Mesenchymal Stem Cell Extracellular Vesicles as Adjuvant to Bone Marrow Stimulation in Chondral Defect Repair in a Minipig Model

Objective This study evaluated the effects of mesenchymal stem cell-extracellular vesicles (MSC-EVs) on chondrocyte proliferation in vitro and on cartilage repair in vivo following bone marrow stimulation (BMS) of focal chondral defects of the knee. Methods Six adult Göttingen minipigs received 2 chondral defects in each knee. The pigs were randomized to treatment with either BMS combined with MSC-EVs or BMS combined with phosphate-buffered saline (PBS). Intraarticular injections MSC-EVs or PBS were performed immediately after closure of the surgical incisions, and at 2 and 4 weeks postoperatively. Repair was evaluated after 6 months with gross examination, histology, histomorphometry, immunohistochemistry, and micro-computed tomography (µCT) analysis of the trabecular bone beneath the defect. Results Defects treated with MSC-EVs had more bone in the cartilage defect area than the PBS-treated defects (7.9% vs. 1.5%, P = 0.02). Less than 1% of the repair tissue in both groups was hyaline cartilage. International Cartilage and Joint Preservation Society II histological scoring showed that defects treated with MSC-EVs scored lower on “matrix staining” (20.8 vs. 50.0, P = 0.03), “cell morphology” (35.4 vs. 53.8, P = 0.04), and “overall assessment” (30.8 vs. 52.9, P = 0.03). Consistently, defects treated with MSC-EVs had lower collagen II and higher collagen I areal deposition. Defects treated with MSC-EVs had subchondral bone with significantly higher tissue mineral densities than PBS-treated defects (860 mg HA/cm3 vs. 838 mg HA/cm3, P = 0.02). Conclusion Intraarticular injections of MSC-EVs in conjunction with BMS led to osseous ingrowth that impaired optimal cartilage repair, while enhancing subchondral bone healing.

[1]  S. Lim,et al.  Systemic Mesenchymal Stem Cell-Derived Exosomes Reduce Myocardial Infarct Size: Characterization With MRI in a Porcine Model , 2020, Frontiers in Cardiovascular Medicine.

[2]  P. Yarowsky,et al.  Exosomes Isolated From Platelet-Rich Plasma and Mesenchymal Stem Cells Promote Recovery of Function After Muscle Injury , 2020, The American journal of sports medicine.

[3]  J. Hui,et al.  Mesenchymal stem cell exosomes in bone regenerative strategies—a systematic review of preclinical studies , 2020, Materials today. Bio.

[4]  M. Lind,et al.  CARGEL Bioscaffold improves cartilage repair tissue after bone marrow stimulation in a minipig model , 2020, Journal of Experimental Orthopaedics.

[5]  K. L. Wong,et al.  Mesenchymal Stem Cell Exosomes for Cartilage Regeneration: A Systematic Review of Preclinical In Vivo Studies , 2020 .

[6]  Keng Lin Wong,et al.  Intra-articular injections of mesenchymal stem cell exosomes and hyaluronic acid improve structural and mechanical properties of repaired cartilage in a rabbit model. , 2020, Arthroscopy : the journal of arthroscopic & related surgery : official publication of the Arthroscopy Association of North America and the International Arthroscopy Association.

[7]  M. Lind,et al.  No effect of platelet-rich plasma as adjuvant to bone marrow stimulation for the treatment of chondral defects in a large animal model , 2019, Archives of Orthopaedic and Trauma Surgery.

[8]  R. Jakob,et al.  Systematic Review and Meta-Analysis of the Clinical Evidence on the Use of Autologous Matrix-Induced Chondrogenesis in the Knee , 2019, Cartilage.

[9]  K. L. Wong,et al.  Equivalent 10-Year Outcomes After Implantation of Autologous Bone Marrow–Derived Mesenchymal Stem Cells Versus Autologous Chondrocyte Implantation for Chondral Defects of the Knee , 2019, The American journal of sports medicine.

