Combination of losartan with pirfenidone: a protective anti-fibrotic against pulmonary fibrosis induced by bleomycin in rats.

Pirfenidone (PFD), one acceptable medication for treating idiopathic pulmonary fibrosis (IPF), is not well tolerated by patients at full doses. Hence, employing of some approaches such as combination therapy may be applicable for increasing therapeutic efficacy of PFD. Losartan (LOS), an angiotensin II receptor antagonist, could be a suitable candidate for combination therapy because of its stabilizing effect on the pulmonary function of IPF patients. Therefore, this study aimed to investigate the effects of LOS in combination with PFD on bleomycin (BLM)-induced lung fibrosis in rats. BLM-exposed rats were treated with LOS alone or in combination with PFD. The edema, pathological changes, level of transforming growth factor-ß (TGF-ß1), collagen content, and oxidative stress parameters were assessed in the lung tissues. Following BLM exposure, the inflammatory response, collagen levels, and antioxidant markers in rat lung tissues were significantly improved by PFD, and these effects were improved by combination with LOS. The findings of this in vivo study suggest that the combined administration of PFD and LOS may provide more potent protection against IPF than single therapy through boosting its anti-inflammatory, anti-fibrotic, and anti-oxidant effects. These results hold promise in developing a more effective therapeutic strategy for treating of lung fibrosis.

High inflammatory cytokines gene expression can be detected in workers with prolonged exposure to silver and silica nanoparticles in industries.

Occupational health must be strictly considered in industries particularly in nanoparticle factories where workers were exposed to different types of chemicals. We measured the serum levels of inflammatory cytokines in workers who developed skin lesions after exposure to silver and silica nanoparticles. Using a questionnaire in this cross-sectional study, we identified 110 workers in nanoparticle industries who were exposed to silver and silica nanoparticles. We also included 40 healthy subjects as controls from the administrative department of the same factories who were not exposed to nanoparticles. Peripheral blood samples used to measure the mRNA levels of inflammatory cytokines by qRT-PCR. In comparison with the control group, the workers who developed skin lesions had significantly higher levels of interleukin IL4, IL6, IL8, and TNF-α, particularly after two or three decades of exposure to silver and silica nanoparticles. Participants who were exposed to silver had higher levels of IL6 and IL8 compared with those who were exposed to silica. Necessary measures must be considered to protect workers in nanoparticle industries against the potential toxic effects of these compounds. Our network pharmacology study suggests corresponding biochemical pathways for these disorders.

Oral delivery of posaconazole-loaded phospholipid-based nanoformulation: Preparation and optimization using design of experiments, machine learning, and TOPSIS.

Phospholipid-based nanosystems show promising potentials for oral administration of hydrophobic drugs. The study introduced a novel approach to optimize posaconazole-loaded phospholipid-based nanoformulation using the design of experiments, machine learning, and Technique for Order of Preference by Similarity to the Ideal Solution. These approaches were used to investigate the impact of various variables on the encapsulation efficiency (EE), particle size, and polydispersity index (PDI). The optimized formulation, with %EE of âˆ¼ 74 %, demonstrated a particle size and PDI of 107.7 nm and 0.174, respectively. The oral pharmacokinetic profiles of the posaconazole suspension, empty nanoformulation + drug suspension, and drug-loaded nanoformulation were evaluated. The nanoformulation significantly increased maximum plasma concentration and the area under the drug plasma concentration-time curve (∼3.9- and 6.2-fold, respectively) and could be administered without regard to meals. MTT and histopathological examinations were carried out to evaluate the safety of the nanoformulation and results exhibited no significant toxicity. Lymphatic transport was found to be the main mechanism of oral delivery. Caco-2 cell studies demonstrated that the mechanism of delivery was not based on an increase in cellular uptake. Our study represents a promising strategy for the development of phospholipid-based nanoformulations as efficient and safe oral delivery systems.

Evaluation of Antioxidant and Antibacterial Effects of Lyophilized Cell-Free Probiotic Supernatants of Three Lactobacillus spp. and Their Cytocompatibility Against Periodontal Ligament Stem Cells.

