Department of Pharmacy
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Item Lipopolymeric nanoplex-mediated CRISPR/Cas9 delivery for VEGF-A knockdown in psoriatic angiogenesis(ACS, 2025-10) Yadav, Sushil; Mittal, Anupama; Chitkara, DeepakPsoriasis is a chronic, incurable inflammatory skin disease characterized by immune cell infiltration, aberrant keratinocyte differentiation, and enhanced angiogenesis. Overexpression of the vascular endothelial growth factor-A (VEGF-A) gene promotes angiogenesis and is essential for endothelial cell migration, adhesion, and proliferation. Therefore, downregulating VEGF-A represents a promising therapeutic strategy for angiogenesis-related disorders. We investigated the application of Clustered Regularly Interspaced Short Palindromic Repeats (CRISPR)/CRISPR-associated protein 9 (Cas9) ribonucleoprotein complexes (sgRNA/eGFP-Cas9 RNPs) targeting VEGF-A in psoriasis. To enable efficient delivery in vitro and in vivo, we developed lipopolymeric nanoplexes (NPXs) encapsulating sgRNA/eGFP-Cas9 RNPs. These NPXs exhibited a particle size of 142.2 nm (polydispersity index: 0.144), a zeta potential of +4.27 mV, and achieved >70% transfection efficiency in HaCaT (human immortalized keratinocyte) cells. Ex vivo skin permeation studies demonstrated 66% of permeation after 24 h. The optimized NPX formulation was incorporated into a Carbopol-based gel, which displayed non-Newtonian, shear-thinning behavior with variable thixotropy and achieved 48% of skin permeation after 24 h. In vivo efficacy assessment in an imiquimod-induced psoriasis model in Swiss albino mice showed significantly improved Psoriasis Area and Severity Index (PASI) scores, reduced epidermal damage, and suppressed keratinocyte proliferation compared to naked RNPs and blank gel controls. Gene editing analysis revealed an indel frequency of 40.7% by T7 endonuclease I assay and 14% by Sanger sequencing. Enhanced cellular uptake, efficient skin permeation and retention, and improved therapeutic efficacy collectively highlight the potential of NPX-mediated CRISPR/Cas9 delivery as a noninvasive strategy for psoriasis treatment.Item Eugenol-loaded lipid nanoparticles-derived hydrogels ameliorate psoriasis-like skin lesions by lowering oxidative stress and modulating inflammation(ACS, 2024-10) Chitkara, DeepakPsoriasis is a chronic T-cell-mediated autoimmune skin disorder characterized by excessive epidermal thickening, overproliferation of keratinocyte, disruption of epidermal cell differentiation, and increased blood vessel growth in the dermal layer. Despite the common use of corticosteroids in psoriasis treatment, their limited efficacy and numerous side effects pose significant challenges. This research introduces a promising alternative approach by encapsulating eugenol (EU) in soya phosphatidylcholine (SPC) nanoparticles (EUNPs) which showed spherical shape nanoparticles with a hydrodynamic size of approximately 200 nm, polydispersity index 0.23, encapsulation efficiency of 85% having good colloidal stability indicated by ζ-potential of −27 mV. Later on, these EUNPs were formulated into a topical hydrogel system by using Carbopol 974P (EUNPGel), which exhibited superior drug loading, enhanced release kinetics for 48 h, long-term stability, and the ability to scavenge reactive oxygen species (ROS). Furthermore, EUNPs inhibited keratinocyte proliferation, induced apoptosis, and augmented the uptake of IL-6-mediated inflammation in human keratinocyte cells. Application of EUNPs-loaded gels (EUNPGel) to imiquimod-induced psoriatic lesions demonstrated effective dermal penetration, suppressed keratinocyte hyperplasia and restored epidermal growth. This led to a remarkable reduction in the Psoriasis Area and Severity Index (PASI) score from 3.75 to 0.5 within 5 days. This novel approach enhances ROS scavenging capacity, improves cellular uptake, facilitates skin penetration and retention, reduces the activity of hyperactive immune cells, and suggests potential applications for treating other immune-related disorders such as acne and atopic dermatitis.Item Preclinical safety of tetrahydrocurcumin loaded lipidic nanoparticles incorporated into tacrolimus ointment: In vitro and in vivo evaluation(Elsevier, 2022-09) Chitkara, DeepakPreclinical safety and proof of concept studies for a topical ointment comprising of concentrated tetrahydrocurcumin loaded lipidic nanoparticles (THC-LNs) and tacrolimus ointment (TTO) is proposed in the present investigation. The skin irritation potential and acute dermal toxicity were performed in rats in compliance with the Organization for Economic Cooperation and Development (OECD) guidelines (402, 404 and 410) while the cytotoxic potential was performed in HaCaT cells. Finally, in vivo evaluation was performed in Imiquimod mice model of psoriasis. In primary skin irritation assessment, TTO formulation, marketed formulation (Tacroz® Forte), THC-LNs, and blank LNs were topically applied on intact skin sites in rats while another group served as a negative control group for 72 h. TTO did not induce any adverse reactions. Repeated 28 days dermal toxicity followed by biochemical and histopathological assessment showed negligible alternations and skin lesions. THC-LNs revealed negligible cytotoxic potential in HaCaT cells. TTO showed significantly high anti-psoriatic activity in comparison to marketed ointment. This was also confirmed via histopathological evaluation. Based on these findings, it can be ascertained that TTO showed minimal toxicity and has ample potential for further clinical analysis. The above studies affirm the potential of TTO as an alternative for psoriasis.Item Topical delivery of Anti-VEGF nanomedicines for treating psoriasis(Elsevier, 2023-05) Pandey, Murali Monohar; Chitkara, Deepak; Mittal, AnupamaPsoriasis is a chronic skin disease characterized by inflammation caused by the activation of the immune system. Although the etiology of psoriasis is still unknown, the crosstalk between immune cells, dermal vascular cells, and epidermal keratinocytes has been shown as a critical process. Curiously, the