Faculty Opinions recommendation of Attenuative Effects of Platelet-Rich Plasma on 30 kDa Fibronectin Fragment-Induced MMP-13 Expression Associated with TLR2 Signaling in Osteoarthritic Chondrocytes and Synovial Fibroblasts.

Proteolytic fragments of fibronectin can have catabolic effects on cartilage, menisci, and synovium. Previous studies have reported that Toll-like receptor (TLR) signaling pathways might be associated with joint inflammation and joint destruction. Platelet-rich plasma (PRP) is increasingly being used to treat a range of joint conditions; however, it has yet to be determined whether PRP influences fibronectin fragment (FN-f) procatabolic activity and TLRs. In this study, human primary culture cells were treated with 30 kDa FN-f with/without PRP co-incubation, and then analyzed using real-time PCR to determine gene expression levels in articular chondrocytes, meniscal fibrochondrocytes, and synovial fibroblasts. Protein levels were evaluated by Western immunoblotting. This study observed an increase in the protein expression of matrix metalloproteinases (MMPs), Toll-like receptor 2 (TLR2), nitric oxide synthase 2 (NOS2), prostaglandin-endoperoxide synthase (PTGS2), and cyclooxygenase 2 (COX2) in articular chondrocytes, meniscal fibrochondrocytes, and synovial fibroblasts following insult with 30 kDa FN-f. Upregulation of these genes was significantly attenuated by PRP treatment. TLR2 and matrix metalloproteinase 13 (MMP-13) were also significantly attenuated by cotreatment with 30 kDa FN-f + PRP + TLR2 inhibitor. PRP treatment was shown to attenuate the 30 kDa FN-f-induced MMP-13 expression associated with the decreased expression of TLR2 in osteoarthritic chondrocytes and synovial fibroblasts. PRP treatment was also shown to attenuate procatabolic activity associated with MMP-13 expression via the TLR2 signaling pathway.

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260 PLATELET RICH PLASMA HAS ANTI-INFLAMMATORY EFFECTS ON OSTEOARTHRITIC CHONDROCYTES

Osteoarthritis and Cartilage ◽

10.1016/s1063-4584(11)60287-2 ◽

2011 ◽

Vol 19 ◽

pp. S124-S125

Author(s):

G.M. van Buul ◽

W.L. Koevoet ◽

N. Kops ◽

P.K. Bos ◽

J.A. Verhaar ◽

...

Keyword(s):

Platelet Rich Plasma ◽

Osteoarthritic Chondrocytes ◽

Anti Inflammatory

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Platelet-Rich Plasma Releasate Inhibits Inflammatory Processes in Osteoarthritic Chondrocytes

The American Journal of Sports Medicine ◽

10.1177/0363546511419278 ◽

2011 ◽

Vol 39 (11) ◽

pp. 2362-2370 ◽

Cited By ~ 192

Author(s):

Gerben M. van Buul ◽

Wendy L.M. Koevoet ◽

Nicole Kops ◽

P. Koen Bos ◽

Jan A.N. Verhaar ◽

...

Keyword(s):

Gene Expression ◽

Matrix Metalloproteinase ◽

Platelet Rich Plasma ◽

Interleukin 1 ◽

Cartilage Degeneration ◽

No Production ◽

Major Pathway ◽

Mediated Effects ◽

Osteoarthritic Chondrocytes ◽

A Disintegrin And Metalloproteinase

Background: Platelet-rich plasma (PRP) has recently been postulated as a treatment for osteoarthritis (OA). Although anabolic effects of PRP on chondrocytes are well documented, no reports are known addressing effects on cartilage degeneration. Since OA is characterized by a catabolic and inflammatory joint environment, the authors investigated whether PRP was able to counteract the effects of such an environment on human osteoarthritic chondrocytes. Hypothesis: Platelet-rich plasma inhibits inflammatory effects of interleukin-1 (IL-1) beta on human osteoarthritic chondrocytes. Study Design: Controlled laboratory study. Methods: Human osteoarthritic chondrocytes were cultured in the presence of IL-1 beta to mimic an osteoarthritic environment. Medium was supplemented with 0%, 1%, or 10% PRP releasate (PRPr, the active releasate of PRP). After 48 hours, gene expression of collagen type II alpha 1 (COL2A1), aggrecan (ACAN), a disintegrin and metalloproteinase with thrombospondin motifs (ADAMTS)4, ADAMTS5, matrix metalloproteinase (MMP)13, and prostaglandin-endoperoxide synthase (PTGS)2 was analyzed. Additionally, glycosaminoglycan (GAG) content, nitric oxide (NO) production, and nuclear factor kappa B (NFκB) activation were studied. Results: Platelet-rich plasma releasate diminished IL-1 beta–induced inhibition of COL2A1 and ACAN gene expression. The PRPr also reduced IL-1 beta–induced increase of ADAMTS4 and PTGS2 gene expression. ADAMTS5 gene expression and GAG content were not influenced by IL-1 beta or additional PRPr. Matrix metalloproteinase 13 gene expression and NO production were upregulated by IL-1 beta but not affected by added PRPr. Finally, PRPr reduced IL-1 beta–induced NFκB activation to control levels containing no IL-1 beta. Conclusion: Platelet-rich plasma releasate diminished multiple inflammatory IL-1 beta–mediated effects on human osteoarthritic chondrocytes, including inhibition of NFκB activation. Clinical Relevance: Platelet-rich plasma releasate counteracts effects of an inflammatory environment on genes regulating matrix degradation and formation in human chondrocytes. Platelet-rich plasma releasate decreases NFκB activation, a major pathway involved in the pathogenesis of OA. These results encourage further study of PRP as a treatment for OA.

