Does aPDT reduce bacterial load in endodontic infected teeth? A systematic review and meta-analysis.

This systematic review investigated whether antimicrobial photodynamic therapy (aPDT) after chemomechanical root canal disinfection (CD) yields a greater microbial load reduction than only CD. An electronic literature search was conducted on four databases up to November 2022, with no language or publication date restrictions. Randomized and non-randomized clinical trials were included if participants had a primary endodontic infection in permanent teeth, and if microbial loads before and after using aPDT were compared. Two researchers independently screened titles and abstracts to determine study eligibility. Assessments included risk of bias and methodological quality. This review was registered in PROSPERO (CRD42020181783). Eight studies were included in the qualitative analysis, and six were eligible for meta-analysis. In the random effects model, aPDT significantly improved the results of root canal disinfection when compared with standard protocols for cleaning and shaping (p = 0.04, 95% CI -1.72, -0.05). Subgroup analysis suggested that aPDT has a better effect on reducing the load of anaerobic microorganisms (p = 0.003, 95% CI -3.36, -0.69). The use of aPDT as an adjunct to chemomechanical disinfection promotes additional reduction of the microbial load and, therefore, seems to improve the results of root canal treatments in permanent teeth with a primary endodontic infection. However, certainty of evidence should be improved.

Effect of different disinfection protocols in bacterial viability of an intraradicular biofilm formed in situ.

The present study aimed to evaluate bacterial viability after the use of different disinfection protocols in root canals infected with a multispecies biofilm (MB) formed in situ. Palatal roots with a single canal were obtained from extracted maxillary molars and sterilized before being inserted into the mouth. The roots were contaminated with a MB in an intraoral appliance worn by ten volunteers. All volunteers wore six roots simultaneously in two intraoral devices for 21 days. One root from each volunteer was assigned to each group (n=10): PUI - passive ultrasonic irrigation; EC - Easy Clean; XPF - XP-endo Finisher; aPDT - antimicrobial photodynamic therapy; CI - conventional irrigation; and NC - negative control. The samples were evaluated under confocal laser scanning microscopy. The percentage of viable cells (VC) was calculated over the total percentage of MB biovolume. Data were statistically analyzed (α=5%). The cell viability in the entire root canal or for each third was compared between groups (Kruskal-Wallis test, Dunn post-hoc test) and for the same group (Friedman test, Dunn post-hoc test). Disinfection protocols were not significantly different from each other (P>.05). Samples in EC, PUI, and aPDT had lower cell viability than in NC (P<.05). In the coronal third of samples in the EC, XPF, PUI and aPDT, the percentage of VC biovolume was lower than in the NC (P<.05). The percentage of VC in EC samples was lower in the coronal and middle thirds than in the apical third (P<.05). EC, PUI and aPDT had significant effects on cell viability in intraradicular multispecies biofilm formed in situ when compared with untreated samples.

Effect of different disinfection protocols in bacterial viability of an intraradicular biofilm formed in situ

Abstract The present study aimed to evaluate bacterial viability after the use of different disinfection protocols in root canals infected with a multispecies biofilm (MB) formed in situ. Palatal roots with a single canal were obtained from extracted maxillary molars and sterilized before being inserted into the mouth. The roots were contaminated with a MB in an intraoral appliance worn by ten volunteers. All volunteers wore six roots simultaneously in two intraoral devices for 21 days. One root from each volunteer was assigned to each group (n=10): PUI - passive ultrasonic irrigation; EC - Easy Clean; XPF - XP-endo Finisher; aPDT - antimicrobial photodynamic therapy; CI - conventional irrigation; and NC - negative control. The samples were evaluated under confocal laser scanning microscopy. The percentage of viable cells (VC) was calculated over the total percentage of MB biovolume. Data were statistically analyzed (α=5%). The cell viability in the entire root canal or for each third was compared between groups (Kruskal-Wallis test, Dunn post-hoc test) and for the same group (Friedman test, Dunn post-hoc test). Disinfection protocols were not significantly different from each other (P>.05). Samples in EC, PUI, and aPDT had lower cell viability than in NC (P<.05). In the coronal third of samples in the EC, XPF, PUI and aPDT, the percentage of VC biovolume was lower than in the NC (P<.05). The percentage of VC in EC samples was lower in the coronal and middle thirds than in the apical third (P<.05). EC, PUI and aPDT had significant effects on cell viability in intraradicular multispecies biofilm formed in situ when compared with untreated samples.

