Browsing by Author "Ashish Kumar Agrawal"

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A Bacteriophage Microgel Effectively Treats the Multidrug-Resistant Acinetobacter baumannii Bacterial Infections in Burn Wounds
(Multidisciplinary Digital Publishing Institute (MDPI), 2023) Deepa Dehari; Aiswarya Chaudhuri; Dulla Naveen Kumar; Rohit Patil; Mayank Gangwar; Sonam Rastogi; Dinesh Kumar; Gopal Nath; Ashish Kumar Agrawal
Multidrug-resistant (MDR) Acinetobacter baumannii (A. baumannii) is one of the major pathogens present in burn wound infections. Biofilm formation makes it further challenging to treat with clinically available antibiotics. In the current work, we isolated the A. baumannii-specific bacteriophages (BPABΦ1), loaded into the chitosan microparticles followed by dispersion in gel, and evaluated therapeutic efficacy against MDR A. baumannii clinical strains. Isolated BPABΦ1 were found to belong to the Corticoviridae family, with burst size 102.12 ± 2.65 PFUs per infected host cell. The BPABΦ1 loaded chitosan microparticles were evaluated for quality attributes viz. size, PDI, surface morphology, in vitro release, etc. The developed formulation exhibited excellent antibiofilm eradication potential in vitro and effective wound healing after topical application. © 2023 by the authors.
A Bacteriophage-Loaded Microparticle Laden Topical Gel for the Treatment of Multidrug-Resistant Biofilm-Mediated Burn Wound Infection
(Springer Science and Business Media Deutschland GmbH, 2023) Deepa Dehari; Aiswarya Chaudhuri; Dulla Naveen Kumar; Meraj Anjum; Rajesh Kumar; Akshay Kumar; Dinesh Kumar; Gopal Nath; Ashish Kumar Agrawal
Klebsiella pneumoniae is regarded as one of the most profound bacteria isolated from the debilitating injuries caused by burn wounds. In addition, the multidrug resistance (MDR) and biofilm formation make treating burn patients with clinically available antibiotics difficult. Bacteriophage therapy has been proven an effective alternative against biofilm-mediated wound infections caused by MDR bacterial strains. In the current study, the bacteriophage (BPKPФ1) against MDR Klebsiella pneumoniae was isolated and loaded into the chitosan microparticles (CHMPs), which was later incorporated into the Sepineo P 600 to convert into a gel (BPKPФ1-CHMP-gel). BPKPФ1 was characterized for lytic profile, morphological class, and burst size, which revealed that the BPKPФ1 belongs to the family Siphoviridae. Moreover, BPKPФ1 exhibited a narrow host range with 128 PFU/host cell of burst size. The BPKPФ1-loaded CHMPs showed an average particle size of 1.96 ± 0.51 μm, zeta potential 32.16 ± 0.41 mV, and entrapment efficiency in the range of 82.44 ± 1.31%. Further, the in vitro antibacterial and antibiofilm effectiveness of BPKPФ1-CHMPs-gel were examined. The in vivo potential of the BPKPФ1-CHMPs-gel was assessed using a rat model with MDR Klebsiella pneumoniae infected burn wound, which exhibited improved wound contraction (89.22 ± 0.48%) in 28 days with reduced inflammation, in comparison with different controls. Data in hand suggest the potential of bacteriophage therapy to be developed as personalized therapy in case of difficult-to-treat bacterial infections. Graphical abstract: [Figure not available: see fulltext.]. © 2023, The Author(s), under exclusive licence to American Association of Pharmaceutical Scientists.
