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OBJECTIVES Assessing the evidence and comparing the levels of cytokines in gingival crevicular fluid (GCF) of periodontal healthy sites of smokers and nonsmokers. MATERIALS AND METHODS Seven databases were surveyed for observational studies up to April 8, 2021. Studies comparing cytokine levels on GCF in periodontally healthy sites of smokers vs. nonsmokers were included in the study. The risk of bias was evaluated using NIH (2014) tool. For meta-analyses, levels in GCF were analyzed, followed by evidence certainty assessment using the GRADE approach. RESULTS Eighteen studies were included for qualitative evaluation, and eight were included in meta-analysis. Qualitatively, despite high heterogeneity and risk of bias observed among the studies, most of them presented no significant difference in the gingival crevicular cytokine fluid levels between groups. Regarding meta-analyses, interleukin-8 (IL-8) and superoxide dismutase (SOD) levels in GCF were analyzed. The significant difference was observed only in SOD levels, where heavy smokers had lower levels compared to nonsmokers (MD - 30.06 [- 40.17, - 19.96], p = 0.07, 95%CI), as well as light smokers had lower levels compared to nonsmokers (MD - 15.22 [- 16.05, - 14.39], p

Dental adhesives hydrolyze in the mouth. This study investigated the water sorption (SOR), solubility (SOL) and cytotoxicity (CYTO) of experimental adhesives containing nitrogen-doped titanium dioxide nanoparticles (N_TiO2). Specimens (n = 15/group [SOR, SOL]; n = 10/group [CYTO]) of unaltered Clearfil SE Protect (CSP), OptiBond Solo Plus (OSP), Adper Scotchbond (ASB) and experimental adhesives (OSP + 25% or 30% of N_TiO2) were fabricated, desiccated (37 °C) and tested for SOR and SOL according to ISO Specification 4049 (2009). CYTO specimens were UV-sterilized (8 J/cm2) and monomer extracted in growth medium (1, 3 or 7 days). Human pulp cells were isolated and seeded (0.5 × 104) for MTT assay. SOR and SOL data was analyzed using GLM and SNK (α = 0.05) and CYTO data was analyzed with Kruskal–Wallis and SNK tests (α = 0.05). SOR and SOL values ranged from 25.80 μg/mm3 (30% N_TiO2) to 28.01 μg/mm3 (OSP) and 23.88 μg/mm3 (30% N_TiO2) to 25.39 μg/mm3 (25% N_TiO2). CYTO results indicated that pulp cells exposed to experimental materials displayed comparable viabilities (p > 0.05) to those of OSP. Experimental materials displayed comparable SOR, SOL and CYTO values (p > 0.05) when compared to unaltered materials. N_TiO2 incorporation have not adversely impacted SOR, SOL and CYTO properties of unaltered adhesives.

BACKGROUND This study aimed to evaluate, histomorphometrically, the use of collagen matrix (CM) and/or enamel matrix derivative (EMD) for the treatment of dehiscence-type recession defects in minipigs. METHODS Eight healthy, male, young BR-1 minipigs, with no periodontal disease were treated. Bilateral dehiscence-type defects were surgically created on the buccal of the mandibular premolars (PI and PII). After 30 days, the defects were randomly assigned to four groups: coronally advanced flap (CAF); CAF + CM; CAF + EMD; and CAF + CM + EMD (split-mouth design). The evaluated parameters (mm): total defect length; new cementum (NC); new bone (NB); gingival margin position; total epithelium length; epithelium on the root; connective tissue adaptation; and soft tissue thickness (STT). RESULTS The EMD-treated groups showed a superior length of NC [4.13 ± 1.22 (CAF + EMD); 3.95 ± 1.11 (CAF + CM + EMD); 2.94 ± 0.77 (CAF + CM); 2.72 ± 0.81 (CAF), P = 0.02] and NB [3.21 ± 0.68 (CAF + CM + EMD); 3.01 ± 0.56 (CAF + EMD); 2.15 ± 0.47 (CAF + CM); 2.29 ± 0.82 (CAF), P = 0.005]. The CAF and CAF + CM groups showed a superior epithelial length when compared to EMD-treated groups after 3 months. A superior STT was observed for CAF + CM + EMD group (1.5 ± 0.33) when compared with the other groups [1.09 ± 0.26 (CAF + EMD); 1.04 ± 0.34 (CAF + CM); and 1.14 ± 0.29 (CAF), P = 0.03]. CONCLUSION(S) The results of the present study indicate that EMD application, irrespective of the combination with CM, may improve the periodontal regeneration of dehiscence-type defects in this animal model.

Made available in DSpace on 2022-04-29T08:34:56Z (GMT). No. of bitstreams: 0 Previous issue date: 2022-01-01 Background and objective: Previous studies have demonstrated an association between the IL10 promoter rs6667202 (C > A) single-nucleotide polymorphism (SNP) and grade C, stage 3 or 4 periodontitis (Perio4C) in the Brazilian population, where the altered A allele was detected more frequently in these patients. However, no functional analysis of this variation has yet been performed. Thus, the objective of this preliminary study was to evaluate the functionality of rs6667202 in gingival fibroblasts (GFs) of individuals with Perio4C and with periodontal health (PH) stimulated with Aggregatibacter actinomycetencomitans protein extract (AaPE). Methods: Patients with PH and Perio4C were segregated according to their genotype (AA, AC, or CC), and a biopsy was performed to establish the culture of the GFs. After GFs exposure to AaPE at 5 µg/ml for 1.5 h, RNA was extracted to analyze IL10 expression by qPCR. Aliquots of the cell's supernatant were subjected to immunoenzymatic analysis (MAGpix) to detect interleukin-10 (IL-10). Results: In PH, the genotypes AA and AC are related to less expression of IL10 (p = 0.027 and p 0.05), although a lower IL-10 expression and release compared with PH CC was seen (p = 0.033 and p

