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Cloves (Syzygium aromaticum) as been used as traditional medicine for many years and they possess antibacterial, antifungal and antiviral properties. Clove is known for its anticancer property on various cancer cell lines and is well established, but its anticancer effect on OSCC cell lines is less known. Aim of the study was to determine the anticancer and antioxidant effect of Syzygium aromaticum extract on OSCC cell lines (KB cell lines) and compare the same with normal mouse fibroblasts cell lines (L292 cell lines). KB cell lines and L292 cell lines were commercially obtained. Clove was obtained from local market and ethanolic extract (EC) of clove was prepared. Anticancer activity was assessed by MTT, neutral red, DAPI and Double staining assay and antioxidant assay was carried out by FRAP, PM and DPPH assay. The antioxidant property of EC of clove increased with increase in the concentration in a dose dependent manner. Both MTT and Neutral Red assay showed increase in cell death with increase in concentration of EC of clove. Double staining and DAPI showed increase in cell death when treated with EC of clove. The anticancer and antioxidant activity of EC of clove was comparable with standard drug used in the assay. This in vitro study demonstrates effective anticancer and antioxidant activity on KB cell lines when compared to standard control. However, further studies are to be conducted in order to characterize other potential antitumor components of the clove, so that it can be used as therapeutic agent in treating oral carcinoma.

Oral hygiene maintenance is important to maintain optimal oral health. Oral health is affected by dysbiotic oral microflora in the dental plaque. Virulent factors of pathogenic organisms, such as gingipain, are responsible for tissue degradation and host tissue invasion in periodontal disease. We sought to investigate the distribution of gingipain genotypes (rgpA and kgp) of P. gingivalis in patients with chronic periodontitis and healthy individuals. The study included individuals positive for P. gingivalis, with 95 samples in the chronic periodontitis (CP) group and 35 samples in the healthy (H) group. We found that kgp-I and kgp-II types were prevalent in 67.36% and 32.64% of the samples in the CP group, respectively. In the H group, kgp-II was highly prevalent (97.14%). The rgpA genotype, type A was found in 78.95% and 82.85% of the samples in the CP and H group, respectively. The mean level of PD and CAL were increased in the presence of kgp-I and decreased in the presence of kgp-II. The mean level of P. gingivalis was increased in the presence of kgp-I and rgpA, type A. Our results show that kgp-I and kgp-II are strongly associated with disease and health condition, respectively.

Aim:To evaluate and compare the effect of curcumin (CUR) and Nano-curcumin (N-CUR) on human-derived mesenchymal stem cells (MSCs) in a dose-dependent manner.Materials and Methods:An experimental study performed with putative MSCs from a total of five systemically healthy subjects with chronic periodontitis. These putative MSCs were isolated by cell culture and were further characterized and identified by colony-forming unit assay and immunocytochemical analysis using cell surface markers CD105, CD146, CD45 and CD73. The identified MSCs were treated with different doses of CUR and N-CUR, and compared with α-minimum essential medium (α -MEM) for its cell viability by performing MTT (3-[4,5-dimethylthiazol-2-yl]-2,5-diphenyl tetrazolium bromide) assay for 48 and 72 hr. The statistically analysis was performed using one-way analysis of variance (ANOVA) followed by Tukey’s post hoc test and Bonferroni’s post hoc test.Results:Compared to the α-MEM group, both CUR and N-CUR treated cells have shown significantly ( P = .029) higher survival rate at lower concentration (0.1 and 0.5 µM/L), at 48 hr incubation. However, there was no statistically significant difference between the CUR and N-CUR groups on cell survival rate at both 48 and 72 hr incubation. When compared between the concentrations of the same group, significantly higher cell viability ( P = .001) was observed at lower concentrations (0.1, 0.5 µM/L) in both test groups after incubation for 48 and 72 hr.Conclusion:Both CUR and N-CUR have a dose-dependent effect on human derived MSCs survival when incubated for 48 hr, whereas N-CUR shows increased cell survival rate even at 72 hr of incubation. Although, the cautious use of CUR and N-CUR at higher concentrations is recommended.