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Release kinetic of pro- and anti-inflammatory biomolecules from platelet-rich plasma and functional study on osteoarthritis synovial fibroblasts

Cytotherapy ◽

10.1016/j.jcyt.2020.02.006 ◽

2020 ◽

Vol 22 (7) ◽

pp. 344-353 ◽

Cited By ~ 2

Author(s):

Erminia Mariani ◽

Alice Roffi ◽

Luca Cattini ◽

Lia Pulsatelli ◽

Elisa Assirelli ◽

...

Keyword(s):

Platelet Rich Plasma ◽

Synovial Fibroblasts ◽

Functional Study ◽

Release Kinetic ◽

Anti Inflammatory

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Optimization of Lyophilized Hyperacute Serum (HAS) as a Regenerative Therapeutic in Osteoarthritis

International Journal of Molecular Sciences ◽

10.3390/ijms22147496 ◽

2021 ◽

Vol 22 (14) ◽

pp. 7496

Author(s):

Isabel Olmos Calvo ◽

Olga Kuten-Pella ◽

Karina Kramer ◽

Ágnes Madár ◽

Szilvia Takács ◽

...

Keyword(s):

Gene Expression ◽

Gene Expression Analysis ◽

Platelet Rich Plasma ◽

Cell Types ◽

Inflammatory Conditions ◽

Osteoarthritic Chondrocytes ◽

Prp Gene ◽

Human Explants ◽

Cytokine Quantification

Hyperacute serum (HAS) is a blood derivative product that promotes the proliferation of various cell types and controls inflammation in vitro. The aim of this study is to investigate the regenerative potential of different formulations of HAS, including lyophilized and hyaluronic acid combined versions, to obtain a stable and standardized therapeutic in osteoarthritis (OA), which may be able to overcome the variability limitations of platelet-rich plasma (PRP). Primary human osteoarthritic chondrocytes were used for testing cellular viability and gene expression of OA-related genes. Moreover, a co-culture of human explants of cartilage, bone and synovium under inflammatory conditions was used for investigating the inflammatory control capacities of the different therapeutics. In this study, one formulation of lyophilized HAS achieved the high cell viability rates of liquid HAS and PRP. Gene expression analysis showed that HAS induced higher Col1a1 expression than PRP. Cytokine quantification from supernatant fluids revealed that HAS treatment of inflamed co-cultures significantly reduced levels of IL-5, IL-15, IL-2, TNFα, IL-7 and IL-12. To conclude, lyophilized HAS is a stable and standardized therapeutic with high potential in joint regeneration.

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Moving toward targeting the right phenotype with the right platelet-rich plasma (PRP) formulation for knee osteoarthritis

Therapeutic Advances in Musculoskeletal Disease ◽

10.1177/1759720x211004336 ◽

2021 ◽

Vol 13 ◽

pp. 1759720X2110043

Author(s):

Isabel Andia ◽

Leire Atilano ◽

Nicola Maffulli

Keyword(s):