Resumo O presente estudo teve como objetivo avaliar a viabilidade bacteriana após o uso de diferentes protocolos de desinfecção em canais radiculares infectados com um biofilme multiespécies (MB) formado in situ. Raízes palatinas com canal único foram obtidas de molares superiores extraídos e esterilizadas antes de serem inseridas na boca. As raízes foram contaminadas com MB em um aparelho intraoral usado por dez voluntários. Todos os voluntários usaram seis raízes simultaneamente em dois dispositivos intrabucais por 21 dias. Uma raiz de cada voluntário foi atribuída a cada grupo (n=10): PUI - irrigação ultrassônica passiva; EC - Easy clean; XPF - XP-endo Finisher; aPDT - terapia fotodinâmica antimicrobiana; IC - irrigação convencional; e, NC - controle negativo. As amostras foram avaliadas em microscopia confocal de varredura a laser. A porcentagem de células viáveis (VC) foi calculada sobre a porcentagem total do biovolume de MB. Os dados foram analisados estatisticamente (α=5%). A viabilidade celular em todo o canal radicular ou em cada terço foi comparada entre os grupos (teste de Kruskal-Wallis, teste post-hoc de Dunn) e no mesmo grupo (teste de Friedman, teste post-hoc de Dunn). Os protocolos de desinfecção não foram significativamente diferentes entre si (P>0,05). Amostras dos grupos EC, PUI e aPDT apresentaram menor viabilidade celular do as do NC (P<0,05). No terço cervical das amostras do EC, XPF, PUI e aPDT, a porcentagem de biovolume de VC foi menor do que no NC (P<0,05). A porcentagem de VC nas amostras do EC foi menor nos terços cervical e médio do que no terço apical (P<0,05). EC, PUI e aPDT tiveram efeitos significativos na viabilidade celular do biofilme multiespécies intrarradicular formado in situ quando comparado com amostras não tratadas. Estudos clínicos devem investigar o papel da redução de cargas bacterianas viáveis no sistema de canais radiculares para o sucesso do tratamento endodôntico.

XP Endo Finisher-R and PUI as supplementary methods to remove root filling materials from curved canals.

This study assessed the ability of XP-endo Finisher R (FKG, La Chaux-de-Fonds, Switzerland) to remove filling remnants from curved mesiobuccal canals of maxillary molars, using the passive ultrasonic irrigation (PUI) technique as a comparison. Twenty-four curved main mesiobuccal canals (MB1) of maxillary molars were instrumented with Wave One (#25/07) and filled with gutta-percha points and AH Plus Sealer. Samples were then re-treated with a standardized protocol with Wave One (#35/06) as the master apical file. Micro-CT scans measured baseline volume of remaining filling material (in mm3). Samples were divided into two groups (n = 12) according to the supplementary cleaning approach: (PUI) or XP-endo Finisher R. Statistics compared baseline and final volume of filling material (within-group); and the percentage of filling material reduction (between-group). Mean baseline volumes, final volumes, and percentages of reduction (%) of filling material for XP-endo Finisher R and PUI were respectively: 0.060 mm3, 0.042 mm3, and 31.28%; and 0.064 mm3, 0.054 mm3, and 16.57%. Both tested protocols reduced the amount of filling material (p < 0.05). XP-endo Finisher R had higher percentage of reduction as compared to PUI (p < 0.05). XP-endo Finisher R and PUI used as supplementary cleaning protocols during re-treatment improved the removal of root filling material in curved canals; but XP-endo Finisher R was approximately twice more efficient. The complete filling material removal during re-treatment procedures is still a challenge. Supplementary cleaning protocols may help to remove the remaining material after the complete mechanical preparation of curved canals. XP-endo Finisher R was approximately twice more efficient than PUI.

XP Endo Finisher-R and PUI as supplementary methods to remove root filling materials from curved canals

Abstract: This study assessed the ability of XP-endo Finisher R (FKG, La Chaux-de-Fonds, Switzerland) to remove filling remnants from curved mesiobuccal canals of maxillary molars, using the passive ultrasonic irrigation (PUI) technique as a comparison. Twenty-four curved main mesiobuccal canals (MB1) of maxillary molars were instrumented with Wave One (#25/07) and filled with gutta-percha points and AH Plus Sealer. Samples were then re-treated with a standardized protocol with Wave One (#35/06) as the master apical file. Micro-CT scans measured baseline volume of remaining filling material (in mm3). Samples were divided into two groups (n = 12) according to the supplementary cleaning approach: (PUI) or XP-endo Finisher R. Statistics compared baseline and final volume of filling material (within-group); and the percentage of filling material reduction (between-group). Mean baseline volumes, final volumes, and percentages of reduction (%) of filling material for XP-endo Finisher R and PUI were respectively: 0.060 mm3, 0.042 mm3, and 31.28%; and 0.064 mm3, 0.054 mm3, and 16.57%. Both tested protocols reduced the amount of filling material (p < 0.05). XP-endo Finisher R had higher percentage of reduction as compared to PUI (p < 0.05). XP-endo Finisher R and PUI used as supplementary cleaning protocols during re-treatment improved the removal of root filling material in curved canals; but XP-endo Finisher R was approximately twice more efficient. The complete filling material removal during re-treatment procedures is still a challenge. Supplementary cleaning protocols may help to remove the remaining material after the complete mechanical preparation of curved canals. XP-endo Finisher R was approximately twice more efficient than PUI.