Aim: The cancer stem cells (CSCs) are known to be responsible for drug resistance and cancer relapse in the treatment of cancer. Identification and isolation of CSCs and study of their properties will play a crucial role in developing an effective drug against these targets. The aim of the study was to isolate CSCs from primary cancer by the tumorspheres enrichment method, to confirm by indirect immunofluorescence and gene expression of stem cell markers by using real-time polymerase chain reaction (RT-PCR) technique. Materials and Methods: In this in vitro study, we enriched oral CSCs through tumorsphere formation assay from seven primary cultures of OSCC patients with defined serum media. The expression and localization of the cell surface markers of CD133 and CD44 were tested by indirect immunofluorescence. Gene expression of stem cell markers such as CD44, CD133, Oct4, Sox2, and Nanog were quantified by RT-PCR technique. One-way analysis of variance was applied to analyze gene expression. Results: Tumorsphere formation has been used to isolate the CSCs from the OSCC tissue culture. Both CD133 and CD44 antibody confirmed the presence of CSCs through indirect immunofluorescence. In comparison to parental cell lines, the expression levels of CD133, CD44, Oct4, Sox2, and Nanog stem cell were significantly higher in CSC-enriched subpopulations. Conclusions: The cost-effective spheroid enrichment and the indirect immunofluorescence methods are useful for the isolation of CSCs from the primary tumor.

Porphyromonas gingivalis is a keystone pathogen and major colonizer in host tissue which plays a pivotal role in periodontitis among the other polymicrobial infections. Increasing facts demonstrate that curcumin has antibacterial activity and anti-biofilm effect against the periodontopathogens through diverse mechanisms that have a positive impact on periodontal health. The present study was aimed to elucidate the effect of curcumin on biofilm formation and virulence factor gene expression of P. gingivalis. By using gene expression studies, we exploited the mechanism of anti-biofilm effects of curcumin on P. gingivalis. The minimum inhibitory concentration and minimum bactericidal concentration of curcumin for both ATCC and clinical strains of P. gingivalis were found to be 62.5 and 125 µg ml−1 respectively. Curcumin prevented bacterial adhesion and biofilm formation in a dose-dependent manner. Further, curcumin attenuated the virulence of P. gingivalis by reducing the expression of genes coding for major virulence factors, including adhesions (fimA, hagA, and hagB) and proteinases (rgpA, rgpB, and kgp). The results indicated that curcumin has shown anti-biofilm as well as antibacterial activity against P. gingivalis. Further, curcumin because of its pleiotropic actions could be a simple and inexpensive therapeutic strategy in the treatment of periodontal disease.

Background and Aims:Porphyromonas gingivalis ( P. gingivalis) is considered as an important pathogen responsible for periodontal disease which is characterized by inflammation of gingiva and destruction of periodontal ligament and alveolar bone leading to loss of tooth. Along with clinical investigations, suitable microbiological analysis needs to be performed which could provide more insight into the disease severity. We aim to quantify P. gingivalis by real-time PCR (RT-PCR) and analyze its association with demographic data including clinical parameters.Materials and Methods:The study consisted of chronic periodontitis patients (CP group) and healthy subjects (H group) with 120 samples in each group. RT-PCR was carried out by the SYBR Green assay to target 16S ribosomal ribonucleic acid species-specific region of P. gingivalis. Standard strain of P. gingivalis ATCC 33277 was used as a control.Results:In the CP group, 79.16% samples were found positive for P. gingivalis, whereas 29.17% samples were positive in the H group. A significant difference was found when the prevalence was compared within males and females ( P < .001 for both). In the older age groups, we found a higher rate of detection of P. gingivalis. As analyzed by Spearman’s correlation test, the number of cells of P. gingivalis was significantly associated with probing depth ( P = .02) and clinical attachment level ( P = .01) in the CP group. The mean cell number of P. gingivalis was found to be increasing with increasing levels of probing depth and clinical attachment level ( P < .001 and P = .01, respectively).Conclusion:The present study reaffirms that the P. gingivalis microbe is significantly associated with the chronic periodontitis and that its level varies with the severity of the disease. Colonization of the bacterium is significantly associated with severe forms of the disease.

