Browsing by Author "Roy, Aniruddha"

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Anticancer and immunostimulatory activity by conjugate of paclitaxel and non-toxic derivative of lps for combined chemo-immunotherapy
(Springer, 2012-05) Roy, Aniruddha
Cancer is a multifactorial syndrome; hence, multidimensional therapy with a chemo-immunotherapeutic conjugate could be more effective in curing the disease.
Apremilast loaded lyotropic liquid crystalline nanoparticles embedded hydrogel for improved permeation and skin retention: An effective approach for psoriasis treatment
(Elsevier, 2023-06) Singhvi, Gautam; Roy, Aniruddha
The present work aimed to prepare and evaluate Apremilast loaded lyotropic liquid crystalline nanoparticles (LCNPs) formulation for skin delivery to enhance the efficacy with reduced adverse effects of the oral therapy in psoriasis treatment. The LCNPs were prepared using the emulsification using a high shear homogenizer for size reduction and optimized with Box Behnken design to achieve desired particle size and entrapment efficiency. The selected LCNPs formulation was evaluated for in-vitro release, in-vitro psoriasis efficacy, skin retention, dermatokinetic, in-vivo skin retention, and skin irritation study. The selected formulation exhibited 173.25 ± 2.192 nm (polydispersity 0.273 ± 0.008) particle size and 75.028 ± 0.235% entrapment efficiency. The in-vitro drug release showed the prolonged-release for 18 h. The ex-vivo studies revealed that LCNPs formulation exhibited drug retention up to 3.2 and 11.9-fold higher, in stratum corneum and viable epidermis compared to conventional gel preparation. In-vitro cell line studies performed on immortal keratinocyte cells (HaCaT cells) demonstrated non-toxicity of selected excipients used in designed LCNPs. The dermatokinetic study revealed the AUC0–24 of the LCNPs loaded gel was 8.4 fold higher in epidermis and 2.06 fold in dermis, respectively compared to plain gel. Further, in-vivo animal studies showed enhanced skin permeation and retention of Apremilast compared to conventional gel.
Autophagy inhibition potentiates SAHA‑mediated apoptosis in glioblastoma cells by accumulation of damaged mitochondria
(Spandidos, 2018) Chowdhury, Rajdeep; Roy, Aniruddha; Mukherjee, Sudeshna
Glioblastoma multiforme (GBM), often referred to as a grade IV astrocytoma, is the most invasive type of tumor arising from glial cells. The main treatment options for GBM include surgery, radiation and chemotherapy. However, these treatments tend to be only palliative rather than curative. Poor prognosis of GBM is due to its marked resistance to standard therapy. Currently, temozolomide (TMZ), an alkylating agent is used for treatment of GBM. However, GBM cells can repair TMZ‑induced DNA damage and therefore diminish the therapeutic efficacy of TMZ. The potential to evade apoptosis by GBM cells accentuates the need to target the non‑apoptotic pathway and/or inhibition of pro‑survival strategies that contribute to its high resistance to conventional therapies. In recent studies, it has been demonstrated that HDAC inhibitors, such as vorinostat (suberoyl anilide hydroxamic acid; SAHA) can induce autophagy in cancer cells, thereby stimulating autophagosome formation. In addition, a lysosomotropic agent such as chloroquine (CQ) can result in hyper‑accumulation of autophagic vacuoles by inhibiting autophagosome‑lysosome fusion, which can drive the cell towards apoptosis. Hence, we postulated that combination treatment with SAHA and CQ may lead to increased formation of autophagosomes, resulting in its hyper‑accumulation and ultimately inducing cell death in GBM cells. In the present study, we demonstrated that CQ co‑treatment enhanced SAHA‑mediated GBM cell apoptosis. Inhibition of the early stage of autophagy by 3‑methyladenine pre‑treatment reduced cell death confirming that apoptosis induced by CQ and SAHA was dependent on autophagosome accumulation. We also demonstrated that autophagy inhibition led to enhanced ROS, mitochondria accumulation and reduced mitochondrial membrane potential resulting in cell death. The present study provides cellular and molecular evidence concerning the combined effect of SAHA and CQ which can be developed as a therapeutic strategy for the treatment of glioblastoma in the future.