Background: Periodontitis is a chronic disease characterized by the inflammation of the periodontium and leads to progressive damage, such as gingival atrophy, alveolar bone loss, and tooth loss. Streptococcus mutans and Aggregatibacter actinomycetemcomitans are bacteria that support the occurrence of periodontitis via the ability to form biofilms or damage the alveolar bone and periodontal ligaments. On the other hand, periodontal ligament stem cells (PDLSCs) are cells with differentiation capability into osteoblasts or osteoblasts. Due to their role in periodontal homeostasis and regeneration, PDLSCs are considered to control periodontitis progression. However, probiotics are helpful microorganisms known to have antimicrobial and immune-regulating effects. Objectives: This study aimed to evaluate the antioxidant activity and antimicrobial effects of lyophilized cell-free supernatants (LCFSs) derived from three probiotic strains of Lactobacillus on S. mutans and A. actinomycetemcomitans. Moreover, the effect of these lyophilized supernatants was investigated on the viability and migration capability of PDLSCs. Methods: The antibacterial effects of LCFSs of three probiotic bacteria were investigated by determining the minimum inhibitory concentration and minimum bactericidal concentration. Then, the effect of LCFSs on the survival and migration of PDLSCs was investigated by the MTT method (at 24 and 72 hours) and scratch test (at 0, 24, and 48 hours), respectively. Finally, the antioxidant effect of LCFSs was assessed by the 2,2-diphenyl-1-picrylhydrazyl (DPPH) assay and ferric reducing/antioxidant power methods. Results: The antibacterial properties of different concentrations of acidic and neutral LCFSs derived from three studied probiotic bacteria on S. mutans and A. actinomycetemcomitans were observed within the range of 12.5 - 50% (v/v) (1/8 - 1/2 dilutions with culture medium). Although there were no significant toxic (~ 100% viability) and wound healing effects on PDLSCs when the cells were exposed to either acidic or neutral studied LCFSs in a concentration of 5% (v/v), they showed significant antioxidant activity (~ 90% DPPH inhibition and 0.5 mM Fe2+/L). Conclusions: The results revealed that 5% (v/v) 48-hour acidic and neutral supernatants of three studied probiotics might play a beneficial role in controlling periodontitis.

Sterility and bioactivity evaluation of two types of bone graft substitutes after removing the original packaging.

Background: Xenograft and allograft bone substitutes are widely used to replace the missing bone in defects. Since removing the packaging of these grafts can nullify their sterilization, this study aimed to evaluate the sterility and bioactivity changes of an allograft and a xenograft following uncapping/recap. Methods: Two types of commercial allograft and xenograft vials were unpacked and further exposed to operating room air, where implant surgery was performed for one second, ten minutes, and one hour. After three repetitions, samples were analyzed using microbiological tests and scanning electron microscopy (SEM) with energy dispersive x-ray analysis (EDX) for sterility and bioactivity evaluation. Results: None of the bone graft samples showed microbial growth or bioactivity-negative changes after seven days of unpacking the vials. Conclusion: Despite the positive results of this study, future studies and more analysis considering influential factors are required. Also, disinfection and air exchange must still be observed during biomaterial application and bone grafting procedures.

Inhalation of multi-wall carbon nanotubes changes the expression of apoptosis and cancer genes in rat brain and lungs.