Alginate lyase immobilized chitosan nanoparticles of ciprofloxacin for the improved antimicrobial activity against the biofilm associated mucoid P. aeruginosa infection in cystic fibrosis
(Elsevier B.V., 2019) Krishna Kumar Patel; Muktanand Tripathi; Nidhi Pandey; Ashish Kumar Agrawal; Shilpkala Gade; Md. Meraj Anjum; Ragini Tilak; Sanjay Singh
Dense colonization of mucoid Pseudomonas aeruginosa within the self-secreted extracellular matrix (mainly alginate), called biofilm, is a principal reason for the failure of antimicrobial therapy in cystic fibrotic patients. Alginate is a key component in the biofilm of mucoid P. aeruginosa and responsible for surface adhesion and stabilization of biofilm. To overcome this problem, alginate lyase functionalized chitosan nanoparticles of ciprofloxacin were developed for the effective treatment of P. aeruginosa infection in cystic fibrosis patients. The developed nanoparticles were found to have desired quality attributes and demonstrated sustained release following the Higuchi release kinetics. Drug compatibility with the chitosan was confirmed by FTIR while powder X-ray diffraction analysis confirmed the entrapment of drug within the nanoparticle matrix. Lactose adsorbed NPs showed promising aerodynamic property. Nanoparticles showed prolonged MIC and significant reduction in biofilm aggregation and formation in planktonic bacterial suspension. Nanoparticles exhibited significantly higher inhibitory effect against biofilm of P. aeruginosa and reduced the biomass, thickness and density confirmed by confocal microscopy. Furthermore, developed nanoparticles were haemocompatible and did not exhibit any toxicity in vitro MTT assay and in vivo on lungs male Wistar rats. The data in hand collectively suggest the proposed strategy a better alternative for the effective treatment of cystic fibrosis infections. © 2019
Anacardic acid encapsulated solid lipid nanoparticles for Staphylococcus aureus biofilm therapy: chitosan and DNase coating improves antimicrobial activity
(Springer, 2021) Md Meraj Anjum; Krishna Kumar Patel; Deepa Dehari; Nidhi Pandey; Ragini Tilak; Ashish Kumar Agrawal; Sanjay Singh
Biofilm mediated bacterial infections are the key factors in the progression of infectious diseases due to the evolution of antimicrobial resistance. Traditional therapy involving antibiotics is not adequate enough for treatment of such infections due to the increased resistance triggered by biofilm. To overcome this challenge, we developed anacardic acid (Ana) loaded solid lipid nanoparticles (SLNs), further coated with chitosan and DNase (Ana-SLNs-CH-DNase). The DNase coating was hypothesized to degrade the e-DNA, while chitosan was coated to yield positively charged SLNs with additional adhesion to biofilms. The SLNs were developed using homogenization method and further evaluated for particle size, polydispersity index, zeta potential, and entrapment efficiency. Drug excipient compatibility was confirmed by using FT-IR study, while encapsulation of Ana in SLNs was confirmed by X-ray diffraction study. The SLNs demonstrated sustained release for up to 24 h and excellent stability at room temperature for up to 3 months. The developed SLNs were found non-toxic against human immortalized keratinocyte (HaCaT) cells while demonstrated remarkably higher antimicrobial efficacy against Staphylococcus aureus. Excellent effect of the developed SLNs on minimum biofilm inhibition concentration and minimum biofilm eradication concentration further confirmed the superiority of the developed formulation strategy. A significant (p < 0.05) reduction in biofilm thickness and biomass, as confirmed by confocal laser scanning microscopy, was observed in the case of developed SLNs in comparison with control. Cumulatively, the results suggest the enhanced efficacy of the developed formulation strategy to overcome the biofilm-mediated antimicrobial resistance. [Figure not available: see fulltext.] © 2020, Controlled Release Society.
Antibiofilm Potential of Silver Sulfadiazine-Loaded Nanoparticle Formulations: A Study on the Effect of DNase-I on Microbial Biofilm and Wound Healing Activity
(American Chemical Society, 2019) Krishna Kumar Patel; D. Bhavya Surekha; Muktanand Tripathi; Md. Meraj Anjum; M.S. Muthu; Ragini Tilak; Ashish Kumar Agrawal; Sanjay Singh
Biofilm resistance is one of the severe complications associated with chronic wound infections, which impose extreme microbial tolerance against antibiotic therapy. Interestingly, deoxyribonuclease-I (DNase-I) has been empirically proved to be efficacious in improving the antibiotic susceptibility against biofilm-associated infections. DNase-I hydrolyzes the extracellular DNA, a key component of the biofilm responsible for the cell adhesion and strength. Moreover, silver sulfadiazine, a frontline therapy in burn wound infections, exhibits delayed wound healing due to fibroblast toxicity. In this study, a chitosan gel loaded with solid lipid nanoparticles of silver sulfadiazine (SSD-SLNs) and supplemented with DNase-I has been developed to reduce the fibroblast cytotoxicity and overcome the biofilm-imposed resistance. The extensive optimization using the Box-Behnken design (BBD) resulted in the formation of SSD-SLNs with a smooth surface as confirmed by scanning electron microscopy and controlled release (83%) for up to 24 h. The compatibility between the SSD and other formulation excipients was confirmed by Fourier transform infrared, differential scanning calorimetry, and powder X-ray diffraction studies. Developed SSD-SLNs in combination with DNase-I inhibited around 96.8% of biofilm of Pseudomonas aeruginosa as compared to SSD with DNase-I (82.9%). In line with our hypothesis, SSD-SLNs were found to be less toxic (cell viability 90.3 ± 3.8% at 100 μg/mL) in comparison with SSD (Cell viability 76.9 ± 4.2%) against human dermal fibroblast cell line. Eventually, the results of the in vivo wound healing study showed complete wound healing after 21 days' treatment with SSD-SLNs along with DNase-I, whereas marketed formulations SSD and SSD-LSNs showed incomplete healing after 21 days. Data in hand suggest that the combination of SSD-SLNs with DNase-I is an effective treatment strategy against the biofilm-associated wound infections and accelerates wound healing. © 2019 American Chemical Society.