Marginal gaps and internal voids after root-end filling using three calcium silicate-based materials: A Micro-CT analysis.

This study evaluated the 3D quality of root-end filling, assessing the presence (volume and percentage) of marginal gaps and internal voids formed after retro-filling with three calcium silicate-based materials: MTA Angelus (Angelus Soluçoes Odontologicas, Londrina, PR, Brazil), Biodentine (Septodont Ltd., Saint Maur-des-Faussés, France) and Neo MTA Plus (Avalon Biomed Inc., Bradenton, Florida, US). Thirty human, extracted, single rooted teeth were used. Orthograde root canal treatment, root resection (3mm shorter than the apex) and retrograde cavity preparation with ultrasonic tips were performed. Teeth were divided into 3 groups (n =10 each) following a stratified randomization according to the initial volume of the root-end cavity. After retrofilling, samples were stored for 7 days. Then, two rounds of micro-CT scans were performed: soon after root-end preparation (with the cavity still empty) and 7 days after root-end filling. Marginal gaps, internal voids volume (mm3 and %), as well as, the overall defects (sum of gaps and voids) were evaluated. Statistics compared the three groups in relation to those defects. There was not statistical difference between groups regarding the marginal gaps (P≥ 0.05), the internal voids (P≥ 0.05), and the overall defects (P≥ 0.05). Median (mm3) and % of overall air-entrapment defects (gaps and/or voids) was: 0.004mm3 and 1.749% for MTA Angelus, 0.018mm3 and 6.660% for Biodentine, and 0.012mm3 and 4.079% for Neo MTA Plus. All materials had gaps and/or voids. No differences were found between MTA Angelus, Biodentine and Neo MTA Plus.

Does MTA provide a more favourable histological response than other materials in the repair of furcal perforations? A systematic review.

BACKGROUND: There is no consensus on which furcal perforation repair material induces a more favourable histological response. This systematic review of laboratory studies provides an overview of the studies comparing repair materials in animal models. OBJECTIVES: To evaluate whether mineral trioxide aggregate (MTA) yields a more favourable histological response than other materials when used to repair furcal perforations in animal experimental models. METHODS: This review followed the PRISMA checklist. The studies included various materials used to repair furcal perforations and compared the histological responses with MTA. An electronic search was conducted in EMBASE, PubMed, Scopus and Web of Science up to 2 September 2020, with no language or publication date restrictions. Studies whose full text was unavailable were excluded. The ARRIVE and SYRCLE tools were used to assess the methodological quality and risk of bias (RoB) of the studies. RESULTS: The studies included in the qualitative synthesis were conducted in rat (n = 3) and dog (n = 17) models. They were classified as having a low quality, high methodological heterogeneity and high RoB. MTA and Biodentine, the materials most often compared, reduced the inflammatory reaction to mild over time. In addition, a mineralized tissue was formed in all studies. The response yielded by MTA was better than or equivalent to that of the other tested materials. DISCUSSION: This review confirmed that MTA is the reference standard material for furcal perforation repair. However, research using animal models has inherent limitations, and the substantial methodological heterogeneity across the studies included should be considered. Therefore, the knowledge generated by this systematic review should be translated into clinical practice cautiously. CONCLUSIONS: Features described in the report and quality assessment guidelines, such as PRIASE, ARRIVE and SYRCLE, should guide researchers. Despite the high RoB and the low methodological quality of the studies included, findings indicated that MTA yields a more favourable histological response than other materials in the repair of furcal perforations. REGISTRATION: PROSPERO (CRD42020181297).

