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Glaucoma is one of the leading causes of loss of vision. The problems associated with the marketed formulations of anti-glaucoma drugs are low bioavailability, unwanted side effects, and low patient compliance. Hydrogels are an important class of soft materials that play a crucial role in developing an ocular drug delivery system. They assume a special significance in addressing the problems associated with the marketed formulations of eyedrops. An appropriate design of the hydrogel system capable of encapsulating single or multiple drugs for glaucoma has emerged in recent times to overcome such challenges. Although various modes of imaging play critical roles in assessing the efficacy of these formulations, evaluating hydrogels for drug permeation and retention remains challenging. Especially, the assessment of dual drugs in the hydrogel system is not straightforward due to the complexity in measuring drug penetration and retention for in vivo or ex vivo systems. There is a need to develop tools for the fabrication and validation of hydrogel-based systems that give insight into precorneal retention, biocompatibility, cellular uptake, and cell permeation. The current review highlights some of the complexities in formulating hydrogel and benchmarking technologies, including confocal laser scanning microscopy, fluorescent microscopy, slit-lamp biomicroscopy, and camera-based imaging. This review also summarizes recent evaluations of various hydrogel formulations using in vitro and in vivo models. Further the article will help researchers from various disciplines, including formulation scientists and biologists, set up preclinical protocols for evaluating polymeric hydrogels. © 2022 American Chemical Society. All rights reserved.

Gelatin has been the biomaterial of choice for decades now. Its low cost, renewable, nontoxic and biodegradable properties make it one of the most desirable materials for controlled release applications. However, the usage of gelatin is limited by its poor mechanical/thermal stability and high water solubility. Chemical cross-linkers and hydrophobic modifications of gelatin have solved this problem but they lead to the problem of toxicity and/ or high processing cost. This research attempts to employ a nontoxic hydrophobic drug molecule to curb early degradation of gelatin in an aqueous environment. We report the design of non-cross-linked gelatin capsules with high dissolution resistance in an aqueous medium. Piperine, a hydrophobic drug (Solubility: 40mg/L in water) was coated on the gelatin capsules to enhance its stability in an aqueous environment. The hydrophobic piperine molecules repelled the water molecules to intensify its dissolution resistance. This stabilization was used to control the release of naproxen sodium, encapsulated inside the gelatin matrix. Piperine, in this case, acts as a placebo i.e it has zero therapeutic effect but its presence was necessary to control the early degradation of gelatin matrix. The deposition of piperine was done using the solvent evaporation method where ethanol was used as the solvent. The wettability studies revealed the hydrophobic nature of surface after the deposition of piperine while SEM analysis showed the presence of long cylindrical (fiber-like) structures over the gelatin surface. Further investigation (FTIR/ATR and molecular dynamics) revealed that the long fiber structures were due to the crystallization of piperine over the surface of gelatin. This crystallization was triggered by the intermolecular association (hydrogen bond) of ethanol and piperine. These observations enabled us to optimize the piperine loading protocol over the gelatin capsules that helped in achieving a zero order naproxen release for 3