Periodontal disease is an oral inflammatory disease that destroys the tooth supporting periodontal tissues resulting in tooth loss. Porphyromonas gingivalis is a keystone pathogen that plays a significant role in periodontitis. In previous studies, resveratrol has shown significant results by targeting inflammatory and adhesive markers. Virulence factors of P. gingivalis play an important role in the bacterial adhesion and colonization. In this study, we aimed to demonstrate the anti-biofilm and anti-bacterial activity of resveratrol and also study the effect of resveratrol on the expression of virulence factor genes of P. gingivalis using reverse transcriptase polymerase chain reaction (RT-PCR). The anti-microbial and anti-biofilm activity of resveratrol on P. gingivalis was carried out by broth microdilution assay and biofilm adhesion reduction-crystal violet assay, respectively. We carried out the gene expression analysis by RT-PCR with the P. gingivalis treated compound to analyze the change in the expression of virulence factors: fimbriae and gingipain. Minimal inhibitory concentrations (MIC) of resveratrol against P. gingivalis and other clinical strains are in the range of 78.12-156.25 μg/mL. Resveratrol dose-dependently prevented the biofilm formation and also attenuated the virulence of P. gingivalis by reducing the expression of virulence factor genes such as fimbriae (type II and IV) and proteinases (kgp and rgpA). Resveratrol demonstrated superior anti-bacterial and anti-biofilm activity against P. gingivalis. There was significant reduction in the expression of fimbriae and gingipain with the resveratrol-treated compound. The results suggest that resveratrol, due to its multiple actions, may become a simple and inexpensive therapeutic strategy for treating periodontal disease.

Melanoma is the most deadly and life-threatening form of skin cancer with progressively higher rates of incidence worldwide. The objective of the present investigation is to develop and to statisti...

Objective: Cancer stem cells (CSCs) belong to a subpopulation of undifferentiated cells present within tumors that have the potential to regenerate, differentiate, maintenance of pluripotency, drug resistance, and tumorigenicity when transplanted into an innate host. These can influence the growth and behavior of these tumors and are used to investigate the initiation, progression, and treatment strategies of laryngeal cancer. Research on CSC science and targeted therapies were hinge on their isolation and/or enrichment procedures. The object of the study is to isolate cancer stem cells from primary laryngeal carcinoma (CSCPLC) by tumor spheres enrichment. We checked the properties of self-renewal, stemness, clonogenicity, and chemotherapeutic resistance. Materials and Methods: We performed tumor sphere formation assay (primary, secondary, and tertiary) chemotherapy resistance by 3-(4,5-dimethylthiazol-2-yl)-2,5-diphenyltetrazolium bromide assay were performed to evaluate the CSC cells. Immunofluorescence for stem cell markers (CD133+, CD44+) and gene expression of stem cell markers for CD133+, CD44+, OCT4, SOX2, and NANOG was done using the real-time polymerase chain reaction technique. Results: We were able to isolated CSC subpopulations from PLC cell lines by the tumor sphere method. These cells exhibited good primary, secondary, and tertiary tumor sphere formation efficiency and also disclosed a resistant index of more than 2. Immunofluorescence for stem cell markers (CD133+ and CD44+) confirms the presence of CSC. There was significantly higher mRNA expression of stem cell markers in CSC enriched subpopulations compared to the parental cell lines. Conclusion: We conclude that tumor spheres enrichment is an efficient, economical, and reliable approach for the isolation and characterization of CSC from PLC cell lines. These cells demonstrated the properties of self-renewal, stemness, clonogenicity, and chemotherapeutic resistance.

Porphyromonas gingivalis is regarded as a &ldquo, keystone pathogen&rdquo, in periodontitis. The fimbria assists in the initial attachment, biofilm organization, and bacterial adhesion leading to the invasion and colonization of host epithelial cells. The present study aimed to investigate the occurrence of fimA genotypes in patients with chronic periodontitis and healthy individuals in the Indian population, and to study their association with the number of P. gingivalis cells obtained in subgingival plaque samples of these subjects. The study comprised 95 samples from the chronic periodontitis (CP) group and 35 samples from the healthy (H) group, which were detected positive for P. gingivalis in our previous study. Fimbrial genotyping was done by PCR and PCR-restriction fragment length polymorphism (RFLP). The fimA type II was more prevalent in the CP group (55.89%), followed by type IV (30.52%), whereas in the H group, type I was the most prevalent fimbria (51.42%). The quantity of P. gingivalis cells increased with the presence of fimA types II and III. Our results suggest a strong relationship between fimA types II and IV and periodontitis, and between type I and the healthy condition. The colonization of organisms was increased with the occurrence of type II in deep periodontal sites, which could play an important role in the progression of the disease.

