The Effect of Photobiomodulation on Nitrite and Inflammatory Activity in Radiotherapy-Induced Oral Mucositis: A Randomized Clinical Trial.

BACKGROUND AND OBJECTIVES: To investigate pro- and anti-inflammatory cytokines and nitrite salivary levels in patients with head and neck cancer receiving photobiomodulation therapy (PBMT) associated with a Preventive Oral Care Program (POCP), for prevention and control of oral mucositis (OM) during radiotherapy (RT) associated or not with chemotherapy protocol. STUDY DESIGN/MATERIALS AND METHODS: In this randomized double-blinded clinical trial, 48 patients were randomly assigned to two groups: PBMT (n = 25) and Control (n = 23). In the PBMT group, patients were submitted to PBMT associated with the POCP. In the Control group, patients were submitted only to the POCP. Saliva samples were collected in the 1st (baseline), 7th, 14th, 21st, and 30th sessions of RT, and the levels of interleukin-6 (IL-6), IL-8, IL-10, IL-12p70, IL-1ß, and tumoral necrosis factor-α (TNF-α) were measured using the cytometric bead array. Nitrite levels were measured by colorimetric method. OM was assessed using the World Health Organization and the National Cancer Institute scales. RESULTS: Patients in the PBMT group presented less severe OM. PBMT tended to stabilize nitrite concentration levels during the RT regimen. The IL-1ß concentration was associated with higher OM scores. PBMT promoted an increase in IL-12p70, TNF-α, and IL-10 concentration. CONCLUSION: PBMT was effective in the prevention and control of severe OM, and its mechanism of action may be related to a better balance of inflammatory response that may favor injury control. Lasers Surg. Med. © 2020 Wiley Periodicals LLC.

Calcifying epithelial odontogenic tumours: Collaborative study of 32 cases and review of literature.

OBJECTIVES: To analyse the occurrence of calcifying epithelial odontogenic tumours (CEOT) based on biopsy records from different Brazilian geographic regions and to contrast the data with a review of the literature. MATERIALS AND METHODS: A 2-step study was conducted. Step 1 consisted of a collaborative study of biopsies obtained from 1953 to 2017 at six Brazilian oral and maxillofacial pathology centres. Evaluation of 86,268 biopsy records was performed. Demographic and histopathological diagnosis data were assessed. In Step 2, a review of the literature of case reports and cases series of CEOT identified across five electronic databases was conducted. RESULTS: In the collaborative study, 32 cases of CEOT were evaluated. This figure represented 0.03% of the oral and maxillofacial lesions and 1.7% of all odontogenic tumours across the centres. Women in the fourth decade of life were more affected. CEOT occurred more in the mandible than in the maxilla (ratio 1.9:1). The review of the literature showed that Asian individuals were more affected by this neoplasm. CONCLUSIONS: Useful knowledge on the epidemiology, treatment and follow-up of CEOT has been provided. Demographic data and clinical features of the cases presented in this collaborative study were quite similar to those of studies reported worldwide.

A multicentre study of 268 cases of calcifying odontogenic cysts and a literature review.

OBJECTIVES: To investigate the frequency of calcifying odontogenic cysts (COCs) that have been submitted for microscopic examination from representative geographic regions of Brazil and to compare it with literature data. MATERIALS AND METHODS: A retrospective study was conducted on biopsies obtained from 1953 to 2016 at 10 Brazilian oral and maxillofacial pathology centres. A total of 198,350 biopsy specimens were analysed. Demographic data and histopathological diagnosis were evaluated descriptively and statistically. In addition, a literature review of case series was carried out in four electronic databases. RESULTS: A total of 268 cases of COC were surveyed, representing 0.1% of the oral lesions at the centres studied. Female patients in their second decade of life and the maxilla were more affected. The mean lesion size of symptomatic individuals was larger than that of cases without symptoms (p = 0.026). The literature review showed a higher frequency in Asia and Europe, mainly affecting men in the third decade of life. CONCLUSIONS: COC is a rare lesion. Novel data on the clinicopathological features of 268 cases have been added to the literature. Data regarding gender and age of the Brazilian patients reported herein contrast with findings of case series and retrospective studies reported elsewhere.

