Intra-articular TSG-6 delivery from heparin-based microparticles reduces cartilage damage in a rat model of osteoarthritis.

As a potential treatment for osteoarthritis (OA), we have developed injectable and hydrolytically degradable heparin-based biomaterials with tunable sulfation for the intra-articular delivery of tumor necrosis factor-alpha stimulated gene-6 (TSG-6), a protein known to inhibit plasmin which may degrade extracellular matrix within OA joints. We first assessed the effect of heparin sulfation on TSG-6 anti-plasmin activity and found that while fully sulfated (Hep) and heparin desulfated at only the N position (Hep-N) significantly enhanced TSG-6 bioactivity in vitro, fully desulfated heparin (Hep-) had no effect, indicating that heparin sulfation plays a significant role in modulating TSG-6 bioactivity. Next, TSG-6 loaded, degradable 10 wt% Hep-N microparticles (MPs) were delivered via intra-articular injection into the knee at 1, 7, and 15 days following medial meniscal transection (MMT) injury in a rat model. After 21 days, cartilage thickness, volume, and attenuation were significantly increased with soluble TSG-6, indicating degenerative changes. In contrast, no significant differences were observed with TSG-6 loaded MP treatment, demonstrating that TSG-6 loaded MPs reduced cartilage damage following MMT injury. Ultimately, our results indicate that Hep-N can enhance TSG-6 anti-plasmin activity and that Hep-N-based biomaterials may be an effective method for TSG-6 delivery to treat OA.

[1]  P. Roughley,et al.  Cartilage-specific constitutive expression of TSG-6 protein (product of tumor necrosis factor alpha-stimulated gene 6) provides a chondroprotective, but not antiinflammatory, effect in antigen-induced arthritis. , 2002, Arthritis and rheumatism.

[2]  Aaron D Baldwin,et al.  Production of heparin-functionalized hydrogels for the development of responsive and controlled growth factor delivery systems. , 2007, Journal of controlled release : official journal of the Controlled Release Society.

[3]  A. Parkar,et al.  Overlapping sites on the Link module of human TSG‐6 mediate binding to hyaluronan and chondroitin‐4‐sulphate , 1997, FEBS letters.

[4]  R. Guldberg,et al.  Quantitative pre-clinical screening of therapeutics for joint diseases using contrast enhanced micro-computed tomography. , 2016, Osteoarthritis and cartilage.

[5]  T. Glant,et al.  Anti-inflammatory and chondroprotective effect of TSG-6 (tumor necrosis factor-alpha-stimulated gene-6) in murine models of experimental arthritis. , 2001, The American journal of pathology.

[6]  S. Anand,et al.  A novel chondroprotective property of TSG-6 has therapeutic potential for OA , 2016 .

[7]  Shingo Nakamura,et al.  Controlled release of fibroblast growth factor-2 from an injectable 6-O-desulfated heparin hydrogel and subsequent effect on in vivo vascularization. , 2006, Journal of biomedical materials research. Part A.

[8]  H. Ronday,et al.  Difference in expression of the plasminogen activation system in synovial tissue of patients with rheumatoid arthritis and osteoarthritis. , 1996, British journal of rheumatology.

[9]  G. Borschel,et al.  Controlled release of glial-derived neurotrophic factor from fibrin matrices containing an affinity-based delivery system. , 2009, Journal of biomedical materials research. Part A.

[10]  J. Temenoff,et al.  Dual Affinity Heparin-Based Hydrogels Achieve Pro-Regenerative Immunomodulation and Microvascular Remodeling. , 2017, ACS biomaterials science & engineering.

[11]  R E Guldberg,et al.  Quantitative assessment of articular cartilage morphology via EPIC-microCT. , 2009, Osteoarthritis and cartilage.

[12]  A. Imberty,et al.  Heparan Sulfate/Heparin Oligosaccharides Protect Stromal Cell-derived Factor-1 (SDF-1)/CXCL12 against Proteolysis Induced by CD26/Dipeptidyl Peptidase IV* , 2004, Journal of Biological Chemistry.