Asparagus racemosus root-derived carbon nanodots as a nano-probe for biomedical applications
(Springer, 2022) Gaurav Gopal Naik; Tarun Minocha; Abhineet Verma; Sanjeev Kumar Yadav; Satyen Saha; Ashish Kumar Agrawal; Sanjay Singh; Alakh N. Sahu
Nowadays, green nanotechnology has emerged as a crucial and eco-friendly approach to combat the problems affecting human health or the environment. It deals with the extensive utilization of plants owing to their cocktail of natural and chemical substances to eliminate adverse pollutants involved in the synthesis of nanomaterials. Herein, we report a facile one-step hydrothermal carbonization approach synthesizing fluorescent bluish-green carbon nanodots (CNDs) using Asparagus racemosus roots termed ARCD. The fluorescence quantum yield of ARCD was further improved by surface passivating it with a 1:1 ratio of Carrageenan (CAR) and Polyethylenimine (PEI), termed ARCCD. First, we demonstrate the application of ARCD and ARCCD as a nano-probe for sensing As3+ and Ag+. Second, we show an inhibitory effect on cancer cell proliferation against breast (MDA-MB-231) and cervical (SiHa) cancer. Also, no significant cytotoxicity was observed in the normal kidney (HEK 293) cells, indicating cytotoxic specificity of the developed CNDs. Third, we demonstrate antibacterial activity against clinically isolated multi-drug-resistant strains and free radicals scavenging potential. Next, we present the in vivo toxicity evaluation of these CNDs in Swiss albino mice, which exhibited minimal toxicity as confirmed by biochemical, hematological, and histological parameters. This study deals with an innovative approach to constructing a multifunctional nano-probe for potential biomedical and environmental applications. Graphical abstract: [Figure not available: see fulltext.]. © 2022, The Author(s), under exclusive licence to Springer Science+Business Media, LLC, part of Springer Nature.
Bacteriophage entrapped chitosan microgel for the treatment of biofilm-mediated polybacterial infection in burn wounds
(Elsevier B.V., 2023) Deepa Dehari; Dulla Naveen Kumar; Aiswarya Chaudhuri; Akshay Kumar; Rajesh Kumar; Dinesh Kumar; Sanjay Singh; Gopal Nath; Ashish Kumar Agrawal
Staphylococcus aureus (S. aureus) and Pseudomonas aeruginosa (P. aeruginosa) bacteria are most commonly present in burn wound infections. Multidrug resistance (MDR) and biofilm formation make it difficult to treat these infections. Bacteriophages (BPs) are proven as an effective therapy against MDR as well as biofilm-associated wound infections. In the present work, a naturally inspired bacteriophage cocktail loaded chitosan microparticles-laden topical gel has been developed for the effective treatment of these infections. Bacteriophages against MDR S. aureus (BPSAФ1) and P. aeruginosa (BPPAФ1) were isolated and loaded separately and in combination into the chitosan microparticles (BPSAФ1-CHMPs, BPPAФ1-CHMPs, and MBP-CHMPs), which were later incorporated into the SEPINEO™ P 600 gel (BPSAФ1-CHMPs-gel, BPPAФ1-CHMPs-gel, and MBP-CHMPs-gel). BPs were characterized for their morphology, lytic activity, burst size, and hemocompatibility, and BPs belongs to Caudoviricetes class. Furthermore, BPSAФ1-CHMPs, BPPAФ1-CHMPs, and MBP-CHMPs had an average particle size of 1.19 ± 0.11, 1.42 ± 0.21, and 2.84 ± 0.28 μm, respectively, and expressed promising in vitro antibiofilm eradication potency. The ultrasound and photoacoustic imaging in infected burn wounds demonstrated improved wound healing reduced inflammation and increased oxygen saturation following treatment with BPs formulations. The obtained results suggested that the incorporation of the BPs in the MP-gel protected the BPs, sustained the BPs release, and improved the antibacterial activity. © 2023 Elsevier B.V.
