Developing a Novel biomimetic hydrogel sourced from Human Tenons Fascia for stromal replacement and 3D bioprinting corneal constructs

Abstract

Purpose : Corneal scarring due to infection, injury or inflammation is a significant cause of blindness affecting millions worldwide. Human tenons fascia is an abundant extra ocular connective tissue in the orbit, which is harvested to seal corneal perforations because of its healing, bio-integration and remodelling properties. In this study we tried to explore developing a biomimetic hydrogel from human tenons fascia as a potential alternative to corneal stromal regeneration and developing 3d bio-printed constructs.

Methods : This study reports the development of a novel biomimetic hydrogel composed of decellularized extracellular matrix (dECM) derived from human tenons fascia using a simple detergent free decellularization technique. A thorough in-vitro physical and chemical characterization was performed comparing it with native and decellularized cornea hydrogel. The hydrogel was then optimised by assessing its rheological properties to develop a bioink for 3d bioprinting. Finally we have used customised CAD models and moulds to bioprint curved corneal stromal constructs.

Results : This is the first time a dECM hydrogel was developed using human tenons fascia which retained the major extra-cellular matrix components, demonstrated physical, chemical, and environmental stability, and was tested as non-pyrogenic and non-immunogenic. It also showed presence of Collagen Type 1 Type 5 alongwith other extracellular matrix proteins lumican, decorin keratocan similar to that of native cornea and additionally presence of Collagen type 6, type 3. The human tenons fascia dECM hydrogel showed similar rheology cell viability in comparison with the native human cornea derived dECM hydrogel. It also demonstrated stability at 37 degrees Celsius, good injectability and shape retention ideal for a bioink to facilitate 3D bioprinting of curved corneal stromal constructs.

Conclusions : Human tenons fascia is available in abundance is used by corneal surgeons for sealing perforations. It is a potential alternative to human cadaveric corneal stroma to develop bioengineered cornea. A dECM tenons fascia hydrogel sourced from human tissues holds many advantages over various animal, plant derived natural and synthetic biomaterials and has potential applications in corneal bioengineering and regeneration.

This abstract was presented at the 2024 ARVO Annual Meeting, held in Seattle, WA, May 5-9, 2024.