Association between host defence peptide IDR-1002 and ciprofloxacin: Effects on human dental pulp cells.

To evaluate the effects of the association of host defence peptide IDR-1002 and ciprofloxacin on human dental pulp cells (hDPSCs). hDPSCs were stimulated with ciprofloxacin and IDR-1002. Cell viability (by MTT assay), migration capacity (by scratch assay), production of inflammatory and anti-inflammatory mediators by hDPSCs (RT-PCR) and osteogenic differentiation (alizarin red staining) were evaluated. Phenotypic profile of hDPSCs demonstrated 97% for positive marked mesenchymal stem cell. Increased pulp cell migration and proliferation were observed after 24 and 48 h of exposure to IDR-1002 with ciprofloxacin. Mineral matrix formation by hDPSCs was observed of the association while its reduction was observed in the presence of peptide. After 24 h, the association between ciprofloxacin and IDR-1002 significantly downregulated TNFRSF-1, IL-1ß, IL-8, IL-6 and IL-10 gene expression (p ≤ 0.0001). The association between the IDR-1002 and ciprofloxacin showed favourable immunomodulatory potential, emerging as a promising option for pulp revascularisation processes.

Host defense peptides combined with MTA extract increase the repair in dental pulp cells: in vitro and ex vivo study.

Host Defense Peptides (HDPs) have, in previous studies, been demonstrating antimicrobial, anti-inflammatory, and immunomodulatory capacity, important factors in the repair process. Knowing these characteristics, this article aims to evaluate the potential of HDPs IDR1018 and DJK-6 associated with MTA extract in the repair process of human pulp cells. Antibacterial activity of HDPs, MTA and HDPs combined with MTA in Streptococcus mutans planktonic bacteria and antibiofilm activity was evaluated. Cell toxicity was assayed with MTT and cell morphology was observed by scanning electron microscopy (SEM). Proliferation and migration of pulp cells were evaluated by trypan blue and wound healing assay. Inflammatory and mineralization related genes were evaluated by qPCR (IL-6, TNFRSF, DSPP, TGF-ß). Alkaline phosphatase, phosphate quantification and alizarin red staining were also verified. The assays were performed in technical and biological triplicate (n = 9). Results were submitted for the calculation of the mean and standard deviation. Then, normality verification by Kolmogorov Smirnov test, analyzing one-way ANOVA. Analyses were considered at a 95% significance level, with a p-value < 0.05. Our study demonstrated that HDPs combined with MTA were able to reduce biofilms performed in 24 h and biofilm performed over 7 days S. mutans biofilm (p < 0.05). IDR1018 and MTA, as well as their combination, down-regulated IL-6 expression (p < 0.05). Tested materials were not cytotoxic to pulp cells. IDR1018 induced high cell proliferation and combined with MTA induced high cellular migration rates in 48 h (p < 0.05). Furthermore, the combination of IDR1018 and MTA also induced high expression levels of DSPP, ALP activity, and the production of calcification nodules. So, IDR-1018 and its combination with MTA could assist in pulp-dentine complex repair process in vitro.

Concentrated MTA Repair HP reduced biofilm and can cause reparative action at a distance.

AIM: To evaluate in vitro whether MTA Repair HP can induce repair processes at a distance, including its effects on biofilm, cell viability, migration, production of TGF-ß, phosphate and ALP, evaluated through MTA diluted extracts. METHODOLOGY: Initially, antibacterial tests were performed with the bacterium Streptococcus mutans (ATCC 25175) in the presence of MTA extracts (dilutions of 1:1, 1:2 and 1:4). Growth inhibition assay by microdilution in broth, antibiofilm plate assay of young biofilm and antibiofilm assay in confocal microscopy of mature biofilm were carried out. Then, pulp cells were stimulated in the presence of several MTA dilutions, and cell viability (MTT assay), proliferation and migration capacity (scratch assay) were evaluated. To evaluate the capacity of 1:1, 1:2 and 1:4 dilutions of MTA Repair HP to promote the production of important agents of odontogenic differentiation and mineralization, ALP activity, TGF-ß secretion and phosphate quantification were measured. Statistical differences were verified using one-way and two-way anova and Tukey's post-tests. RESULTS: The test dilutions of MTA Repair HP did not inhibit planktonic S. mutans growth but were able to reduce young and mature S. mutans biofilm (p < 0.001). In addition, none of the MTA Repair HP dilutions was cytotoxic for pulp cells. The 1:2 and 1:4 dilutions of MTA Repair HP induced migration and proliferation of pulp cells (p < 0.05). ALP activity and TGF-ß secretion were independent of the tested dilution (p < 0.001). Diluted 1:4 MTA Repair HP produced less phosphate than the more concentrated 1:1 and 1:2 MTA dilutions (p < 0.001). CONCLUSIONS: Undiluted MTA Repair HP reduced S. mutans biofilm, when compared to 1:2 and 1:4 MTA dilutions. Furthermore, none of the tested dilutions was cytotoxic to pulp cells. MTA Repair HP promoted cell migration and proliferation at a distance, assessed through the dilution of the MTA. Even from a distance, MTA Repair HP has the ability to participate in some events related to repair, such as migration, proliferation and TGF production.

Proteomic analysis of human dental pulp in different clinical diagnosis.