Effect of low-level laser therapy on chemoradiotherapy-induced oral mucositis and salivary inflammatory mediators in head and neck cancer patients.

BACKGROUND AND OBJECTIVE: Oral mucositis (OM) is considered a painful and debilitating side effect in patients receiving head and neck cancer treatment. Low-level laser therapy (LLLT) proved to be effective to prevent and treat chemoradiotherapy-induced OM. The aim of this study was to evaluate the effect of LLLT in the severity of OM in patients with head and neck cancer and on the release of salivary inflammatory mediators. Clinical (score of OM severity) and biochemical parameters (concentration of inflammatory mediators, growth factors, and enzymes in saliva) were used. MATERIALS AND METHODS: Thirty patients were randomized into two groups: control and laser. LLLT was performed three times a week in the laser group, while control group received sham irradiation. OM severity was assessed according to the World Health Organization (WHO) and National Cancer Institute (NCI) scales. Pro-inflammatory and anti-inflammatory cytokines (TNF-α, IL-6, IL-1ß, IL-10, TGF-ß), growth factors (EGF, FGF, VEGF), and metalloproteinases (MMP2/TIMP2, MMP9/TIMP2) concentrations were assessed using ELISA test. Saliva samples were collected on admission, and at the 7th, 21st, and 35th sessions of radiotherapy. RESULTS: The laser group showed a reduction in the severity of OM, which coursed with significantly diminished salivary concentration of EGF and VEGF in the 7th radiotherapy session and of IL-6 and FGF in the 35th. There was a trend for reduced levels of IL-1ß, TNF-α, IL-10, TGF-ß, MMP2/TIMP2, MMP9/TIMP2 in the laser group compared to the control, however, no statistically significant differences were found. CONCLUSIONS: These findings demonstrated that LLLT was effective in reducing the severity of chemoradiotherapy-induced OM and was associated with the reduction of inflammation and repair.

Macrophage migration inhibitory factor and oral cancer.

BACKGROUND: Macrophage migration inhibitory factor (MIF) is a pleiotropic cytokine with pro-inflammatory functions and involved in tumorigenesis. The aim of this study was to evaluate the expression and localization of the macrophage MIF in oral squamous carcinoma (OSC). In addition, the relationship between MIF expression and clinicopathological parameters such as survival data, tobacco use, alcohol habits, TNM stage, tumor graduation, and peritumoral inflammatory infiltrate were evaluated. METHODS: Using immunohistochemistry, expression and localization of MIF was detected in 44 specimens of OSC. The absolute number and relative proportions of MIF-positive cells detected were also determined separately for tumor parenchyma vs. stroma. All counts were determined from 10 consecutive high-power fields using an integration graticule. Moreover, some parameters were analyzed separately for lip and intra-oral cancers. RESULTS: Migration inhibitory factor-positive cells were observed in both the tumor parenchyma and in inflammatory cells of all specimens. In contrast, MIF expression was not detected in tumoral nests associated with poorly differentiated tumors. In specimens of lip cancer, a greater number of MIF-positive stromal immune cells were detected than in intra-oral cancer specimens (Mann-Whitney test, P = 0.049). CONCLUSIONS: Oral squamous carcinoma cells consistently express MIF independent of their location. Lip tumors presented more MIF-positive peritumoral inflammatory cells, similar to control, suggesting that immunological differences in leukocyte activation exist between in lip and intra-oral cancers.

Oral lymphatic malformations: A multicenter study of 208 cases and literature review.

BACKGROUND: To evaluate the frequency and analyze demographic and clinical characteristics of individuals with a histopathological diagnosis of oral lymphatic malformations (OLMs). METHODS: A multicenter study was performed, collecting biopsy record data from a consortium of Brazilian Oral and Maxillofacial Pathology Centers. A review was also conducted to compare this data with cases already available in the literature. RESULTS: This study retrieved 208 cases of OLM in the multicenter study and 1035 cases in the literature review. In both, OLMs affected male and female individuals equally, with the most affected site being the tongue. Individuals ≥60 years of age were uncommonly affected. Symptomatic and larger lesions were more commonly reported in the literature review. CONCLUSIONS: This study comprises the largest sample of OLMs to date. OLMs are rare conditions, without sex predilection. The elderly proved to be less frequently affected, and the tongue is the most commonly affected site.