Chemical intervention for enhancing growth and reducing grain arsenic accumulation in rice
(Elsevier Ltd, 2021) Ashish Kumar Srivastava; Manish Pandey; Tejashree Ghate; Vikash Kumar; Munish Kumar Upadhyay; Arnab Majumdar; Abhay Kumar Sanjukta; Ashish Kumar Agrawal; Sutapa Bose; Sudhakar Srivastava; Penna Suprasanna
Thiourea supplementation downregulated aquaporins to restrict arsenic accumulation in seedlings. Foliar-applied thiourea enhanced yield and reduced grain arsenic accumulation. © 2021 Elsevier Ltd; Arsenic (As) is a ubiquitous environmental carcinogen that enters the human food chain mainly through rice grains. In the present study, we evaluated the potential of thiourea (TU; non-physiological reactive oxygen species scavenger) in mitigating the negative effects of arsenic (As) stress in indica rice variety IR64, with the overall aim to reduce grain As accumulation. At seedling stage, As + TU treatment induced the formation of more numerous and longer crown roots compared with As alone. The As accumulation in main root, crown root, lower leaf and upper leaf was significantly reduced to 0.1-, 0.14-, 0.16-, 0.14-fold, respectively in As + TU treated seedlings compared with those of As alone. This reduced As accumulation was also coincided with light-dependent suppression in the expression levels of aquaporins and photosynthesis-related genes in As + TU treated roots. In addition, the foliar-supplemented TU under As-stress maintained reducing redox conditions which decreased the rate of As accumulation in flag leaves and, eventually grain As by 0.53-fold compared with those of As treatment. The agronomic feasibility of TU was validated under naturally As contaminated sites of Nadia (West Bengal, India). The tiller numbers and crop productivity (kg seed/ha) of TU-sprayed plants were increased by 1.5- and 1.18-fold, respectively; while, grain As accumulation was reduced by 0.36-fold compared with those of water-sprayed control. Thus, this study established TU application as a sustainable solution for cultivating rice in As-contaminated field conditions. © 2021 Elsevier Ltd
Combination Therapy Comprising Paclitaxel and 5-Fluorouracil by Using Folic Acid Functionalized Bovine Milk Exosomes Improves the Therapeutic Efficacy against Breast Cancer
(MDPI, 2022) Dulla Naveen Kumar; Aiswarya Chaudhuri; Deepa Dehari; Anusmita Shekher; Subash C. Gupta; Shreyasi Majumdar; Sairam Krishnamurthy; Sanjay Singh; Dinesh Kumar; Ashish Kumar Agrawal
Paclitaxel (PAC) has been approved by FDA for clinical use (Taxol®), yet dose-dependent severe toxicity due to the adjuvant Cremophor EL® in combination with ethanol is a major drawback. The drawbacks of the current therapy can be overcome by (i) finding a suitable vehicle that cannot only bypass the above adjuvant but also be used to deliver drugs orally and (ii) combining the PAC with some other chemotherapeutics to have the enhanced therapeutic efficacy. In the current work, we have used folic acid (FA) functionalized bovine milk-derived exosomes for oral delivery of PAC in combination with 5-fluorouracil (5-FU). Exosomes before and after the drug loading were found to have a particle size in the range of 80–100 nm, polydispersity index (PDI ~0.20), zeta potential (~−25 mV), entrapment efficiency (~82%), practical drug loading (~28%) and sustained drug release for 48 h. Significant decreases in IC50 were observed in the case of exosomes loaded drugs which further improved following the FA functionalization. FA functionalized coumarin-6-loaded exosomes showed remarkably higher cellular uptake in comparison with free coumarin-6. Moreover, FA-functionalized drug-loaded exosomes showed a higher apoptotic index with better control over cell migration. Collectively, data suggested the enhanced efficacy of the combination following its loading to the folic acid functionalized exosomes against breast cancer. © 2022 by the authors.