OBJECTIVES: The present study aimed to identify proteins obtained from pulp tissue and correlate with each clinical diagnosis (healthy pulp, inflamed pulp, and necrotic pulp). MATERIALS AND METHODS: A total of forty-five molars were used. Three biological replicas were evaluated. Lysis and sonication were used for protein extraction. Protein quantification was assessed by using the Bradford technique, and shotgun proteome analysis was performed by nanoUPLC-MSE using a Synapt G2 mass spectrometer. Mass spectra data were processed using the Waters PLGS software, and protein identification was done using the human Uniprot database appended to the PLGS search engine. RESULTS: A total of 123 different proteins were identified in all evaluated pulp conditions. Among these, 66 proteins were observed for healthy pulp, 66 for inflamed pulp, and 91 for necrotic pulp. Most protein identification was related to immune response, multi-organism process, platelet activation, and stress in inflamed pulp samples compared to healthy pulp. Proteins related to cellular component organization or biogenesis, developmental process, growth, immune response, multi-organism process, response to stimulus, signaling, stress, and transport were identified in cases of apical periodontitis compared to inflamed pulp. CONCLUSIONS: The progression of the disease to inflamed pulp promoted a high abundance of proteins related to the immune system and stress. Comparing the necrotic pulp with inflamed pulp conditions, a high abundance of proteins was noticed related to metabolism, transport, and response between organisms. CLINICAL RELEVANCE: This finding may assist in future studies of new markers, understanding of tissue engineering, and development of future products.

Systemic conditions of diabetic patients diagnosed with apical periodontitis / Condições sistêmicas de pacientes diabéticos diagnosticados com periodontite apical

ABSTRACT Objective: This study aimed to evaluate the association between glycemic control status in type 2 diabetes mellitus (T2DM) patients and apical periodontitis. Methods: Twenty-seven patients were involved in this study. The survey was based on anamnesis, intra and extra oral examination and radiographic evaluation. Diabetes mellitus information involved type of diabetes and blood glucose analysis. Patients were divided according to their metabolic control status (glycemic controlled and poorly controlled T2DM patients). Results: A higher fasting blood glucose level (p = 0.004) and a higher percentage of HbA1c (p = 0.0001) were demonstrated in poorly controlled T2DM patients when compared to glycemic controlled T2DM. However, the frequency of apical periodontitis and the elapsed time since diabetes mellitus diagnosis were higher in controlled T2DM patients, reaching 64%. Nevertheless, controlled T2DM patients presented a higher number of apical periodontitis cases (p < 0.05). Findings support that controlled patients T2DM presented higher presence of apical periodontitis than poorly controlled T2DM ones. In these patients, the time elapsed since the diagnosis was higher, which may have provided a longer period of oscillation and/or uncontrolled metabolism. Conclusions: Therefore, it might contribute to the development and maintenance of apical periodontitis in glycemic controlled patients of this study.

RESUMO Objetivo: Este estudo objetivou avaliar a associação entre o estado de controle glicêmico em pacientes com diabetes mellitus tipo 2 (DM2) e a periodontite apical. Métodos: Vinte e sete pacientes foram envolvidos neste estudo. A pesquisa baseou-se na anamnese, exame intra e extraoral e avaliação radiográfica. As informações sobre o diabetes mellitus envolveram o tipo de diabetes e a análise da glicose sanguínea. Os pacientes foram divididos de acordo com seu estado de controle metabólico (pacientes com DM2 com controle glicêmico e pacientes com DM2 mal controlados). Resultados: Um maior nível de glicose em jejum (p = 0,004) e uma maior porcentagem de HbA1c (p = 0,0001) foram demonstrados em pacientes com DM2 mal controlada quando comparados com DM2 com controle glicêmico. Porém, a frequência de periodontite apical e o tempo decorrido desde o diagnóstico de diabetes mellitus foram maiores nos pacientes com DM2 controlado, chegando a 64%. No entanto, os pacientes com DM2 controlada apresentaram um maior número de casos de periodontite apical (p < 0,05). Os achados suportam que pacientes controlados com DM2 apresentam maior presença de periodontite apical do que pacientes com DM2 mal controlada. Nesses pacientes, o tempo decorrido desde o diagnóstico foi maior, o que pode ter proporcionado um período maior de oscilação e/ou metabolismo descontrolado. Conclusão: Portanto, pode contribuir para o desenvolvimento e manutenção da periodontite apical nos pacientes com controle glicêmico deste estudo.

Percepções de pacientes sobre a relação entre diabetes mellitus e saúde oral: um estudo transversal / Patients perceptions towards diabetes mellitus and oral health: a cross-sectional study

Purpose: To evaluate the knowledge of diabetic patients towards the connection between diabetes mellitus (DM) and oral diseases. Oral status was also assessed in order to evaluate the correlation among patients' perception and their oral health. Material and Methods: A sample of 132 diabetic subjects answered a questionnaire containing 12 questions addressing their knowledge, attitudes and practices related to oral health. Oral examination accessed the presence of cavity carious lesions, residual roots, dental biofilm, calculus, gingivitis, tooth mobility and gingival recession. Results: Fifty-four percent of the sample had never been instructed by their health professionals that DM could cause oral diseases. However, 66% presumed being more vulnerable to develop oral illnesses and 57.5% answered that they assumed having no oral disturbances at that moment. In contrast, intraoral clinical examination showed that 99% presented at least one oral injury such as caries lesions, plaque/calculus, gingival inflammation, tooth mobility, residual root and xerostomia. Conclusions: Health care professionals usually neglect oral status in diabetic patients. Above all, there is a huge gap between patients' perceptions towards oral health and their real oral status. This study highlights the need of developing new models of prevention that properly address the important clinical relation between oral diseases and DM.(AU)