V S N Malleswara Rao Peram
@chips.ac.in
Associate Professor, Department of Pharmaceutics
Chebrolu Hanumaiah Institute of Pharmaceutical Sciences
RESEARCH INTERESTS
Topical and Transdermal drug delivery, metallic nanoparticles, natural products, analytical method development and validation, quality by design
Scopus Publications
Scholar Citations
Scholar h-index
Scholar i10-index
Scopus Publications
Arzoo Toyeba Jamadar, Malleswara Rao Peram, Nagesh Chandrasekhar, Ankita Kanshide, Vijay M. Kumbar, and Prakash V. Diwan
Springer Science and Business Media LLC
Malleswara Rao Peram, Chandrakant Dhananjay, Nagesh Chandrasekhar, Vijay M Kumbar, Vidyadhara Suryadevara, Sachin R Patil, and Sally A El-Zahaby
Informa UK Limited
Vijay M. Kumbar, Uday Muddapur, Abdullatif Bin Muhsinah, Saad Ali Alshehri, Mohammed Merae Alshahrani, Ibrahim Abdullah Almazni, Manohar S. Kugaji, Kishore Bhat, Malleswara Rao Peram, Mater H. Mahnashi,et al.
MDPI AG
Oral cancer has a high mortality rate, which is mostly determined by the stage of the disease at the time of admission. Around half of all patients with oral cancer report with advanced illness. Hitherto, chemotherapy is preferred to treat oral cancer, but the emergence of resistance to anti-cancer drugs is likely to occur after a sequence of treatments. Curcumin is renowned for its anticancer potential but its marred water solubility and poor bioavailability limit its use in treating multidrug-resistant cancers. As part of this investigation, we prepared and characterized Curcumin nanomicelles (CUR-NMs) using DSPE-PEG-2000 and evaluated the anticancer properties of cisplatin-resistant cancer cell lines. The prepared CUR-NMs were sphere-shaped and unilamellar in structure, with a size of 32.60 ± 4.2 nm. CUR-NMs exhibited high entrapment efficiency (82.2%), entrapment content (147.96 µg/mL), and a mean zeta potential of −17.5ζ which is considered moderately stable. The cellular uptake and cytotoxicity studies revealed that CUR-NMs had significantly higher cytotoxicity and cellular uptake in cisplatin drug-resistant oral cancer cell lines and parental oral cancer cells compared to plain curcumin (CUR). The DAPI and FACS analysis corroborated a high percentage of apoptotic cells with CUR-NMs (31.14%) compared to neat CUR (19.72%) treatment. Conclusively, CUR-NMs can potentially be used as an alternative carrier system to improve the therapeutic effects of curcumin in the treatment of cisplatin-resistant human oral cancer.
Ankita Kanshide, Malleswara Rao Peram, Nagesh Chandrasekhar, Arzoo Jamadar, Vijay Kumbar, and Manohar Kugaji
Springer Science and Business Media LLC
KishoreG Bhat, VijayMahadev Kumbar, UdayM Muddapur, HR Shwetha, ManoharS Kugaji, MalleswaraRao Peram, and Santosh Dindawar
Medknow
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.
Malleswara Rao Peram, Sachin R. Patil, Vijay M. Kumbar, Manohar S. Kugaji, Kishore G. Bhat, Prakash V. Diwan, and Sunil Jalalpure
Bentham Science Publishers Ltd.