Development of Anacardic Acid/hydroxypropyl-β-cyclodextrin inclusion complex with enhanced solubility and antimicrobial activity
(Elsevier B.V., 2019) Md Meraj Anjum; Krishna Kumar Patel; Nidhi Pandey; Ragini Tilak; Ashish Kumar Agrawal; Sanjay Singh
Anacardic Acid (AnAc), a key constituent of cashew nut shell liquid (CNSL), has been reported to exhibit antibacterial activity against S. aureus but its bioactivity and clinical applicability are limited owing to its poor water solubility and physicochemical stability. The aim of the present study was to develop inclusion complex of AnAc (C15:3) and hydroxypropyl-β-cyclodextrin (HP-β-CD) to improve its aqueous solubility and antimicrobial activity. The inclusion complex was prepared using the co-evaporation method considering different molar ratios of AnAc (C15:3) and HP-β-CD. The prepared inclusion complex was characterized using FT-IR, DSC, XRD, SEM, and 1H NMR, which provided appropriate evidence to confirm the formation of the inclusion complex. The prepared inclusion complex demonstrated ~2009 times improved aqueous solubility. The developed complex maintained the antimicrobial activity of pure AnAc against S. aureus. AnAc/HP-β-CD complex exhibited excellent biofilm dispersal potential against mature biofilms. Confocal laser scanning microscopy (CLSM) demonstrated a remarkable decrease in bacterial biomass and thickness upon treatment with the inclusion complex. Conclusively, the developed inclusion complex of Anacardic Acid (C15:3) can be effectively used to overcome the poor aqueous solubility and improve the therapeutic efficacy for many other indications. © 2019
DNase-I functionalization of ciprofloxacin-loaded chitosan nanoparticles overcomes the biofilm-mediated resistance of Pseudomonas aeruginosa
(Springer Science and Business Media Deutschland GmbH, 2020) Krishna Kumar Patel; Ashish Kumar Agrawal; Md. Meraj Anjum; Muktanand Tripathi; Nidhi Pandey; Sankha Bhattacharya; Ragini Tilak; Sanjay Singh
Pseudomonas aeruginosa infection in cystic fibrosis (CF) is a major complication which aggravates the disease complexity and progression. The improvement of antimicrobial therapy against the P. aeruginosa biofilm infection in CF by dissembling the extracellular matrix is a fundamental concept of this study. Chitosan nanoparticle was successfully fabricated and characterized. Furthermore, various in vitro antimicrobial efficacy, biofilm dispersal potential, and in vitro as well as in vivo toxicity were assessed. The chitosan nanoparticles of ciprofloxacin functionalized with DNase-I had spherical shape with desired quality attributes (particle size—212.3 ± 8.9; polydispersity index—0.288 ± 0.06, zeta potential—14.6 ± 1.3, and entrapment efficiency—53.7 ± 3.8%). The developed DNase-I functionalized chitosan nanoparticles laden with ciprofloxacin demonstrated substantial and prolonged microbial inhibition, efficiently prevented the biofilm development, and possessed the excellent biofilm dispersal potential. Moreover, the confocal study demonstrated that the biofilm treated with DNase-I functionalized chitosan nanoparticles of ciprofloxacin had minimum biofilm thickness, biomass, and microbial density compared to the other treatment groups. Additionally, the developed formulation was found to be safe and had minimal in vitro as well as in vivo toxicity. Thus, the findings of this study suggests ciprofloxacin-loaded chitosan nanoparticles functionalized with DNase-I as an effective and safe treatment approach for the P. aeruginosa infection in CF. © 2019, King Abdulaziz City for Science and Technology.