overexpression of vascular endothelium growth factor (VEGF), which further propagates angiogenesis and cell proliferation, helps in the progression of the psoriasis plaque. Literature evidence shows the betterment of the psoriasis plaque when administering anti-VEGFA treatment. In our opinion, several other therapies targeting VEGF/VEGFR axis, such as employing peptides, RNAi and CRISPRi with nanocarriers as delivery strategy could redirect the therapeutic approach for the treatment of psoriasis. Collectively, this short communication highlights the significance of VEGFA, its downstream targets, and nanomedicines in the treatment of psoriasis. Nevertheless, continued research in this area could enable novel therapeutics and strategies to enter the market with a substantial social impact.Item Coenzyme Q10 loaded lipid-polymer hybrid nanoparticles in gel for the treatment of psoriasis like skin condition(Elsevier, 2022-10) Chitkara, Deepak; Mittal, AnupamaPsoriasis is characterized by severe dermal inflammation caused by pre-oxidants that dominate the skin's antioxidant system. The available therapeutic strategies seeks betterment in terms of cost effectiveness, long-term usage and safety. This research work highlights the use of monolithic polymer-lipid hybrid nanoparticles to deliver coenzyme Q10, a prominent antioxidant and anti-inflammatory biomolecule, to the skin. The system has both polymeric and lipidic characteristics, so each could compensate the other's shortcomings. The particle size, polydispersity index, and encapsulation efficiency of the nanoformulation were found to be 121 ± 11.61 nm, 0.252 ± 0.073, and 78.57 ± 3.88%, respectively, when produced employing the hot homogenization technique. The nanoparticles released CoQ10 steadily and consistently for up to 3 days. Further, the nanoformulation was used as a topical gel (CoQ10–0.06% w/w), and demonstrating non-newtonian rheological characteristics. An imiquimod-induced psoriatic mouse model was used to evaluate the in vivo efficacy of the nanoformulation gel, which produced a significant increase in CoQ10 efficacy when applied topically compared to free CoQ10 gels. Overall, the CoQ10 loaded NanoHybrid gel was found to be efficient for the in vivo application for the treatment of psoriasis like skin conditions.Item Nano-enabled topical delivery of anti-psoriatic small molecules(Elsevier, 2021-04) Chitkara, Deepak; Mittal, AnupamaPsoriasis is chronic autoimmune disorder mediated via T cell activation affecting 3% of population globally and is characterized by various features including erythema, scaling, hyperkeratosis, hyperplasia, plaques etc. Inspite of on going extensive researches in this disease area there is not a single therapy which claims complete cure and their primary focus is based on ameliorating the major symptoms underlying this pathophysiology and improving the medical condition. Out of the various currently assessable therapies, topical drug delivery approaches are mostly preferred because of their several advantages including direct targeting to desired site eliminating exposure to remaining body organs and hence minimizing the adverse side effects. The conventional drug delivery approaches (gels, creams and ointments) available as over the counter (OTC) products are associated with several disadvanatges due to local toxicity (irritation, burning, atropy etc), systemic toxicity (associated with drug leaching), reduced treatment efficacy (due to lower penetration of active in the viable epidermal region and poor retentive capability) and high dosing frequency with poor patient compliance because of reduced benefit to risk ratio. To address the above challenges and improve the benefit to risk ratio, nanoformulation based strategies are adopted due to several advantages including slow and sustained drug release (eliminating local toxicity due to sudden exposure of drug), deeper skin penetration, higher cellular uptake with localized retention with negligible systemic leaching. This review emphasizes on various types of nano-carriers (liposomes, niosomes, nano-emulsions, solid lipid nanoparticles, polymeric nanoparticles, etc) loaded with different anti-psoriatic small molecules in psoriasis treatment.Item Multi-component clobetasol-loaded monolithic lipid-polymer hybrid nanoparticles ameliorate imiquimod-induced psoriasis-like skin inflammation in Swiss albino mice(Elsevier, 2020-10) Chitkara, Deepak; Mittal, AnupamaLipid-polymer hybrid nanoparticles (LPNs) exhibit several advantages over polymeric and non-polymeric systems in terms of improved drug loading, controlled release, stability, and cellular uptake. Herein we report a scalable and stable monolithic lipid-polymer hybrid nanoparticles (LPNs) consisting of a combination of lipids (solid and liquid) and an amphiphilic copolymer, mPEG-PLA. Clobetasol propionate, a topical corticosteroid, was encapsulated in the hydrophobic core of these LPNs that showed spherical shaped particles with a z-average size of 94.8 nm (PDI = 0.213) and encapsulation efficiency of 84.3%. These clobetasol loaded LPNs (CP/LPNs) were formulated into a topical hydrogel using carbopol 974P. CP/LPNs gel showed a sustained in vitro clobetasol release for 7 days with no burst release and 6 month stability at 2-8°C and room temperature. Further, CP/LPNs showed an improved cellular uptake with significant growth inhibition of HaCaT cells. In ex vivo studies, these LPNs penetrated into the viable epidermis and dermis region of the psoriatic skin with undetectable quantities leaching to the reservoir. Further, the topical application of CP/LPNs gel on Swiss albino mice with psoriasis-like inflammation showed negligible leaching of clobetasol into the systemic circulation. Efficacy assessment showed significantly improved PASI score, reduced skin damage and proliferation after treatment with CP/LPNs gel as compared to marketed product (Clobetamos™). Collectively, the enhanced cellular uptake, high skin penetration with increased skin retention, and improved efficacy demonstrate the potential of these LPNs for future clinical application.