A new corneal epithelial biomimetic 3D model for in vitro eye toxicity assessment: Development, characterization and applicability.

In vitro eye toxicity assessment using reconstructed corneal epithelial models has emerged highlighting its applicability domain for Classification and Labeling of products and chemicals. However, due to bureaucratic issues, such models are not commercially available in Brazil and Latin America. In this work, we developed, characterized and evaluated the applicability of a new corneal epithelial biomimetic model using a cell lineage for in vitro eye toxicity assessment. The reconstructed tissue was obtained through the cultivation of HaCaT cells in an air-liquid interface, which presented morphology and biomarkers expression such as cytokeratin, CD44, and Ki-67 similar to human tissue. Furthermore, tissue viability was evaluated after exposure of the epithelial model to isolated chemicals from different Globally Harmonized System (GHS) eye irritation categories, and it has been demonstrated to be a suitable endpoint for classification of test materials, allowing discrimination between irritant and non-irritant chemicals. Furthermore, the model showed suitability for testing "real-life mixtures", once it identified irritant products between the analyzed eyebrow henna samples commercially labeled as non-irritants. This reproducible and low-cost epithelial corneal model presents features very important for Brazil and South America for R&D&I with no unnecessary animal experimentation.

Thermoreversible mucoadhesive polymer-drug dispersion for sustained local delivery of budesonide to treat inflammatory disorders of the GI tract.

Mucoadhesive drug formulations have been studied and used as alternatives to conventional formulations in order to achieve prolonged retention at the intended site. In addition to providing a controlled drug release, several drugs and disease conditions might benefit from mucoadhesive formulations, contributing to better therapeutic outcomes. Here, we describe the development and the in vitro/in vivo characterization of a mucoadhesive in situ gellifying formulation using PF127, a thermo reversible polymer, entrapping budesonide (BUD), a potent corticosteroid used for the treatment of a wide range of inflammatory diseases, including those affecting mucosas, such as in the GI tract. PF127 formulations (15-17%) were successfully prepared by a cold method as a thermo reversible in situ gelling dispersion for mucosal drug delivery, as confirmed by DSC. Sol-gel temperatures of PF127 formulations (25-39 °C) were observed by dynamic gelation and determined by microrheology and oscillatory rheometry. X-ray diffractograms and TEM images showed that BUD was completely solubilized within the polymeric micelles. In vitro, the gels showed 5-14 g force of mucoadhesion, and the ex vivo studies confirmed that the formulation efficiently adhered to the mucosa. Histopathological analysis combined with fluorescence images and ex vivo intestinal permeation confirmed that the formulation remained on the TGI mucosa for at least 4 h after administration. In vivo studies conducted in a murine model of intestinal mucositis demonstrated that the 16% PF127 BUD formulation was able to resolve the inflammatory injury in the intestinal mucosa. Results demonstrate that fine-tuning of PF127 formulations along with adequate selection of the drug agent, thorough characterization of the dispersions and their interactions with biological interfaces leads to the development of effective controlled drug delivery systems targeted to GI inflammatory diseases.

Intravenous delivery of a liposomal formulation of voriconazole improves drug pharmacokinetics, tissue distribution, and enhances antifungal activity.