Autophagy inhibition potentiates SAHA‑mediated apoptosis in glioblastoma cells by accumulation of damaged mitochondria
(Spandidos Publications, 2018-04) Chowdhury, Rajdeep; Roy, Aniruddha
Glioblastoma multiforme (GBM), often referred to as a grade IV astrocytoma, is the most invasive type of tumor arising from glial cells. The main treatment options for GBM include surgery, radiation and chemotherapy. However, these treatments tend to be only palliative rather than curative. Poor prognosis of GBM is due to its marked resistance to standard therapy. Currently, temozolomide (TMZ), an alkylating agent is used for treatment of GBM. However, GBM cells can repair TMZ‑induced DNA damage and therefore diminish the therapeutic efficacy of TMZ. The potential to evade apoptosis by GBM cells accentuates the need to target the non‑apoptotic pathway and/or inhibition of pro‑survival strategies that contribute to its high resistance to conventional therapies. In recent studies, it has been demonstrated that HDAC inhibitors, such as vorinostat (suberoyl anilide hydroxamic acid; SAHA) can induce autophagy in cancer cells, thereby stimulating autophagosome formation. In addition, a lysosomotropic agent such as chloroquine (CQ) can result in hyper‑accumulation of autophagic vacuoles by inhibiting autophagosome‑lysosome fusion, which can drive the cell towards apoptosis. Hence, we postulated that combination treatment with SAHA and CQ may lead to increased formation of autophagosomes, resulting in its hyper‑accumulation and ultimately inducing cell death in GBM cells. In the present study, we demonstrated that CQ co‑treatment enhanced SAHA‑mediated GBM cell apoptosis. Inhibition of the early stage of autophagy by 3‑methyladenine pre‑treatment reduced cell death confirming that apoptosis induced by CQ and SAHA was dependent on autophagosome accumulation. We also demonstrated that autophagy inhibition led to enhanced ROS, mitochondria accumulation and reduced mitochondrial membrane potential resulting in cell death. The present study provides cellular and molecular evidence concerning the combined effect of SAHA and CQ which can be developed as a therapeutic strategy for the treatment of glioblastoma in the future.
Bio-hydrogel for Prolonged Controlled Gastro-retentive Drug Dispenser
(IEEE, 2019) Belgamwar, Sachin U.; Roy, Aniruddha
A carbohydrate based polyacrylamide (Paam) modified with guar gum (GG) bio-hydrogel was developed for triggered, controlled gastro-retentive drug dispenser (GRDD) application. Temperature was applied as external stimuli and acidic medium (pH4) was used to release the drug from gel network. The release phenomenon was observed until 60 hours to understand the suitability for prolonged gastro retentive drug dispensing vehicle. The cytotoxicity was assessed by XTT assay with MCF-7 cell line.
Breaking the niche: multidimensional nanotherapeutics for tumor microenvironment modulation
(Springer, 2022-06) Roy, Aniruddha
Most of the current antitumor therapeutics were developed targeting the cancer cells only. Unfortunately, in the majority of tumors, this single-dimensional therapy is found to be ineffective. Advanced research has shown that cancer is a multicellular disorder. The tumor microenvironment (TME), which is made by a complex network of the bulk tumor cells and other supporting cells, plays a crucial role in tumor progression. Understanding the importance of the TME in tumor growth, different treatment modalities have been developed targeting these supporting cells. Recent clinical results suggest that simultaneously targeting multiple components of the tumor ecosystem with drug combinations can be highly effective. This type of “multidimensional” therapy has a high potential for cancer treatment. However, tumor-specific delivery of such multi-drug combinations remains a challenge. Nanomedicine could be utilized for the tumor-targeted delivery of such multidimensional therapeutics. In this review, we first give a brief overview of the major components of TME. We then highlight the latest developments in nanoparticle-based combination therapies, where one drug targets cancer cells and other drug targets tumor-supporting components in the TME for a synergistic effect. We include the latest preclinical and clinical studies and discuss innovative nanoparticle-mediated targeting strategies.
