The participation of tumor residing pericytes in oral squamous cell carcinoma.

Pericytes are perivascular cells related to vessel structure and angiogenesis that can interact with neoplastic cells, interfering with cancer progression and outcomes. This study focused on the characterization of pericytes in oral squamous cell carcinoma (OSCC) using clinical samples and a transgenic mouse model of oral carcinogenesis. Nestin-/NG2+ (type-1) and nestin+/NG2+ (type-2) pericytes were analyzed by direct fluorescence after induction of oral carcinogenesis (4-nitroquinoline-1-oxide). Gene expression of neuron glial antigen-2 (NG2), platelet-derived growth factor receptor beta (PDGFR-ß), and cluster of differentiation 31 (CD31) was examined in human OSCC tissues. The protein expression of von Willebrand factor and NG2 was assessed in oral leukoplakia (i.e., oral potentially malignant disorders) and OSCC samples. Additionally, clinicopathological aspects and survival data were correlated and validated by bioinformatics using The Cancer Genome Atlas (TCGA). Induction of carcinogenesis in mice produced an increase in both NG2+ pericyte subsets. In human OSCC, advanced-stage tumors showed a significant reduction in CD31 mRNA and von Willebrand factor-positive vessels. Low PDGFR-ß expression was related to a shorter disease-free survival time, while NG2 mRNA overexpression was associated with a reduction in overall survival, consistent with the TCGA data. Herein, oral carcinogenesis resulted in an increase in NG2+ pericytes, which negatively affected survival outcomes.

Efficacy of antiseptics and chemomechanical methods for dentin caries lesions: A systematic review with GRADE approach.

Objectives: Selective caries removal aims to remove carious tissue in deep dentin lesions. However, a discussion stands on the value of antiseptics and chemomechanical adjuvant methods to reduce the bacterial load on residual caries lesions. This systematic review has addressed two main clinical questions to compare the antimicrobial efficacy of available methods using (1) antiseptic or (2) chemomechanical agents before restoring dentin carious lesions. Methods: We included randomized and non-randomized controlled trials (RCTs/ NRCTs). We searched eight databases from inception to October 2021. Paired reviewers independently screened studies, extracted data, and assessed the risk of bias. The primary outcome was the reduction in the number of total bacterial in dentin, whereas secondary outcomes were reduction in the number of Lactobacillus and Streptococcus. We used the ratio of ratio of post-treatment to baseline means between two interventions in the logarithmic scale as a proper effect measure. Certainty of evidence was assessed with the Grading of Recommendations, Assessment, Development and Evaluation approach. Results: We included 14 RCTs and 9 NRCTs, with nine interventions. Regardless the method, the number of bacteria at baseline was similar or exceeded that after the intervention, particularly in NRCTs. The evidence was inconclusive for most comparisons. Among antiseptic agents, chlorhexidine (CHX) resulted in an average of 1.14 times [95% confidence interval (CI): 1.08-1.21] more total bacterial than photodynamic therapy in RCTs. Among NRCTS, the natural agents resulted in five times more total bacterial than CHX (95% CI: 2-11). For chemomechanical methods, the control resulted in eight times (95% CI: 4-17) more total bacterial than Carisolv (SHAA). Conclusions: The certainty of the evidence was very low for all comparisons showing uncertainty whether one treatment could be more effective than another for dentin disinfection. So far, exclusively removing soft carious dentin would be enough to reduce the bacterial count.

Equisetum hyemale-derived unprecedented bioactive composite for hard and soft tissues engineering.

Although Bioactive Glasses (BGs) have been progressively optimized, their preparation often still involves the use of toxic reagents and high calcination temperatures to remove organic solvents. In the present work, these synthesis related drawbacks were overcome by treating the ashes from the Equisetum hyemale plant in an ethanol/water solution to develop a bioactive composite [glass/carbon (BG-Carb)]. The BG-Carb was characterized by scanning electron microscopy, and transmission electron microscopy; and its chemical composition was assessed by inductively coupled plasma-optical emission spectroscopy. Brunauer-Emmett-Teller gas adsorption analysis showed a specific surface area of 121 m2 g-1. The formation of hydroxyapatite (HA) surface layer in vitro was confirmed by Fourier-transform infrared spectroscopy analysis before and after immersion in simulated body fluid (SBF) solution. The Rietveld refinement of the XRD patterns and selected area electron diffraction analyses confirmed HA in the sample even before immersing it in SBF solution. However, stronger evidences of the presence of HA were observed after immersion in SBF solution due to the surface mineralization. The BG-Carb samples showed no cytotoxicity on MC3T3-E1 cells and osteo-differentiation capacity similar to the positive control. Altogether, the BG-Carb material data reveals a promising plant waste-based candidate for hard and soft tissue engineering.