Voriconazole (VCZ), a triazole with a large spectrum of action is one of the most recommended antifungal agents as the first line therapy against several clinically important systemic fungal infections, including those by Candida albicans. This antifungal has moderate water solubility and exhibits a nonlinear pharmacokinetic (PK) profile. By entrapping VCZ into liposomes, it is possible to circumvent certain downsides of the currently available product such as a reduction in the rate of its metabolization into an inactive form, avoidance of the toxicity of the sulfobutyl ether-beta-cyclodextrin (SBECD), vehicle used to increase its solubility. PKs and biodistribution of VCZ modified by encapsulation into liposomes resulted in improved antifungal activity, due to increased specificity and tissue penetration. In this work, liposomal VCZ resulted in AUC0-24/MIC ratio of 53.51 ± 11.12, whereas VFEND® resulted in a 2.5-fold lower AUC0-24/MIC ratio (21.51 ± 2.88), indicating favorable antimicrobial systemic activity. VCZ accumulation in the liver and kidneys was significantly higher when the liposomal form was used. Protection of the drug from biological degradation and reduced rate of metabolism leads to a 30% reduction of AUC of the inactive metabolite voriconazole-N-oxide (VNO) when the liposomal drug was administered. Liposomal VCZ presents an alternative therapeutic platform, leading to a safe and effective treatment against systemic fungal infections.

Thioredoxin and metallothionein: Homeostasis-related proteins in lip carcinogenesis.

OBJECTIVE: Thioredoxin (Trx) and metallothionein (MT) are involved in the development of some carcinomas; however, the role of these proteins in labial carcinogenesis has not yet been tested. The aims of the study were to evaluate and to correlate the immunoexpression of Trx and MT in actinic cheilitis, lip squamous cell carcinoma, and normal vermillion lip mucosa. DESIGN: Immunohistochemistry was undertaken for Trx and MT in samples of actinic cheilitis, lip squamous cell carcinoma, and normal lip mucosa. Qualitative and semi-quantitative evaluations were conducted. The proportion of stained cells, intensity of staining, and the cell compartment labeled were evaluated. A quickscore index was also calculated by multiplying the values of extension and intensity of nuclear and cytoplasmic staining, respectively, giving a maximum value of 9. Statistics were performed. RESULTS: A remarkable nuclear Trx staining was seen in normal lip mucosa and cheilitis, not in carcinoma (p<0.05). Cytoplasmic Trx expression was widely detected in all lesions (p>0.05). MT was broadly expressed in nuclei and cytoplasm of carcinoma, but not in normal lip mucosa and cheilitis (p<0.05). Quickscores were in accordance with the qualitative results. CONCLUSIONS: The current study showed a different immunopattern of Trx and MT between normal lip mucosa, actinic cheilitis and lip squamous cell carcinoma. The cellular compartment-based analyses evidenced differences that can be related to the proteins function. Considering the relevant roles of these proteins in cellular homeostasis, they seem to have an important role in lip carcinogenesis.

Clinical manifestations due to severe plasminogen deficiency: a case report.

Plasminogen deficiency is a rare, destructive, and badly defined disorder. Recurrent and progressive gingival nodular hyperplasia with ulceration would appear to be an unreported complication caused by this deficiency. In some of the reported cases, gingival hyperplasia occurred in association with an eye disease called ligneous conjunctivitis. Including this case report, only 11 patients with proven functional plasminogen and oral lesions have been reported in the literature in English. The purpose of this paper was to present the case of a child patient with recurrent clinical manifestations caused by severe plasminogen deficiency who responded positively to corticosteroid treatment.

Traumatic neuroma of the mandible: A case report with spontaneous remission.

Traumatic neuroma is a well-known disorder involving peripheral nerves, which occurs following trauma or surgery. The lesion develops most commonly in the soft tissues of the mental foramen area, lower lip and tongue. Intra-osseous lesions arising in jawbones are very uncommon. In this paper, we report a new case of an intra-osseous traumatic neuroma, discovered incidentally on a panoramic radiograph obtained for orthodontic documentation. In addition, the case herein described developed spontaneous remission, a situation not previously reported in the literature. Finally, we discuss relevant demographic, clinical, microscopic, immunohistochemical and treatment aspects of traumatic neuromas. Key words:Amputation neuroma, traumatic neuroma, mandible, spontaneous remission.

Expression of midkine in ameloblastomas and its correlation with clinicopathologic parameters.