Carboxymethylcellulose-Based and Docetaxel-Loaded Nanoparticles Circumvent P-Glycoprotein-Mediated Multidrug Resistance
(ACS, 2014-02) Roy, Aniruddha
Taxanes are a class of anticancer agents with a broad spectrum and have been widely used to treat a variety of cancer. However, its long-term use has been hampered by accumulating toxicity and development of drug resistance. The most extensively reported mechanism of resistance is the overexpression of P-glycoprotein (Pgp). We have developed a PEGylated carboxymethylcellulose conjugate of docetaxel (Cellax), which condenses into ∼120 nm nanoparticles. Here we demonstrated that Cellax therapy did not upregulate Pgp expression in MDA-MB-231 and EMT-6 breast tumor cells, whereas a significant increase in Pgp expression was measured with native docetaxel (DTX) treatment. Treatment with DTX led to 4–7-fold higher Pgp mRNA expression and 2-fold higher Pgp protein expression compared with Cellax treatment in the in vitro and in vivo system, respectively. Cellax also exhibited significantly increased efficacy compared with that of DTX in a taxane-resistant breast tumor model. Against the highly Pgp expressing EMT6/AR1 cells, Cellax exhibited a 6.5 times lower IC50 compared with that of native DTX, and in the in vivo model, Cellax exhibited 90% tumor growth inhibition, while native DTX had no significant antitumor activity.
Chitosan, chondroitin sulfate, and hyaluronic acid based in-situ forming scaffold for efficient cell grafting
(Elsevier, 2023-01) Roy, Aniruddha
Current cell grafting techniques are majorly dependent on seeding cells on a pre-formed scaffold. However, cells grow in a 2-dimensional (2D) space in such constructs, not mimicking the tissue's 3-dimensional (3D) architecture. The present study evaluated a unique poly-electrolyte complexation (PEC) based strategy for the 3D engraftment of cells in a porous polymeric scaffold. The scaffold was synthesized using a positively charged polysaccharide chitosan (CH) and negatively charged glycosaminoglycans chondroitin sulfate (CS) and hyaluronic acid (HA). Two different scaffolds were synthesized, one using CH and CS [CH-CS] and another using CH and CS + HA [CH-(CS-HA)]. The physicochemical characterization of both the PECs confirmed electrostatic interactions, leading to a porous and viscoelastic PEC formation. Fibroblast cells were grafted and seeded in both scaffolds to evaluate the effect of different scaffold compositions and the difference between seeded and grafted cells. Imaging studies confirmed that grafting of the fibroblast cells supports cellular proliferation. The qPCR studies demonstrated increased expression of functional markers TGF-β, α-SMA, collagen-I, and fibronectin in the CH-(CS-HA) grafted cells. In summary, it was demonstrated that an in-situ forming PEC of CH, CS, and HA had good physicochemical properties for cell grafting and supported grafted cells with improved function
Chitosan-Chondroitin sulfate based polyelectrolyte complex for effective management of chronic wounds
(Elsevier, 2019-07) Roy, Aniruddha
Acute and chronic wound remain an unresolved clinical problem among various demographic groups. Traditional marketed products focus mainly on inhibition of bacterial growth at the wound site neglecting the tissue repair, which significantly affect the healing rate. It would be highly beneficial if a wound healing material can be developed which has both antibacterial as well as tissue regenerating potential. We have prepared a polyelectrolyte complex (PEC) using chitosan (CH) and chondroitin sulfate (CS) which can form an in-situ scaffold by spontaneous mixing. The fabrication of CH-CS PEC was optimized using Quality-By-Design (QbD) approach. The prepared PEC showed very high swelling and porosity property. It was found to be non-hemolytic with good blood compatibility and low blood clotting index. It also exhibited good antibacterial activity against both gram-positive and gram-negative bacteria. The cell proliferation study exhibited good cytocompatibility and almost four-fold increase in cell density when treated with CH-CS PEC compared to control. In summary, we demonstrated that the prepared CH-CS PEC showed good blood compatibility, high antibacterial effect, and promoted wound healing potentially by stimulating fibroblast growth, making it an ideal wound dressing material.
