De-fibrinated Platelet Lysate Preparation via Fibrinogen Extraction
Solution Overview
Problem
Current methods for preparing platelet-rich lysate face challenges such as variability in platelet concentration, use of bovine thrombin, and the formation of fibrin clots that trap growth factors, leading to inefficiencies and potential immune reactions, along with the need for animal-derived anticoagulants.
Innovation Solution
A method is developed to remove fibrinogen from plasma using a fibrinogen-selective binding agent like polyethylene glycol (PEG), allowing for the creation of a platelet lysate depleted of fibrinogen, which preserves growth factors and avoids the use of bovine thrombin and animal-derived anticoagulants.
Engineering Contradictions & Design Principles
Engineering Contradiction Analysis
1Quantity of substance
If freeze/thaw cycles are used to lyse platelets, then platelet lysis is achieved, but fibrin clots form that trap growth factors
Solution Approach 1:
The patent applies preliminary action by adding a fibrinogen-selective binding agent to plasma before platelet lysis occurs. This removes fibrinogen from the plasma in advance, preventing fibrin clot formation during subsequent freeze/thaw cycles. The binding agent is added to plasma prior to platelet activation and lysis, ensuring that when growth factors are released, they remain soluble and available rather than being trapped in clots.
2Quantity of substance
If bovine thrombin is used to activate platelets, then platelet activation is achieved, but immune reactions and disease transmission risks increase
Solution Approach 1:
The patent replaces bovine thrombin with calcium chloride, which is a small inorganic salt without immuneogenic properties. Calcium chloride serves the same function of activating platelets by triggering the coagulation cascade, but unlike bovine thrombin, it does not carry risks of immune reactions or disease transmission. This substitution uses a simple, non-biological reagent that achieves the same activation purpose without the harmful associations.
3Reliability
If anticoagulants are added to cell culture media to prevent clotting, then clotting is prevented, but animal-derived products are used which carry disease transmission risks
Solution Approach 1:
The patent extracts fibrinogen from plasma using a fibrinogen-selective binding agent, thereby removing the substrate required for fibrin clot formation. By taking out fibrinogen before platelet lysis, the need for anticoagulants in the final product is eliminated. The resulting platelet lysate is inherently resistant to clotting because the essential component (fibrinogen) has been removed, not because anticoagulants are present.
4Reliability
If heparin is used to control clotting, then clotting is controlled, but the final product becomes more expensive
Solution Approach 1:
The patent extracts fibrinogen from plasma using a fibrinogen-selective binding agent, thereby removing the substrate required for fibrin clot formation. By taking out fibrinogen before platelet lysis, the need for anticoagulants in the final product is eliminated. The resulting platelet lysate is inherently resistant to clotting because the essential component (fibrinogen) has been removed, not because anticoagulants are present.
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 results in a standardized growth factor concentrate that enhances therapeutic applications by maintaining the efficacy of growth factors while reducing variability and immune reaction risks, providing a more reliable and safer product for tissue regeneration and cell culture.
Implementation Method 1
A method is developed to remove fibrinogen from plasma using a fibrinogen-selective binding agent like polyethylene glycol (PEG)
Implementation Method 2
This was termed Platelet Lysate (PL). Freezing, however, has been reported to cause the formation of a fibrin clot
Data Source
AI summary
This invention relates to the methods of preparation of a growth factor concentrate derived from platelet lysate and modified to be devoid of fibrinogen. Moreover, the present invention concerns therapeutic applications of said growth factor concentrate to enhance mitogenic, osteogenic, chondrogenic and angiogenic abilities. The disclosed preparation can be used immediately, or stored frozen until use, or lyophilized, or mixed with pharmaceutical formulation, such as biodegradable polymer or other delivery vehicles, to make products.