Fiber and textile in drug delivery to combat multidrug resistance microbial infection
(Elsevier, 2022) Deepa Dehari; Aiswarya Chaudhuri; Dulla Naveen Kumar; Gopal Nath; Ashish Kumar Agrawal
Medical textile is a booming area within the textile sector that has significantly increased research focus over the past 10 years. It is rapidly becoming a vital part of the textile business. Antimicrobial resistance (AMR) or multidrug resistance (MDR) has emerged as a major health concern and a big challenge issue throughout the world in recent years. After decades of neglect, the AMR has now gained international notice. Antibiotic-resistant strains have posed a danger to science and medicine’s achievements since they render traditional antimicrobial therapies ineffective. Various fiber and textile fabrication methods may be utilized to develop and create fibrous components, needed for targeted and controlled medication delivery systems, therefore contributing to the fighting strategies against AMR. Antimicrobial agents of several types, such as quaternary ammonium compounds (QACs), polybiguanides, triclosan, metals, and their oxides, natural dyes, natural polymers, herbal extracts, and essential oils, have been utilized and administered with textile fibers. Because of technological breakthroughs, particularly but not primarily in nanotechnology, the specialized healthcare textile sector is undergoing tremendous evolution and extension, especially in fields such as wound and tissue repair, personal protective clothing or equipment (PPE), sterile gauze, surgical hosiery, and implantable textiles (e.g., vascular grafts, surgical suture, reabsorbable polymers). Additionally, to be a vital force against microbes, every antimicrobial therapy applied to the textile must be nontoxic both for human health and the ecosystem. The book chapters introduce antimicrobial textiles, antimicrobial nanofibers, and metal-integrated nanoparticles-based fabrics as solutions to the AMR and MDR problems. © 2023 Elsevier Ltd. All rights reserved.
Impact of the Drug Loading Method on the Drug Distribution and Biological Efficacy of Exosomes
(Springer Science and Business Media Deutschland GmbH, 2023) Dulla Naveen Kumar; Aiswarya Chaudhuri; Dinesh Kumar; Sanjay Singh; Ashish Kumar Agrawal
Exosomes are biological nanovesicles that are intrinsically loaded with thousands of biomacromolecules and are principally responsible for cell-to-cell communication. Inspired by the natural payload, they have been extensively investigated as drug delivery vehicles; however, the drug distribution, whether into or onto exosomes, is still debatable. In the present work, we have tried to investigate it systemically by selecting 5-fluorouracil (5-FU) (hydrophilic) and paclitaxel (PAC) (hydrophobic), drugs with very different physicochemical characteristics, for the loading to the exosomes. Exosomes were obtained from bovine milk, and the drugs were loaded using three different methods: incubation, sonication, and triton x-100. The particle size was found to be approximately 100 nm in all the cases; however, the highest drug loading was found in the sonication method. Fluorescence spectrophotometer, EDX analysis, EDX mapping, XPS, and XRD analysis indicated the possible presence of more drugs over the surface in the case of the incubation method. Drugs loaded by the sonication method had more controlled release than simple incubation and triton x-100. The method of drug loading had an insignificant effect on the cytotoxicity while in line with our previous observation, the combination (PAC and 5-FU) exhibited synergism as evidenced by ROS assay, colony formation assay, and mitochondrial membrane potential assay. Graphical Abstract: [Figure not available: see fulltext.]. © 2023, The Author(s), under exclusive licence to American Association of Pharmaceutical Scientists.
Introduction to smart polymers and their application
(Elsevier, 2022) Aiswarya Chaudhuri; Kamalpreet Kaur Sandha; Ashish Kumar Agrawal; Prem N. Gupta
Adaptation is the key feature that permits the living system to respond to the external environment and get accustomed to the changing conditions. In a similar way, scientists have tried to develop polymers that could mold themselves according to the external/surrounding changes (stimulus) and form a more compatible system. This leads to the emergence of “smart polymers” that undergo physical or chemical changes in response to small environmental changes (temperature, light, pH, etc.), which proves the versatility as well as the sensitivity of the smart polymers. It could be stated that smart polymers lead to the development of much tunable, programmable, and accurate systems. Smart polymers were extensively used in the field of biomedicine which includes tissue engineering, the cell culture system, drug delivery system, sensors or actuators, and biomedical devices. This chapter provides a wide overview of smart polymers and the most interesting applications developed so far. © 2023 Elsevier Inc. All rights reserved.