Combined Chemo-immunotherapy as a Prospective Strategy To Combat Cancer: A Nanoparticle Based Approach
(ACS, 2010-09) Roy, Aniruddha
The prime objective of this study was to develop a combined chemo-immunotherapeutic formulation which could directly kill cancer cells as well as activate the immunosuppressed tumor microenvironment to mount a robust antitumor immune response. Paclitaxel (PTX) and SP-LPS (nontoxic derivative of lipopolysaccharide) were selected as anticancer drug and immunostimulant respectively. Poly(lactic-co-glycolic acid) (PLGA) based PTX and SP-LPS containing nanoparticles (TLNP) were prepared by the double-emulsion method (w/o/w) and characterized in terms of size, zeta potential and transmission electron microscopy (TEM). The release behavior of PTX and SP-LPS from the TLNP exhibited a biphasic pattern characterized by an initial burst followed by slow continuous release. In vitro anticancer activity of TLNP was found to be higher compared to PTX when studied in a tumor cell−splenocyte coculture system. TLNP activated murine monocytes induced the secretion of various proinflammatory cytokines. After iv administration of TLNP in tumor bearing C57BL/6 mice, the amount of PTX in the tumor mass was found to be higher in TLNP treated mice as compared to commercial Taxol group at all time points studied. In vitro studies suggest that nanoparticles containing PTX and SP-LPS have both direct cytotoxicity and immunostimulatory activity. Hence this might have potential as a chemo-immunotherapeutic formulation against cancer with advantage over present day chemotherapy with Taxol, in terms of tumor targeting, less toxicity and immunostimulation.
Comparison of Tumor Penetration of Podophyllotoxin–Carboxymethylcellulose Conjugates with Various Chemical Compositions in Tumor Spheroid Culture and In Vivo Solid Tumor
(ACS, 2017-04) Roy, Aniruddha
Polymer conjugation is an attractive approach for delivering insoluble and highly toxic drugs to tumors. However, most reports in the literature only disclose the optimal composition without emphasizing rational design or composition optimization to achieve maximized biological effects. In this study, we aimed to demonstrate that composition of a polymer conjugate would determine its physiochemical characteristics, tumor penetration, and, ultimately, the in vivo efficacy. We also aimed to examine whether the tumor spheroid model could generate comparable results with the in vivo tumor model in terms of tumor penetration and efficacy of the various polymer conjugates. We have designed a polymer conjugate delivery system for a chemotherapeutic drug podophyllotoxin (PPT) by covalently conjugating PPT and polyethylene glycol (PEG) with acetylated carboxymethyl cellulose to yield conjugates containing various amounts of PPT and PEG. Depending on the composition, these conjugates self-assembled into nanoparticles (NPs) with different physicochemical properties. Conjugates with an increased PPT content formed particles with an increased diameter. In the present study, we selected three conjugates representing compositions containing high, medium, and low drug content, and compared their particle formation, drug release kinetics, their ability to penetrate tumor spheroid and in vivo s.c. tumor, and finally their antitumor efficacy in spheroid culture and an in vivo s.c. tumor model. We found that the low drug content conjugate formed smaller NPs (20 nm) compared to the high drug content conjugates (30–120 nm), and displayed faster drug release kinetics (5%/day vs 1–3%/day), improved tumor penetration, and enhanced antitumor efficacy in both the spheroid model and s.c. tumor model. In particular, the low drug content conjugate preferentially accumulated in the hypovascular region within the tumor, inducing complete regression of s.c. tumors and the metastasis to the lungs. Our data indicate composition optimization is needed to select the optimal conjugate, and tumor spheroid culture is a robust screening tool to help select the optimal formulation
A comprehensive review of the strategies to improve oral drug absorption with special emphasis on the cellular and molecular mechanisms
(Elsevier, 2021-02) Roy, Aniruddha
Oral drug delivery is the most convenient and cost-effective route of administration, due to which most of the drugs are administered through the oral route. Apparently, oral delivery is the easiest approach for drug administration; however, in reality, it is one of the most complicated routes. Although the gastrointestinal (GI) tract offers a large surface area for absorption, the GI epithelium is designed for the permeation of molecules with a specific set of physicochemical properties only. Due to this, different drugs exhibit considerable variabilities in absorption; one can show more than 90% absorption, while others may show less than 10%. To improve the oral absorption of low bioavailable drugs, multiple strategies have been developed, with variable success. To design a successful strategy, a thorough understanding of the GI physiology and the mechanism of drug absorption is pivotal. Different parts of the GI tract have significant variations, all of which have a profound impact on drug absorption. In this comprehensive review, we have done a cross-disciplinary exploration of the cellular physiology of different barriers present in the GI tract, how they impact drug absorption, and also discussed the diverse strategies developed to overcome absorption barriers present in the GI tract