One of the important issues in urban areas is air pollution which causes respiratory disorders. A significant association between exposure to inhaled particulate matter (PM), mainly ultrafine particles, and increased neurological and pulmonary morbidity and mortality was observed in some research. This study aimed to demonstrate the relation between multi-wall carbon nanotubes (MWCNTs) inhalation and the carcinogenic effect of these materials in the brain and lungs. For this purpose, we investigated gene expression in rat brain and lung tissues induced by exposure to MWCNTs. Rats were exposed to MWCNTs in diameters of 10 and 100 nm (pure and impure) at a concentration of 5 mg/m3. Exposure was done through a whole-body exposure chamber for 5 h/day, 5 days/week for 14 days. After exposure, both brain and lung tissues were isolated to evaluate certain gene expressions including Bax, Bcl2, Rac1, Tp53, Mmp12, and Arc. The results showed that exposure to impure and pure MWCNTs (10 and 100 nm) at a concentration of 5 mg/m3 causes up-regulation or down-regulation of some of these genes. The results suggest that impure and pure MWCNTs (10 and 100 nm) can increase the risk of central nervous system disorders such as Alzheimer's disease and increase the risk of carcinogenesis in the lung tissues of rats exposed to MWCNTs (Tab. 2, Fig. 2, Ref. 64). Text in PDF www.elis.sk Keywords: multi-wall carbon nanotube, inhalation, gene expression, carcinogenicity, brain, lung.

Losartan enhances the suppressive effect of pirfenidone on the bleomycin-induced epithelial-mesenchymal transition and oxidative stress in A549 cell line.

Objectives: Idiopathic pulmonary fibrosis (IPF) is a fatal lung disease. Despite the promising anti-fibrotic effect, the toleration of pirfenidone (PFD) by the patients in full dose is low. Combination therapy is a method for enhancing the therapeutic efficiency of PFD and decreasing its dose. Therefore, the present study evaluated the effect of a combination of losartan (LOS) and PFD on oxidative stress parameters and the epithelial-mesenchymal transition (EMT) process induced by bleomycin (BLM) in human lung adenocarcinoma A549 cells. Materials and Methods: The non-toxic concentrations of BLM, LOS, and PFD were assessed by the MTT assay. Malondialdehyde (MDA) and anti-oxidant enzyme activity including catalase (CAT) and superoxide dismutase (SOD) were assessed after co-treatment. Migration and western blot assays were used to evaluate EMT in BLM-exposed A549 after single or combined treatments. Results: The combination treatment exhibited a remarkable decrease in cellular migration compared with both single and BLM-exposed groups. Furthermore, the combination treatment significantly improved cellular anti-oxidant markers compared with the BLM-treated group. Moreover, combined therapy markedly increased epithelial markers while decreasing mesenchymal markers. Conclusion: This in vitro study revealed that the combination of PFD with LOS might be more protective in pulmonary fibrosis (PF) than single therapy because of its greater efficacy in regulating the EMT process and oxidative stress. The current results might offer a promising therapeutic strategy for the future clinical therapy of lung fibrosis.

Preparation of Citrus reticulata peel nano-encapsulated essential oil and in vitro assessment of its biological properties.

Dental caries is the most common biofilm-dependent oral disease. Streptococcus mutans is among the main microorganisms responsible for the development of dental caries. Nano-suspension of Citrus reticulata (tangerine) peel essential oil in 0.5% (v/v) concentration was prepared and its antibacterial effect on S. mutans in planktonic and biofilm forms as well as its cytotoxic and antioxidant effects were assessed and compared with chlorhexidine (CHX). The minimum inhibitory concentration (MIC) of free essential oil, nano-encapsulated essential oil, and CHX was 5.6% (v/v), 0.0005% (v/v), and 0.0002% (w/v), respectively. The percentage of biofilm inhibition by the free essential oil, nano-encapsulated essential oil, and CHX at half-MIC was 67.3%, 24%, and 90.6%, respectively. The nano-encapsulated essential oil had no cytotoxicity and showed significant antioxidant effects in different concentrations. Nano-encapsulation of tangerine peel essential oil significantly enhanced its biological activities in much lower concentrations than the free essential oil (11,000 times diluted). It also showed lower cytotoxicity and higher antibiofilm effects in sub-MICs compared with CHX, indicating the optimal potential of tangerine nano-encapsulated essential oil for incorporation in the composition of organic antibacterial and antioxidant mouth rinses.

Comparative effect of anthocyanin on proliferation and migration of human gingival fibroblasts in the absence or presence of nicotine.

