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Abstract

Platelet rich plasma (PRP) is a popular autologous therapy in clinic for tissue repair. Upon activation, platelets produce growth factors, which aid in tissue regeneration. However, previous literature shows that vascular endothelial growth factor (VEGF) can be detrimental to cartilage repair and regeneration due to the avascular nature of the tissue. Removal of VEGF from PRP via Avastin conjugated magnetic beads has been shown to promote improved cartilage formation. However, removal of VEGF via the VEGF inhibitor, Avastin, can produce adverse side effects if the dose is dispensed incorrectly or if it is delivered to patients who do not require the chemotherapeutic drug. Thus, we aim to investigate whether VEGF removal in PRP via Avastin conjugated magnetic beads promotes chondrogenesis of the chondrogenic mouse cell line, ATDC5. In this study, Avastin (Bevacizumab) is conjugated to DynabeadsTM M-270 Epoxy beads using the DynabeadsTM Antibody Coupling Kit. The beads were used to treat activated human PRP samples. VEGF and TGF-š¯›½ ELISAs were utilized to analyze the concentration of growth factors before and after bead treatment. In vitro ATDC5 experiments were conducted to determine PRPsā€™ effect on inducing chondrogenesis before and after treatment of VEGF removal. Our study suggests that the Avastin conjugated magnetic beads can selectively remove VEGF from PRP and, additionally, that the PRP with removed VEGF promotes the upregulation of key chondrogenesis factors such as Collagen 2 (Col2a1), while key hypertrophic cartilage factors such as Alp and VEGF are downregulated. Moreover, our research shows that Avastin conjugated beads can successfully and specifically remove VEGF from patient PRP samples. This proof-of-concept investigation shows that this technology could be used to remove specific growth factors, such as VEGF, in order to create specialized PRP therapies. Further investigation is needed using a more effective chondrogenesis model such as bone marrow-derived mesenchymal stem cells (BMSCs) or animal models.

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