Periodontal dental ligament (PDL) is composed of heterogeneous population of mesenchymal progenitor cells. The mechanisms that regulate the differentiation of these cells towards osteoblast/cementoblast phenotype are not fully understood. Some studies have demonstrated that is possible to change the pattern of cell differentiation via epigenetic mechanisms. The proposal of this study was to investigate whether 5-aza-2'-deoxycytidine (5-aza-dC) treatment would stimulate the osteoblast/cementoblast differentiation of periodontal ligament mesenchymal progenitor cells (PDL-CD105

Background Cellular cementum, a mineralized tissue covering apical tooth roots, grows by apposition to maintain the tooth in its occlusal position. We hypothesized that resident cementocytes would show morphological changes in response to cementum apposition, possibly implicating a role in cementum biology. Methods Mandibular first molars were induced to super-erupt (EIA) by extraction of maxillary molars, promoting rapid new cementum formation. Tissue and cell responses were analyzed at 6 and/or 21 days post-procedure (dpp). Results High-resolution micro-computed tomography (micro-CT) and confocal laser scanning microscopy showed increased cellular cementum by 21 dpp. Transmission electron microscopy (TEM) revealed that cementocytes under EIA were 50% larger than control cells, supported by larger pore sizes detected by micro-CT. Cementocytes under EIA displayed ultrastructural changes consistent with increased activity, including increased cytoplasm and nuclear size. We applied EIA to Hyp mutant mice, where cementocytes have perilacunar hypomineralization defects, to test cell and tissue responses in an altered mechanoresponsive milieu. Hyp and WT molars displayed similar super-eruption, with Hyp molars exhibiting 28% increased cellular cementum area versus 22% in WT mice at 21 dpp. Compared to control, Hyp cementocytes featured well-defined, disperse euchromatin and a thick layer of peripherally condensed heterochromatin in nuclei, indicating cellular activity. Immunohistochemistry (IHC) for cementum markers revealed intense dentin matrix protein-1 expression and abnormal osteopontin deposition in Hyp mice. Both WT and Hyp cementocytes expressed gap junction protein, connexin 43. Conclusion This study provides new insights into the EIA model and cementocyte activity in association with new cementum formation.

Made available in DSpace on 2021-06-25T12:41:06Z (GMT). No. of bitstreams: 0 Previous issue date: 2021-03-22 Fundação de Amparo à Pesquisa do Estado de São Paulo (FAPESP) University of Birmingham, United Kingdom Periodontal ligament cells (PDLCs) have well documented osteogenic potential; however, this commitment can be highly heterogenous, limiting their applications in tissue regeneration. In this study, we use PDLC populations characterized by high and low osteogenic potential (h-PDLCs and l-PDLCs, respectively) to identify possible sources of such heterogeneity and to investigate whether the osteogenic differentiation can be enhanced by epigenetic modulation. In h-PDLCs, low basal expression levels of pluripotency markers (NANOG, OCT4), DNA methyltransferases (DNMT1, DNMT3B), and enzymes involved in active DNA demethylation (TET1, TET3) were prerequisite to high osteogenic potential. Furthermore, these genes were downregulated upon early osteogenesis, possibly allowing for the increase in expression of the key osteogenic transcription factors, Runt-related transcription factor 2 (RUNX2) and SP7, and ultimately, mineral nodule formation. l-PDLCs appeared locked in the multipotent state and this was further enhanced upon early osteogenic stimulation, correlating with low RUNX2 expression and impaired mineralization. Further upregulation of DNMTs was also evident, while pretreatment with RG108, the DNMTs' inhibitor, enhanced the osteogenic program in l-PDLCs through downregulation of DNMTs, increased RUNX2 expression and nuclear localization, accelerated expression of osteogenic markers, and increased mineralization. These findings point toward the role of DNMTs and Ten Eleven Translocations (TETs) in osteogenic commitment and support application of epigenetic approaches to modulate biomineralization in PDLCs. Univ Estadual Campinas, Piracicaba Dent Sch, Dept Prosthodont & Periodont, Piracicaba, Brazil Paulista Univ UNIP, Sch Dent, Hlth Sci Inst, Dr Bacelar St 1212, BR-04026002 Sao Paulo, Brazil Paulista Univ UNIP, Program Environm & Expt Pathol, Sao Paulo, Brazil Sao Paulo State Univ, Biosci Inst, Dept Chem & Biochem, Botucatu, SP, Brazil Univ Birmingham, Sch Dent, Inst Canc & Genom Sci, Inst Clin Sci, 5 Mill Pool Way, Birmingham B5 7EG, W Midlands, England Sao Paulo State Univ, Biosci Inst, Dept Chem & Biochem, Botucatu, SP, Brazil FAPESP: 2013/09650-8 FAPESP: 2017/07944-5 University of Birmingham, United Kingdom: 2017/07944-5 FAPESP: 2015/02160-0

Grade C periodontitis in youngers is characterized by a severe form of periodontitis, and IL10 rs6667202 single nucleotide polymorphism (SNP) has been described as an important feature in this disease etiology. Aim: This study aimed to evaluate, in vivo, the functionality of IL10 rs6667202 SNP on IL-10 gingival fluid levels. Methods: Thirty patients with Perio4C were selected, 15 with the IL10 AA genotype (rs6667202) and 15 with AC/CC genotypes. The gingival fluid was collected from two sites with probing depth ≥ 7 mm and bleeding on probing, and two healthy sites. The IL-10 concentration was determined by Luminex/MAGpix platform. Results: In deep pockets, the IL10 AA genotype presented a lower concentration of IL-10 when compared with AC or CC genotypes (p0.05). Conclusion: IL10 rs6667202 SNP decreases the production of IL-10 in crevicular fluid, potentially affecting this disease progression.