Background: Oral fibroblast malfunction can result in periodontal diseases. Nicotine can prolong the healing process as an irritant of oral tissues. Anthocyanins have been demonstrated to have potential benefits in preventing or treating smoking-related periodontal diseases. Cyanidin chloride's (CC's) potential in oral wound healing and the viability, proliferation, and migration of human gingival fibroblasts (HGFs) were examined in the presence and absence of nicotine by an in vitro study. Methods: The effects of different nicotine concentrations (1, 2, 3, 4, and 5 mM) on the viability and proliferation of HGF cells were evaluated in the presence and absence of different CC concentrations (5, 10, 25, and 50 µM) using the quantitative MTT assay. The scratch test was performed to evaluate the migration of CC-treated cells in the presence of 2.5-mM nicotine. Results: No cytotoxicity was observed at 1‒100 µM CC concentrations after 24, 48, and 72 hours of exposure to HGF cells. However, a concentration of 200 µM significantly reduced cell viability by about 20% at all the three-time intervals (P<0.05). Also, 3‒5-mM concentrations of nicotine significantly reduced cell viability in a dose- and time-dependent manner. Moreover, the understudied CC concentrations decreased nicotine's adverse effects on cell migration to some extent. Conclusion: Although the understudied CC concentrations could not significantly reduce the adverse effects of understudied nicotine concentrations on the viability and proliferation of HGF cells, they were able to reduce the detrimental effects of nicotine on cell migration significantly.

Synthesis and characterization of a calcium phosphate bone cement with quercetin-containing PEEK/PLGA microparticles.

This study aimed to describe the synthesis and characterization of a calcium phosphate cement (CPC) with polyetheretherketone/poly (lactic-co-glycolic) acid (PEEK/PLGA) micro-particles containing quercetin. CPC powder was synthesized by mixing dicalcium phosphate anhydrate and tetracalcium phosphate. To synthesize PEEK/PLGA microparticles, PLGA85:15 was mixed with 90 wt% PEEK. The weight ratio of quercetin/PLGA/PEEK was 1:9:90 wt%. PEEK/PLGA/quercetin microparticles with 3, 5, and 6 wt% was added to CPC. The setting time, compressive strength, drug release profile, solubility, pH, and porosity of synthesized cement were evaluated. The morphology and physicochemical properties of particles was analyzed by scanning electron microscopy, Fourier-transform infrared spectroscopy (FTIR), x-ray diffraction (XRD), and inductively coupled plasma. Cytotoxicity was assessed by the methyl thiazolyl tetrazolium assay using dental pulp stem cells. Expression of osteoblastic differentiation genes was evaluated by real-time polymerase chain reaction. Data were analyzed by one-way ANOVA and Tukey's test (alpha = 0.05). The setting time of 3 wt% CPC was significantly longer than 5 and 6 wt% CPC (P< 0.001). The 6 wt% CPC had significantly higher compressive strength than other groups (P= 0.001). The release of quercetin from CPCs increased for 5 d, and then reached a plateau. XRD and FTIR confirmed the presence of hydroxyapatite in cement composition. Significantly higher expression of osteocalcin (OCN) and osteopontin (OPN) was noted in 3 wt% and 6 wt% CPCs. Addition of quercetin-containing PEEK/PLGA microparticles to CPC enhanced its compressive strength, decreased its setting time, enabled controlled drug release, and up-regulated OPN and OCN.

Applications of Novel and Nanostructured Drug Delivery Systems for the Treatment of Oral Cavity Diseases.