The present study deals with the formulation and evaluation of Solid Lipid Nanoparticles (SLNs) based transdermal patch of colchicine. Colchicine loaded SLNs (C-SLNs) were prepared by ultrasonication method and optimized with respect to the concentration of lipid and surfactant by 32 full factorial design. Properties like particle size, shape, zeta potential, polydispersity index and percentage entrapment efficiency were determined. The transdermal potential of C-SLNs was evaluated by ex-vivo drug permeation using Wistar rat skin and fluorescence microscopy. The optimized formulation was fabricated into the transdermal patch and in-vivo pharmacokinetic and pharmacodynamic study (subcutaneous air pouch model) was carried out. Particle size, zeta potential and entrapment efficiency of the optimized formulation were found to be 107 nm, (−) 17.4 mV and 37.25% respectively. Formulation F5 showed highest transdermal flux (8.76 μg/cm2/hrs). Fluorescence study confirms that SLNs have more penetration potential through the skin. AUC0→∞ of C-SLNs patch was improved by 2.84 times more than colchicine patch. Significant (p

Since time immemorial, turmeric has been widely marketed and consumed as dietary supplement due to its diverse medicinal properties. Curcuminoids—comprising a mixture of curcumin (CUR), demethoxycurcumin (DMC), and bisdemethoxycurcumin (BDMC)—are the prime bioactive constituents of turmeric. However, the usage of curcuminoids is limited by their chemical instability. The lack of information on comparative stability profiles of curcuminoids (in pure and mixture form) prompted us to study how pure curcuminoids and their mixtures behave under different stress degradation conditions. The order of stability of curcuminoids when exposed to acidic, alkaline, and oxidative degradation was found to be as follows: BDMC > DMC > CUR. While the pure and mixture forms of curcuminoids were stable against heat, they completely degraded upon exposure to sunlight. The degradation extent of curcuminoids (in mixture form) was substantially less as compared to their pure form; therefore, this suggested the synergistic stabilizing influence of DMC and BDMC in the curcuminoids’ mixture.

Porphyromonas gingivalis is a keystone pathogen and major colonizer in host tissue which plays a pivotal role in periodontitis among the other polymicrobial infections. Increasing facts demonstrate that curcumin has antibacterial activity and anti-biofilm effect against the periodontopathogens through diverse mechanisms that have a positive impact on periodontal health. The present study was aimed to elucidate the effect of curcumin on biofilm formation and virulence factor gene expression of P. gingivalis. By using gene expression studies, we exploited the mechanism of anti-biofilm effects of curcumin on P. gingivalis. The minimum inhibitory concentration and minimum bactericidal concentration of curcumin for both ATCC and clinical strains of P. gingivalis were found to be 62.5 and 125 µg ml

Melanoma is the most deadly and life-threatening form of skin cancer with progressively higher rates of incidence worldwide. The objective of the present investigation is to develop and to statistically optimize and characterize curcumin (CUR) loaded ethosomes for treatment of melanoma. A two factor, three level (3

A single robust RP-HPLC method was developed and validated as per ICH guidelines for the accurate quantification of curcuminoids in commercial turmeric products, ayurvedic medicines, and nanovesicular systems. The proposed chromatographic method was found to be specific, linear (r2 ≥ 0.999), precise at intra- and inter-day levels (%RSD −1 for curcumin (CUR), 9.24 and 30.80 n g mL−1 for demethoxycurcumin (DMC), and 6.48 and 21.61 n g mL−1 for bisdemethoxycurcumin (BDMC) respectively. Amongst different commercial turmeric products and ayurvedic medicines tested, the contents of curcumin (3.54 ± 0.06 to 25.8 ± 0.08 mg g −1), demethoxycurcumin (1.28 ± 0.02 to 9.97 ± 0.03 mg g −1) and bisdemethoxycurcumin (0.50 ± 0.01 to 5.97 ± 0.01 mg g −1) varied significantly. The developed method was effectively applied to the determination of encapsulation efficiency (EE) of curcuminoids (ranged between 84.33 ± 3.50% to 96.59 ± 2.53%) in the nanovesicular systems. In conclusion, the reported method is suitable for analysis of curcuminoids in a wide variety of turmeric products and used for the quality control of products that contain curcuminoids.

