Research Projects

Fungal keratitis is a serious infection of the cornea caused by several types of fungi. It can lead to severe complications, including blindness. Fungal keratitis most commonly affects young adults and middle-aged individuals, particularly those between 20 and 50 years old. Several existing gaps persist in the management of fungal keratitis, including delayed diagnosis, limited antifungal therapeutics, diagnostic tools, treatment efficacy, and prevention strategies, among others. The management sometimes necessitates the painful delivery of therapeutics using hypodermic needles at the target site of infection. However, this can be overcome by fabricating an advanced, minimally invasive device containing antifungal agents. Moreover, the study can be more realistic when performed in bioprinted tissues that can mimic the native human structures. To this end, this research aims to fabricate a dissolvable microneedle ocular contact lens patch containing sertaconazole nanosuspension for the treatment of fungal keratitis. Sertaconazole (SN), a model antifungal drug that belongs to BCS Class II, is anticipated for the current proposed work. Also, 3D bioprinted cornea will be produced, mimicking the native corneal tissue to prove the substantial efficacy of the prepared formulations. The solubility of SN (0.0063 mg/mL) will be enhanced by nanosuspension preparation (SN@NSPs). The compatibility studies will be evaluated using FT-IR, DSC, and powder XRD. The final optimized formulation will be loaded in a polymeric solution to cast onto the microneedle male molds to obtain a dissolvable polymeric-based ocular contact lens patch impregnated with micron-sized needles. The microneedle-containing drug-loaded nanosuspension (SN@NSPs-MN) will be further evaluated for its size and shape, DSC, FT-IR, XRD, mechanical strength, dissolution time, and in vitro studies, among others. Antifungal disc diffusion studies will be performed using various strains of fungi. Ex vivo and in vivo studies will be performed in excised porcine cornea and healthy rabbits, respectively. In both studies, the amount of drug-loaded in the nanosuspension permeated and retained will be calculated. Moreover, a bioprinted corneal model will be developed. Briefly, human adipose stem cell-derived corneal stromal keratocytes (hASC-CSKs) or human pluripotent stem cell-derived corneal endothelial cells (hPSC-CEnCs) will be explored, which will be integrated into the 3D bioprinted cornea to mimic human corneal structure. The fabricated model will be further characterized using techniques including SEM, mechanical strength, in vitro degradation studies, optical transmittance, and cytocompatibility, among others. After characterization, the model will be used to study the influence of the SN-loaded NSPs and SN@NSPs-MN formulations. To simulate infection, the model will be exposed to fungal strains of Candida albicans. Ex vivo studies will be conducted for these formulations across the 3D bioprinted cornea with and without infections (as a control), to understand the drug release kinetics. In in vivo studies, animals induced with fungal keratitis will be exposed to the application of hypodermic needles, SN@NSPs, and SN@NSPs-MN. TEWL, TEER, LDF, and Draize tests will be performed to evaluate the integrity and irritancy of the cornea after the application of various formulations. After 48 h of study, the animals will be euthanized, and the corneal tissues will be collected to quantify the drug. Overall , the proposed work will delve into the pathophysiological and mechanistic insights with drug diffusion dynamics using 3D printed microneedles and a corneal scaffold. Notably, the fabricated system aims at targeted delivery of antifungal nanosuspension with reduced invasive procedures. 3D printed microneedle contact lens, and corneal scaffold could be a momentous change in personalized ocular therapy and corneal transplant; therefore, it is translational.