Early acquisition of a pathogenic microbiota and the presence of dysbiosis in childhood is associated with susceptibility to and the familial aggregation of periodontitis. This longitudinal interventional case–control study aimed to evaluate the impact of parental periodontal disease on the acquisition of oral pathogens in their offspring. Subgingival plaque and clinical periodontal metrics were collected from 18 parents with a history of generalized aggressive periodontitis and their children (6–12 years of age), and 18 periodontally healthy parents and their parents at baseline and following professional oral prophylaxis. 16S rRNA amplicon sequencing revealed that parents were the primary source of the child's microbiome, affecting their microbial acquisition and diversity. Children of periodontitis parents were preferentially colonized by Filifactor alocis, Porphyromonas gingivalis, Aggregatibacter actinomycetemcomitans, Streptococcus parasanguinis, Fusobacterium nucleatum and several species belonging to the genus Selenomonas even in the absence of periodontitis, and these species controlled inter-bacterial interactions. These pathogens also emerged as robust discriminators of the microbial signatures of children of parents with periodontitis. Plaque control did not modulate this pathogenic pattern, attesting to the microbiome's resistance to change once it has been established. This study highlights the critical role played by parental disease in microbial colonization patterns in their offspring and the early acquisition of periodontitis-related species and underscores the need for greater surveillance and preventive measures in families of periodontitis patients.

Objectives The imbalanced host response in front of a dysbiotic biofilm is one of the major aspects of severe periodontitis, which also presents a strong familial aggregation related to the susceptibility factors transmission within family members. This study hypothesized that aggressive periodontitis (GAgP) patients and their descendants could present a similar trend of a local inflammatory response that is different from healthy controls. Methods Fifteen GAgP subjects and their children and fifteen healthy subjects and their children were clinically assessed, and the concentration of interferon (IFN)-γ, interleukin (IL)-10, IL-17, IL-1β, IL-4, IL-6, IL-8, and tumor necrosis factor (TNF)-α was evaluated in the gingival fluid using the multiplexed bead immunoassay. Results Children from the GAgP group presented lower IL-10 and IFN-γ subgingival concentration than Health children, despite no difference in the clinical parameters. GAgP parents showed a lower IFN-γ, IL-10, and IL-6 than healthy subjects. IL-10/IL-1β and IFN-γ/IL-4 ratios were reduced in GAgP dyads, suggesting a familial trend in the subgingival cytokine's profile. The cytokines correlated to the clinical data and were predictors of probing depth increase. Conclusion GAgP parents and their children presented a similar cytokine profile and an imbalance in the subgingival response characterized by decreased IFN-γ/IL-4 and IL10/IL-1β ratios.

Background Heterogeneous cell populations of osteo/cementoblastic (O/C) or fibroblastic phenotypes constitute the periodontal dental ligament (PDL). A better understanding of these PDL cell subpopulations is essential to propose regenerative approaches based on a sound biological rationale. Objective Our study aimed to clarify the differential transcriptome profile of PDL cells poised to differentiate into the O/C cell lineage. Methodology To characterize periodontal-derived cells with distinct differentiation capacities, single-cell-derived clones were isolated from adult human PDL progenitor cells and their potential to differentiate into osteo/cementoblastic (O/C) phenotype (C-O clones) or fibroblastic phenotype (C-F clones) was assessed in vitro. The transcriptome profile of the clonal cell lines in standard medium cultivation was evaluated using next-generation sequencing technology (RNA-seq). Over 230 differentially expressed genes (DEG) were identified, in which C-O clones showed a higher number of upregulated genes (193) and 42 downregulated genes. Results The upregulated genes were associated with the Cadherin and Wnt signaling pathways as well as annotated biological processes, including "anatomical structure development" and "cell adhesion." Both transcriptome and RT-qPCR showed up-regulation of WNT2, WNT16, and WIF1 in C-O clones. Conclusions This comprehensive transcriptomic assessment of human PDL progenitor cells revealed that expression of transcripts related to the biological process "anatomical structure development," Cadherin signaling, and Wnt signaling can identify PDL cells with a higher potential to commit to the O/C phenotype. A better understanding of these pathways and their function in O/C differentiation will help to improve protocols for periodontal regenerative therapies.

Made available in DSpace on 2021-06-25T12:22:54Z (GMT). No. of bitstreams: 0 Previous issue date: 2020-10-23 Fundação de Amparo à Pesquisa do Estado de São Paulo (FAPESP) Coordenação de Aperfeiçoamento de Pessoal de Nível Superior (CAPES) Background Periodontal disease and diabetes mellitus (DM) are highly prevalent and interrelated diseases, resulting in altered host response microbiota. Thus, this study aimed to evaluate the impact of DM on local levels of lipopolysaccharide (LPS) and lipoteichoic acid (LTA) and their relationship with cytokines and matrix metalloproteinases' (MMPs) profile. Methods This case-control study included diabetic (n = 15) and non-diabetic (n = 15) subjects presenting Stage 3-4, Grade C, Periodontitis. Gingival crevicular fluid (GCF) was collected, and LPS and LTA levels were analyzed by enzyme-linked immunosorbent assay (ELISA), while IFN-gamma, IL-10, IL-17, IL-1 beta, IL-4, MMP-2, and MMP-9 were measured by LUMINEX/MAGpix. Mann-Whitney and Spearman's correlation tests were used to compared and to correlate variables (p < 0.05). Results Higher levels of LTA, LPS, IL-10, IL-1 beta, and MMP-2 (p < 0.05) and lower levels of IL-17 were found in the DM group (p < 0.05). Non-diabetic subjects presented higher LPS, IFN-gamma, IL-17, and MMP-2 levels and lower IL-10 concentration (p < 0.05). No significant correlation was seen between LPS and cytokine profile in non-diabetic. Local levels of LTA were positively correlated with IL-17 and MMP-2 and negatively with IL-10. Conclusion LTA and LPS drove the inflammatory profile through the modulation of cytokines and MMPs in a different manner in DM and non-diabetic subjects. Univ Estadual Campinas, Dent Sch Piracicaba, Dept Prosthodont & Periodont, Campinas, Brazil Sao Paulo State Univ, Div Periodontol, Coll Dent, FOSJC, Sao Paulo, Brazil Sao Paulo State Univ, Div Periodontol, Coll Dent, FOSJC, Sao Paulo, Brazil FAPESP: 2015/502640 FAPESP: 2018/12208-9 CAPES: 001