Nestin and Neuron-glial antigen 2 transgenes unveil progenitor units in murine salivary glands.

OBJECTIVE: Undifferentiated cells play pivotal roles in sustaining tissue homeostasis during physiological turnovers and after tissue impairment. Nestin and Neuron-glial antigen 2 (NG2) are markers frequently deployed to distinguish progenitor populations. In the salivary gland scenario, these markers remain largely unknown. Particularly for a double-labeled group of progenitor cells (NG2+Nestin+), their phenotype and distribution have never been explored in freshly isolated tissues. Herein, we analyzed a subset of plastic cells that express Nestin and NG2 near the ducts and in the periacinar region of the major salivary glands of murine samples. DESIGN: The major salivary glands tissues of Nestin-GFP/NG2-DsRed mice were analyzed under a fluorescence microscope. The cells marked by GFP and DsRed were counted in the merged image component of random representative images obtained for each gland sample at × 20 magnification. RESULTS: In the parotid, submandibular, and sublingual glands, the population of cells exclusively expressing Nestin was more abundant. There was a predominance of Nestin, NG2, and double-labeled cells in the submandibular gland compared to the parotid gland, mainly near the ductal system. Of note, the sublingual and parotid glands had similar populations of Nestin+ and NG2+, especially in acini, and some positive cells were observed surrounding ducts. CONCLUSIONS: Collectively, our study revealed differential expression patterns of Nestin and NG2, alone or in combination, in the salivary gland subset during homeostasis.

Photobiomodulation drives pericyte mobilization towards skin regeneration.

Photobiomodulation is being widely applied for improving dermal or mucosal wound healing. However, the underlying cellular and molecular processes that directly contribute to its effects remain poorly understood. Pericytes are relevant cells involved in the wound microenvironment and could be one of the main targets of photobiomodulation due to their plasticity and perivascular localization. Herein, we investigate tissue repair under the photobiomodulation stimulus using a pericyte labeled (or reporter) transgenic mice. Using a model of two contralateral back wounds, one the control and the other photoactivated daily (660 nm, 20 mW, 0.71 W/cm2, 5 J/cm2, 7 s, 0.14 J), we showed an overall influx of immune and undifferentiated cells and higher mobilization of a potent pericyte subpopulation (Type-2 pericytes) in the photoactivated wounds in comparison to the controls. Doppler analysis showed a significant increase in the blood flow in the photoactivated wounds, while marked vascular supply was observed histologically. Histochemical analysis has indicated more advanced stages of tissue repair after photoactivation. These data suggest that photobiomodulation significantly accelerates tissue repair through its vascular effects with direct recruitment of pericytes to the injury site.

Fotoativação mobiliza pericitos e células de origem neural no reparo da polpa dentária / Photoactivation mobilizes pericytes and cells of neural origin in the repair of dental pulp