Curcumin loaded nanostructured lipid carriers for enhanced skin retained topical delivery: optimization, scale-up, in-vitro characterization and assessment of ex-vivo skin deposition
(Elsevier, 2020-09) Singhvi, Gautam; Roy, Aniruddha
Nanostructured lipid carriers (NLC) have become a promising drug delivery system for topical delivery of drugs. Delivery of lipophilic drugs with improved stability and entrapment efficiency is one of the foremost benefits of NLC based formulations. The objective of the present study was to improve the permeation of poorly soluble curcumin into topical skin layers for the treatment of chronic inflammatory disorder psoriasis and microbial mediated acne vulgaris. Hot emulsification followed by probe sonication method was employed for the preparation of the curcumin loaded NLC. Further, in-vitro and ex-vivo characterization was performed for designed NLC. The designed NLC showed a mean particle size 96.2 ± 0.9 nm, entrapment efficiency of 70.5 ± 1.65% and zeta potential of -15.2 ± 0.566 mV. Curcumin-NLC showed extended in-vitro release upto 48 hours, whereas free curcumin showed 100% drug release within 4 hours. Ex-vivo skin permeation studies revealed 3.24 fold improved permeation and skin retention in the case of curcumin loaded NLC gel compared to free curcumin gel. The cell viability studies demonstrated the formulation components showed no toxicity towards keratinocyte cells. In keratinocyte cells, improved cell uptake was observed for curcumin-NLC compared to free curcumin dispersion. The results suggested that the NLC based formulation had potential to improve the efficacy of curcumin.
Design and dermatokinetic evaluation of Apremilast loaded nanostructured lipid carriers embedded gel for topical delivery: A potential approach for improved permeation and prolong skin deposition
(Elsevier, 2021-10) Singhvi, Gautam; Roy, Aniruddha
The present study aimed to develop Apremilast loaded nanostructured lipid carriers (NLCs) for topical delivery to overcome the limitations of oral therapy and increase the efficacy. Apremilast loaded NLCs were prepared by hot emulsification technique. The developed formulation was optimized by Box Behnken design and characterized for size, entrapment efficiency, and zeta potential. The selected formulation was investigated for in-vitro release, ex-vivo skin retention, dermatokinetic, psoriasis efficacy, in-vivo skin retention and skin irritation study. The NLCs characterization results showed its spherical shape with the particle size of 157.91 ± 1.267 nm (0.165 ± 0.017 PDI). The entrapment efficiency and zeta potential were found to be 69.144 ± 0.278% and -16.75 ± 1.40 mV, respectively. The in-vitro release study revealed a controlled release of Apremilast from NLCs up to 24 h. The ex-vivo study showed 3-fold enhanced skin retention compared to conventional gel preparation. The formulation depicted improved psoriasis efficacy indicating reduced TNF-α mRNA expression. The cytotoxicity and skin irritation study revealed the prepared formulation has no toxicity or irritation. The study depicts the NLCs loaded Apremilast can be explored for the topical delivery for treatment of psoriasis with improved skin retention and efficacy.
Design, synthesis and in vitro evaluation of primaquine and diaminoquinazoline hybrid molecules against the malaria parasite
(Wiley, 2025-01) Sundriyal, Sandeep; Roy, Aniruddha
In this study, we built on the known inhibitory potential of diaminoquinazolines (DAQs) against different stages of Plasmodium development and designed a convenient two-step synthesis to combine DAQ with primaquine (PQ) pharmacophore. The PQ-DAQ hybrids displayed potent in vitro activities in the low nanomolar range (IC50 135.20–398.80 nM) against all intra-erythrocytic stages of the drug-sensitive 3D7 strain, with significant potency enhancement compared to PQ alone (IC50 9370 nM). These hybrids were also potent at killing drug-resistant strains (Dd2, Dd2 R539T, IPC4912, CamWT C580Y, and 7G8) in the nanomolar range, with 11 f being the most effective compound (IC50 172.20–396.60 nM). Notably, for the first time, we present evidence that the DAQ-based compound 8 and its hybrids can inhibit β-hematin formation in vitro with potency (IC50 0.90–27.80 μM), suggesting hemozoin formation to be one of the potential targets of this series. Lastly, two hybrids with potent antiplasmodial activity were also found to be safe up to 10 μM against human HepG2 cells, suggesting the possibility of achieving host vs parasite selectivity with this series.