A single robust reversed-phase high-performance liquid chromatography (RP-HPLC) method was developed and validated as per International Conference on Harmonization guidelines for the accurate quantification of curcuminoids in commercial turmeric products, Ayurvedic medicines, and nanovesicular systems. The proposed chromatographic method was found to be specific, linear (r2 ≥ 0.999), precise at intra- and inter-day levels (percentage relative standard deviation −1 for curcumin, 9.24 and 30.80 ng mL−1 for demethoxycurcumin, and 6.48 and 21.61 ng mL−1 for bisdemethoxycurcumin, respectively. Among different commercial turmeric products and Ayurvedic medicines tested, the contents of curcumin (3.54 ± 0.06–25.8 ± 0.08 mg g−1), demethoxycurcumin (1.28 ± 0.02–9.97 ± 0.03 mg g−1), and bisdemethoxycurcumin (0.50 ± 0.01–5.97 ± 0.01 mg g−1) varied significantly. The developed method was effectively applied to the determination of encapsulation efficiency of curcuminoids (ranged between 84.33 ± 3.50 and 96.59 ± 2.53%) in the nanovesicular systems. In conclusion, the reported method is suitable for the analysis of curcuminoids in a wide variety of turmeric products and used for the quality control of products that contain curcuminoids.

Saraca asoca (Roxb.) De Wilde (Ashoka) is a highly valued endangered medicinal tree species from Western Ghats of India. Besides treating cardiac and circulatory problems, S. asoca provides immense relief in gynecological disorders. Higher price and demand, in contrast to the smaller population size of the plant, have motivated adulteration with other plants such as Polyalthia longifolia (Sonnerat) Thwaites. The fundamental concerns in quality control of S. asoca arise due to its part of medicinal value (Bark) and the chemical composition. Phytochemical fingerprinting with proper selection of analytical markers is a promising method in addressing quality control issues. In the present study, high-performance liquid chromatography of phenolic compounds (gallic acid, catechin, and epicatechin) coupled to multivariate analysis was used. Five samples each of S. asoca, P. longifolia from two localities alongside five commercial market samples showed evidence of adulteration. Subsequently, multivariate hierarchical cluster analysis and principal component analysis was established to discriminate the adulterants of S. asoca. The proposed method ascertains identification of S. asoca from its putative adulterant P. longifolia and commercial market samples. The data generated may also serve as baseline data to form a quality standard for pharmacopoeias. SUMMARY Simultaneous quantification of gallic acid, catechin, epicatechin from Saraca asoca by high-performance liquid chromatographyDetection of S. asoca from adulterant and commercial samplesUse of analytical method along with a statistical tool for addressing quality issues. Abbreviations used: HPLC: High Performance Liquid Chromatography; RP-HPLC: Reverse Phase High Performance Liquid Chromatography; CAT: Catechin; EPI: Epicatechin; GA: Gallic acid; PCA: Principal Component Analysis.

Saraca asoca (Roxb.) De Wilde (Ashoka) is a highly valued endangered medicinal tree species from Western Ghats of India. Besides treating cardiac and circulatory problems, S. asoca provides immense relief in gynecological disorders. Higher price and demand, in contrast to the smaller population size of the plant, have motivated adulteration with other plants such as Polyalthia longifolia (Sonnerat) Thwaites. The fundamental concerns in quality control of S. asoca arise due to its part of medicinal value (Bark) and the chemical composition. Phytochemical fingerprinting with proper selection of analytical markers is a promising method in addressing quality control issues. In the present study, high-performance liquid chromatography of phenolic compounds (gallic acid, catechin, and epicatechin) coupled to multivariate analysis was used. Five samples each of S. asoca, P. longifolia from two localities alongside five commercial market samples showed evidence of adulteration. Subsequently, multivariate hierarchical cluster analysis and principal component analysis was established to discriminate the adulterants of S. asoca. The proposed method ascertains identification of S. asoca from its putative adulterant P. longifolia and commercial market samples. The data generated may also serve as baseline data to form a quality standard for pharmacopoeias. Abbreviations used: HPLC: High Performance Liquid Chromatography; RP-HPLC: Reverse Phase High Performance Liquid Chromatography; CAT: Catechin; EPI: Epicatechin; GA: Gallic acid; PCA: Principal Component Analysis.