PURPOSE: Novel drug delivery systems (DDSs) hold great promise for the treatment of oral cavity diseases. The main objective of this article was to provide a detailed overview regarding recent advances in the use of novel and nanostructured DDSs in alleviating and treating unpleasant conditions of the oral cavity. Strategies to maximize the benefits of these systems in the treatment of oral conditions and future directions to overcome these issues are also discussed. METHODS: Publications from the last 10 years investigating novel and nanostructured DDSs for pathologic oral conditions were browsed in a systematic search using the PubMed/MEDLINE, Web of Science, and Scopus databases. Research on applications of novel DDSs for periodontitis, oral carcinomas, oral candidiasis, xerostomia, lichen planus, aphthous stomatitis, and oral mucositis is summarized. A narrative exploratory review of the most recent literature was undertaken. FINDINGS: Conventional systemic administration of therapeutic agents could exhibit high clearance of drugs from the bloodstream and low accumulation at the target site. In contrast, conventional topical systems face problems such as short residence time in the affected region and low patient compliance. Novel and nanostructured DDSs are among the most effective and commonly used methods for overcoming the problems of conventional DDSs. The main advantages of these systems are that they possess the ability to protect active agents from systemic and local clearance, enhance bioavailability and cellular uptake, and provide immediate or modified release of therapeutic agents after administration. In the design of local drug delivery devices such as nanofiber mats, films, and patches, components and excipients can significantly affect factors such as drug release rate, residence time in the oral cavity, and taste in the mouth. Choosing appropriate additives is therefore essential. IMPLICATIONS: Local drug delivery devices such as nanofiber mats, nanoparticles, liposomes, hydrogels, films, and patches for oral conditions can significantly affect drug efficacy and safety. However, more precise clinical studies should be designed and conducted to confirm promising in vitro and in vivo results. In recent years, novel and nanostructured DDSs increasingly attracted the attention of researchers as a means of treatment and alleviation of oral diseases and unpleasant conditions. However, more clinical studies should be performed to confirm promising in vitro and in vivo results. To transform a successful laboratory model into a marketable product, the long-term stability of prepared formulations is essential. Also, proper scale-up methods with optimum preparation costs should be addressed.

The effect of naproxen patches on relieving orthodontic pain by evaluation of VAS and IL-1β inflammatory factor: a split-mouth study

ABSTRACT Introduction: Pain related to orthodontic tooth movement is common and cause dissatisfaction and discomfort. Objective: The present study aimed to compare the efficacy of naproxen patches in pain control during orthodontic tooth separation, by means of visual analogue scale (VAS) and interleukin 1β (IL-1β) levels in gingival crevicular fluid (GCF). Methods: In this split-mouth triple-blind clinical trial, with 40 patients following separation, 5% naproxen or placebo patches were randomly placed on the upper right or left first molars every 8 hours. Pain intensity scores were determined after 2 and 6 hours, sleep time, 24 hours, days 2, 3 and 7 by the patients using a 100-mm VAS ruler. IL-1β levels in GCF were evaluated by ELISA at baseline, 1 and 24 hours and 7 days. Paired samples t-tests and two-way repeated measures ANOVA analysis of variance with a significance level of 0.05 were applied. Results: A total number of 30 patients (13 males and 17 females) finished the trial. Significant differences were found in pain scores (p< 0.0001) and IL-1β levels (p= 0.047) between naproxen and placebo groups. Lower pain scores were reported for the patients using naproxen patches at all time points, except 1 hour after separation. IL-1β levels were lower for the patients using naproxen patches only 1 hour after separation (p= 0.047). The peak of pain scores and IL-1β levels were calculated at 24 hours. Conclusion: In the light of VAS scores and IL-1β levels, naproxen patches reduced the pain caused by separator placement.

