Blood-derived anti-inflammatory protein solution blocks the effect of IL-1β on human macrophages in vitro

Abstract

Objective The purpose of this research was to determine if an autologous protein solution (APS), prepared from platelet-rich plasma (PRP), could reduce the deleterious effects of inflammatory cytokines in vitro. Methods APS was prepared by processing human blood in a tuned density buoy separation device (Platelet Separation System, Biomet Biologics, LLC) to produce platelet-rich plasma (PRP) and processing the PRP in a concentration device containing polyacrylimide beads to produce a highly concentrated anti-inflammatory solution. A functional assay was designed using recombinant interleukin (IL)-1b to upregulate IL-8 production by human monocytes. Either recombinant human interleukin-1 receptor antagonist (rhIL-1ra) or APS was added to some samples to determine if a reduced inflammatory response could be identified in vitro. The enzyme-linked immunosorbent assay (ELISA) method was employed to perform cytokine analyses, and Student's t test (a = 0.05) was used for all statistical analyses. Results Both the rhIL-1ra and the APS reduced the effect of IL-1b on human macrophages in vitro. This was measured by the reduced production of IL-8 and tumor necrosis factor (TNF)-a. Further analysis of the supernatants confirmed the presence of high concentrations of IL-1ra and soluble TNF receptor I (sTNF-RI) with the APS treatment. Conclusion The ability of the APS to reduce the effect of IL-1b and limit the expression of other inflammatory cytokines in vitro validates its potential use as an autologous treatment for osteoarthritis.

Autologous solution protects bovine cartilage explants from IL‐1α‐ and TNFα‐induced cartilage degradation

“…The results of this study demonstrated for the first time APS's protective effects on cartilage tissue and proliferative effects on cells inside the cartilage tissue. These observations provide a “bridge” between previous APS studies using cultured cells involved in OA progression and the positive results of an equine OA clinical trial . Demonstrating that APS has OA modifying‐effects on cells, tissues, and entire organisms could be crucial in the development of future human clinical trials.…”

“…Previous research has demonstrated that APS inhibits the inflammatory responses of monocytes and chondrocytes to IL‐1 and/or TNFα in vitro. The outcome measures of those studies, IL‐8 and MMP‐13 concentrations produced by the cultured cells, indicated that APS may act to inhibit cartilage degradation . APS also has demonstrated improved gate biomechanics, lameness, and pain in a prospective and double‐blinded clinical trial to treat equine OA .…”

Human blood‐based anti‐inflammatory solution inhibits osteoarthritis progression in a meniscal‐tear rat study

“…Cell culture studies, animal trials, and human clinical trials have supported the anti‐inflammatory mechanism of action for APS therapy. Cell culture studies have demonstrated that APS inhibits the IL‐1β and TNFα inflammatory signaling cascade, as well as the secretion of MMPs . In a cartilage tissue explant study, APS reduced inflammatory cytokine‐prompted GAG and collagen release and had a proliferative effect on chondrocytes .…”

Platelet-Rich Plasma Increases Matrix Metalloproteinases in Cultures of Human Synovial Fibroblasts

“…The molecular complexity of PRP preparations necessitates further study to determine the net overall effect for each clinical application 4,7 . For intra-articular applications, the effect of PRP on chondrocytes has been studied in cell and tissue culture 8,9 .…”