Background: Linagliptin (LNG) is an oral hypoglycemic agent that acts by inhibiting the enzyme dipeptidyl peptidase - 4 (DPP-4) and reduces blood sugar levels in type-II diabetic patients. To date, the literature presents few analytical methods for the determination of LNG. However, no reversed phase-high performance liquid chromatography (RP-HPLC) method has been reported for the determination of LNG in nanotransfersomes and in vitro skin permeation samples. Objective: The present study involves the development and validation of RP-HPLC method to quantify LNG in both nanotransfersomes and in vitro skin permeation and deposition samples. Methods: The chromatographic analysis was performed on Luna C18 (2) column (250 x 4.6 mm, 5μm particle size) with a mobile phase consisting of a mixture of methanol: 0.2% orthophosphoric acid (50:50, v/v) at a flow rate of 1.0 mL/min, detection wavelength of 227 nm, and column temperature of 40 °C. Results: The method was found to be specific, linear (r2 ≥ 0.999; 2-12 μg/mL), precise at both intra and inter-day levels (percentage relative standard deviation; % RSD < 2.00), accurate (percentage recovery 100.21-103.83%), and robust. The detection and quantification limits were 0.27 and 0.82 μg/mL, respectively. The mean % entrapment efficiency and the cumulative amount of LNG permeated across the rat skin from different transfersomal formulations ranged between 40.78 ± 2.54 % to 52.26 ± 2.15 % and 79.54 ± 16.67 to 200.74 ± 35.13 μg/cm2 respectively. Conclusion: The method was successfully applied to determine the entThe method was successfully applied to determine the entrapment efficiency, in vitro skin permeation and deposition behavior of LNG-nanotransfersomes.rapment efficiency, in vitro skin permeation and deposition behavior of LNG-nanotransfersomes.
Vijay M. Kumbar, Malleswara Rao Peram, Manohar S. Kugaji, Tejas Shah, Sanjivani P. Patil, Uday M. Muddapur, and Kishore G. Bhat
Springer Science and Business Media LLC
Vijay M. Kumbar, Uday M. Muddapur, Kishore G. Bhat, Shwetha H.R., Manohar S. Kugaji, and Malleswara Rao Peram
SAGE Publications
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.
Pinjari Hameeda, Sandeep Katti, Rajkishore Jammalamadugu, Kishore Bhatt, Malleswara Rao Peram, and Vijay Kumbar
SAGE Publications
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.
Manohar Kugaji, Uday Muddapur, Kishore Bhat, Vinayak Joshi, Manjunath Manubolu, Kavitha Pathakoti, Malleswara Rao Peram, and Vijay Kumbar
MDPI AG
Porphyromonas gingivalis is regarded as a “keystone pathogen” 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.
Manohar S. Kugaji, Uday M. Muddapur, Kishore G. Bhat, Vinayak M. Joshi, Vijay M. Kumbar, and Malleswara Rao Peram
SAGE Publications
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.
Malleswara Rao Peram, Sunil Jalalpure, Vijay Kumbar, Sachin Patil, Sumit Joshi, Kishore Bhat, and Prakash Diwan
Informa UK Limited
Abstract 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 (32) factorial design approach was employed for the optimization of ethosomes. The prepared ethosomes were evaluated for size, zeta potential, entrapment efficiency, in vitro skin permeation and deposition ability. The optimized ethosomal formulation was evaluated for in vitro cytotoxicity and cellular uptake studies using A375 human melanoma cells. The optimized formulation has imperfect round shaped unilamellar structures with a mean vesicle size of 247 ± 5.25 nm and an entrapment efficiency of 92.24 ± 0.20%. The in vitro skin permeation studies proved the superiority of ethosomes over the traditional liposomes in terms of the amount of drug permeated and deposited in skin layers. Fluorescence microscopy showed the enhanced penetration of ethosomes into the deeper layers of the skin. In vitro cytotoxicity and cellular uptake studies revealed that curcumin ethosomes have significantly improved cytotoxicity and cellular uptake in A375 human melanoma cell lines. The colony formation assay results showed that curcumin ethosomes have a superior antiproliferative effect as they effectively inhibit the clonogenic ability of A375 cells. The flow cytometry results indicate that curcumin ethosomes induce cell death in A375 cells by apoptosis mechanism. The present study provides a strong rationale and motivation for further investigation of newly developed curcumin ethosomes as a potential therapeutic strategy for melanoma treatment.
Manohar S. Kugaji, Vijay M. Kumbar, Malleswara Rao Peram, Sanjivini Patil, Kishore G. Bhat, and Prakash V. Diwan
Wiley
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.
