Enzyme-Free Hydrogel Production Without Proteolytic Contamination

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Solution Overview

Problem

Existing hydrogel production processes involve enzymatic digestion, which can leave proteolytic enzymes in the final products, making them unsuitable for human application due to potential tissue degradation.

Innovation Solution

An enzyme-free process is developed to produce tissue-specific extracellular matrix solutions and hydrogels by decellularizing tissues and hydrolyzing the matrix in an acidic environment without proteolytic enzymes, followed by pH neutralization and cross-linking to create stronger and faster-crosslinking hydrogels.

Engineering Contradictions & Design Principles

VSEngineering Contradiction Analysis

1Ease of manufacture

If enzymatic digestion is used to produce hydrogels, then the hydrogel production process is simplified, but proteolytic enzymes remain in the final product causing tissue degradation

Engineering Contradiction:
Improvehydrogel production processVSAvoidproteolytic enzyme contamination
Core Design Contradiction:
Ease of manufactureVSObject-generated harmful factors

Solution Approach 1:

The patent removes the harmful proteolytic enzymes from the hydrogel production process by using a chemical hydrolysis method instead of enzymatic digestion. This extraction eliminates the source of contamination while maintaining the ability to break down extracellular matrix proteins for hydrogel formation.

Inventive Principle:
Principle #2Taking out (Extraction)

Solution Approach 2:

The patent replaces the biological enzymatic system with a chemical hydrolysis system. Instead of using proteolytic enzymes to break down extracellular matrix proteins, the process uses chemical hydrolysis in an acidic environment, substituting a mechanical/chemical process for a biological one to eliminate enzyme contamination.

Inventive Principle:
Principle #28Mechanics substitution (Replace mechanical system)

2Productivity

If proteolytic enzymes are used for extracellular matrix digestion, then protein breakdown is efficient, but the final hydrogel becomes unsuitable for human application

Engineering Contradiction:
Improveprotein breakdown efficiencyVSAvoidsuitability for human application
Core Design Contradiction:
ProductivityVSReliability

Solution Approach 1:

The patent changes the fundamental parameter of the digestion method from enzymatic to chemical hydrolysis. By using acidic hydrolysis conditions instead of enzymatic processes, the patent achieves protein breakdown efficiency while eliminating the reliability issue of enzyme contamination in human applications.

Inventive Principle:
Principle #35Parameter changes

3Reliability

If enzyme-free hydrolysis is used, then the hydrogel is suitable for human application, but the cross-linking speed and strength are reduced

Engineering Contradiction:
Improvesuitability for human applicationVSAvoidcross-linking speed
Core Design Contradiction:
ReliabilityVSSpeed

Solution Approach 1:

The patent performs preliminary hydrolysis of the extracellular matrix proteins in an acidic environment before the cross-linking step. This preliminary action prepares the protein structure to enable faster and stronger cross-linking in the subsequent step, compensating for the enzyme-free approach and achieving both human applicability and cross-linking performance.

Inventive Principle:
Principle #10Preliminary action

4Ease of manufacture

If enzymatic digestion is used, then the extracellular matrix is broken down effectively, but the process leaves residual enzymes that cause unwanted proteolysis

Engineering Contradiction:
Improveextracellular matrix digestionVSAvoidunwanted proteolysis
Core Design Contradiction:
Ease of manufactureVSObject-affected harmful factors

Solution Approach 1:

The patent substitutes the enzymatic digestion system with a chemical hydrolysis system that does not leave residual active enzymes. The chemical hydrolysis process breaks down the extracellular matrix effectively while not introducing proteolytic activity that could cause unwanted proteolysis in the final hydrogel product.

Inventive Principle:
Principle #28Mechanics substitution (Replace mechanical system)

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 enzyme-free process results in hydrogels suitable for human application, with enhanced strength and speed of cross-linking, reducing the risk of tissue rejection and enabling therapeutic uses for tissue regeneration.

Implementation Method 1

hydrolyze the isolated extracellular matrix in an acidic environment, in the absence of proteolytic enzymes

Methodology Applied
Scientific EffectHydrolysis: Hydrolysis

Implementation Method 2

cross-linking to create stronger and faster-crosslinking hydrogels

Methodology Applied
Scientific EffectCross-linking: Chemical Bonding

Data Source

PatentUS20250230402A1Enzyme-free processes to produce hydrogels
Publication Date: 2025.07.17 TISSUELABS PESQUISA E DESENVOLVIMENTO LTDA

AI summary

The present invention is in the field of tissue and organ bioengineering and refers to an enzyme-free process to produce a tissue-specific extracellular matrix solution, based on a decellularized extracellular matrix, comprising: (a) decellularizing a tissue or organ to obtain the isolated extracellular matrix; and (b) hydrolyzing the isolated extracellular matrix in an acidic environment, in the absence of proteolytic enzymes, optionally followed by purification of the solution. Still, the present invention refers to an enzyme-free process to produce a tissue-specific hydrogel. Thus, the present invention also relates to hydrogels produced according to the current processes, and to ancillary inventions, having the tissue-specific extracellular matrix solution, the hydrogel, and/or the products derived from the tissue-specific extracellular matrix solution as the core that connects the inventive aspects disclosed in this document.