Introdução: O laser tem se mostrado capaz de fotoativar células-tronco endógenas induzindo sua diferenciação em múltiplos tecidos, além de modular processos inflamatórios e imunes em microambientes injuriados. Essa terapia é hoje chamada de terapia por fotobiomodulação (PBM) ou simplesmente fotoativação. Células da glia e pericitos - células perivasculares -, por sua vez, têm sido sugeridas como as principais células de reparo da polpa dentária. No entanto, os efeitos da PBM sobre essas células nunca foram explorados utilizando técnicas elegantes (state-of-art) de rastreamento, as quais poderiam prover informações relevantes sobre as interações do laser com os componentes celulares marcados. Objetivos: Este estudo visou verificar a capacidade estimuladora da PBM na mobilização de pericitos e células de origem neural no reparo da polpa dentária após injúria tecidual in vivo. Para isso, foi utilizado um modelo murino(NG2-DsRed/Nestin-GFP) com transgene para pericitos (NG2) e células neurais indiferenciadas (Nestina). Métodos: As polpas dentárias dos primeiros molares superiores dos camundongos (n =12) foram expostas utilizando broca #1190F e lima K #20. O grupo PBM (n = 6) foi tratado imediatamente, 24h, 48h e 72h após a injúria com laser de diodo (InGaAlP; 660nm; 20mW; 5J/cm2; 0,71 W/cm2; 7s; modo contínuo e em contato) e o outro grupo foi mantido como controle(n = 6) sem qualquer tratamento. No 4º dia, os animais foram eutanasiados e os efeitos da aplicação da PBM sobre a mobilização dos pericitos e no reparo da polpa dentária foram verificados por microscopia confocal e por análises histológicas (H&E e azul de toluidina). Polpas dentárias saudáveis foram utilizadas como parâmetro de normalidade (n = 6). Os dados foram analisados pelo teste de ANOVA seguido pelo post-hoc de Tukey (¿ = 0,05). Resultados: A terapia promoveu a mobilização significativa de pericitos e células indiferenciadas nos cornos pulpares coronários contíguos à região injuriada em relação ao controle (p<0,05). Além disso a PBM mostrou intensa proliferação de capilares terminais nos dentes fotoativados (p<0,05), enquanto manteve sinais de vitalidade pulpar nos terços coronários adjacentes à injúria. Embora não identificados na polpa dentária em nenhum grupo estudado, mastócitos puderam ser observados intactos ou degranulados nos tecidos orais moles adjacentes ao dente fotoativado (p<0,05). Conclusão: A PBM estimulou a a neoformação da microvasculatura tecidual local e contribuiu para o influxo de células potentes para a região injuriada. A PBM pode ser considerada uma terapia adjuvante promissora em tratamentos endodônticos regenerativos da polpa dentária.

Introduction: Laser light has proven to be capable of photoactivate endogenous stem cells inducing their differentiation into multiple tissues, in addition to modulating inflammatoryand immune processes in injured microenvironments. This therapy is now called photobiomodulation therapy (PBM). Glial cells and pericytes ­ the perivascular cells ­ have been identified as the true stem cells of our body. However, the effects of PBM on these cells have never been explored using elegant (state-of-art) tracking techniques, which could providerelevant information about the laser's interactions with these labeled cellular components. Objectives: The goal of this study is to verify the stimulating capacity of PBM in the mobilization of pericytes and other endogenous cells in the repair of the dental pulp upon tissueinjury in vivo using a murine model (NG2-DsRed/Nestin-GFP) with transgenes for pericytes (NG2) and undifferentiated cells (Nestin). Methods: The dental pulps of the animals' first upper molars (n = 12) were exposed using drill 3195 and K#20 file. The PBM group (n = 6) was treated for 3 consecutive days with a diode laser (InGalP; 660nm; 20mW; 5J/cm2 ; 0.71 W/cm2 ;7s; continuous and in contact) and the other group was maintained as control (n = 6) without any treatment. On the 4 th day, the animals were euthanized and the effects of the application ofPBM on the mobilization of pericytes and on the dental pulp repair were verified by confocal microscopy and histological analysis (H&E and toluidine blue), respectively. Healthy dental pulps were used as a normality parameter (n = 6). The data were analyzed by the ANOVA testfollowed by the Tukey's poshoc (α = 0.05). Results: PBM showed an intense proliferation of terminal capillaries in the photoactivated teeth (p<0.05), while signs of pup vitality were observed in the coronal thirds adjacent to the injury site. In addition, the therapy promoted significant mobilization of pericytes and undifferentiated cells in the coronary pulp horns contiguous to the injury concerning the control group (p<0.05). Mast cells could not be identified in the dental pulp of any of the studied groups but could be seen mostly degranulated in the smooth tissues adjacent to the mesial root of the teeth of the photoactivated group (p<0.05). Conclusion: PBM stimulates the pulp tissue microvasculature neoformation and contributes to the influx of potent cells into the injured site. PBM may be a promise adjunct tool in regenerative endodontic procedures of the pulp tissue