Objective To evaluate clinical and radiographic characteristics in peri-implant marginal tissues in patients with a history of chronic periodontitis, rehabilitated using tissue-level or bone-level implants. Material and methods Using a split-mouth design, 20 patients with a history of chronic periodontitis were selected and received two different implants, tissue-level group (n = 20) and the bone-level group (n = 20). Peri-implant probing depth, relative peri-implant mucosal margin position, relative peri-implant clinical attachment level, peri-implant plaque index and peri-implant bleeding on probing were evaluated at prosthesis installation, 1, 3, 6, 12 and 24 months after implant loading. Radiographic marginal bone level was evaluated at implant insertion, prosthesis installation, 6 and 24 months after implant loading. Results The mean difference of peri-implant marginal bone resorption from implant installation to 24 months in function was 0.75 ± 1.12 mm for the tissue-level group and 0.70 ± 0.72 mm for the bone-level group. No statistically significant difference was found between groups at all assessment periods for clinical and radiographic peri-implant evaluation. Conclusion Under a rigid supportive therapy, both approaches performed likewise regarding clinical and radiographic parameters for rehabilitation of patients with a history of chronic periodontitis.

BACKGROUND Aggressive periodontitis (AgP) is influenced by genetic factors. Recently, the single nucleotide polymorphisms (SNPs) rs1537415 (GLT6D1), rs6667202 (IL10), and rs1333048 (ANRIL) were associated with AgP in different European populations. However, these specific SNPs have not yet been determined in Brazilians. Therefore, this study investigated whether these SNPs previously associated with AgP could be replicated among Brazilians. METHODS The SNPs rs1537415, rs6667202, and rs1333048 were genotyped using 5'-nuclease allelic discrimination assay in AgP (n = 200), chronic periodontitis (CP, n = 190), and healthy patients (H, n = 196). Differences in allele and genotype frequencies were analyzed using chi-square tests and stepwise logistic regression. RESULTS The minor C allele of rs6667202 was less frequently detected in AgP patients (23.5%) when compared to non-AgP groups (H = 34.2% and CP = 30.3%; p

Periodontal ligament (PDL) is constituted by heterogeneous cell populations with different cell differentiation capacities, here we investigated the effect of an epigenetic modulator on the induction of osteo/cementoblastic differentiation of human periodontal ligament mesenchymal progenitor cell clones with positive surface markers for CD105 (hPDLCD105+) and with low osteo/cementoblastic potential (LOP). These clones were pretreated with 5-aza-2'-deoxycytidine (5-aza-dC) and subjected to mineralization assays (Alizarin Red and Xylenol Orange), transcript levels’ analysis of ALP, ANXA2, ASPN, BMP2, FGF7, OCN, OPN and RUNX2, using qRT-PCR. As results, 5-aza-dC enhances the capacity of mineralized matrix deposition of two LOP clones (L1 and L2), however different results regarding to the quantity of calcium nodules production was found, where L1 mineralization and osteogenic-related gene expression was greater than L2. The data show that the augmentation of BMP2, FGF7 and ANXA2 transcript levels may lead to a better osteo/cementogenic response, while the elevation of ASPN gene expression may block the mineralization performance of LOP clones. These results showed that pre-treatment with 5-aza-dC can positively modulate the osteo/cementogenic differentiation of hPDL-CD105+ cell clones with low osteo/ emento-blastic potential. Those findings contribute to better understand the complex mechanisms that can lead PDL stem cells to regenerate the periodontal tissues.

Tissue engineering is a contemporary field of science, which aims to create conditions based on principles of cell and molecular biology, bioengineering and biomaterials to regenerate tissues. Mesenchymal stem cells present high proliferation rates and are able to differentiate into multilineages under certain conditions, suggesting that they have great potential to act in regeneration field. Tooth derived stem cells are a suitable alternative source of mesenchymal cells once they are easily accessible and have poor morbidity to the donor. Studies showed that they have been isolated and characterized from diverse tissues such as dental pulp, exfoliated deciduous teeth, periodontal ligament, gingiva, dental follicle and apical papilla. However studies show that there is heterogeneity among these populations and there is no standard method to select the most appropriate tooth derived stem cells for regenerative procedures. The aim of this review is to present the current perspective of the multiple types of tooth-derived stem cells and to discuss the basis for their use in periodontal tissue engineering. RESUMO A engenharia de tecidos é um campo contemporâneo da ciência, que visa criar condições baseadas em princípios de biologia celular e molecular, bioengenharia e biomateriais para regenerar tecidos. As células tronco mesenquimais apresentam altas taxas de proliferação e são capazes de se diferenciar, sob certas condições, em multi-linhagens, sugerindo que elas têm grande potencial para atuar no campo da regeneração. As células tronco derivadas de tecidos dentais são uma fonte alternativa adequada de células mesenquimais uma vez que são de fácil acesso e têm baixa morbidade para o doador. Estudos demonstraram que elas já foram isoladas e caracterizadas a partir de diversos tecidos tais como polpa dentária, dentes decíduos esfoliados, ligamento periodontal, gengiva, folículo dental e papila apical. Entretanto, os estudos demonstram que há heterogeneidade entre essas populações e não existe um método padrão para selecionar as células-tronco dentais mais apropriadas para procedimentos regenerativos. O objetivo desta revisão é apresentar o conhecimento atual dos vários tipos de células-tronco derivadas de dentes e discutir as novas perspectivas para seu uso na engenharia de tecidos periodontais.

