Life Sciences & Biotechnology
Title : | 3D Bioprinting Biomimetic Dermo-Epidermal Construct using Engineered Silk Spidroin with Vasculature Guidance for Skin Tissue Regeneration and Organotypic Tissue Model |
Area of research : | Life Sciences & Biotechnology |
Focus area : | 3D Bio printing, Skin tissue engineering |
Principal Investigator : | Dr. Janani Radhakrishnan, National Institute Of Animal Biotechnology, Uttar Pradesh |
Timeline Start Year : | 2022 |
Timeline End Year : | 2025 |
Contact info : | jananiradhakrishnan313@gmail.com |
Details
Executive Summary : | The emergence of 3D bioprinting technology has revolutionized the tissue engineering strategies by personalization and precise designing of tissue mimetic constructs customized to meet the patients’ need. The study aims to fabricate dermo-epidermal skin construct 3D bioprinting with layer specific characteristics, guided vasculature for dermis to regenerate skin tissue and serve as organotypic tissue model. Tissue analogous scaffolds with bioactive biomaterials contribute to scaffolding for repopulation of cells, matrix secretion and achieve regenerative potential. Nature-inspired biomaterials exhibit versatile properties for tissue engineering, however demerits such as availability, difficulty in processing hinders its applicability. Genetic code expansion enables manipulation of amino acid sequences and strategies that yields diverse congener proteins incorporated with desired functional peptides. Silk spidroin from spider dragline silk demonstrated regenerative potential with desired characteristics including biocompatibility, biodegradability, low density, mechanical strength and less immunogenic. Engineered silk spidroin with favorable cell interaction moieties will be tailored through unnatural amino acids (UAAs) incorporation and amino groups of congener protein interacts with aldehydes of oxidized polysaccharides to form schiff’s base hydrogel. Hydrogel blended with synthetic polymers serves as medium for suspending cells to develop dermo and epidermal-specific bioinks. Hydrogel precursor solution will be dispersed with keratinocytes to serve as epidermal bioink. To enhance the vasculature of dermis, the dermo-specific bioink will be printed using dual extruders for fibroblasts and endothelial cells with porous micro-structure developed by phase-separating the engineered silk protein. Printability of dermo and epidermal bioinks will be investigated by rheological measurements and various patterning of bioprint will be achieved. Various patterning of co-culturing endothelial cells with dual extrusion for dermal layer with optimal neo-vascularization guidance will be evaluated. The physicochemical characterization of printable bioink and printed constructs including viscoelasticity, swelling behavior, biodegradation, composition and microstructure (electron microscopy) analysis will be recorded. Skin regenerative potential in vitro will be assessed by cell viability, cell proliferation, neo-vasculogenesis, specific gene and protein expression for matrix deposition. In vitro regenerative potential under simulated diabetic culturing conditions will also be evaluated. Finally, 3D bioprinted biomimetic dermo-epidermal construct will be printed with mesenchymal stem cells for skin tissue regeneration in vivo in a diabetes induced rat critical sized defect model. In addition, the developed vascularized dermo-epidermal construct will be evaluated for physiological relevance as organotypic skin equivalent for preclinical studies. |
Co-PI: | Dr. Ayyadurai Niraikulam, CSIR- Central Leather Research Institute, Tamil Nadu-600020 |
Total Budget (INR): | 36,30,954 |
Organizations involved