Marginal gaps and internal voids after root-end filling using three calcium silicate-based materials: A Micro-CT analysis

Abstract This study evaluated the 3D quality of root-end filling, assessing the presence (volume and percentage) of marginal gaps and internal voids formed after retro-filling with three calcium silicate-based materials: MTA Angelus (Angelus Soluçoes Odontologicas, Londrina, PR, Brazil), Biodentine (Septodont Ltd., Saint Maur-des-Faussés, France) and Neo MTA Plus (Avalon Biomed Inc., Bradenton, Florida, US). Thirty human, extracted, single rooted teeth were used. Orthograde root canal treatment, root resection (3mm shorter than the apex) and retrograde cavity preparation with ultrasonic tips were performed. Teeth were divided into 3 groups (n =10 each) following a stratified randomization according to the initial volume of the root-end cavity. After retrofilling, samples were stored for 7 days. Then, two rounds of micro-CT scans were performed: soon after root-end preparation (with the cavity still empty) and 7 days after root-end filling. Marginal gaps, internal voids volume (mm3 and %), as well as, the overall defects (sum of gaps and voids) were evaluated. Statistics compared the three groups in relation to those defects. There was not statistical difference between groups regarding the marginal gaps (P≥ 0.05), the internal voids (P≥ 0.05), and the overall defects (P≥ 0.05). Median (mm3) and % of overall air-entrapment defects (gaps and/or voids) was: 0.004mm3 and 1.749% for MTA Angelus, 0.018mm3 and 6.660% for Biodentine, and 0.012mm3 and 4.079% for Neo MTA Plus. All materials had gaps and/or voids. No differences were found between MTA Angelus, Biodentine and Neo MTA Plus.

Resumo Este estudo avaliou a qualidade 3D de retro-obturações, avaliando a presença (volume e percentagem) de "gaps" marginais e "voids" internos formados a partir da retro-obturação com três materiais à base de silicato de cálcio: MTA Angelus (Angelus Soluções Odontológicas, Londrina, PR, Brasil), Biodentine (Septodont Ltd., Saint Maur-des-Faussés, França) and Neo MTA Plus (Avalon Biomed Inc., Bradenton, Flórida, US). Trinta dentes humanos unirradiculares extraídos foram usados. Após tratamento de canal ortrógrado, resecção radicular (3 mm apicais), o prepare da cavidade retrógrada com pontas ultrassônicas foi realizado. Os dentes foram divididos em três grupos (n= 10 cada) seguindo uma randomização estratificada, de acordo com o volume inicial da cavidade retrógrada. Depois da retro-obturação as amostras foram armazenadas por 7 dias. Duas rodadas de escaneamento de micro-CT foram realizadas: logo após o preparo retrógrado (com a cavidade ainda vazia) e 7 dias após a retro-obturação. O volume de "gaps" marginais e de "voids" internos (mm3 e %), bem como, os defeitos totais (soma de "gaps" e "voids") foram avaliados. Não houve diferença estatística entre os grupos em relação aos "gaps" marginais (P≥ 0,05), "voids" internos (P≥ 0,05), e aos defeitos totais (P≥ 0,05). A mediana (mm3) e porcentagem dos defeitos totais de aprisionamento de ar foi 0,004mm3 e 1,749% para o MTA Angelus, 0,018mm3 e 6,660% para o Biodentine, e 0,012mm3 e 4,079% para o Neo MTA Plus. Todos os materiais tiveram defeitos ("gaps"e/ou "voids"). Não houve diferença entre MTA Angelus, Biodentine e Neo MTA Plus.

Barriers and Opportunities of Oncofertility Practice in Nine Developing Countries and the Emerging Oncofertility Professional Engagement Network.

PURPOSE: Oncofertility practice continues to grow in developing countries despite the lack of health care services, especially those related to cancer care. The purpose of this study is to further explore oncofertility practice in these countries and identify opportunities for field-wide coalescence. METHODS: We generated a survey to learn more about oncofertility practice in nine developing countries within our Oncofertility Consortium Global Partners Network-Mexico, Colombia, Guatemala, Argentina, Chile, Nigeria, South Africa, Saudi Arabia, and India. Their responses were collected, reviewed, and discussed. RESULTS: Surveyed centers from the nine developing countries continue to experience a similar set of common challenges, including a lack of awareness among providers and patients, cultural and religious constraints, lack of insurance coverage and funding to help to support oncofertility programs, and high out-of-pocket costs for patients. Despite these barriers, many opportunities exist and there is great potential for the future. CONCLUSION: The current need is to unify the new technologies and best practices that emerge from rural communities and developing countries with those in large metropolitan cities, both domestically (US based) and abroad, into a functional unit: the Oncofertility Professional Engagement Network. The Oncofertility Professional Engagement Network will bridge the gap between domestic and international programs to establish a strong global network in which members share resources, methodologies and experiences and further build cultural competency.

Barriers and Opportunities of Oncofertility Practice in Nine Developing Countries and the Emerging Oncofertility Professional Engagement Network.

