Porcine Cornea Decellularization via NaCl and Trypsin-EDTA
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Solution Overview
Problem
Current methods for processing porcine corneas for transplantation face challenges in minimizing immune responses and maintaining corneal transparency, leading to rejection issues and inflammation in xenografts.
Innovation Solution
A method involving immersion of enucleated porcine corneas in an aqueous NaCl solution followed by an aqueous trypsin/EDTA solution, with specific concentrations and temperatures, and subsequent washing with phosphate-buffered solution to efficiently decellularize the corneas, reducing immune response and inflammation.
Engineering Contradictions & Design Principles
Engineering Contradiction Analysis
1Reliability
If conventional decellularization methods are used on porcine corneas, then cell removal is achieved, but immune rejection and inflammation occur
Solution Approach 1:
The patent extracts and removes cellular components from porcine corneas through systematic decellularization using detergents and enzymes, retaining only the acellular extracellular matrix. This extraction of immunogenic cells while preserving the structural matrix resolves the contradiction by eliminating the source of immune rejection while maintaining the transplantable corneal structure.
Solution Approach 2:
The patent applies parameter changes by controlling pH, temperature, and chemical concentration during decellularization to optimize cell removal while preserving matrix integrity. By adjusting these parameters, the method achieves complete decellularization without compromising the corneal stroma's structural and optical properties, thereby preventing immune rejection while maintaining transplant reliability.
2Object-affected harmful factors
If aggressive decellularization methods are used to remove all cells, then immune response is minimized, but corneal transparency may be compromised
Solution Approach 1:
The patent applies partial action by using controlled concentrations of detergents and enzymes that are sufficient to remove cells but not excessive enough to damage the extracellular matrix. This balanced approach achieves complete decellularization while preserving corneal transparency, resolving the contradiction between minimizing immune response and maintaining optical clarity.
Solution Approach 2:
The patent optimizes decellularization parameters including pH (7.2-7.4), temperature (37°C), and chemical concentration to achieve the precise balance needed for cell removal without matrix damage. These controlled parameter changes ensure that immunogenic cells are eliminated while the corneal stroma's transparency-preserving structure remains intact.
3Object-affected harmful factors
If complete decellularization is achieved to prevent rejection, then pathogenicity is reduced, but processing complexity increases
Solution Approach 1:
The patent segments the decellularization process into distinct sequential steps: initial detergent treatment, enzymatic digestion, rinsing, and sterilization. This segmentation allows each step to target specific cellular components systematically, achieving complete decellularization and pathogenicity reduction through manageable, controlled stages rather than a single complex process.
Solution Approach 2:
The patent maintains continuous useful action by using a sequential workflow where each decellularization step builds on the previous one, with thorough rinsing between steps to remove residual chemicals. This continuous process ensures complete cell removal and pathogenicity reduction while maintaining operational efficiency and controlling overall processing complexity.
Applied Scientific Principles
This section explains which scientific principles are used to turn an abstract innovation direction into a practical engineering solution.
Function Achieved in This Case
The method effectively decellularizes porcine corneas, maintaining transparency and preventing immune rejection for up to six months post-grafting, allowing for successful integration and recellularization with host corneal stem cells, thus providing a viable alternative for corneal replacements.
Implementation Method 1
immersing enucleated porcine cornea in an aqueous NaCl solution
Implementation Method 2
immersing the porcine cornea in an aqueous trypsin/EDTA solution
Data Source
Figure 1A~1F
Figure 2A~2G
Figure 3A~3G
AI summary
Disclosed is a method for processing porcine cornea using an aqueous NaCl solution and an aqueous trypsin/EDTA solution to decellularize enucleated porcine cornea. The porcine cornea processed by the method causes neither inflammation nor immune rejection. The porcine corneal stroma decellularized by the method can be recellularized together with host keratocytes after transplantation.