RESUMO Introdução: a dor relacionada à movimentação dentária ortodôntica é comum e causa insatisfação e desconforto. Objetivo: o presente estudo teve como objetivo avaliar a eficácia de curativos de naproxeno no controle da dor durante a separação ortodôntica dos dentes, por meio de escalas visuais analógicas (EVA) e dos níveis de interleucina 1β (IL-1β) no fluido crevicular gengival (FCG). Métodos: neste ensaio clínico, triplo-cego, boca dividida, com 40 pacientes após a separação dos dentes, foram aplicados, de forma aleatória, curativos com naproxeno a 5% ou placebo, nos primeiros molares superiores, direito ou esquerdo, a cada 8 horas. Os escores de intensidade da dor foram registrados pelos pacientes após 2 e 6 horas, durante o sono, após 24 horas, 2, 3 e 7 dias, usando uma EVA de 100 mm. Os níveis de IL-1β no FCG foram avaliados pelo ELISA no momento inicial, e após 1 e 24 horas e 7 dias. Foram aplicados testes t para amostras pareadas e ANOVA de duas vias para medidas repetidas, com nível de significância de 0,05. Resultados: no total, 30 pacientes (13 homens e 17 mulheres) terminaram o ensaio clínico. Diferenças significativas foram encontradas nos escores de dor (p< 0,0001) e níveis de IL-1β (p= 0,047) entre os grupos naproxeno e placebo. Índices mais baixos de dor foram relatados pelos pacientes que usaram curativos de naproxeno em todos os tempos avaliados, com exceção de 1 hora após a separação. Os níveis de IL-1β foram menores nos pacientes que usaram os curativos de naproxeno apenas 1 'hora após a separação (p= 0,047). Os picos dos escores de dor e dos níveis de IL-1β foram registrados 24 horas após a separação. Conclusão: considerando-se os escores das EVAs e os níveis de IL-1β, pode-se concluir que os curativos de naproxeno reduziram a dor causada pela instalação dos separadores ortodônticos.

Effect of sodium chloride on gene expression of Streptococcus mutans and zeta potential of demineralized dentin.

PURPOSE: In this work, the effects of sodium chloride (NaCl) on gene expression of planktonic Streptococcus mutans cells are investigated. Also assessed are the effects of NaCl on zeta potential of sound and demineralized dentin. METHODS: The relative level of glucosyltransferase B (gtfB), gtfC and gtfD transcription of S. mutans in the presence of NaCl was evaluated by quantitative polymerase chain reaction (qPCR). The osmolality of varying salt (NaCl) concentrations and their influence on the zeta potential of sound and demineralized dentin was investigated as well. RESULTS: NaCl significantly reduced the expression of gtfB and C genes in planktonic S. mutans; whereas, gtf D gene expression significantly increased in the presence of NaCl (P < 0.05). NaCl at concentrations of 37.5 mg/ml reduced zeta potential of demineralized dentin, while no significant decrease of zeta potential was found when sound dentin was exposed to this concentration. CONCLUSION: NaCl reduces the expression of some gtfs in S. mutans and increases negative potential charge of demineralized dentin.

Effect of one and two sessions of antimicrobial photodynamic therapy on clinical and microbial outcomes of non-surgical management of chronic periodontitis: A clinical study.

Background: This study aimed to compare the effect of one and two sessions of antimicrobial photodynamic therapy (aPDT) as an adjunct to scaling and root planing (SRP) on clinical and microbial parameters in patients with chronic periodontitis. Methods: This study was conducted on 20 patients. The dental quadrants of patients were randomly assigned to SRP at baseline (group 1), SRP at baseline and one month (group 2), SRP plus aPDT at baseline (group 3) and SRP plus aPDT at baseline and one month (group 4). Probing depth (PD), clinical attachment level (CAL) gain, and bleeding on probing (BoP) were measured at baseline, and one and three months later. F. nucleatum counts were determined by PCR. ANOVA was used for the comparison of these variables between the groups. Results: In all the groups, PD reduction and CAL gain increased significantly at 1- and 3-month intervals compared to baseline (P=0.001). At three months, the difference in PD between groups 1 and 3 was statistically significant (P=0.014). CAL gain between groups 2 and 4 at one month (P=0.016) and three months (P=0.001) wasstatistically significant. Reduction in F. nucleatum counts was not significant between the four study groups (P>0.05). Conclusion: A combination of two sessions of aPDT and SRP could improve CAL gain; however, further long-term studies are necessary in this regard.

The effect of naproxen patches on relieving orthodontic pain by evaluation of VAS and IL-1ß inflammatory factor: a split-mouth study.