PURPOSE: Oncofertility practice continues to grow in developing countries despite the lack of health care services, especially those related to cancer care. The purpose of this study is to further explore oncofertility practice in these countries and identify opportunities for field-wide coalescence. METHODS: We generated a survey to learn more about oncofertility practice in nine developing countries within our Oncofertility Consortium Global Partners Network-Mexico, Colombia, Guatemala, Argentina, Chile, Nigeria, South Africa, Saudi Arabia, and India. Their responses were collected, reviewed, and discussed. RESULTS: Surveyed centers from the nine developing countries continue to experience a similar set of common challenges, including a lack of awareness among providers and patients, cultural and religious constraints, lack of insurance coverage and funding to help to support oncofertility programs, and high out-of-pocket costs for patients. Despite these barriers, many opportunities exist and there is great potential for the future. CONCLUSION: The current need is to unify the new technologies and best practices that emerge from rural communities and developing countries with those in large metropolitan cities, both domestically (US based) and abroad, into a functional unit: the Oncofertility Professional Engagement Network. The Oncofertility Professional Engagement Network will bridge the gap between domestic and international programs to establish a strong global network in which members share resources, methodologies and experiences and further build cultural competency.

Influence of photobiomodulation therapy on root development of rat molars with open apex and pulp necrosis.

This study aimed to evaluate the role of photobiomodulation (PBM) in apexification and apexogenesis of necrotic rat molars with an open apex. Rat molars were exposed to the oral environment for 3 weeks. Canals were rinsed with 2.5% NaOCl and 17% EDTA, filled with antibiotic paste and sealed. After 7 days, canals were rinsed and divided into six groups (n=6): mineral trioxide aggregate (MTA); blood clot (BC); human dental pulp stem cells (hDPSC); MTA+PBM; BC+PBM; and hDPSC+PBM. In hDPSC groups, a 1% agarose gel scaffold was used. Two groups were not exposed: healthy tooth+PBM (n = 6), healthy tooth (n = 3); and one was exposed throughout the experiment: necrotic tooth (n = 3). In PBM groups, irradiation was performed with aluminum gallium indium phosphide (InGaAlP) diode laser for 30 days within 24-h intervals. After that, the specimens were processed for histological and immunohistochemical analyses. Necrotic tooth showed greater neutrophil infiltrate (p < 0.05). Necrotic tooth, healthy tooth, and healthy tooth+PBM groups showed absence of a thin layer of fibrous condensation in the periapical area. All the other groups stimulated the formation of a thicker layer of fibers (p < 0.05). All groups formed more mineralized tissue than necrotic tooth (p < 0.05). PBM associated with MTA, BC, or hDPSC formed more mineralized tissue (p < 0.05). MTA+PBM induced apexification (p < 0.05). Rabbit polyclonal anti-bone sialoprotein (BSP) antibody confirmed the histological findings of mineralized tissue formation, and hDPSC groups exhibited higher percentage of BSP-positive cells. It can be concluded that PBM improved apexification and favored apexogenesis in necrotic rat molars with an open apex.

Antimicrobial effect of bioceramic cements on multispecies microcosm biofilm: a confocal laser microscopy study.

OBJECTIVES: To assess the viability of multispecies microcosm biofilm after contact with NeoMTA Plus, Biodentine, and MTA Angelus. MATERIALS AND METHODS: Fifty-four human dentin blocks (4 × 5 × 4 mm) were allocated to Hawley retainers, worn by six volunteers for 72 h. The blocks were then individually incubated in BHI broth for 21 days at 37 °C. At the end of experimental time for biofilm growth, the samples were randomly divided into four groups (n = 12): NeoMTA Plus, Biodentine, MTA Angelus, and negative control. The materials were placed in contact with the blocks. All samples were placed in cell-culture plate wells and incubated in BHI broth for 7 days at 37 °C. One sample from each volunteer (n = 6) was analyzed by SEM to describe the biofilm morphology. CLSM was performed to determine the percentage of viable biofilm biovolume. The data were statistically analyzed by one-way ANOVA and Tukey's multiple comparison test (α = 5%). RESULTS: SEM showed biofilm formed by spherical and rod-shaped bacteria surrounded by an extracellular matrix. No material was able to kill all biofilm cells, and all groups had more than 50% of viable bacteria. NeoMTA Plus was significantly different from the negative control group (P < .05). CONCLUSIONS: All tested materials were not effective against multispecies microcosm biofilm. CLINICAL RELEVANCE: NeoMTA Plus, Biodentine, and MTA Angelus were not effective against multispecies microcosm biofilm. It is essential to understand that these bioceramic cements are indicated for infected clinical situations. Thus, complementary disinfection procedures should be conducted prior to filling with these materials.

Bone tissue reaction, setting time, solubility, and pH of root repair materials.