This study evaluates the transcriptome of healthy gingival tissue from edentulous sites in patients with a history of generalized aggressive periodontitis (GAgP), chronic periodontitis (CP), and in patients with no history of periodontitis (H), using microarray and quantitative reverse transcription polymerase chain reaction (qRT-PCR) analysis.Healthy gingival tissue from edentulous sites was taken from patients from the GAgP (n = 12), CP (n = 12), and H (n = 12) groups. Initially, total RNA from four tissue samples per group was used in transcriptomic microarray analysis. Differential gene expression (fold-change), gene ontology (GO; biologic process), and pathway analyses were performed. Genes that were differentially expressed and showing a significant role on altered pathways were validated by qRT-PCR analysis on 12 samples per group.In total, 270 probe sets and 50 GO groups were differentially expressed (upregulated or downregulated) between GAgP and H. Natural killer cell receptors and other genes related to the immune system were upregulated in GAgP, whereas genes with functions in neural processes and in proliferation/differentiation of keratinocytes were underexpressed. There were 220 probe sets and 75 GO groups that were differentially expressed when comparing CP and GAgP. CP was characterized by increased expression of genes related to responses to external stimuli and an underexpression of immune system-related genes. qRT-PCR analysis confirmed microarray results, that killer cell immunoglobulin (Ig)-like receptor, two Ig domains and long cytoplasmic tail 4; interleukin-6; and selectin E were more highly expressed in patients from the GAgP group compared with the CP and H groups.This study demonstrates differences in the transcriptome of healthy gingival tissue from edentulous sites in patients with GAgP compared with patients from H or CP groups.

To evaluate, in vitro, the effect of two glucose concentrations (0.1 mM and 1.0 mM, simulating glucose concentration in saliva of healthy and diabetic individuals) on Candida albicans biofilm grown on epithelial monolayer.C. albicans was inoculated on epithelial monolayers supplemented with 0.1 mM, 1.0 mM or no glucose. Control groups without C. albicans were also evaluated. Tissue response was assessed through the production of Interleukin-1α, Interleukin-8, Interleukin-6, Interleukin-10 and tumor necrosis factor-α. The complex of monolayer and biofilms were evaluated by quantitative reverse transcription polymerase chain reaction for expression of E-cadherin (CDH1), Caspase-3 (CASP3), β-defensin-1 (DEFB-1) and β-defensin-3 (DEFB-3). The biofilm architecture was visualized by confocal laser scanning microscopy.The production of Interleukin-1α and Interleukin-8 were increased in the presence of C. albicans (p 0.05). Glucose did not interfere in the release of any cytokine evaluated. C. albicans downregulated transcripts for CDH1 (p 0.05). Glucose did not induce a significant change in CDH1, CASP3, DEFB-1 and DEFB-3 messenger RNA expression. The biofilms were more structured in the presence of glucose, but no difference in the diffusion of hyphae through the epithelial cells were observed.The data suggest that glucose concentration does not affect the behavior of C. albicans during tissue invasion and other mechanisms must be related to the greater susceptibility of diabetic individuals to candidiasis.

Células multipotentes do ligamento periodontal de humanos (PDLSCs) podem dar origem ao tecido ósseo e são candidatas promissoras ao reparo periodontal e em terapias regenerativas em doenças ósseas. No entanto, algumas PDLSCs apresentam diferentes potenciais osteogênicos, o que pode estar relacionado à baixa previsibilidade e alta variação em resultados de regeneração óssea. O fenótipo celular é determinado por mecanismos, entre eles destacam-se a regulação de genes linhagem-específica, marcadores de multipotencialidade e mecanismos epigenéticos. Porém, esses mecanismos não estão completamente elucidados e ainda não se sabe as razões biológicas para que PDLSCs apresentem potenciais osteogênicos distintos. Portanto, para a aplicação segura das PDLSCs em ensaios clínicos futuros, é preciso melhor entender a osteogênese nestas células. O objetivo foi produzir resultados que preenchessem lacunas no conhecimento da diferenciação osteogênica, avaliando o impacto da osteogênese em marcadores de multipotencialidade e de linhagem específica, em PDLSCs com potenciais osteogênicos distintos.

BACKGROUND Mesenchymal stem cells differentiate into distinct mesenchymal cell lineages and regulate the immune response. The aim of this study was to determine whether periodontal ligament-derived mesenchymal stem cells (PDLSCs) have the ability to modulate neutrophil responses via paracrine mechanisms. METHODS CD105-enriched PDLSCs were seeded for 24 h and challenged with Porphyromonas gingivalis total protein extract (PgPE) (0 or 2 ug/mL) for 3 h. Cells were then washed and further cultured for 18 h and the supernatants were collected and stored. Next, neutrophil-differentiated human promyelocytic leukemia HL-60 cells (HL60D) were treated with PDLSCs supernatants and HL-60D activation and functional responses were determined. RESULTS PgPE treatment induced higher secretion of inflammatory markers and chemokines by PDLSCs, including RANTES, eotaxin, interferon (IFN)-γ- inducible protein 10 (IP-10), monocyte chemoattractant protein-1 (MCP-1), IFN-γ, interleukin (IL)-6, IL-8 and IL-1ra (P

Studies have demonstrated that children from aggressive periodontitis (AgP) parents presented precocious alterations in their periodontal condition, and the use of chemical agents in association to plaque control could be useful to control these alterations. This study aimed to evaluate the effect of Triclosan toothpaste to modulate the clinical and subgingival condition in children from AgP parents. Fifteen children from AgP parents and 15 from periodontally healthy parents were included in this crossover placebo study. Children were randomly allocated into triclosan or placebo therapy, using selected toothpaste for 45 days. After 15 days of wash-out, groups were crossed, changing the used toothpaste. Clinical examination and saliva, crevicular gingival fluid (GCF), and subgingival biofilm collection were performed at baseline and 45 days of each phase. GCF cytokines’ levels were analyzed by Luminex/MAGpix platform and subgingival and salivary periodontal pathogens’ levels by qPCR. At baseline, AgP group presented higher plaque index (PI), gingival index (GI), and bleeding on probing (BoP), higher Aggregatibacter actinomycetemcomitans (Aa) abundance in saliva and subgingival biofilm, and lower levels of INF-ɣ, IL-4, and IL-17 in GCF. Placebo therapy only reduced PI in both groups. Triclosan toothpaste reduced PI and GI in both groups. Triclosan promoted reduction of BoP and probing depth (PD), Aa salivary, and IL-1β levels in AgP group. In health group, triclosan reduced INF-ɣ and IL-4 concentration. Triclosan toothpaste demonstrated to be more effective than placebo toothpaste to control the periodontal condition in children from AgP parents, by reducing the BoP, PD, salivary Aa, and IL-1β. Triclosan toothpaste can improve oral conditions in higher-risk population for AgP. This study was registered at ClinicalTrials.gov with the identifier NCT03642353.