[10]  M. Lind,et al.  No Effect of Platelet-Rich Plasma Injections as an Adjuvant to Autologous Cartilage Chips Implantation for the Treatment of Chondral Defects , 2019, Cartilage.

[11]  M. Akagi,et al.  Transplantation of autologous bone marrow-derived mesenchymal stem cells under arthroscopic surgery with microfracture versus microfracture alone for articular cartilage lesions in the knee: A multicenter prospective randomized control clinical trial , 2019, Regenerative therapy.

[12]  A. Hill,et al.  Defining mesenchymal stromal cell (MSC)-derived small extracellular vesicles for therapeutic applications , 2019, Journal of extracellular vesicles.

[13]  S. Lim,et al.  MSC exosomes alleviate temporomandibular joint osteoarthritis by attenuating inflammation and restoring matrix homeostasis. , 2019, Biomaterials.

[14]  Jing Xu,et al.  Minimal information for studies of extracellular vesicles 2018 (MISEV2018): a position statement of the International Society for Extracellular Vesicles and update of the MISEV2014 guidelines , 2018, Journal of Extracellular Vesicles.

[15]  N. Iwasaki,et al.  Bone Marrow Stimulation Technique Augmented by an Ultrapurified Alginate Gel Enhances Cartilage Repair in a Canine Model , 2018, The American journal of sports medicine.

[16]  S. Lim,et al.  MSC exosomes mediate cartilage repair by enhancing proliferation, attenuating apoptosis and modulating immune reactivity. , 2018, Biomaterials.

[17]  Yin Hu,et al.  Exosomes from human umbilical cord blood accelerate cutaneous wound healing through miR-21-3p-mediated promotion of angiogenesis and fibroblast function , 2018, Theranostics.

[18]  H. Lurås,et al.  Microfracture is more cost-effective than autologous chondrocyte implantation: a review of level 1 and level 2 studies with 5 year follow-up , 2017, Knee Surgery, Sports Traumatology, Arthroscopy.

[19]  S. Lim,et al.  MSC exosome as a cell-free MSC therapy for cartilage regeneration: Implications for osteoarthritis treatment. , 2017, Seminars in cell & developmental biology.

[20]  M. Lind,et al.  Autologous Cartilage Chip Transplantation Improves Repair Tissue Composition Compared With Marrow Stimulation , 2017, The American journal of sports medicine.

[21]  Peter Zioupos,et al.  Localized tissue mineralization regulated by bone remodelling: A computational approach , 2017, PloS one.

[22]  S. Lim,et al.  Exosomes derived from human embryonic mesenchymal stem cells promote osteochondral regeneration. , 2016, Osteoarthritis and cartilage.

[23]  C. Şen,et al.  Can chondral healing be improved following microfracture? The effect of adipocyte tissue derived stem cell therapy. , 2016, The Knee.

[24]  M. Lind,et al.  Implantation of Autologous Cartilage Chips Improves Cartilage Repair Tissue Quality in Osteochondral Defects , 2016, American Journal of Sports Medicine.

[25]  S. Lim,et al.  MSC secretes at least 3 EV types each with a unique permutation of membrane lipid, protein and RNA , 2016, Journal of extracellular vesicles.

[26]  S. Tadano,et al.  A Novel Bone Marrow Stimulation Technique Augmented by Administration of Ultrapurified Alginate Gel Enhances Osteochondral Repair in a Rabbit Model. , 2015, Tissue engineering. Part C, Methods.

[27]  B. FisherMatthew,et al.  Fibrous Scaffolds with Varied Fiber Chemistry and Growth Factor Delivery Promote Repair in a Porcine Cartilage Defect Model , 2015 .

[28]  D. Howard,et al.  Delivering rhFGF‐18 via a bilayer collagen membrane to enhance microfracture treatment of chondral defects in a large animal model , 2015, Journal of orthopaedic research : official publication of the Orthopaedic Research Society.