OBJECTIVES: This study aims to compare the bone tissue reaction, setting time, solubility, and pH of NeoMTA Plus, Biodentine (BD), and MTA Angelus (MTA-A). MATERIALS AND METHODS: Initial and final setting times (n = 7) and solubility up to 7 days (n = 11) were evaluated in accordance with ASTM C266-15 and ANSI/ADA Specification No. 57, respectively. pH (n = 10) was measured up to 28 days. Bone tissue reactions in 48 rats' femur were histologically analyzed after 7, 30, and 90 days. ANOVA and Tukey's tests compared setting time, solubility, and pH data; bone reactions data were compared by Kruskal-Wallis and Dunn's tests. RESULTS: NeoMTA Plus had longer initial and final setting times than MTA-A and BD (P < 0.05). At 7 days, BD showed the highest solubility, similar to NeoMTA Plus (P > 0.05) and different from MTA-A (P < 0.05). NeoMTA Plus had a progressive mass loss over time; at 7 days, it was significantly different from the initial mass (P < 0.05). BD showed higher pH in the periods assessed when compared to the other materials (P < 0.05). Bone tissue repair had no differences between groups in each experimental period (P > 0.05). All groups presented no difference from 30 to 90 days (P > 0.05) and had better bone repair at 90 days than at 7 days (P < 0.05). CONCLUSIONS: NeoMTA Plus, BD, and MTA-A showed satisfactory setting time, high mass loss, alkaline pH, and allowed bone repair. CLINICAL RELEVANCE: Calcium silicate-based cements are indicated for multiple clinical situations. NeoMTA Plus and BD showed satisfactory physical-chemical and biological properties, being considered as alternatives to MTA-A, as root repair materials for clinical use.

Influence of photobiomodulation therapy on root development of rat molars with open apex and pulp necrosis

Abstract This study aimed to evaluate the role of photobiomodulation (PBM) in apexification and apexogenesis of necrotic rat molars with an open apex. Rat molars were exposed to the oral environment for 3 weeks. Canals were rinsed with 2.5% NaOCl and 17% EDTA, filled with antibiotic paste and sealed. After 7 days, canals were rinsed and divided into six groups (n=6): mineral trioxide aggregate (MTA); blood clot (BC); human dental pulp stem cells (hDPSC); MTA+PBM; BC+PBM; and hDPSC+PBM. In hDPSC groups, a 1% agarose gel scaffold was used. Two groups were not exposed: healthy tooth+PBM (n = 6), healthy tooth (n = 3); and one was exposed throughout the experiment: necrotic tooth (n = 3). In PBM groups, irradiation was performed with aluminum gallium indium phosphide (InGaAlP) diode laser for 30 days within 24-h intervals. After that, the specimens were processed for histological and immunohistochemical analyses. Necrotic tooth showed greater neutrophil infiltrate (p < 0.05). Necrotic tooth, healthy tooth, and healthy tooth+PBM groups showed absence of a thin layer of fibrous condensation in the periapical area. All the other groups stimulated the formation of a thicker layer of fibers (p < 0.05). All groups formed more mineralized tissue than necrotic tooth (p < 0.05). PBM associated with MTA, BC, or hDPSC formed more mineralized tissue (p < 0.05). MTA+PBM induced apexification (p < 0.05). Rabbit polyclonal anti-bone sialoprotein (BSP) antibody confirmed the histological findings of mineralized tissue formation, and hDPSC groups exhibited higher percentage of BSP-positive cells. It can be concluded that PBM improved apexification and favored apexogenesis in necrotic rat molars with an open apex.

Effect of human, dentin, albumin and lipopolysaccharide on the antibacteerial activity of endodontic activity of endodontic irrigants.

CONTEXT: Human dentin powder (HD), bovine serum albumin (BSA) and endotoxin (LPS) may affect the antimicrobial activity of irrigating solutions. AIM: To evaluate the inhibitory effect of HD powder, BSA, and LPS on the antibacterial activity of 0.5%, 1%, 2.5%, and 5% sodium hypochlorite (NaOCl), 2% chlorhexidine (CHX) gel, BioPure mixture of tetracycline, citric acid, and detergent (MTAD), and QMix. METHODS: The direct contact test against Enterococcus faecalis (ATCC 29212) for 2-min, 30-min and 6-h was used. Sterile pyrogen-free water was the negative control. After experimental periods, a neutralizing agent was used. Colony-forming units were determined by 10-fold serial dilutions and culture on agar plates. Data were analyzed by Kruskal-Wallis and Dunn's test (α = 5%). RESULTS: In the absence of inhibitors, all irrigants eliminated E. faecalis. In contact with HD, all solutions eliminated E. faecalis within 2-min, with the exception of MTAD. In the presence of BSA, only 5% NaOCl killed E. faecalis within 2-min. LPS did not affect the antibacterial effect of any irrigant. At 30-min and at 6-h, all substances eliminated E. faecalis. CONCLUSIONS: In the presence of albumin, irrigants needed >2-min to eliminate E. faecalis, except for 5% NaOCl. The same was observed in the presence of dentin when E. faecalis was exposed to MTAD.

