Bone Tissue Decellularization via Sequential Chemical Immersion

Resolve Bottlenecks,
Find Innovative Solutions
Generate Solutions

Solution Overview

Problem

Current methods for decellularizing bone tissue are inefficient and incomplete, particularly due to the difficulty in removing cells from tightly packed bone structures, which limits the use of bone grafts in load-bearing applications and poses risks of rejection and infection in transplantation.

Innovation Solution

A process involving sequential immersion of bone tissue samples in solutions containing chelating agents, anionic detergents, non-ionic detergents, enzymes, and antibiotics effectively removes cellular components without damaging the three-dimensional structure, using a specific sequence and washing steps to ensure complete decellularization.

Engineering Contradictions & Design Principles

VSEngineering Contradiction Analysis

1Reliability

If conventional decellularization methods are used on bone tissue, then some cellular components are removed, but complete decellularization cannot be achieved due to the tightly packed bone structure

Engineering Contradiction:
Improvedecellularization completenessVSAvoidprocess complexity
Core Design Contradiction:
ReliabilityVSDevice complexity

Solution Approach 1:

The decellularization process is divided into multiple sequential steps, each targeting different cellular components. The method segments the complex task of complete decellularization into manageable stages: initial detergent treatment for membrane removal, followed by enzyme treatment for matrix degradation, and final cleaning steps, allowing comprehensive cell removal from tightly packed bone structures

Inventive Principle:
Principle #1Segmentation

Solution Approach 2:

The method applies preliminary detergent treatment to bone tissue before transplantation to pre-remove cellular membranes and components. This preliminary action prepares the bone graft by eliminating immunogenic elements in advance, reducing rejection risks before the tissue encounters the recipient's immune system

Inventive Principle:
Principle #10Preliminary action

2Reliability

If multiple treatment solutions are used sequentially to achieve complete decellularization, then decellularization effectiveness is improved, but processing time increases

Engineering Contradiction:
Improvedecellularization effectivenessVSAvoidprocessing time
Core Design Contradiction:
ReliabilityVSLoss of time

Solution Approach 1:

The decellularization process employs continuous sequential treatment with different solutions without idle interruptions. Each treatment step flows into the next: detergent solution treatment continues into enzyme solution treatment, which transitions to alcohol rinsing, maintaining continuous useful action throughout the process to efficiently achieve complete decellularization

Inventive Principle:
Principle #20Continuity of useful action

Solution Approach 2:

The method uses periodic application of different treatment solutions with specific durations. Each solution is applied for an optimized time period: initial detergent treatment for a set duration, followed by enzyme treatment for another period, then alcohol rinsing in periodic cycles, allowing systematic removal of cellular components while managing overall processing time

Inventive Principle:
Principle #19Periodic action

3Strength

If bone tissue is used as graft without complete decellularization, then structural integrity is maintained, but rejection and infection risks increase

Engineering Contradiction:
Improvestructural integrityVSAvoidrejection and infection risks
Core Design Contradiction:
StrengthVSObject-affected harmful factors

Solution Approach 1:

The method selectively extracts cellular components from bone tissue while leaving the mineral and organic matrix intact. Through sequential detergent and enzyme treatments, cellular membranes, cytoplasm, and nuclear materials are removed and extracted from the bone structure, eliminating immunogenic elements while preserving the mechanical framework for successful grafting

Inventive Principle:
Principle #2Taking out (Extraction)

Solution Approach 2:

Various chemical solutions serve as intermediaries between the bone tissue and the decellularization goal. Detergents act as intermediaries to solubilize and remove cellular membranes, enzymes serve as intermediaries to digest cellular proteins and matrices, and alcohol solutions act as intermediaries for final cleaning and sterilization, each facilitating specific aspects of cell removal while protecting the bone structure

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 process allows for the complete and effective decellularization of bone tissue, preserving its structural integrity, which can be used as biologic scaffolds for tissue engineering applications, such as grafts, without the risks associated with traditional transplantation.

Implementation Method 1

treating said sample with: i. a solution comprising at least one chelating agent

Methodology Applied
Scientific EffectChelation:

Implementation Method 2

a solution comprising at least one anionic detergent

Methodology Applied
Scientific EffectSurfactant action: Surfactant

Implementation Method 3

a solution comprising at least one non-ionic detergent

Methodology Applied
Scientific EffectMicelle formation:

Implementation Method 4

a solution comprising at least one enzyme

Methodology Applied
Scientific EffectEnzymatic degradation: Enzyme

Data Source

PatentEP3095469B1Process for bone tissue decellularization
Publication Date: 2019.09.11 INCI ILYAS

AI summary

A process for preparing decellularized isolated bone tissue samples, comprises the steps of a) providing an isolated bone tissue sample, b) treating said sample by immersions in: a solution comprising at least one chelating agent, a solution comprising at least one anionic detergent, a solution comprising at least one non-ionic detergent, a solution comprising at least one enzyme, a solution comprising at least one antibiotic. Decellularized bone tissue samples as obtainable by a process according to the invention can be used as grafts.