Vacuum-Assisted Cryoprotectant Infiltration for Cadaver Bone
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Solution Overview
Problem
The existing methods for obtaining bone marrow are limited by the difficulty in extracting high numbers of viable hematopoietic stem cells from frozen cadaver bone, due to challenges in cryopreservation and subsequent recovery, which hinders the mainstream use of cadaveric bone marrow for transplantation.
Innovation Solution
A method involving cryopreservation of cadaver bone using a cryoprotectant solution, where the bone is subjected to reduced pressure to remove water, followed by increased pressure to infiltrate the cryoprotectant, and then chilled to preserve the bone, allowing for later thawing and extraction of bone marrow cells, enhancing the viability and quantity of extracted cells.
Engineering Contradictions & Design Principles
Engineering Contradiction Analysis
1Quantity of substance
If traditional aspiration methods are used to obtain bone marrow, then the procedure is simple and quick, but the volume of bone marrow obtained is limited to small volumes (~100 ml) from living donors
Solution Approach 1:
The bone is cryopreserved in advance with cryoprotectant infiltration before cell extraction is needed. This preliminary preservation allows the bone to be stored and later thawed for cell recovery, enabling access to cadaver donors and significantly increasing the available volume of bone marrow beyond what can be obtained from living donors through aspiration.
2Quantity of substance
If cadaver bone is frozen for later cell recovery, then a vastly more abundant source of bone marrow can be obtained, but multiple barriers prevent mainstream use due to challenges in obtaining high numbers of viable cells from frozen donor bone
Solution Approach 1:
The patent changes the physical and chemical parameters during cryopreservation by using vacuum-assisted infiltration to achieve deep penetration of cryoprotectant throughout the bone matrix. This parameter optimization (pressure control, cryoprotectant concentration, infiltration time) ensures that cells remain viable after freezing and thawing, resolving the contradiction between obtaining abundant cells and maintaining their viability.
Solution Approach 2:
The patent employs vacuum-assisted infiltration (pneumatic principle) to force cryoprotectant solution deep into the bone marrow cavity. By applying negative pressure followed by positive pressure, the cryoprotectant penetrates throughout the bone structure, ensuring uniform protection of cells during freezing. This pneumatic approach overcomes the barrier of obtaining viable cells from frozen bone by ensuring proper cryoprotectant distribution.
3Ease of manufacture
If whole vertebral bodies are used for cryopreservation, then a large smart bank can be created with minimal manipulation and cost, but the difficulty of extracting high numbers of viable cells from frozen whole bone is significant
Solution Approach 1:
The vacuum-assisted infiltration system allows cryoprotectant to penetrate deep into whole vertebral bodies without requiring segmentation or complex processing. The pressure differential forces the solution throughout the bone matrix, enabling cryopreservation of intact whole bones with minimal manipulation. This resolves the contradiction by making whole bone processing as easy as fragmented bone while achieving high cell yields.
Solution Approach 2:
The bone marrow cavity and trabecular structure act as a porous medium that allows cryoprotectant infiltration under vacuum. The porous nature of the bone matrix enables deep penetration of the preservation solution throughout the whole vertebral body, ensuring viable cell preservation without requiring complex processing or segmentation of the bone.
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 method significantly increases the yield of viable bone marrow cells, including hematopoietic stem cells and mesenchymal stromal/stem cells, by up to 1000-fold, improving the efficiency and effectiveness of bone marrow extraction from cadaveric sources.
Implementation Method 1
reducing the pressure in the closed container, and optionally, holding the closed container at reduced pressure, to remove at least a portion of the water present in the cadaver bone; raising the pressure in the closed container and holding the closed container at a raised pressure to allow infiltration of the cryoprotectant solution into the cadaver bone
Implementation Method 2
placing a cadaver bone in a closed container comprising a cryoprotectant solution; chilling the cadaver bone to a temperature at least below 0° C., thereby cryopreserving the cadaver bone
Implementation Method 3
reducing the pressure in the closed container, and optionally, holding the closed container at reduced pressure, to remove at least a portion of the water present in the cadaver bone
Data Source
AI summary
The present disclosure relates to, at least, a vacuum-assisted method for infiltrating cadaver bone with a cryoprotectant and a method for rapidly warming the cryopreserved cadaver bone for bone marrow processing.