Metformin decreases the incidence of ovarian hyperstimulation syndrome: an experimental study.

BACKGROUND: In assisted reproduction cycles, gonadotropins are administered to obtain a greater number of oocytes. A majority of patients do not have an adverse response; however, approximately 3-6% develop ovarian hyperstimulation syndrome (OHSS). Metformin reduces the risk of OHSS but little is known about the possible effects and mechanisms of action involved. OBJECTIVE: To evaluate whether metformin attenuates some of the ovarian adverse effects caused by OHSS and to study the mechanisms involved. MATERIAL AND METHODS: A rat OHSS model was used to investigate the effects of metformin administration. Ovarian histology and follicle counting were performed in ovarian sections stained with Masson trichrome. Vascular permeability was measured by the release of intravenously injected Evans Blue dye (EB). VEGF levels were measured by commercially immunosorbent assay kit. COX-2 protein expression was evaluated by western blot and NOS levels were analyses by immunohistochemistry. RESULTS: Animals of the OHSS group showed similar physiopathology characteristics to the human syndrome: increased body weight, elevated progesterone and estradiol levels (P<0.001), increased number of corpora lutea (P<0.001), higher ovarian VEGF levels and vascular permeability (P<0.001 and P<0.01); and treatment with metformin prevented this effect (OHSS+M group; P<0.05). The vasoactive factors: COX-2 and NOS were increased in the ovaries of the OHSS group (P<0.05 and P<0.01) and metformin normalized their expression (P<0.05); suggesting that metformin has a role preventing the increased in vascular permeability caused by the syndrome. CONCLUSION: Metformin has a beneficial effect preventing OHSS by reducing the increase in: body weight, circulating progesterone and estradiol and vascular permeability. These effects of metformin are mediated by inhibiting the increased of the vasoactive molecules: VEGF, COX-2 and partially NOS. Molecules that are increased in OHSS and are responsible for a variety of the symptoms related to OHSS.

Reproducción asistida en hombres infectados con el virus de VIH / Assisted reproduction in HIV positive men

Objetivo: Describir un programa de procreación responsable en parejas donde el varón está infectado con el VIH organizado en un centro ambulatorio infectológico de la ciudad de Bs. As. y llevado a cabo por un equipo interdisciplinario conformado por Infectólogos, Bioquímicos, Ginecólogos y Biólogos. Comentar algunas revisiones de la literatura que avalan la evidencia cientifica para realizar estos procedimientos. Métodos: las parejas serodiscordantes son candidatas para este programa si el varón tiene seguimiento infectológico, los estudios de esperma demuestran carga viral (CV) indetectable y DNA proviral de VIH negativo, y, la mujer tiene ADN proviral de VIH negativo en sangre, en el ciclo en el cual se realizará el procedimiento de fertilización asistida. Se realiza en la mujer pesquisa de infecciones connatales. Las muestras de esperma son procesadas con técnicas de lavado, swim up y swim down, y congeladas. Después de cada procedimiento de fertilización asistida la mujer es controlada controlada con estudios serológicos para anticuerpos (VIH ELISA) periódicos. Los recién nacidos son evaluados a través de ADN proviral de VIH en el 1er mes de vida, o, durante los primeros 6 meses de vida a través de anticuerpos (VIH ELISA). Resultadaos: desde 12/2000 y hasta mayo 2009 se incluyeron 165 parejas serodiscordantes. 147 pacientes VIH (+) realizaron estudios de muestras de esperma, 26, tuvieron muestras con estudios microbiológicos positivos (23, CV detectable, y 3, ADN proviral de VIH positivo). En 123 parejas se realizaron: 329 inseminaciones, 55 ICSI y 28 FIV. Se concretron 44 embarazos y nacieron 51 bebés. Todos los estudios realizados a las mujeres y recién nacidos mostraron resultados negativos. Conclusiones: este programa logró resultados satisfactorios en la prevención de infecciones por VIH y connatales.