Luliconazole Nail Lacquer for the Treatment of Onychomycosis: Formulation, Characterization and In Vitro and Ex Vivo Evaluation
(Springer Science and Business Media Deutschland GmbH, 2022) Deepa Dehari; Abhishesh Kumar Mehata; Vishnu Priya; Dharmnath Parbat; Deepak Kumar; Anand Kumar Srivastava; Sanjay Singh; Ashish Kumar Agrawal
Onychomycosis is the most common fungal infection of the nail affecting the skin under the fingertips and the toes. Currently, available therapy for onychomycosis includes oral and topical therapies, either alone or in combination. Oral antifungal medication has been associated with poor drug bioavailability and potential gastrointestinal and systemic side effects. The objective of this study was to prepare and evaluate the luliconazole nail lacquer (LCZ-NL) for the effective treatment of onychomycosis. In the current work, LCZ-NL was formulated in combination with penetration enhancers to overcome poor penetration. A 32 full factorial formulation design of experiment (DOE) was applied for optimization of batches with consideration of dependent (drying time, viscosity, and rate of drug diffusion) and independent (solvent ratio and film former ratio) variables. The optimized formulation was selected based on drying time, viscosity, and rate of drug diffusion. The optimized formulation was further evaluated for % non-volatile content assay, smoothness of flow, water resistance, drug content, scanning electron microscope (SEM), atomic force microscope (AFM), X-ray diffraction (XRD), differential scanning calorimetry (DSC), in vitro drug release, ex vivo transungual permeation, antifungal efficacy, and stability study. The optimized LCZ-NL contained 70:30 solvent ratio and 1:1 film former ratio and was found to have ~ 1.79-fold higher rate of drug diffusion in comparison with LULY™. DSC and XRD studies confirmed that luliconazole retains its crystalline property in the prepared formulation. Antifungal study against Trichophyton spp. showed that LCZ-NL has comparatively higher growth inhibition than LULY™. Hence, developed LCZ-NL can be a promising topical drug delivery system for treating onychomycosis. Graphical abstract: [Figure not available: see fulltext.]. © 2022, The Author(s), under exclusive licence to American Association of Pharmaceutical Scientists.
Piperine loaded metal organic frameworks reverse doxorubicin induced chemobrain in adult zebrafish
(Elsevier B.V., 2023) Sanchit Arora; Vishal Kumar; Lakshay Kapil; Ashish Kumar Agrawal; Arti Singh; Charan Singh
The study's primary goal was to enhance medicinal potential of piperine (PIP)-loaded zeolitic imidazolate frameworks-8 (PIP@ZIF-8) against doxorubicin (DOX)-induced cognitive impairments in zebrafish. Herein, PIP@ZIF-8 was synthesized via easy, economical and reproducible ultrasonication method followed by spray drying technology. ZIF-8's structural integrity has been confirmed by PXRD, and even after PIP was encapsulated, the structure of ZIF-8 remained unchanged. Pure ZIF-8 and PIP@ZIF-8 were subjected to TEM analysis, which revealed hexagonal morphology with a nanosize range. FTIR and UV–Visible spectroscopy studies confirmed the drug loading of ZIF-8. Studies on in vitro release revealed 71.48 ± 7.21% and 34.56 ± 5.35% PIP release from PIP@ZIF-8 and unformulated PIP, respectively in pH 7.4. The highest antioxidant scavenging results were obtained with vitamin C (73.77 ± 6.7%) at an intensity of 200 μg/ml, though it was 65.09 ± 2.5% and 57.99 ± 3.1% for PIP@ZIF-8 and PIP, respectively. In vivo studies on zebrafish showed that DOX administration remarkably impaired cognitive activity in T-Maze, and downregulated spatial memory and locomotor activity in the open field test. In addition, DOX administration caused a downregulation in GSH and SOD levels and increase in LPO, AChE and TNF-α levels compared to the vehicle group along with changes in brain histopathology. Further, PIP@ZIF-8 reversed the DOX-induced cognitive impairments by its antioxidant and neuroprotective properties. It can be concluded that PIP@ZIF-8 has a promising therapeutic potential against the chemotherapy-induced cognitive impairments in zebrafish. © 2023 Elsevier B.V.
Preformulation Challenges: The Concept Behind the Selection, Design and Preparation of Nanoformulations
(Springer International Publishing, 2020) Krishna Kumar Patel; Ashish Kumar Agrawal; Sanjay Singh
Nano-drug delivery carriers have fascinated researchers worldwide for the last two to three decades. The nanoformulations are preferred over conventional dosage forms as they provide improved drug solubility, bioavailability, drug stability under adverse external or physiological conditions, controlled drug release for prolonged action and target specificity using ligand binding and many more. Multiple essential aspects must be carefully studied and implemented for the development of efficient drug delivery vehicles. These aspects include rationale of nanoparticle preparation; use of polymeric or lipid nanoparticle; selection of polymer, lipid and excipients; method of preparation; screening of critical formulation or process parameters that affect the critical quality attributes; optimization of process to obtain the desired formulation characteristics; characterization of nanoparticles; strategy to improve the low entrapment efficiency and stability, etc. This chapter comprehensively summarizes all these aspects of nanoformulation development and proposes solutions for these challenges. Although a variety of nanoformulations have been described in literature, this chapter is restricted to discuss in detail the liposomes and polymeric and lipid nanoparticles. © Springer Nature Switzerland AG 2020.