Cementum is the mineralized tissue covering the tooth root that functions in tooth attachment and post-eruptive adjustment of tooth position. It has been reported to be highly similar to bone in several respects but remains poorly understood in terms of development and regeneration. Here, we investigate whether cementocytes, the residing cells in cellular cementum, have the potential to be protagonist in cementum homeostasis, responding to endocrine signals and directing local cementum metabolism. Cells from healthy erupted human teeth were isolated using sequential collagenase/EDTA digestions, and maintained in standard cell culture conditions. A cementocyte-like cell line was cloned (HCY-23, for human cementocyte clone 23), which presented a cementocyte compatible gene expression signature, including the expression of dentin matrix protein 1 ( DMP1 ), sclerostin ( SOST ), and E11/gp38/podoplanin ( E11 ). In contrast, these cells did not express the odontoblast/dentin marker dentin sialoprotein ( DSPP ). HCY-23 cells produced mineral-like nodules in vitro under differentiation conditions, and were highly responsive to inorganic phosphate (Pi). Within the limits of the present study, it can be concluded that cementocytes are phosphate-responsive cells, and have the potential do play a key role in periodontal homeostasis and regeneration.

Periodontal ligament (PDL) has been reported to be a source of mesenchymal stem cells (MSCs).New vascular networks from undifferentiated cells are essential for repair/regeneration of specialized tissues, including PDL. The current study aims to determine potential of CD105(+)-enriched cell subsets of periodontal ligament cells (PDLSCs) to differentiate into endothelial cell (EC)-like cells and to give insights into the mechanism involved.CD105(+)-enriched PDLSCs were induced to EC differentiation by endothelial growth medium 2 (EGM-2) for 3, 7, 14, and 21 days, with mRNA/protein levels and functional activity assessed by: 1) real-time polymerase chain reaction; 2) Western blotting; 3) fluorescence-activated cell sorting; 4) immunohistochemistry; 5) immunofluorescence; 6) matrigel; and 7) small interfering RNA assays.Data analyses demonstrated that EGM-2 treated PDLSCs presented increased expression of EC markers, including: 1) CD105; 2) kinase domain-containing receptor; and 3) Ulex europaeus agglutinin 1, and were able to form cord/tube-like structures. Gene and protein expression analysis showed that neuropilin 2 (NRP2), a key factor for vascular development, was significantly downregulated during EC differentiation. NRP2 was constitutively expressed in mouse PDL tissues by immunohistochemistry analysis, and NRP2 knockdown in CD105(+)-enriched PDLSCs resulted in increased cord/tube-like structures in a matrigel assay.These findings demonstrated the potential of CD105(+)-enriched PDLSCs to support angiogenesis, and NRP2 as a pivotal factor regulating this process.

Aim: This study assessed the prevalence of periodontal disease in the adult and elderly populations from Jundiaí City, and its association with individual social inequalities in a conceptual framework approach. Methods: The survey was conducted with a sample of 342 adults and 145 elderly, and periodontal disease was assessed based on the Community Periodontal Index (CPI) and Clinical Attachment Loss (CAL). A questionnaire addressing socio-demographic and behavioral variables, smoking and diabetes was included. Bivariate and multivariate analyses, using binary regression analyses, were carried out in a hierarchical approach with conceptual framework to reveal association among periodontal disease and social-demographic, smoking and diabetes variables. Results: One adult and fifty-six elderly who had lost all teeth were excluded from the sample. Mild periodontal disease (CAL ≤3 mm) was the condition more prevalent in the adult (74%) and elderly populations (60.6%). Adjusted analysis revealed that low educational level (OR 2.65, 95% CI 1.19-5.88), irregular use of tooth floss (OR 1.9, 95% CI 1.06-3.40), and smoking (OR 2.14, 95% CI 1.04-4.42) were independently associated with moderate/severe periodontal disease (CAL and Probing Depth ≥4 mm) in the adult group. For the elderly group, low educational levels (OR 0.16, 95% CI 0.04-0.58), use of public dental service (OR 5.32, 95% CI 1.23-23.03), and diabetes condition (OR 3.78, 95% CI 1.20-11.91) were significantly associated with periodontal disease. Conclusion: In conclusion, the data showed that education level, smoking habits, diabetes, use of dental floss and type of dental service are factors associated to moderate/severe periodontal disease among Brazilians from Jundiaí City.

Aim Generalized aggressive periodontitis (GAP) is a severe and multifactorial disease in which a familial aggregation and a specific microbiological profile have been suggested. Thus, this case–control study evaluated the clinical and subgingival microbial profile of GAP subjects and their families compared to healthy families. Methods Fifteen families with parents presenting periodontal health and 15 with parents with a history of GAP were selected. Each family should have at least one child between 6 and 12 years old. Plaque index (PI), gingival index (GI), and periodontal probing depth (PPD), as well as Porphyromonas gingivalis, Tannerella forsythia, and Aggregatibacter actinomycetemcomitans (Aa) amounts (by qPCR), were assessed from all subjects. Results Children of GAP families showed a higher PI, GI, and PPD when compared to children of healthy families (p ≤ 0.05). A higher frequency of detection and amounts of Aa was observed in GAP children compared to children of healthy families (p ≤ 0.05). Moreover, a significant association between Aa amounts and gingival bleeding was observed in children (p ≤ 0.05, r = 0.37). Conclusion Children from GAP families have worst clinical conditions, i.e. higher levels of PI, GI, and PPD, a more pathogenic microbiological profile, and the amount of Aa are associated with a higher marginal inflammation.