Development and evaluation of a simvastatin-loaded biopolymeric scaffold for improved angiogenesis and healing of diabetic wounds
(Elsevier, 2023-09) Roy, Aniruddha; Majumder, Syamantak
In diabetic wounds, lack of angiogenesis limits the supply of oxygen and nutrients at the wound site, resulting in poor healing. A well-known lipid-lowering drug, simvastatin (SIM), exhibited pleiotropic effects in wound healing, including promotion of angiogenesis. However, its clinical application is limited due to its poor physicochemical properties, including low solubility. In this study, a Soluplus and TPGS-based mixed micelle was developed for loading SIM in an in-situ forming chitosan-chondroitin sulfate-based poly-electrolyte complex hydrogel (CH-CS PEC). The hypothesis was that CH-CS PEC would improve overall wound healing due to the favorable viscoelasticity and porosity, whereas SIM would assist neoangiogenesis. SIM-loaded CH-CS PEC exhibited good mechanical stability and viscoelastic properties and demonstrated prolonged release of SIM. The formulation promoted endothelial cell sprouting in an ex-vivo rat aortic ring assay. Applying SIM-loaded CH-CS PEC in a diabetes-induced rat wound model resulted in faster wound closure, increased collagen deposition, and enhanced neovascularization with up-regulation of vascular endothelial growth factor (VEGF) expression. In summary, we have developed a drug-loaded, in-situ forming scaffold that can be directly applied at the wound site and can improve wound healing by promoting angiogenesis and collagen deposition at the wound site. This study demonstrated the combined efficacy of a viscoelastic scaffold and a proangiogenic drug for enhanced wound healing. The easy and simple fabrication method of the drug-loaded scaffold makes it suitable for clinical translation.
Development of a size-tunable paclitaxel micelle using a microfluidic-based system and evaluation of its in-vitro efficacy and intracellular delivery
(Elsevier, 2020-12) Roy, Aniruddha
Many challenges still remain for the successful translation of nanodelivery systems from the discovery to clinical practice. The primary concerns include easy scale-up process, uniformity, low size variation, and the ability to control critical physicochemical properties, including size and shape. In the current study, we have developed an easy-to-scale, size-tunable polymeric micelle formulation of paclitaxel using Soluplus, an amphiphilic graft copolymer with the help of a microfluidic platform. Two variable-sized polymeric micelles, ~90 nm, and ~180 nm were prepared by the variation of the drug: polymer ratio. Physicochemical characterization of the micelles was performed in terms of size, polydispersity, solubility, solubility factor, drug loading, and drug release. In-vitro efficacy was analyzed in terms of cytotoxicity in both 2D and 3D spheroid models, along with penetration of the micelles in the 3D spheroid. Intracellular uptake and lysosomal colocalization were also studied. Results demonstrated that the smaller micelles have better encapsulation efficiency with controlled drug release and enhanced cytotoxicity in both 2D and 3D models. Cellular internalization studies indicated that the smaller micelles have higher cellular uptake. In the 3D spheroid model, the smaller micelles exhibited almost complete penetration in 24 h, whereas the larger micelles showed negligible penetration. Altogether, this study indicated that this process could be used for the development of clinically successful nanoformulations.
Development of a tumor extracellular pH-responsive nanocarrier by terminal histidine conjugation in a star shaped poly(lactic-co-glycolic acid)
(Elsiever, 2021) Chowdhury, Rajdeep; Roy, Aniruddha
After reaching the tumor site, nanoparticles (NPs) mostly accumulate in the periphery of the tumor, as their intra-tumoral penetration is prevented due to the low perfusion, high interstitial fluid pressure, and dense matrix present in the tumor. A pH-responsive carrier can improve tumor permeation by releasing the drug quickly in the acidic tumor pH, helping its uniform tumor distribution through diffusion. In the current study, we have developed a histidine modified star-shaped PLGA (sPLGA-His) for the tumor-targeted delivery of the drug combination of docetaxel and disulfiram. The sPLGA-His NPs exhibited a rapid pH-responsive drug release behavior, with significantly increased drug release at pH 6.5 compared to pH 7.4 in 12 h. In-vitro cytotoxicity analysis showed that the pH-sensitive sPLGA-His NPs had enhanced efficacy in both 2D and 3D cell culture models. In the cell uptake study, the sPLGA-His NPs exhibited endosomal escape and uniform cellular distribution, whereas sPLGA NPs were found to be accumulated in the endosomes. In the tumor spheroid model, deep penetration was observed with the sPLGA-His NPs, while sPLGA NPs were found to be accumulated in the periphery. Using fluorescent colocalization as well as FRET analysis, increased release of the encapsulated cargo was noticed with the sPLGA-His NPs, compared to sPLGA NPs. Altogether, the sPLGA-His NPs can be used as a tumor extracellular pH-responsive nanocarrier for efficient drug delivery to the tumor.