Sumit Ashok Joshi, Sunil Satyappa Jalalpure, Amolkumar Ashok Kempwade, and Malleswara Rao Peram
Bentham Science Publishers Ltd.
Sumit Ashok Joshi, Sunil Satyappa Jalalpure, Amolkumar Ashok Kempwade, and Malleswara Rao Peram
Elsevier BV
Malleswara Rao Peram, Sunil S. Jalalpure, Sumit A. Joshi, Mahesh B. Palkar, and Prakash V. Diwan
Informa UK Limited
ABSTRACT 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 <2.0%), accurate (percentage recovery 99.14–102.29%), and robust. The limits of detection and quantification were found to be 7.40 and 24.70 ng mL−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. GRAPHICAL ABSTRACT
Malleswara R. Peram, Sunil S. Jalalpure, Mahesh B. Palkar, and Prakash V. Diwan
Springer Science and Business Media LLC
SandeepRamachandra Pai, Subarna Roy, Satisha Hegde, HarshaVasudev Hegde, SunilSatyappa Jalalpure, and MalleswaraRao Peram
Medknow
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.
Simon M. Loveday, Malleswara R. Peram, Harjinder Singh, Aiqian Ye, and Geoffrey B. Jameson
Royal Society of Chemistry (RSC)
We investigated the relationship between structure andin vitropeptic digestibility of heated and unheated β-lactoglobulin. Surprisingly, the native protein was digested in two distinct phases, which we hypothesise is due to the binding of an inhibitory peptide to the active site of pepsin, followed by a pH-gated transition that releases the inhibitory peptide.
Malleswara R. Peram, Simon M. Loveday, Aiqian Ye, and Harjinder Singh
American Dairy Science Association
In vitro gastric digestion of heat-induced aggregates of β-lactoglobulin (β-LG) in simulated gastric fluid was investigated using sodium dodecyl sulfate-PAGE (under nonreducing and reducing conditions), native PAGE, 2-dimensional electrophoresis, and size exclusion chromatography. Heating at 90°C significantly increased the digestibility of β-LG, with a high initial digestion rate followed by a relatively constant rate of digestion at a high enzyme:substrate (E:S) ratio of 3:1. At a low E:S ratio (1:6), the rate of digestion of β-LG was slower, and intermediate- and low-molecular-weight species could be seen. The high-molecular-weight nonnative aggregates (e.g., pentamers, tetramers, and trimers) were digested relatively rapidly, whereas some of the nonnative dimers were resistant to digestion and others were digested rapidly. The intermediate-molecular-weight species (21 to 23 kDa) were digested slowly. The digestibility of nonnative β-LG aggregates varied significantly depending on the E:S ratio and the types of aggregate. Further investigation is necessary to identify and characterize slowly digested dimers and species of intermediate molecular weight.
Jean-Christophe Colas, Wanlong Shi, V.S.N. Malleswara Rao, Abdelwahab Omri, M. Reza Mozafari, and Harjinder Singh
Elsevier BV
Kianoush Khosravi-Darani, Abbas Pardakhty, Hamid Honarpisheh, V.S.N. Malleswara Rao, and M. Reza Mozafari
Elsevier BV
RC Doijad, FV Manvi, V.S.N Malleswara Rao, and Prajakta Alase
OMICS Publishing Group
The poor bioavailability and therapeutic response exhibited by conventional ophthalmic solutions due to rapid precorneal elimination of drug may be overcome by the use of in situ gel-forming systems that are instilled as drops into the eye and undergo a sol-gel transition in the cul-de-sac. The present work describes the formulation and evaluation of an ophthalmic delivery system of an antibacterial agent, gatifloxacin, based on the concept of ion-activated systems. Sodium alginate was used as the gelling agent in combination with hydroxy propyl methyl cellulose (Methocel E50LV), which acted as a viscosity enhancing agent. The developed formulations were therapeutically efficacious, stable, non-irritant and provided sustained release of the drug over an eight hour period. The developed system is thus a viable alternative to conventional eye drops.