OBJECTIVE: Midkine (MK) is a heparin-binding growth factor that is overexpressed in various human cancers. The aim of this study was to investigate the expression of MK in ameloblastomas and correlate the results with clinicopathologic parameters. STUDY DESIGN: Cases of ameloblastoma seen between 1999 and 2010 were identified. Clinical information was collected regarding age, gender, race, and location of tumor. Cases were classified as solid/multicystic, unicystic, and peripheral. The expression of midkine was assessed using immunohistochemistry. A significant difference was considered present at P < 0.05. RESULTS: A total of 34 cases of ameloblastoma and 4 cases of ameloblastic carcinomas were identified. MK was expressed in 67% of lesions (23.5% weak expression; 14.7% moderate expression; 29.4% strong expression). A significant difference was seen between solid/multicystic and unicystic lesions. CONCLUSIONS: MK is expressed in the majority of ameloblastomas, suggesting a role of the protein in the tumor's development, progression, and behavior.

A new nitrosyl ruthenium complex nitric oxide donor presents higher efficacy than sodium nitroprusside on relaxation of airway smooth muscle.

Nitric oxide (NO) has been demonstrated to be the primary agent in relaxing airways in humans and animals. We investigated the mechanisms involved in the relaxation induced by NO-donors, ruthenium complex [Ru(terpy)(bdq)NO(+)](3+) (TERPY) and sodium nitroprusside (SNP) in isolated trachea of rats contracted with carbachol in an isolated organs chamber. For instance, we verified the contribution of K(+) channels, the importance of sGC/cGMP pathway, the influence of the extra and intracellular Ca(2+) sources and the contribution of the epithelium on the relaxing response. Additionally, we have used confocal microscopy in order to analyze the action of the NO-donors on cytosolic Ca(2+) concentration. The results demonstrated that both compounds led to the relaxation of trachea in a dependent-concentration way. However, the maximum effect (E(max)) of TERPY is higher than the SNP. The relaxation induced by SNP (but not TERPY) was significantly reduced by pretreatment with ODQ (sGC inhibitor). Only TERPY-induced relaxation was reduced by tetraethylammonium (K(+) channels blocker) and by pre-contraction with 75mM KCl (membrane depolarization). The response to both NO-donors was not altered by the presence of thapsigargin (sarcoplasmic reticulum Ca(2+)-ATPase inhibitor). The epithelium removal has reduced the relaxation only to SNP, and it has no effect on TERPY. The both NO-donors reduced the contraction evoked by Ca(2+) influx, while TERPY have shown a higher inhibitory effect on contraction. Moreover, the TERPY was more effective than SNP in reducing the cytosolic Ca(2+) concentration measured by confocal microscopy. In conclusion, these results show that TERPY induces airway smooth muscle relaxation by cGMP-independent mechanisms, it involves the fluxes of Ca(2+) and K(+) across the membrane, it is more effective in reducing cytosolic Ca(2+) concentration and inducing relaxation in the rat trachea than the standard drug, SNP.

Analysis of cell proliferation and pattern of invasion in oral squamous cell carcinoma.

Cell proliferation markers play an important role in the biological behavior of neoplasms. This study investigated the immunohistochemical expression of PCNA, Ki-67 and Cyclin B1 proteins based on the pattern of cell invasion in oral squamous cell carcinoma (OSCC). A total of 39 OSCC specimens and 13 samples of normal oral mucosa (control) were immunohistochemically analyzed. Protein expression was evaluated according to World Health Organization - Histological Malignancy Grading (WHO-HMG) and a specific grading system for invasion, graded from 1 to 4, varying from a consistently well-defined "pushing" border to diffuse infiltration and cellular dissociation, and was then correlated with clinical features. We found higher expression of Ki-67 and Cyclin B1 in OSCC when compared with the control group. High Ki-67 expression levels were more commonly seen in the floor of the mouth than in the tongue (P = 0.009). Cyclin B1 showed a positive correlation with histological grade, according to WHO-HMG criteria (P = 0.01). Our results suggest that Cyclin B1 is a reliable proliferation marker for indicating degree of tumor proliferation. Correlations between PCNA, Ki-67, Cyclin B1 and invasive tumor front with overall survival were not observed. Further studies are needed in order to elucidate whether cell proliferation activity at the tumor invasion front is related to prognosis.

