Tissue Substitute Production Using Macromolecular Crowders

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Solution Overview

Problem

Current tissue engineering methods face challenges in producing tissue substitutes quickly and efficiently, with limitations in collagen matrix deposition in vitro, immune responses to animal-derived collagens, and slow production times for cell-based therapies.

Innovation Solution

The use of poly-dispersed macromolecular crowders, such as carrageenan, in cell culture media to create a crowded extracellular environment that accelerates collagen matrix deposition and extracellular matrix production, allowing for rapid production of tissue substitutes within 48 hours without animal-derived compounds.

Engineering Contradictions & Design Principles

VSEngineering Contradiction Analysis

1Loss of time

If conventional cell culture methods are used for tissue substitute production, then production time is extended, but productivity is reduced

Engineering Contradiction:
Improveproduction timeVSAvoidtissue substitute production rate
Core Design Contradiction:
Loss of timeVSProductivity

Solution Approach 1:

The patent applies macromolecular crowding by adding high molecular weight polymers (dextran, Ficoll, or polyethylene glycol) to the cell culture medium at specific concentrations (e.g., 10-40% v/v). This changes the physical-chemical parameters of the culture environment, creating excluded volume effects that accelerate procollagen conversion to collagen and enhance extracellular matrix production, thereby reducing production time from weeks to days while increasing productivity

Inventive Principle:
Principle #35Parameter changes

Solution Approach 2:

The patent uses macromolecular crowders as intermediary substances that mediate the acceleration of collagen matrix deposition. These crowders (dextran, Ficoll, or polyethylene glycol) act as intermediaries that create steric hindrance and excluded volume effects, facilitating the conversion of procollagen to collagen and enhancing cell metabolism without being consumed in the process

Inventive Principle:
Principle #24Intermediary (Mediator)

2Ease of manufacture

If animal-derived collagen is used in tissue engineering, then scaffold fabrication is improved, but immune response and disease transmission risk increase

Engineering Contradiction:
Improvescaffold fabricationVSAvoidimmune response and disease transmission
Core Design Contradiction:
Ease of manufactureVSObject-affected harmful factors

Solution Approach 1:

The patent employs human fibroblasts to self-produce human collagen and extracellular matrix components within the culture system. The cells naturally synthesize and deposit human-specific collagen types (I, III, V) and other matrix proteins, eliminating the need for animal-derived collagens. This self-service approach ensures biocompatibility, reduces immune response, and prevents interspecies disease transmission while maintaining ease of manufacture through standardized cell culture protocols

Inventive Principle:
Principle #25Self-service

3Object-affected harmful factors

If recombinant collagen is used for scaffold fabrication, then animal disease transmission is avoided, but expression levels are low and commercialisation is prohibited

Engineering Contradiction:
Improvedisease transmission riskVSAvoidcollagen expression level
Core Design Contradiction:
Object-affected harmful factorsVSProductivity

Solution Approach 1:

The patent uses macromolecular crowders (dextran, Ficoll, or polyethylene glycol) as intermediaries to enhance the in vitro production of human collagen by human fibroblasts. These crowders create excluded volume effects that accelerate procollagen conversion to collagen and increase overall collagen expression levels by several-fold compared to conventional culture conditions, making the system commercially viable while maintaining freedom from animal disease transmission risks

Inventive Principle:
Principle #24Intermediary (Mediator)

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

This approach enables the rapid production of biologically active, host-specific tissue substitutes that can be used for various tissue engineering applications, avoiding immune rejection and interspecies transmission risks, with enhanced collagen and fibronectin deposition, and maintaining cell viability and morphology.

Implementation Method 1

The use of poly-dispersed macromolecular crowders, such as carrageenan, in cell culture media to create a crowded extracellular environment that accelerates collagen matrix deposition and extracellular matrix production

Methodology Applied
Scientific EffectMacromolecular crowding:

Data Source

PatentEP3453752B1Engineered living tissue substitute
Publication Date: 2022.10.26 THE NAT UNIV OF IRELAND GALWAY
  • EP3453752B1 patent drawingFigure 1~2
  • EP3453752B1 patent drawingFigure 3~4
  • EP3453752B1 patent drawingFigure 5

AI summary

The present invention relates generally to the field of tissue engineering and in particular to the production of tissue films or cell matrices, which can be used as a living tissue substitute or an artificial tissue construct in tissue repair or replacement.