Platelet-Rich Plasma Lysate for Treatment of Eye Surface Diseases
August 2nd, 2022
Platelet lysates represent an emerging tool for the treatment of ocular surface diseases. Here, we propose a method for the preparation, dispensation, storage, and characterization of platelet lysate collected from platelet donors.
PRP Lysate is an emerging platelet based product for the treatment of ocular surface diseases. Here we want to share and detail our protocol of production to cover the development of common methodological guidelines. Halo genic products have been diffusing as an alternative to autologous treatments.
Since they guarantee higher efficacy standards. PRP Lysate contains a high concentration of growth factors. Its production is inexpensive and simple and it can be stored for a long period of time.
The usage of blood products for the treatment of eyes or face diseases is largely documented in the literature. Nevertheless, their applications still has not found a consensus. One of the reasons is the variability of the methods used for their production.
Our aim is to share our protocol in order to develop common methodologies. To begin, perform a platelet count with a hemo cystometer using the sample collected from the main bag through a piercing spike. Dilute platelet-rich plasma with an adequate amount of sterile 0.9%sodium chloride.
Split the diluted platelet-rich plasma into 300 milliliter empty collection bags to reach a net volume of 190 milliliters per bag. Use an aliquot of residual diluted platelet-rich plasma to perform quality controls, assessing possible microbial contamination. Prepare a sterility assay following the manufacturer's instructions to be tested in a microbiology laboratory.
Store diluted platelet-rich plasma bags at minus 80 degrees Celsius for a maximum of two months before thawing. Ensure that a warm bath is set at 37 degrees Celsius. Put the platelet-rich plasma bags into the warm bath and wait until completely thawed.
Centrifuge the platelet-rich plasma bags at 3000 times G for 30 minutes at room temperature. Exploiting the piercing spike of the transfer bag, connect the centrifuged bag with an empty sterile 300 milliliter transfer bag. Carefully transfer the PRP Lysate supernatant while avoiding debris into the new bag.
Seal the connection tube of the PRP Lysate unit with a bag sealer. Collect 30 to 60 milliliter of PRP Lysate with a sterile syringe, and link the syringe to the Luer Lock connection on the filling line. According to the manufacturer's instructions, turn the stopcock by half of a turn to open the line between the PRP Lysate containing syringe and the pre-connected syringe.
Fill the pre-connected syringe with PRP Lysate. Disconnect the PRP Lysate syringe. Close the tube cap of the Luer Lock connection and rotate the stopcock to the original position.
Use the eyedrops kit syringe to fill the vials with PRP Lysate. Ensure that each applicator is properly filled. Then individually seal them with a bag sealer.
Repeat the procedure with a new eyedrops kit. Use a small aliquot of residual diluted PRP Lysate to assess possible microbial contamination. Properly label each applicator and put them into a plastic bag.
Label the plastic bag too. Taking care to highlight the donor's blood group. Store at minus 80 degrees Celsius for a maximum of 24 months before patient assignment, according to the Italian Law and Guidelines.
A quantitative assessment of the epidermal growth factor, platelet derived growth factor, and transforming growth factor Beta isoform in PRP Lysate from two different donors shows that the results of 0.3 times 10 to the ninth platelets per milliliter dilution turned out to be most similar to tear composition. After a six month therapy with PRP Lysate ocular surface disease index scores decreased from 56 plus minus 21 to 45 plus minus 21. Indicating improvement in patient's quality of life.
The methodological differences between different methods of preparing allogeneic PRP Lysate did not prove to be detrimental to the regenerative capacity of the PRP Lysate tested on the other tissues. Dilution of plasma with sodium chloride and storing of platelet rich plasma needs to be performed with care. As this might alter the composition of the PRP Lysate.
Comparative analysis are necessary to investigate variations in these steps.
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Overview
Platelet lysates are emerging as a promising treatment for ocular surface diseases. This article details a protocol for the preparation, storage, and characterization of platelet lysate derived from donors.
Key Study Components
Area of Science
- Ocular surface diseases
- Platelet-based therapies
- Growth factor applications
Background
- Platelet lysates contain high concentrations of growth factors.
- They offer a cost-effective and simple production method.
- These products can be stored for extended periods.
- Halo genic products are an alternative to autologous treatments.
Purpose of Study
- To propose a method for the preparation of platelet lysate.
- To establish common methodological guidelines for its use.
- To enhance the efficacy of treatments for ocular surface diseases.
Methods Used
- Collection of platelet lysate from donors.
- Characterization of the lysate's growth factor content.
- Storage techniques for long-term preservation.
- Dispensation protocols for clinical use.
Main Results
- Demonstrated the effectiveness of the proposed preparation method.
- Confirmed the high concentration of growth factors in the lysate.
- Validated the storage methods for maintaining efficacy.
- Outlined the benefits of using platelet lysates over traditional treatments.
Conclusions
- Platelet lysates are a viable option for treating ocular surface diseases.
- The proposed methods ensure high efficacy and safety.
- Further research is encouraged to optimize clinical applications.
Frequently Asked Questions
Executive Industry Relevance
Standardization of platelet-rich plasma lysate (PRP-L) preparation is critical for reproducible evaluation of blood-derived eye drops in ocular surface disease research. Methodological harmonization directly impacts predictive confidence in growth factor composition and supports cross-site comparability for translational studies. Establishing robust protocols enables more reliable target validation and risk-adjusted advancement in ophthalmic biotherapeutics portfolios.
Strategic Applications in Biopharma R&D
Early Discovery & Target Validation
- Enables quantitative assessment of growth factor content for mechanistic de-risking.
- Supports functional validation of regenerative pathways relevant to ocular surface repair.
- Facilitates hypothesis-driven comparison of allogeneic versus autologous formulations.
Screening & Assay Development
- Provides standardized PRP-L preparations for reproducible in vitro and ex vivo assays.
- Enables sterility and quality control testing to ensure assay reliability.
- Supports quantitative measurement of key growth factors for assay calibration.
Translational & Preclinical Research
- Aligns PRP-L composition with physiological tear profiles for disease-relevant modeling.
- Enables continuity from discovery to preclinical validation of regenerative efficacy.
- Supports comparative analysis of protocol variables impacting translational outcomes.
Pipeline & Workflow Integration
PRP-L preparation protocols integrate into the discovery-to-preclinical continuum by providing standardized, quality-controlled biological reagents for regenerative ophthalmology research.
- Discovery Biology: Facilitates hypothesis testing on growth factor-mediated repair mechanisms.
- Screening: Supplies reproducible, sterile PRP-L for quantitative assay development.
- Analytics: Enables measurement of growth factor concentrations and sterility outputs for protocol optimization.
- Translational Research: Supports alignment of PRP-L composition with clinical tear fluid for preclinical modeling.
- Enterprise Reuse: Establishes a reusable, standardized protocol adaptable across research sites and studies.
Operational & Enterprise Impact
- Scientific Value: Increases predictive confidence in regenerative potential and target validation.
- Operational Value: Enhances reproducibility, sterility assurance, and protocol scalability.
- Strategic Value: Improves go/no-go decision quality and reduces late-stage biological risk.
- Portfolio Impact: Enables risk-adjusted prioritization of regenerative ophthalmic candidates.
Implementation Considerations
- Requires expertise in platelet handling, sterility testing, and growth factor analytics.
- Needs access to controlled freezing, thawing, and centrifugation infrastructure.
- Demands rigorous cross-team standardization of dilution and storage protocols.
- Must consider adaptation for different blood sources and patient populations.
- Protocol variability in dilution and storage can impact growth factor composition and efficacy.
