontic irrigant.

Glycol methacrylate: an alternative method for embedding subcutaneous implants.

The aim of this investigation was to study the use of glycol methacrylate (GMA) as an embedding material for rat subcutaneous tissue, which received implants of tubes with endodontic sealer. After fixation, the specimens were dehydrated in a growing alcohol series up to 95%, immersed in infiltration GMA solution and then in embedding solution. The blocks were cut into 3.0 microm sections and stained with hematoxylin-eosin. The quality of cell definition and staining allowed a quantitative analysis of the cells infiltrated in the end of the tubes. It was also possible to identify each type of inflammatory cell. Moreover, it was possible to distinguish clearly chronic from acute inflammatory cells. The GMA technique is easy to execute and reproducible, and provides a better definition of tissue cells, thus permitting definition of the degree of the inflammatory process. Therefore, it is an excellent alternative for the evaluation of the biocompatibility of endodontic sealers.

Regional odontodysplasia. Report of two cases.

Two cases of regional odontodysplasia in girls are reported; one affected the lower incisors, and the other the left maxilla. The first case was radiographically followed over a 6-year period, during which time the ghost teeth exhibited significant dentin formation, along with a resultant decrease in pulp size and relative normalization of the radicular anatomy. The second case involved the deciduous molars and the first permanent molar. In addition to tooth alterations, both cases exhibited many odontogenic epithelial islands and extensive areas of calcification in the mucosa. Diagnosis, causes, and treatment are discussed in the light of recent data.

Oral changes associated with biliary atresia and liver transplantation.

A child is reported with green hypoplastic teeth in both dentitions, resulting from hyperbilirubinemia biliary atresia. Following liver transplantation, labial hirsutism, gingival hyperplasia and herpes labial, were seen as a consequence of cyclosporin therapy and iatrogenic immunosuppression. The oral manifestations and management in biliary atresia and after liver transplantation are reviewed.

Subversion of antimicrobial calprotectin (S100A8/S100A9 complex) in the cytoplasm of TR146 epithelial cells after invasion by Listeria monocytogenes.

Expressed by squamous mucosal keratinocytes, calprotectin is a complex of two EF-hand calcium-binding proteins of the S100 subfamily (S100A8 and S100A9) with significant antimicrobial activity. Calprotectin-expressing cells resist invasion by Porphyromonas gingivalis, Listeria monocytogenes, and Salmonella enterica serovar Typhimurium (S. typhimurium). To understand the interactions between calprotectin and invasive bacteria, we studied the distribution of calprotectin in the cytoplasm of TR146 epithelial cells. In response to L. monocytogenes, calprotectin mobilized from a diffuse cytoplasmic distribution to a filamentous pattern and colocalized with the microtubule network. Listeria more frequently invaded cells with mobilized calprotectin. Calprotectin mobilization was listeriolysin O-dependent and required calcium (extracellular and intracellular) and an intact microtubule network. In the presence of preformed microtubules in vitro, the anti-Listeria activity of calprotectin was abrogated. To facilitate intraepithelial survival, therefore, Listeria mobilizes calprotectin to colocalize with cytoplasmic microtubules, subverting anti-Listeria activity and autonomous cellular immunity.