INTRODUCTION: Pain related to orthodontic tooth movement is common and cause dissatisfaction and discomfort. Objective: The present study aimed to compare the efficacy of naproxen patches in pain control during orthodontic tooth separation, by means of visual analogue scale (VAS) and interleukin 1ß (IL-1ß) levels in gingival crevicular fluid (GCF). METHODS: In this split-mouth triple-blind clinical trial, with 40 patients following separation, 5% naproxen or placebo patches were randomly placed on the upper right or left first molars every 8 hours. Pain intensity scores were determined after 2 and 6 hours, sleep time, 24 hours, days 2, 3 and 7 by the patients using a 100-mm VAS ruler. IL-1ß levels in GCF were evaluated by ELISA at baseline, 1 and 24 hours and 7 days. Paired samples t-tests and two-way repeated measures ANOVA analysis of variance with a significance level of 0.05 were applied. RESULTS: A total number of 30 patients (13 males and 17 females) finished the trial. Significant differences were found in pain scores (p< 0.0001) and IL-1ß levels (p= 0.047) between naproxen and placebo groups. Lower pain scores were reported for the patients using naproxen patches at all time points, except 1 hour after separation. IL-1ß levels were lower for the patients using naproxen patches only 1 hour after separation (p= 0.047). The peak of pain scores and IL-1ß levels were calculated at 24 hours. CONCLUSION: In the light of VAS scores and IL-1ß levels, naproxen patches reduced the pain caused by separator placement.

Truncated forms of RUNX3 Unlike Full Length Protein Alter Cell Proliferation in a TGF-ß Context Dependent Manner.

The Runt related transcription factors (RUNX) are recognized as key players in suppressing or promoting tumor growth. RUNX3, a member of this family, is known as a tumor suppressor in many types of cancers, although such a paradigm was challenged by some researchers. The TGF-ß pathway governs major upstream signals to activate RUNX3. RUNX3 protein consists of several regions and domains. The Runt domain is a conserved DNA binding domain and is considered as the main part of RUNX proteins. Herein, we compared the effects of Runt domains and full-Runx3 in cell viability by designing two constructs of Runx3, including N-terminal region and Runt domain. We investigated the effect of full-Runx3, N-t, and RD on growth inhibition in AGS, MCF-7, A549, and HEK293 cell lines which are different in TGF-ß sensitivity, in the absence and presence of TGF-ß. The full length RUNX3 did not notably inhibit growth of these cell lines while, the N-t and RD truncates showed different trends in these cell lines. Cell proliferation in the TGF-ß impaired context cell lines (AGS and MCF-7) significantly decrease while in the A549 significantly increase. On the other hand, transfection of N-t and RD did not considerably affect the cell proliferation in the HEK293.Our results show that full-lenght RUNX3 did not affect the cell viability. Conversely, the N-t and RD constructs significantly changed cell proliferation. Therefore, therapeutic potentials for these truncated proteins are suggested in tumors with RUNX proteins dysfunction, even in the TGF-ß impair context.

Comparison of Er:YAG Laser and Hand Instrumentation on the Attachment of Cultured Human Gingival Fibroblasts to Periodontally Involved Root Surfaces.

Introduction: The present study compared the effects of erbium-doped yttrium aluminium garnet (Er:YAG) laser and hand instrumentation on the attachment of human gingival fibroblast (HGF) cells to periodontally involved root surfaces. Methods: A total of 40 tooth specimens were collected and treated in four distinct groups: scaled and root planed with hand instruments, scaled with Er:YAG laser, treated with a combination of hand instruments and Er:YAG laser and non-treated control group. The attachment and proliferation rate of HGF were assessed using MTT assay and scanning electron microscope (SEM) examination was used for cell morphological evaluation. Results: The MTT (3-[4, 5-dimethylthiazol-2-yl]-2, 5 diphenyl tetrazolium bromide) assay showed significant decrease in HGF cell viability in both hand instruments only and combination treated teeth specimens compared to control specimens (P<0.05), 24 hours after cell seeding. However, at time 48, the cell viability of attached cells in these 2 treated groups was almost similar to control. In contrast, at 24 and 48 hours after cell seeding, viability of attached cells was higher than control in Er:YAG laser treated only specimens (P<0.05). According to SEM study, the laser treated specimens showed more surface roughness. Conclusion: Er:YAG laser increased attachment and proliferation of HGF cells in comparison to the hand instruments method.