[13]  M. Forster,et al.  Characterization of the Interaction between Tumor Necrosis Factor-stimulated Gene-6 and Heparin , 2005, Journal of Biological Chemistry.

[14]  D. Mooney,et al.  Local delivery of VEGF and SDF enhances endothelial progenitor cell recruitment and resultant recovery from ischemia. , 2015, Tissue engineering. Part A.

[15]  J. Temenoff,et al.  Effect of selective heparin desulfation on preservation of bone morphogenetic protein-2 bioactivity after thermal stress. , 2015, Bioconjugate chemistry.

[16]  T. Kishimoto,et al.  Bioactivity screening of partially desulfated low-molecular-weight heparins: a structure/activity relationship study. , 2011, Glycobiology.

[17]  R. Guldberg,et al.  Intra-articular injection of micronized dehydrated human amnion/chorion membrane attenuates osteoarthritis development , 2014, Arthritis Research & Therapy.

[18]  Robert E. Guldberg,et al.  Analysis of cartilage matrix fixed charge density and three-dimensional morphology via contrast-enhanced microcomputed tomography , 2006, Proceedings of the National Academy of Sciences.

[19]  C. Werner,et al.  A novel, biased-like SDF-1 derivative acts synergistically with starPEG-based heparin hydrogels and improves eEPC migration in vitro. , 2012, Journal of controlled release : official journal of the Controlled Release Society.

[20]  M. Roberts,et al.  Disappearance kinetics of solutes from synovial fluid after intra-articular injection. , 1994, British journal of clinical pharmacology.

[21]  Y. Inoue,et al.  Selective N-desulfation of heparin with dimethyl sulfoxide containing water or methanol. , 1976, Carbohydrate research.

[22]  Y. Inoue,et al.  Solvolytic desulfation of glycosaminoglycuronan sulfates with dimethyl sulfoxide containing water or methanol. , 1977, Carbohydrate research.

[23]  B. Cronstein,et al.  TNF/IL-1-inducible protein TSG-6 potentiates plasmin inhibition by inter-alpha-inhibitor and exerts a strong anti-inflammatory effect in vivo. , 1996, Journal of immunology.

[24]  J. Temenoff,et al.  Core-shell microparticles for protein sequestration and controlled release of a protein-laden core. , 2017, Acta biomaterialia.

[25]  S. Abramson,et al.  TSG-6 activity as a novel biomarker of progression in knee osteoarthritis. , 2014, Osteoarthritis and cartilage.

[26]  Xinqiao Jia,et al.  Heparin-decorated, hyaluronic acid-based hydrogel particles for the controlled release of bone morphogenetic protein 2. , 2011, Acta biomaterialia.

[27]  M. Maitz,et al.  Heparin desulfation modulates VEGF release and angiogenesis in diabetic wounds. , 2015, Journal of controlled release : official journal of the Controlled Release Society.

[28]  M. Doherty,et al.  Plasminogen activators and their inhibitors in synovial fluids from normal, osteoarthritis, and rheumatoid arthritis knees. , 1996, Annals of the rheumatic diseases.

[29]  Andrew D Pearle,et al.  Basic science of articular cartilage and osteoarthritis. , 2005, Clinics in sports medicine.

[30]  S. Torihashi,et al.  Acute and Temporal Expression of Tumor Necrosis Factor (TNF)-α-stimulated Gene 6 Product, TSG6, in Mesenchymal Stem Cells Creates Microenvironments Required for Their Successful Transplantation into Muscle Tissue* , 2015, The Journal of Biological Chemistry.

[31]  L. Wancket,et al.  Anatomical Localization of Cartilage Degradation Markers in a Surgically Induced Rat Osteoarthritis Model , 2005, Toxicologic pathology.