To evaluate the treatment of gingival recessions by semilunar coronally positioned flap plus enamel matrix derivative (SCPF + EMD).Thirty patients with class I localized gingival recession were included. They were randomly allocated in two groups: SCPF + EMD and SCPF. Recession height (RH), recession width (RW), width of keratinized tissue (WKT), thickness of keratinized tissue (TKT), probing depth (PD), and clinical attachment level (CAL) were measured at baseline, 6 and 12 months post-surgery. Patient/professional evaluation of esthetics and root sensitivity was performed.After 12 months, mean root coverage was 1.98 ± 0.33 mm for SCPF + EMD (90.86 ± 14.69%) and 1.85 ± 0.41 mm (79.76 ± 17.44%) for SCPF (p 0.05). The esthetic evaluation by the patient showed preference for SCPF + EMD. According to the professional evaluation (QCE), the use of EMD decreases the appearance of postoperative scar tissue line. There was a significant reduction in root hypersensitivity with no further complaints by the patients.The addition of EMD provides significantly better esthetics to SCPF, according to patient and professional assessments. SCPF + EMD is effective but not superior to SCPF for root coverage, after 12 months.Previous clinical trials showed that the combination of EMD with coronally advanced flaps may enhance the outcome of root coverage. There is a lack of studies testing the combination of EMD with SCPF. The combination SCPF + EMD provides better esthetics when compared to the SCPF and is effective, but not superior, to SCPF for root coverage, after 12 months.NCT02459704.

Made available in DSpace on 2022-04-28T19:07:09Z (GMT). No. of bitstreams: 0 Previous issue date: 2017-12-01 Background: Gingival recession (GR) might be associated with patient discomfort due to cervical dentin hypersensitivity (CDH) and esthetic dissatisfaction. The aim is to evaluate the effect of root coverage procedure with a xenogenous collagen matrix (CM) and/or enamel matrix derivative (EMD) in combination with a coronally advanced flap (CAF) on CDH, esthetics, and oral health-related quality of life (OHRQoL) of patients with GR. Methods: Sixty-eight participants with single Miller Class I/II GRs were treated with CAF (n = 17), CAF + CM (n = 17), CAF + EMD (n = 17), and CAF + CM + EMD (n = 17). CDH was assessed by evaporative stimuli using a visual analog scale (VAS) and a Schiff scale. Esthetics outcome was assessed with VAS and the Questionnaire of Oral Esthetic Satisfaction. Oral Health Impact Profile-14 (OHIP-14) questionnaire was used to assess OHRQoL. All parameters were evaluated at baseline and after 6 months. Results: Intragroup analysis showed statistically significant reduction in CDH and esthetic dissatisfaction with no intergroup significant differences (P >0.05). The impact of oral health on QoL after 6 months was significant for CAF + CM, CAF + EMD, and CAF + CM + EMD (P 0.05). Conclusion: Root coverage procedures improve patient OHRQoL by impacting on a wide range of dimensions, perceived after reduction of CDH and esthetic dissatisfaction of patients with GRs treated with CAF + CM, CAF + EMD, and CAF + CM + EMD. Department of Prosthesis and Periodontology Division of Periodontics Piracicaba Dental School State University of Campinas, Avenida Limeira, 901-Areião, PO Box 52 Department of Diagnosis and Surgery Division of Periodontics State University of São Paulo Campus

Porphyromonas gingivalis (Pg) is a major periodontal pathogen that contains immunostimulatory components. Periodontal ligament mesenchymal stem cells (PDLMSCs) are responsible for regeneration of the periodontium that is lost due to periodontitis. Pathologic factors within the microenvironment that impair resident PDLMSCs are not well understood. The present study investigates in vitro the effects of Pg protein extract (PgPE) on biologic properties of CD105-enriched PDL progenitor cell populations (PDL-CD105Five populations of PDL-CD105PgPE treatment (2 μg/mL) did not affect cell viability or survival but induced a significant increase in IL-1β, TNF-α, and IL-6 messenger RNA (mRNA) expression and positive staining for IL-6. A total of 29 genes from the IL-6/STAT3 pathway were upregulated on PgPE stimulation. These genes are related to biologic processes involved in the control of cell survival (B-cell lymphoma 2 [BCL2]), cell proliferation (hepatocytehepatocyte growth factor), cytokine-mediated signaling pathway (suppressor of cytokine signaling 3, C-X-C ligand 8 [CXCL8]), and response to stress (CXCL8, mitogen-activated protein kinase 3, BCL2-associated X protein, and BCL2). Additionally, PgPE treatment caused an increase in alkaline phosphatase mRNA expression in PDL-CD105These results suggest that the inflammatory profile induced by PgPE treatment in PDL-CD105

Made available in DSpace on 2018-11-26T17:12:14Z (GMT). No. of bitstreams: 0 Previous issue date: 2016-12-01 Department of Prosthodontics and Periodontics, Division of Periodontics, Piracicaba Dental School, State University of Campinas-UNICAMP The objective of this study is to clinically evaluate the outcomes following treatment of single gingival recessions with either coronally advanced flap technique (CAF) alone or combined with a porcine collagen matrix graft (CM). This is a randomized parallel design clinical trial, including forty patients with single Miller Class I or II gingival recession, with a depth ae 2 mm and located at upper canines or premolars. The patients were randomly assigned to receive either CAF or CAF + CM. The primary outcome variable was gingival recession reduction (Rec Red). Baseline recession depth was 3.14 +/- A 0.51 mm for CAF group and 3.16 +/- A 0.65 mm for CAF + CM group (p > 0.05). Both groups showed significant Rec Red (p < 0.05), up to 6 months. Rec Red for CAF + CM was 2.41 +/- A 0.73 mm and was 2.25 +/- A 0.50 mm for CAF alone (p > 0.05). Root coverage was 77.2 % in the CAF + CM group and 72.1 % in the CAF group (p > 0.05). Complete root coverage (CRC) was found in 40 % of the cases in the CAF + CM group and in 35 % of the sites treated with CAF. Keratinized tissue thickness (KTT) was 0.26 mm higher in CAF + CM group (p < 0.05). It can be concluded that CAF + CM does not provide a superior recession reduction when compared to CAF; however, it may offer a small gain in KTT after 6 months. CAF + CM can be suggested as a valid therapeutic option to achieve root coverage and some increase in soft tissue thickness after 6 months. State Univ Campinas UNICAMP, Dept Prosthodont & Periodont, Div Periodont, Piracicaba Dent Sch, Ave Limeira 901, Sao Paulo, Brazil State Univ Sao Paulo, Dept Periodontol, Sao Jose Campos Dent Sch, Sao Paulo, Brazil Univ Bern, Dept Periodontol, Bern, Switzerland State Univ Sao Paulo, Dept Periodontol, Sao Jose Campos Dent Sch, Sao Paulo, Brazil