Long-term effects of nifedipine on human gingival epithelium: a histopathological and immunohistochemical study.

The chronic usage of nifedipine is associated with the appearance of gingival overgrowth (GO). The frequency of GO associated with chronic nifedipine therapy remains controversial and the possible subclinical effects of this drug on the gingival epithelium should be investigated. We investigated the epithelial proliferation index and apoptosis rate, and their association with epithelial enlargement. Proliferation (Ki67 and Cyclin B1) and apoptosis (BCL2, Bax and p53) markers were identified by immunohistochemistry in twenty-one samples of gingival tissue from patients undergoing chronic treatment with nifedipine and in eleven samples of gingival tissue from healthy patients who did not use drugs associated with GO (control). Our results show that the epithelial tissue of nifedipine users has considerably longer rete pegs compared to control (P = 0.01). However, the density of Ki67(+) and Cyclin B1(+) cells was similar in both groups. Regarding apoptosis, we found more BCL2(+) cells in the nifedipine group when compared to controls (P = 0.12). An increase in Bax(+) cells in the nifedipine group compared to control (P = 0.003) was also seen, and slightly lower levels of p53(+) expression were observed (P = 0.51). Our results suggest that the chronic use of nifedipine is not associated with subclinical changes in gingival tissue.

Expression of bone resorption regulators (RANK, RANKL, and OPG) in odontogenic tumors.

OBJECTIVE: To investigate the expression of bone resorption regulators (receptor activator of nuclear factor kappa B [RANK], RANK ligand [RANKL], and osteoprotegerin [OPG]) in calcifying cystic odontogenic tumor (CCOT), adenomatoid odontogenic tumor (AOT), calcifying epithelial odontogenic tumor (CEOT), odontogenic myxoma (OM), and ameloblastic fibroma (AF). STUDY DESIGN: The expression of these mediators was evaluated by means of immunohistochemistry. RESULTS: All specimens demonstrated positive immunoreactivity to RANK, RANKL, and OPG. The quantification of these mediators in epithelium revealed a similar pattern of expression for RANKL and OPG in CCOT, AOT, CEOT, and AF. With regard to stromal/mesenchymal cells, the majority of AOT and CCOT cases showed a higher content of OPG than RANKL, whereas CEOT, OM, and especially AF had a tendency to present a greater content of RANKL than OPG. CONCLUSION: Our data indicate that the CCOT, AOT, CEOT, OM, and AF cell constituents express key regulators of bone metabolism that might locally modulate tumor-associated bone resorption.

Low-grade central osteosarcoma of the mandible: a case study report.

Low-grade central osteosarcoma is a rare type of osteosarcoma with peculiar clinical radiographic and microscopic features. The aim of this article is to report and discuss a case of low-grade central osteosarcoma in the mandible of a 42-year-old woman. The patient reported sensing mild pain and tooth mobility for a period of 4 years, despite continuous dental treatment. Radiographic evaluation showed a mixed radiopaque/radiolucenct lesion in the body, ramus, coronoid process, and condyle of the left side of the mandible. Destruction of the mandibular cortex in the area was also observed. After incisional biopsy, the patient underwent hemimandibulectomy. Microscopic findings showed a tumor exhibiting spindle cells with nuclear hyperchromasia and no mitotic activity, irregular osteoid formation, and soft tissue infiltration. The immunohistochemical analysis of the expression of Ki-67, Cyclin B1, and PCNA proteins (cellular proliferation markers) revealed a very low Ki-67+ and Cyclin B1+ cell index (mean 7% and 3%, respectively), but a moderate number of PCNA+ cells (mean 49%). The 2 years of clinical and imaging postoperative follow-up showed no evidence of recurrence. Clinicians should be aware of these lesions, because histopathologicially low-grade central osteosarcoma may be misinterpreted as fibrous dysplasia.