[32]  M. Perretti,et al.  Inhibitory Effects of TSG‐6 Link Module on Leukocyte–Endothelial Cell Interactions In Vitro and In Vivo , 2004, Microcirculation.

[33]  Nicole Gerwin,et al.  Intraarticular drug delivery in osteoarthritis. , 2006, Advanced drug delivery reviews.

[34]  A. Sappino,et al.  Plasminogen activation in synovial tissues: differences between normal, osteoarthritis, and rheumatoid arthritis joints , 1997, Annals of the rheumatic diseases.

[35]  A. Hoffman,et al.  PEG-cross-linked heparin is an affinity hydrogel for sustained release of vascular endothelial growth factor , 2006, Journal of biomaterials science. Polymer edition.

[36]  C. Hack,et al.  Analysis of intraarticular fibrinolytic pathways in patients with inflammatory and noninflammatory joint diseases. , 1992, Arthritis and rheumatism.

[37]  M. Goldring,et al.  Inflammation in osteoarthritis , 2011, Current opinion in rheumatology.

[38]  J. Temenoff,et al.  Hydrolysis and Sulfation Pattern Effects on Release of Bioactive Bone Morphogenetic Protein-2 from Heparin-Based Microparticles. , 2015, Journal of materials chemistry. B.

[39]  I. Campbell,et al.  The Link Module from Ovulation- and Inflammation-associated Protein TSG-6 Changes Conformation on Hyaluronan Binding* , 2003, Journal of Biological Chemistry.

[40]  C. Werner,et al.  Biohybrid networks of selectively desulfated glycosaminoglycans for tunable growth factor delivery. , 2014, Biomacromolecules.

[41]  B. Huppertz,et al.  Novel regio- and stereoselective O-6-desulfation of the glucosamine moiety of heparin with N-methylpyrrolidinone-water or N,N-dimethylformamide-water mixtures. , 1998, Carbohydrate research.

[42]  J. Vilček,et al.  TSG-6: an IL-1/TNF-inducible protein with anti-inflammatory activity. , 1997, Cytokine & growth factor reviews.

[43]  G. Dooijewaard,et al.  Plasminogen activators in synovial fluid and plasma from patients with arthritis. , 1992, Annals of the rheumatic diseases.

[44]  R. Guldberg,et al.  Localized 3D analysis of cartilage composition and morphology in small animal models of joint degeneration. , 2013, Osteoarthritis and cartilage.

[45]  D. Prockop,et al.  Anti-inflammatory protein TSG-6 secreted by activated MSCs attenuates zymosan-induced mouse peritonitis by decreasing TLR2/NF-κB signaling in resident macrophages. , 2011, Blood.

[46]  F. Barry,et al.  Up-regulation and differential expression of the hyaluronan-binding protein TSG-6 in cartilage and synovium in rheumatoid arthritis and osteoarthritis. , 2001, Osteoarthritis and cartilage.

[47]  M. Goldring,et al.  Osteoarthritis and cartilage: The role of cytokines , 2000, Current rheumatology reports.

[48]  Glenn D Prestwich,et al.  Injectable glycosaminoglycan hydrogels for controlled release of human basic fibroblast growth factor. , 2005, Biomaterials.

[49]  Jie Hu,et al.  Mesenchymal Stem Cells Attenuate Peritoneal Injury through Secretion of TSG-6 , 2012, PloS one.

[50]  Torri E. Rinker,et al.  Localized SDF-1α Delivery Increases Pro-Healing Bone Marrow-Derived Cells in the Supraspinatus Muscle Following Severe Rotator Cuff Injury , 2018, Regenerative Engineering and Translational Medicine.

[51]  J. Temenoff,et al.  Heparin-based hydrogels with tunable sulfation & degradation for anti-inflammatory small molecule delivery. , 2016, Biomaterials science.

[52]  M. Goldring,et al.  Chondrogenesis, chondrocyte differentiation, and articular cartilage metabolism in health and osteoarthritis , 2012, Therapeutic advances in musculoskeletal disease.