P207 - Improving the Efficacy of Platelet Rich Plasma Through the Removal of Unwanted Cytokines
- K. Lydon (Rochester, US)
- K. Lydon (Rochester, US)
- C. Struijk (Rochester, US)
- J. Finnoff (Colorado Springs, US)
- D. Saris (Rochester, US)
Abstract
Purpose
Platelet Rich Plasma (PRP) is commonly used to enhance the repair of musculoskeletal injuries due to its high concentration of platelets and proteins involved in the wound healing process. Due to variation in PRP preparation, and composition between patients, the clinical effectiveness of PRP treatment remains a controversy. Although PRP contains proteins beneficial to the wound healing process, it also contains a significant number of inflammatory molecules that could impair the wound healing process. This project aims to develop a microsphere technology that will specifically remove inflammatory cytokines from PRP, and to assess the changes in regenerative capacity of PRP after the removal of these cytokines. We hypothesize that the removal of these proteins will improve the effect of PRP on musculoskeletal tissue and enhance repair.
Methods and Materials
Polystyrene microspheres (Bang Laboratories) were functionalized with antibodies specific to inflammatory proteins (IL-1β and TNF-α), using carbodiimide coupling. Functionalized microspheres were then added to a single protein solution (1% BSA in PBS) or complex protein solution such as platelet lysate, spiked with target protein, and allowed to react for 1 hour at room temperature. Solutions were then centrifuged, supernatant was analyzed with an ELISA, and microspheres immunofluorescently stained to ensure proteins were bound to microspheres.
Results
15µg of antibody functionalized microspheres (MS) specifically bound and removed more than 1ng of target protein from both simple and complex solutions (Fig.1). Immunofluorescent imaging confirmed that proteins were bound to antibodies on the surface of the microspheres (Fig.2).
Conclusion
A technology has been developed to remove unwanted inflammatory cytokines from PRP. The results of this study confirmed the effectiveness of this technology, and the ability of the functionalized MS to specifically bind to target proteins in complex solutions such as PRP. Ongoing in vitro experiments are analyzing the effect of removing inflammatory proteins from PRP on musculoskeletal co-culture models.