Development of a tumor extracellular pH-responsive nanocarrier by terminal histidine conjugation in a star shaped poly(lactic-co-glycolic acid)
(Elsevier, 2021-06) Roy, Aniruddha; Singhvi, Gautam
The present work aimed to develop topical solid lipid nanocarriers (SLN) loaded hydrogel of apremilast (API) for psoriasis therapy to minimize the systemic adverse effects. The quality by design approach was implemented for the optimization of API loaded SLN using Box-Behnken design. SLN were prepared using hot emulsification followed by size reduction using probe sonication. The size and entrapment were found to be 167.70 nm ± 1.5 (0.238 PDI) and 63.84 ± 0.93%, respectively. The FESEM images of SLN dispersion portrayed the spherical shape of nanocarriers. The in vitro drug release of SLN dispersion showed extended-release up to 18 h and followed the Korsmeyyar-Peppas model with a regression value of 0.958 (n = 0.330), and Akaike index criteria was 63.69. In vitro cell line study, the MTT assay depicted the formulation excipients had minimal effect, and high internalization was observed with SLN dispersion (1.4-fold). The Ct value reduction in the relative expression of TNF-α miRNA was 3-fold higher with SLN dispersion compared to the positive control. The ex vivo skin retention and dermal distribution study by Coumarin-6 dye depicted an increase in permeation and retention with SLN formulation compared to free drug-loaded gel. The dermato-pharmacokinetic study of SLN formulation exhibited 2-fold higher drug retention in the epidermis and 5-fold higher in the dermis compared to free drug. This were stable for 3 months without significant changes. The results suggest that API loaded SLN can be utilized for topical delivery for effective treatment of psoriasis by targeting skin layers. The API loaded SLN based topical gel formulation showed improved permeation, skin deposition and prolonged release compared to conventional preparation. The designed preparation can signify a potential alternative for psoriasis treatment after clinical evaluation in near future.
Development of an in-situ forming, self-healing scaffold for dermal wound healing: in-vitro and in-vivo studies
(Elsevier, 2021-09) Roy, Aniruddha
The importance of the extra-cellular matrix (ECM) for wound healing has been extensively researched. Understanding its importance, multiple ECM mimetic scaffolds have been developed. However, the majority of such scaffolds are prefabricated. Due to their stiffness, prefabricated scaffolds cannot come into direct contact with the basal skin cells at the wound bed, limiting their efficacy. We have developed a unique wound dressing, using chitosan (CH) and chondroitin sulfate (CS), that can form a porous scaffold (CH-CS PEC) in-situ, at the wound site, by simple mixing of the polymer solutions. As CH is positively and CS is negatively charged, mixing these two polymer solutions would lead to electrostatic cross-linking between the polymers, converting them to a porous, viscoelastic scaffold. Owing to the in-situ formation, the scaffold can come in direct contact with the cells at the wound bed, supporting their proliferation and biofunction. In the present study, we confirmed the cross-linked scaffold formation by solid-state NMR, XRD, and TGA analysis. We have demonstrated that the scaffold had a high viscoelastic property, with self-healing capability. Both keratinocyte and fibroblast cells exhibited significantly increased migration and functional markers expression when grown on this scaffold. In the rat skin-excisional wound model, treatment with the in-situ forming CH-CS PEC exhibited enhanced wound healing efficacy. Altogether, this study demonstrated that mixing CH and CS solutions lead to the spontaneous formation of a highly viscoelastic, porous scaffold, which can support epidermal and dermal cell proliferation and bio-function, with an enhanced in-vivo wound healing efficacy.