Efficacy of vitamins E and C for reversing the cytotoxic effects of nicotine and cotinine.

Nicotine has adverse cellular and molecular effects on oral mucosa, bone, and teeth. Vitamin E (α-tocopherol) and vitamin C (ascorbic acid) are biological antioxidants with positive effects on wound healing and bone formation. This in vitro study sought to assess the cytotoxic effects of different concentrations of nicotine and cotinine (a metabolite of nicotine) on MG-63 osteoblast-like cells and human gingival fibroblasts (HGFs) in the presence and absence of antioxidant vitamins E and C (separately and combined). Cell viability and proliferation were assessed using the methyl thiazol tetrazolium (MTT) assay. Cell migration was assessed using the scratch test, and expression of apoptosis-related genes was quantitatively analyzed using real-time PCR. Dose-dependent negative effects of nicotine on the morphology, viability, proliferation, and migration of MG-63 and HGF cells were statistically significantly greater than those of cotinine. Vitamin E (separately and combined with vitamin C) was statistically significantly more effective than vitamin C (at the concentration used in this study) at improving cell viability, proliferation, and migration, and at reducing apoptosis of cells exposed to nicotine or cotinine. Based on the positive results of this study, vitamin C and especially vitamin E (systemically and/or locally) may be useful in the repair and regeneration of oral hard and soft tissues in smokers.

Identification and in vitro characterization of novel nanobodies against human granulocyte colony-stimulating factor receptor to provide inhibition of G-CSF function.

It has been shown that Granulocyte colony-stimulating factor (G-CSF) has a higher expression in malignant tumors, and anti-G-CSF therapy considerably decreases tumor growth, tumor vascularization and metastasis. Thus, blocking the signaling pathway of G-CSF could be beneficial in cancer therapy. This study is aimed at designing and producing a monoclonal nanobody that could act as an antagonist of G-CSF receptor. Nanobodies are the antigen binding fragments of camelid single-chain antibodies, also known as VHH. These fragments have exceptional properties which makes them ideal for tumor imaging and therapeutic applications. We have used our previously built nanobody phage libraries to isolate specific nanobodies to the G-CSF receptor. After a series of cross-reactivity and affinity experiments, two unique nanobodies were selected for functional analysis. Proliferation assay, real-time PCR and immunofluorescence assays were used to characterize these nanobodies. Finally, VHH26 nanobody that was able to specifically bind G-CSF receptor (G-CSF-R) on the surface of NFS60 cells and efficiently block G-CSF-R downstream signaling pathway in a dose-dependent manner was selected. This nanobody could be further developed into a valuable tool in tumor therapy and it forms a basis for additional studies in preclinical animal models.

In Vitro Comparison of Biological Effects of Coe-Pak and Reso-Pac Periodontal Dressings.

OBJECTIVES: The purpose of the present study was to compare the cytotoxicity of Reso-Pac and Coe-Pak periodontal dressing. MATERIAL AND METHODS: According to ISO-10993-12:2012, 1-, 3- and 7-day extracts of the two periodontal dressings were prepared in cell culture medium and exposed to the two cultured cell lines. Cell viability and proliferation at 24 h and 72 h following exposure were evaluated using quantitative MTT assay. RESULTS: The results showed a significant (P < 0.05) reduction in the viability of cells exposed to the 3- and 7-day Coe-Pak extracts at 24 h and 72 h compared to the control group (no exposure to the extract). Reso-Pac extracts slightly decreased cell viability compared to the control group. Understudy materials showed greater cytotoxicity against human osteoblast-like compared to the human gingival fibroblast cells. No significant (P > 0.05) difference was found in the viability of cells exposed to undiluted (100%) one-day extract and diluted (50%) extract of both understudy materials at 24 h and 72 h after exposure. CONCLUSIONS: Based on the results, Reso-Pac periodontal dressing has less cytotoxicity than Coe-Pak.