Gradient Mineralized Bone Matrix via Ultrasound Demineralization
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Solution Overview
Problem
Conventional bone graft materials, particularly those derived from natural bone matrix, face challenges such as irreversible damage during decalcification, loss of biomechanical properties, and reduced regeneration activity due to the use of strong acids and long-term EDTA-2Na treatment, which affects the fusion of fully-mineralized bone ECM with new bone tissue.
Innovation Solution
A method for preparing a gradient mineralized cancellous bone matrix material involves decellularization and ultrasound gradient demineralization of naturally-derived bone tissue to create a scaffold with controlled mineralization degrees, maintaining biomechanical properties and promoting regeneration.
Engineering Contradictions & Design Principles
Engineering Contradiction Analysis
1Loss of substance
If conventional strong acid and long-term EDTA-2Na decalcification is used to prepare fully demineralized bone matrix materials, then mineral removal is achieved, but irreversible damage occurs to the natural bone ECM scaffold material leading to loss of biomechanical properties, minerals, growth active factors, and three-dimensional microstructure
Solution Approach 1:
The patent changes the decalcification parameters from conventional strong acid/long-term treatment to weak acid (pH 4.0-5.0) short-term treatment (4-24 hours), thereby removing excessive minerals while preserving the bone ECM scaffold structure, biomechanical properties, and biological activity
Solution Approach 2:
The patent skips the conventional long-term decalcification process by using weak acid treatment for only 4-24 hours, rapidly achieving partial demineralization without causing irreversible damage to the bone matrix structure
2Strength
If fully-mineralized bone ECM material is used for bone tissue repair, then structural integrity is maintained, but it cannot be well fused with new bone to promote regeneration due to lack of biological activity
Solution Approach 1:
The patent creates local quality differences by achieving gradient mineralization where different regions of the bone ECM scaffold have different mineral contents, with the surface having lower mineral content for better biological activity and fusion with new bone, while the interior maintains higher mineral content for structural support
Solution Approach 2:
The patent applies partial demineralization rather than complete demineralization, removing only the excessive mineral content (achieving 30-70% mineral removal) to balance structural integrity with biological activity and regeneration ability
3Object-affected harmful factors
If conventional decellularization treatment is applied to remove cellular components, then immunogenicity is reduced, but the fusion with new bone tissue during hematoma organizing period is compromised
Solution Approach 1:
The patent applies partial decellularization rather than complete decellularization, removing only the excessive cellular components that cause immunogenicity while preserving sufficient cellular matrix components necessary for fusion with new bone tissue during the hematoma organizing period
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 results in a scaffold with low immunogenicity, rich biologically active components, and enhanced regeneration and vascularization, effectively promoting bone tissue repair and regeneration by maintaining biomechanical properties and mineral enrichment, thereby improving cell adhesion and differentiation.
Implementation Method 1
processing an obtained decellularized bone with an ultrasound gradient demineralization treatment
Implementation Method 2
ultrasound gradient demineralization treatment
Implementation Method 3
rinsing the bone blocks with deionized water containing protease inhibitor
Implementation Method 4
putting the embedding box into a deionized water solution containing acetone and shaking for 1-4 hours
Implementation Method 5
putting the embedding box into a deionized water solution containing tributyl phosphate and shaking for 1-4 hours
Implementation Method 6
freezing and thawing with liquid nitrogen for 2-6 cycles
Implementation Method 7
freezing and thawing with liquid nitrogen for 2-6 cycles, wherein each cycle is from −80° C. to 37° C.
Implementation Method 8
putting the embedding box in a buffer solution containing Triton® X-100, and shaking with a constant temperature shaker for 24 hours
Implementation Method 9
shaking the embedding box in a buffer solution containing SDS with the constant temperature shaker for 36 h
Data Source
AI summary
A gradient mineralized cancellous bone matrix material and a preparation method thereof are provided, and the preparation method includes: processing naturally-derived bone tissue with an immunogenicity removal treatment for decellularization, and processing an obtained decellularzed bone with a gradient demineralization treatment to obtain the gradient mineralized cancellous bone matrix material. The present invention expands a porosity of the bone matrix material and a collagen exposure degree on a surface thereof, which effectively releases growth factors and improves adhesion of the material to the cells, so as to up-regulate genes and proteins related to cell regeneration. The present invention not only retains the biomechanical properties and three-dimensional microstructure of natural bone ECM scaffolds, but also plays an active role for osteogenesis, angiogenesis and collagen mineralization in the early stage of fracture, thereby increasing engraftment adhesion of cells and promoting differentiation induction of cells.