Quality by design–based development and optimization of fourth-generation ternary solid dispersion of standardized Piper longum extract for melanoma therapy
(Springer, 2023) Debadatta Mohapatra; Dulla Naveen Kumar; Singh Shreya; Vivek Pandey; Pawan K. Dubey; Ashish Kumar Agrawal; Alakh N Sahu
The study aimed to enhance the solubility, dissolution, and oral bioavailability of standardized Piper longum fruits ethanolic extract (PLFEE) via fourth-generation ternary solid dispersion (SD) for melanoma therapy. With the use of solvent evaporation method, the standardized PLFEE was formulated into SD, optimized using Box-Wilson’s central composite design (CCD), and evaluated for pharmaceutical performance and in vivo anticancer activity against melanoma (B16F10)–bearing C57BL/6 mice. The optimized SD showed good accelerated stability, high yield, drug content, and content uniformity for bioactive marker piperine (PIP). The X-ray diffraction (XRD), differential scanning calorimetry (DSC), polarized light microscopy (PLM), and selected area electron diffraction (SAED) analysis revealed its amorphous nature. The attenuated total reflectance-Fourier transform infrared spectroscopy (ATR-FTIR) and high-performance thin layer chromatography (HPTLC) revealed the compatibility of excipients with the PLFEE. The contact angle measurement and in vitro dissolution study revealed excellent wetting of SD and improved dissolution profile as compared to the plain PLFEE. The in vivo oral bioavailability of SD reflected a significant (p < 0.05) improvement in bioavailability (F rel = 188.765%) as compared to plain extract. The in vivo tumor regression study revealed the improved therapeutic activity of SD as compared to plain PLFEE. Further, the SD also improved the anticancer activity of dacarbazine (DTIC) as an adjuvant therapy. The overall result revealed the potential of developed SD for melanoma therapy either alone or as an adjuvant therapy with DTIC. Graphical Abstract: [Figure not available: see fulltext.] © 2023, Controlled Release Society.
Recent advancements in nanotechnology-based bacteriophage delivery strategies against bacterial ocular infections
(Elsevier GmbH, 2023) Rohit Patil; Deepa Dehari; Aiswarya Chaudhuri; Dulla Naveen Kumar; Dinesh Kumar; Sanjay Singh; Gopal Nath; Ashish Kumar Agrawal
Antibiotic resistance is growing as a critical challenge in a variety of disease conditions including ocular infections leading to disastrous effects on the human eyes. Staphylococcus aureus (S. aureus) mediated ocular infections are very common affecting different parts of the eye viz. vitreous chamber, conjunctiva, cornea, anterior and posterior chambers, tear duct, and eyelids. Blepharitis, dacryocystitis, conjunctivitis, keratitis, endophthalmitis, and orbital cellulitis are some of the commonly known ocular infections caused by S. aureus. Some of these infections are so fatal that they could cause bilateral blindness like panophthalmitis and orbital cellulitis, which is caused by methicillin-resistant S. aureus (MRSA) and vancomycin-resistance S. aureus (VRSA). The treatment of S. aureus infections with known antibiotics is becoming gradually difficult because of the development of resistance against multiple antibiotics. Apart from the different combinations and formulation strategies, bacteriophage therapy is growing as an effective alternative to treat such infections. Although the superiority of bacteriophage therapy is well established, yet physical factors (high temperatures, acidic pH, UV-rays, and ionic strength) and pharmaceutical barriers (poor stability, low in-vivo retention, controlled and targeted delivery, immune system neutralization, etc.) have the greatest influence on the viability of phage virions (also phage proteins). A variety of Nanotechnology based formulations such as polymeric nanoparticles, liposomes, dendrimers, nanoemulsions, and nanofibres have been recently reported to overcome the above-mentioned obstacles. In this review, we have compiled all these recent reports and discussed bacteriophage-based nanoformulations techniques for the successful treatment of ocular infections caused by multidrug-resistant S. aureus and other bacteria. © 2023 Elsevier GmbH