Preparation of decellularized optic nerve grafts

Area of Science:

• Tissue Engineering
• Biomaterials Science
• Ophthalmology Research

Background:

• Decellularized tissues utilize extracellular matrices (ECMs) for tissue engineering.
• Optic nerve decellularization research is limited.
• Developing effective decellularization protocols is essential for optic nerve regeneration.

Purpose of the Study:

• To investigate and compare different detergent-based decellularization protocols for optic nerve tissues.
• To evaluate the efficacy of various detergents in removing cellular material while preserving ECM integrity.
• To assess the biochemical, mechanical, and cytotoxic properties of decellularized optic nerve scaffolds.

Main Methods:

• Decellularization of optic nerve tissues using various detergents, including Triton X-100, sodium deoxycholate (SD), and sodium dodecyl sulfate (SDS).
• Biochemical analyses to quantify DNA, glycosaminoglycan (GAG), and collagen content.
• Histological staining, scanning electron microscopy (SEM), and transmission electron microscopy (TEM) for structural evaluation.
• Mechanical property testing and cytotoxicity assays using L929 mouse fibroblast cells.

Main Results:

• Triton X-100 alone was insufficient for effective decellularization.
• Combinations of Triton X-100 with sodium deoxycholate preserved 85% of GAG content, whereas SDS preserved only 30%.
• Decellularization protocols minimally impacted ECM structure and mechanical properties.
• All tested methods exhibited minimal cytotoxicity, with over 95% cell viability.

Conclusions:

• The combination of Triton X-100 with sodium deoxycholate (TX-100-SD) and Triton X-100 with sodium dodecyl sulfate (TX-100-SDS) are effective methods for optic nerve decellularization.
• These optimized protocols are suitable for generating acellular optic nerve scaffolds for tissue engineering applications.
• The study provides valuable insights into optimizing decellularization for neural tissue regeneration.