Clinical Outcomes ◽

Platelet Rich Plasma ◽

Synovial Fibroblasts ◽

Imaging Biomarkers ◽

Knee Oa ◽

Provide Pain Relief ◽

Inflammatory Phenotype ◽

Potential Benefits ◽

The Right ◽

Meta Analyses

Intra-articular injections of platelet-rich plasma (PRP) and other novel blood-derived products developed specifically for osteoarthritis (OA) can provide pain relief and potential benefits in disease progression. Meta-analyses show the clinical superiority of PRP compared with other intra-articular injections, but results are modest and the effect sizes are small. PRP injections in knee OA are performed indiscriminately, but the clinical response varies enormously between patients because of an array of mixed OA phenotypes. Subgroup analyses are scarce; some studies stratify patients according to radiographic severity and found better results in early OA, without consensus for more advanced stages of the condition. Parallel identification of soluble and imaging biomarkers is essential to personalise and leverage PRP therapies. The inflammatory phenotype is most interesting from the PRP perspective because PRPs modulate inflammation by releasing a large pool of chemokines and cytokines, which interact with synovial fibroblasts and macrophages; in addition, they can modulate the innate immune response. No soluble biomarkers have been discovered that have implications for OA research and PRP interventions. Clinical examination of patients based on their inflammatory phenotype and imaging identification of pain sources and structural alterations could help discern who will respond to PRP. Synovial inflammation and bone marrow lesions are sources of pain, and intra-articular injections of PRP combined with subchondral bone injection can enhance clinical outcomes. Further refining ultrasound phenotypes may aid in personalising PRP therapies. Intra-articular delivery combined with injections in altered ligamentous structures, medial and coronal ligaments or premeniscal pes anserinus showed positive clinical outcomes. Although the evidence supporting these approaches are weak, they merit further consideration to refine PRP protocols and target the right OA phenotypes.

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270 HYALURONAN SUPPRESSES P38 MITOGEN-ACTIVATED PROTEIN KINASE IN OSTEOARTHRITIC CHONDROCYTES STIMULATED WITH COOH-TERMINAL HEPARIN-BINDING FIBRONECTIN FRAGMENT

Osteoarthritis and Cartilage ◽

10.1016/s1063-4584(09)60292-2 ◽

2009 ◽

Vol 17 ◽

pp. S147

Author(s):

T. Yasuda

Keyword(s):

Protein Kinase ◽

Mitogen Activated Protein Kinase ◽

Heparin Binding ◽

Fibronectin Fragment ◽

Osteoarthritic Chondrocytes ◽

Mitogen Activated Protein

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Components of the nuclear signaling cascade that regulate collagenase gene expression in response to integrin-derived signals.

The Journal of Cell Biology ◽

10.1083/jcb.129.6.1707 ◽

1995 ◽

Vol 129 (6) ◽

pp. 1707-1720 ◽

Cited By ~ 81

Author(s):

P Tremble ◽

C H Damsky ◽

Z Werb

Keyword(s):

Gene Expression ◽

Synovial Fibroblasts ◽

Plasma Fibronectin ◽

Reporter Construct ◽

Fibronectin Receptor ◽

Receptor Antibody ◽

Fibronectin Fragment ◽

Integrin Fibronectin Receptor ◽

Collagenase Gene ◽

In Cells

We have shown previously that the expression of collagenase is upregulated in rabbit synovial fibroblasts cultured on a substrate of antibody to the alpha 5 chain of the alpha 5 beta 1 integrin fibronectin receptor or on the 120-kD cell-binding chymotryptic fragment of plasma fibronectin, but remains at basal levels in cells plated on intact plasma fibronectin. We now have identified some of the components of a signaling pathway that couples the fibronectin receptor to the induction of collagenase transcription. We studied the control of collagenase gene expression in cells adhering to the 120-kD fragment of fibronectin, to antifibronectin receptor antibody, or to plasma fibronectin by transiently introducing promoter-reporter constructs into rabbit synovial fibroblasts before plating cells on these matrices. The constructs contained segments of the human collagenase promoter regulating transcription of chloramphenicol acyl transferase. Expression of constructs containing the -1200/-42-bp segment or the -139/-42-bp segment of the collagenase promoter inserted upstream from the reporter gene was induced to similar extents in cells plated on the 120-kD fragment of fibronectin or on anti-fibronectin receptor antibody, relative to that in fibroblasts plated on fibronectin. The expression of the construct containing the -66/-42-bp segment of the promoter was not regulated and was similar to that of the parent pBLCAT2 plasmid, suggesting that the -139/-67 region of the collagenase promoter, which contains PEA3- and AP1-binding sites, regulates the transcription of collagenase caused by integrin-derived signals. Expression of a reporter construct containing only the PEA3 and AP1 sites in the collagenase promoter (-90/-67) also increased in cells plated on the 120-kD fragment of fibronectin or on anti-fibronectin receptor antibody, relative to that in cells plated on fibronectin. Mutations in either the AP1 or PEA3 site of this minimal promoter abrogated its activity in cells plated on these inductive ligands. Expression of c-fos mRNA increased within 1 h of plating cells on the 120-kD fibronectin fragment or on anti-fibronectin receptor antibody, relative to that in cells plated on fibronectin. c-Fos protein accumulated in the nuclei of fibroblasts within 10 min of plating on the 120-kD fibronectin fragment. The increase in c-Fos was required for the increase in collagenase in cells plated on the 120-kD fibronectin fragment: incubation of cells with antisense, but not sense, c-fos oligonucleotides diminished both basal and induced expression of the -139/-42 collagenase promoter-reporter construct and decreased expression of the endogenous collagenase gene.(ABSTRACT TRUNCATED AT 400 WORDS)

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