Made available in DSpace on 2019-10-04T12:33:03Z (GMT). No. of bitstreams: 0 Previous issue date: 2018-12-03 Fundação de Amparo à Pesquisa do Estado de São Paulo (FAPESP) FP7 Framework Marie Curie Actions Career Integration Grant (methDRE) Human bone marrow-derived mesenchymal stem cells (hBMSCs) are important for tissue regeneration but their epigenetic regulation is not well understood. Here we investigate the ability of a non-nucleoside DNA methylation inhibitor, RG108 to induce epigenetic changes at both global and gene-specific levels in order to enhance mesenchymal cell markers, in hBMSCs. hBMSCs were treated with complete culture medium, 50 mu M RG108 and DMSO for three days and subjected to viability and apoptosis assays, global and gene-specific methylation/hydroxymethylation, transcript levels' analysis of epigenetic machinery enzymes and multipotency markers, protein activities of DNMTs and TETs, immunofluorescence staining and western blot analysis for NANOG and OCT4 and flow cytometry for CD105. The RG108, when used at 50 mu M, did not affect the viability, apoptosis and proliferation rates of hBMSCs or hydroxymethylation global levels while leading to 75% decrease in DNMTs activity and 42% loss of global DNA methylation levels. In addition, DNMT1 was significantly downregulated while TET1 was upregulated, potentially contributing to the substantial loss of methylation observed. Most importantly, the mesenchymal cell markers CD105, NANOG and OCT4 were upregulated being NANOG and OCT4 epigenetically modulated by RG108, at their gene promoters. We propose that RG108 could be used for epigenetic modulation, promoting epigenetic activation of NANOG and OCT4, without affecting the viability of hBMSCs. DMSO can be considered a modulator of epigenetic machinery enzymes, although with milder effect compared to RG108. Univ Estadual Campinas, Piracicaba Dent Sch, Dept Prosthodont & Periodont, Piracicaba, SP, Brazil Univ Birmingham, Sch Canc Sci, Sch Dent, Birmingham, W Midlands, England Sao Paulo State Univ, Biosci Inst, Dept Chem & Biochem, Botucatu, SP, Brazil Univ Paulista, Sch Dent, Div Epigenet, Hlth Sci Inst, Sao Paulo, Brazil Sao Paulo State Univ, Biosci Inst, Dept Chem & Biochem, Botucatu, SP, Brazil FAPESP: 2013/09650-8 FAPESP: 2015/02160-0

Generalized aggressive periodontitis (GAP) is a multifactorial disease that shows a specific microbial profile and a familial aggregation.This study evaluated the salivary microbial profile of families with a history of GAP and compared them with healthy families.Fifteen families with parents presenting periodontal health and 15 with parents with a history of GAP were selected. Each family had a child aged 6-12 years. Stimulated saliva was collected from all subjects, and Porphyromonas gingivalis (Pg), Tannerella forsythia (Tf), and Aggregatibacter actinomycetemcomitans (Aa) amounts were determined.Children of GAP families showed higher detection of Aa (90%) than children of healthy families (45%) (P0.05). Parents with GAP showed a Pg salivary concentration statistically higher than that of healthy parents (P0.05).Children of GAP families, however, exhibited similar Pg concentration than healthy children (P0.05). Tf amounts did not differ either in parents or in children (P0.05) The infection risk calculation indicates that children who have one parent who is positive for Aa have 16.3 times (95% CI 3.1-87.2) more risk of being infected with Aa (P0.05) than children from an Aa-negative family.It may be concluded that children of parents with aggressive periodontitis have higher levels and higher risk of Aa infection.

To clinically evaluate proximal furcations treated with hydroxyapatite/β-tricalcium phosphate (HA/β-TCP) isolated or combined with enamel matrix derivative (EMD).Thirty patients, presenting at least one proximal class II furcation defect, probing pocket depth (PPD) ≥5 mm and bleeding on probing, were included. The defects were assigned to the HA/β-TCP group (n = 15); open-flap debridement (OFD) + HA/β-TCP filling, or, HA/β-TCP-EMD group (n = 15); OFD + HA/β-TCP + EMD filling. Plaque (PI) and gingival index (GI), PPD, relative gingival margin position (RGMP), vertical and horizontal attachment level (RVAL and RHAL), vertical and horizontal bone level (RVBL and RHBL), and furcation diagnosis were evaluated at baseline and at 6 months.Both groups presented improvements after therapies (p 0.05); however, no inter-group differences could be seen in any single parameter (p 0.05). At 6 months, the gains in rVCAL in the HA/β-TCP and HA/β-TCP-EMD groups were 1.47 ± 0.99 and 2.10 ± 0.87 mm, while the RHCAL gains were 1.47 ± 1.46 and 1.57 ± 1.58 mm (p 0.05). The RVBL and RHBL gains for the HA/β-TCP and HA/β-TCP-EMD group were 1.47 ± 1.13 and 1.70 ± 1.26 mm, and 1.90 ± 1.11 and 1.70 ± 1.37 mm respectively (p 0.05). The HA/β-TCP-EMD group showed seven closed furcations versus four in the HA/β-TCP group (p 0.05).Both treatments lead to improvements in all clinical variables studied in the present trial. However, the closure of proximal class II furcation defects is still unpredictable.