Research Projects

Traditional animal models and two-dimensional (2D) cultures lack the physiological complexity of human skin and often fail to accurately predict human responses, prompting regulatory authorities to advocate for validated non-animal methodologies. Regulatory authorities, including the U.S. FDA, ECHA, and EMA, as well as recent amendments to clinical trial regulations by the Government of India in 2023, are emphasizing the adoption of alternatives to animal testing methods, such as 3D models and organ-on-chip technologies. The limitations of animal models, highlight the imperative for advanced, human-relevant models for cosmetic and drug testing. Recent advances in tissue engineering and 3D bioprinting have enabled the fabrication of skin models, yet most existing constructs lack vascular, aged phenotype and immune components, limiting their utility for comprehensive anti-aging research and drug screening. Skin aging is a complex, multifactorial process driven by intrinsic genetic factors and extrinsic influences such as ultraviolet radiation, pollution, and lifestyle, resulting in the accumulation of senescent cells, increased reactive oxygen species (ROS), upregulation of matrix metalloproteinases (MMPs), and decreased synthesis of collagen and elastin. To address these issues, the proposed technology includes, Objective 1: Establishment and validation of human physiomimetic 3D In Vitro Skin model as a preclinical tool for skin permeation, sensitization and hydration assessments. Objective 2: Establishment and validation of 3D In Vitro Aged Skin Model as a preclinical tool for anti-aging therapeutics. This proposal outlines the formulation of a composite bioink comprising silk fibroin methacrylate (SilkMA), gelatin methacrylate (GelMA), and photoactivated platelet releasate (PPR), optimized for printability, mechanical strength, and cytocompatibility. Using extrusion-based 3D bioprinting, multilayered skin constructs incorporating human dermal fibroblasts, keratinocytes, and endothelial cells will be fabricated and matured at an air–liquid interface to promote epidermal differentiation and barrier formation. Aging phenotypes will be induced through mitomycin C treatment to trigger cellular senescence and controlled UV irradiation to simulate photoaging, with subsequent characterization of ECM degradation, MMP-1 activity, and altered gene expression profiles. The model’s utility will be validated by evaluating the efficacy of established antioxidants retinol, L-ascorbic acid, and glutathione on reversing age-associated changes, focusing on cellular viability, ECM protein levels, and key gene expression markers (COL1A1, COL3A1, ELN). Analytical techniques including MTT, ELISA, immunofluorescence, western blotting, qRT-PCR, permeability assessments and mechanical properties will be employed. The engineered 3D skin model, presently validated at Technology Readiness Level (TRL) 3 as a proof of concept (Bhar et al., 2024), will be further developed to incorporate aged tissue phenotypes that emulate senescent skin characteristics. This progression will facilitate targeted screening of anti-aging therapeutic candidates, with the goal of achieving enhanced model maturity, functional validation, and scalability consistent with TRL 5/6 benchmarks for translational readiness. This work addresses critical limitations in current dermatological research platforms by providing a human-relevant system that reduces reliance on animal testing, enhances mechanistic studies of skin aging, and supports regulatory shifts toward alternative testing strategies. Our multidisciplinary team aims to deliver two (02) validated in vitro preclinical models with potential for strategic technology transfer. The integration of advanced biomaterials, 3D bioprinting, and comprehensive analytical methods positions this proposal at the forefront of skin tissue engineering, with significant implications for drug discovery, cosmetic testing, and aging biology.