[29]  Lijun Wu,et al.  HucMSC‐Exosome Mediated‐Wnt4 Signaling Is Required for Cutaneous Wound Healing , 2015, Stem cells.

[30]  M. Lind,et al.  Experimental articular cartilage repair in the Göttingen minipig: the influence of multiple defects per knee , 2015, Journal of Experimental Orthopaedics.

[31]  K. Vehik,et al.  BST-CarGel® Treatment Maintains Cartilage Repair Superiority over Microfracture at 5 Years in a Multicenter Randomized Controlled Trial , 2015, Cartilage.

[32]  Myron Spector,et al.  A Stereological Method for the Quantitative Evaluation of Cartilage Repair Tissue , 2015, Cartilage.

[33]  T. Ogut,et al.  Matrix-induced autologous mesenchymal stem cell implantation versus matrix-induced autologous chondrocyte implantation in the treatment of chondral defects of the knee: a 2-year randomized study , 2015, Archives of Orthopaedic and Trauma Surgery.

[34]  Anup Kumar,et al.  Long-term results after microfracture treatment for full-thickness knee chondral lesions in athletes , 2014, Knee Surgery, Sports Traumatology, Arthroscopy.

[35]  S. Lim,et al.  Mesenchymal stem cell-derived exosomes promote hepatic regeneration in drug-induced liver injury models , 2014, Stem Cell Research & Therapy.

[36]  S. Lim,et al.  Mesenchymal stem cells secrete immunologically active exosomes. , 2014, Stem cells and development.

[37]  A. Getgood,et al.  Intra‐articular injection of rhFGF‐18 improves the healing in microfracture treated chondral defects in an ovine model , 2014, Journal of orthopaedic research : official publication of the Orthopaedic Research Society.

[38]  B. Olsen,et al.  Distribution of Basement Membrane Molecules, Laminin and Collagen Type IV, in Normal and Degenerated Cartilage Tissues , 2014, Cartilage.

[39]  A. A. Shetty,et al.  The effect of platelet rich plasma combined with microfracture for the treatment of chondral defect in a rabbit knee , 2014, Tissue Engineering and Regenerative Medicine.

[40]  M. Shive,et al.  Novel scaffold-based BST-CarGel treatment results in superior cartilage repair compared with microfracture in a randomized controlled trial. , 2013, The Journal of bone and joint surgery. American volume.

[41]  H. Y. Yüksel,et al.  Does platelet-rich plasma enhance microfracture treatment for chronic focal chondral defects? An in-vivo study performed in a rat model. , 2013, Acta orthopaedica et traumatologica turcica.

[42]  S. Lim,et al.  Proteolytic Potential of the MSC Exosome Proteome: Implications for an Exosome-Mediated Delivery of Therapeutic Proteasome , 2012, International journal of proteomics.

[43]  Myron Spector,et al.  Modulation of mesenchymal stem cell chondrogenesis in a tunable hyaluronic acid hydrogel microenvironment. , 2012, Biomaterials.

[44]  A. Tietze,et al.  Changes in 3‐dimensional bone structure indices in hypoparathyroid patients treated with PTH(1‐84): A randomized controlled study , 2012, Journal of bone and mineral research : the official journal of the American Society for Bone and Mineral Research.

[45]  A. Gomoll.  Microfracture and Augments , 2012, The Journal of Knee Surgery.

[46]  J. Steadman,et al.  Evaluation of intra-articular mesenchymal stem cells to augment healing of microfractured chondral defects. , 2011, Arthroscopy : the journal of arthroscopic & related surgery : official publication of the Arthroscopy Association of North America and the International Arthroscopy Association.

[47]  B. Mandelbaum,et al.  Outcomes After a Single-Stage Procedure for Cell-Based Cartilage Repair , 2011, The American journal of sports medicine.

[48]  S. Lim,et al.  Enabling a robust scalable manufacturing process for therapeutic exosomes through oncogenic immortalization of human ESC-derived MSCs , 2011, Journal of Translational Medicine.

