Characterization and Chondroprotective Effects of Extracellular Vesicles From Plasma- and Serum-Based Autologous Blood-Derived Products for Osteoarthritis Therapy.

Alexander Otahal,Stefan Nehrer,Karina Kramer,Zsombor Lacza,Olga Kuten-Pella,Andrea De Luna,Christoph Stotter,René Weiss,Viktoria Weber

doi:10.3389/fbioe.2020.584050

Abstract

Autologous blood products gain increasing interest in the field of regenerative medicine as well as in orthopedics, aesthetic surgery, and cosmetics. Currently, citrate-anticoagulated platelet-rich plasma (CPRP) preparations are often applied in osteoarthritis (OA), but more physiological and cell-free alternatives such as hyperacute serum (hypACT) are under development. Besides growth factors, blood products also bring along extracellular vesicles (EVs) packed with signal molecules, which open up a new level of complexity at evaluating the functional spectrum of blood products. Large proportions of EVs originated from platelets in CPRP and hypACT, whereas very low erythrocyte and monocyte-derived EVs were detected via flow cytometry. EV treatment of chondrocytes enhanced the expression of anabolic markers type II collagen, SRY-box transcription factor 9 (SOX9), and aggrecan compared to full blood products, but also the catabolic marker and tissue remodeling factor matrix metalloproteinase 3, whereas hypACT EVs prevented type I collagen expression. CPRP blood product increased SOX9 protein expression, in contrast to hypACT blood product. However, hypACT EVs induced SOX9 protein expression while preventing interleukin-6 secretion. The results indicate that blood EVs are sufficient to induce chondrogenic gene expression changes in OA chondrocytes, while preventing proinflammatory cytokine release compared to full blood product. This highlights the potential of autologous blood-derived EVs as regulators of cartilage extracellular matrix metabolism and inflammation, as well as candidates for new cell-free therapeutic approaches for OA.

Highlights

Blood-derived products comprise plasma- or serum-based whole blood derivatives
citrate-anticoagulated platelet-rich plasma (CPRP) production enriched platelets by two rounds of centrifugation in presence of the anticoagulant citrate, whereas hyperacute serum (hypACT) was generated in a closed system involving blood clotting and squeezing serum components from the resulting fibrin clot without an anticoagulant
HypACT was in essence devoid of platelets [t(5) = 3.043; p = 0.0287] while containing significantly more erythrocytes [t(5) = 5.998; p = 0.0018] and monocytes [t(5) = 3.816; p = 0.0124] than CPRP

Summary

Introduction

Blood-derived products comprise plasma- or serum-based whole blood derivatives. They have been applied to diseased joints of osteoarthritis (OA) patients via intra-articular injection for over a decade. Blood products are well tolerated and provide reduction of pain and inflammation, as well as increased range of motion (Fotouhi et al, 2018; Tassara et al, 2018; Zarringam et al, 2018; Gato-Calvo et al, 2019). In the context of orthopedics, platelet-rich plasma (PRP) is most commonly used and is characterized by a platelet concentration that exceeds the physiological concentration. At least 21 different PRP production devices are currently available and are applied without standardization (Gato-Calvo et al, 2019; Oudelaar et al, 2019). Donor variability impacts the cell profiles of the final product, and secretory components such as growth factors (Mazzocca et al, 2012)

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