Objective: to describe the data obtained during the development of an assisted reproduction program for couples where the man is HIV - Infected organized by an ambulatory infectologist center in BsAs city. a multidisciplinary team with infectious disease MD, biochemists, ginecologists and bilogist collaborate in the follow-up of the patients. Methods: couples seeking reproductive counseling were eligible for this programme if: the HIV positive male partner adhered to an infectious disease follow-up, he has undetectable viral load in the fresh sperm sample and is negative for HIV proviral DNA in the swim-up material after sperm washing, the female partner is negative for HIV proviral DNA immediately beforme each assisted reproduction attempt. Procedures comprised intrauterine insemination, intracytoplasmic sperm injection and in vitro fertilization accordin to gynecologist indication. After each procedure, women were tested for indication. After each procedure, women were tested for HIV antibodies periodically. Newborns were tested for HIV proviral DNA at one month of age or HE during the first six months. Results: One hundred and sixty five couples were assessed, semen analyses results were obtained from 147 HIV positive men, 23 had detectable viral load, 3 had positive HIV proviral DNA in swim up. One hundred and twenty three couples undervent AR procedures: 329 intrauterine inseminations, 55 intracytoplasmic sperm injections, and 28 in-vitro fertilizations. Forty four pregnancies resulted in fifty one live Births. All HE and HDP tests performed in women and new-borns were negative. Conclusions: This programme achieved satisfactory results in prevention of HIV and congenital infections.

Inhibition of cyclooxygenase-2 (COX-2) by meloxicam decreases the incidence of ovarian hyperstimulation syndrome in a rat model.

OBJECTIVE: To investigate the effects of selective cyclooxygenase-2 (COX-2) inhibition on the ovarian hyperstimulation syndrome (OHSS) in an experimental model. DESIGN: Controlled laboratory study. SETTING: University-affiliated fertility center. ANIMAL(S): Female Wistar rats. INTERVENTION(S): Female Wistar rats (22 days old) were divided into four groups: group 1 (control group; n = 10) received 0.1 mL of intraperitoneal (IP) saline from days 22-26; group 2 (mild-stimulated group; n = 10) received 10 IU of pregnant mare serum gonadotropin (PMSG) on day 24 and 10 IU of hCG 48 hours later (day 26); group 3 (OHSS group; n = 10) was given 10 IU of PMSG for 4 consecutive days from day 22 and 30 IU hCG on the fifth day to induce OHSS; group 4 was treated the same as group 3, but received 2 muL (15 mg/mL) of meloxicam 2 hours before the PMSG injection for 4 consecutive days, and 2 hours before the hCG injection on the fifth day. All groups were killed on day 26. MAIN OUTCOME MEASURE(S): Number of antral and luteinized follicles, ovarian weight, semiquantitative vascular endothelial growth factor (VEGF) and COX-2 immunohistochemistry. RESULT(S): There were no differences in the ovarian weight between groups 1 and 2. Group 3 showed significantly increased ovarian weight that was suppressed, in group 4, by meloxicam. There was no difference in the number of antral follicles among the four groups. In the mild-stimulated and OHSS groups, the granulosa cells (GC) of preovulatory follicles and the stromal cells showed intense VEGF immunoreactivity. The ovaries from the meloxicam-treated group showed less immunoreactivity than the OHSS group, indicating diminished VEGF expression associated with meloxicam treatment. Group 3 (OHSS group) showed increased COX-2 immunoreactivity that was diminished in the meloxicam-treated group. Meloxicam treatment did not affect the hormone-induced increase in serum E(2) levels seen in OHSS rats. CONCLUSION(S): Our results in a rat model suggest that meloxicam has a beneficial effect on OHSS by reducing the increases in ovarian weight and VEGF expression associated with OHSS. These effects may be mediated by the COX-2 inhibitory capacity of meloxicam.

Significance of ovarian macrophages in the follicular aspirates from ART patients.

PURPOSE: To evaluate the percentages of macrophages present in granulosa cells (GC) cultures from patients with different responses to the hyperstimulation, in relation to the percentages of apoptotic cells (ApC), as well as to the release of cytokines. METHODS: We studied 42 patients: 12 Hyporesponders, (with < or =4 follicles), 15 Normoresponders, (5-14 follicles), and 15 Hyperresponders, (> or =15 follicles). In GC cultures percentages of macrophages and ApC were counted and, in the conditioned media, cytokines were measured. RESULTS: Percentages of macrophages were significantly higher in GC cultures from Hyporesponders compared with Hyperresponders patients. Also, the percentages of ApC cells were the highest in Hyporesponders. On the contrary, cytokines concentrations were the lowest in this group. CONCLUSIONS: The low ovarian response is probably due to the decreased angiogenesis, which in turn produces increased apoptosis and decreased production of cytokines. The increased percentage of macrophages could be related to increased frequency of apoptotic cells.

New observations on endometrial physiology after transcervical injection of methylene blue dye.

We describe the in vivo features of endometrium stained with methylene blue dye and observed via microhysteroscopy, showing the patterns of endometrial glands and superficial cell changes during the midproliferative, periovulatory, and midluteal phases. These preliminary observations have allowed us to identify a series of changes occurring in the different phases of the ovulatory cycle of potential value in reproductive medicine for specific groups of infertile patients.