[49]  C. Bünger,et al.  Combined 3D and hypoxic culture improves cartilage-specific gene expression in human chondrocytes , 2011, Acta orthopaedica.

[50]  Guangyi Chen,et al.  Acute Osteoclast Activity following Subchondral Drilling Is Promoted by Chitosan and Associated with Improved Cartilage Repair Tissue Integration , 2011, Cartilage.

[51]  H. Ohgushi,et al.  Safety of autologous bone marrow‐derived mesenchymal stem cell transplantation for cartilage repair in 41 patients with 45 joints followed for up to 11 years and 5 months , 2011, Journal of tissue engineering and regenerative medicine.

[52]  A. Manunta,et al.  The effect of platelet rich plasma combined with microfractures on the treatment of chondral defects: an experimental study in a sheep model. , 2010, Osteoarthritis and cartilage.

[53]  M. Shive,et al.  Scaffold-Guided Subchondral Bone Repair , 2010, The American journal of sports medicine.

[54]  Gerard Pasterkamp,et al.  Exosome secreted by MSC reduces myocardial ischemia/reperfusion injury. , 2010, Stem cell research.

[55]  K. Briggs,et al.  Rehabilitation following microfracture for chondral injury in the knee. , 2010, Clinics in sports medicine.

[56]  Andreas H. Gomoll,et al.  The subchondral bone in articular cartilage repair: current problems in the surgical management , 2010, Knee Surgery, Sports Traumatology, Arthroscopy.

[57]  Johan Bellemans,et al.  Treatment of Symptomatic Cartilage Defects of the Knee , 2009, The American journal of sports medicine.

[58]  Tian Sheng Chen,et al.  Mesenchymal stem cell secretes microparticles enriched in pre-microRNAs , 2009, Nucleic acids research.

[59]  M. Amling,et al.  Long term results after implantation of tissue engineered cartilage for the treatment of osteochondral lesions in a minipig model , 2008, Journal of materials science. Materials in medicine.

[60]  U. Schneider,et al.  The minipig model for experimental chondral and osteochondral defect repair in tissue engineering: Retrospective analysis of 180 defects , 2008, Laboratory animals.

[61]  N. Adachi,et al.  Repair of chronic osteochondral defects in the rat. A bone marrow-stimulating procedure enhanced by cultured allogenic bone marrow mesenchymal stromal cells. , 2006, The Journal of bone and joint surgery. British volume.

[62]  G. Brindley,et al.  Osteochondral Autograft Transplantation in the Porcine Knee , 2006, The American journal of sports medicine.

[63]  J. Steadman,et al.  Microfracture: surgical technique and rehabilitation to treat chondral defects. , 2001, Clinical orthopaedics and related research.

[64]  R P Jakob,et al.  Articular cartilage repair using a tissue-engineered cartilage-like implant: an animal study. , 2001, Osteoarthritis and cartilage.

[65]  H. Imhof,et al.  Subchondral Bone and Cartilage Disease: A Rediscovered Functional Unit , 2000, Investigative radiology.

[66]  J S Thomsen,et al.  Lumbar vertebral body compressive strength evaluated by dual-energy X-ray absorptiometry, quantitative computed tomography, and ashing. , 1999, Bone.

[67]  H. Madry,et al.  Autologous Matrix-Induced Chondrogenesis: A Systematic Review of the Clinical Evidence , 2019, The American journal of sports medicine.

[68]  S. Lim,et al.  Isolation and Characterization of Exosome from Human Embryonic Stem Cell-Derived C-Myc-Immortalized Mesenchymal Stem Cells. , 2016, Methods in molecular biology.

[69]  Hajime Ohgushi,et al.  Repair of articular cartilage defects in the patello‐femoral joint with autologous bone marrow mesenchymal cell transplantation: three case reports involving nine defects in five knees , 2007, Journal of tissue engineering and regenerative medicine.