Kartogenin induces cartilage-like tissue formation in tendon–bone junction

Tendon–bone junctions (TBJs) are frequently injured, especially in athletic settings. Healing of TBJ injuries is slow and is often repaired with scar tissue formation that compromises normal function. This study explored the feasibility of using kartogenin (KGN), a biocompound, to enhance the healing of injured TBJs. We first determined the effects of KGN on the proliferation and chondrogenic differentiation of rabbit bone marrow stromal cells (BMSCs) and patellar tendon stem/progenitor cells (PTSCs) in vitro. KGN enhanced cell proliferation in both cell types in a concentration-dependent manner and induced chondrogenic differentiation of stem cells, as demonstrated by high expression levels of chondrogenic markers aggrecan, collagen II and Sox-9. Besides, KGN induced the formation of cartilage-like tissues in cell cultures, as observed through the staining of abundant proteoglycans, collagen II and osteocalcin. When injected into intact rat patellar tendons in vivo, KGN induced cartilage-like tissue formation in the injected area. Similarly, when KGN was injected into experimentally injured rat Achilles TBJs, wound healing in the TBJs was enhanced, as evidenced by the formation of extensive cartilage-like tissues. These results suggest that KGN may be used as an effective cell-free clinical therapy to enhance the healing of injured TBJs.

[1]  C. Frank,et al.  Compressive compared with tensile loading of medial collateral ligament scar in vitro uniquely influences mRNA levels for aggrecan, collagen type II, and collagenase , 2000, Journal of orthopaedic research : official publication of the Orthopaedic Research Society.

[2]  R. Warren,et al.  Use of Recombinant Human Bone Morphogenetic Protein-2 to Enhance Tendon Healing in a Bone Tunnel , 1999, The American journal of sports medicine.

[3]  H. Takahashi-Iwanaga,et al.  Early anchoring collagen fibers at the bone—tendon interface are conducted by woven bone formation: light microscope and scanning electron microscope observation using a canine model , 2001, Journal of orthopaedic research : official publication of the Orthopaedic Research Society.

[4]  A. Thambyah,et al.  Enhancement of tendon graft osteointegration using mesenchymal stem cells in a rabbit model of anterior cruciate ligament reconstruction. , 2004, Arthroscopy : the journal of arthroscopic & related surgery : official publication of the Arthroscopy Association of North America and the International Arthroscopy Association.

[5]  K. Sairyo,et al.  Callus formation during healing of the repaired tendon-bone junction. A rat experimental model. , 2007, The Journal of bone and joint surgery. British volume.

[6]  I. Siegel,et al.  Injury and Repair of the Musculoskeletal Soft Tissues , 1988 .

[7]  A. Minami,et al.  The effect of transforming growth factor-beta1 on intraosseous healing of flexor tendon autograft replacement of anterior cruciate ligament in dogs. , 2005, Arthroscopy : the journal of arthroscopic & related surgery : official publication of the Arthroscopy Association of North America and the International Arthroscopy Association.

[8]  J. Wang,et al.  The role of engineered tendon matrix in the stemness of tendon stem cells in vitro and the promotion of tendon-like tissue formation in vivo. , 2011, Biomaterials.

[9]  Shin-Yoon Kim,et al.  Tendon-to-bone tunnel healing in a rabbit model: the effect of periosteum augmentation at the tendon-to-bone interface , 2002, Knee Surgery, Sports Traumatology, Arthroscopy.

[10]  C. Shih,et al.  Enveloping the tendon graft with periosteum to enhance tendon-bone healing in a bone tunnel: A biomechanical and histologic study in rabbits. , 2003, Arthroscopy : the journal of arthroscopic & related surgery : official publication of the Arthroscopy Association of North America and the International Arthroscopy Association.

[11]  Superparamagnetic Iron Oxide is Suitable to Label Tendon Stem Cells and Track Them In Vivo with MR Imaging , 2013, Annals of Biomedical Engineering.

[12]  G. Finerman,et al.  MRI and morphology of the insertion of the patellar tendon after graft harvesting. , 1996, The Journal of bone and joint surgery. British volume.

[13]  D. Heymann,et al.  Hamstring insertion site healing after anterior cruciate ligament reconstruction in patients with symptomatic hardware or repeat rupture: a histologic study in 12 patients. , 2003, Arthroscopy : the journal of arthroscopic & related surgery : official publication of the Arthroscopy Association of North America and the International Arthroscopy Association.

[14]  Matthew J. Silva,et al.  Two-portal repair of canine flexor tendon insertion site injuries: histologic and immunohistochemical characterization of healing during the early postoperative period. , 2003, Journal of Hand Surgery-American Volume.

[15]  R. Marx,et al.  Reconstruction of the anterior cruciate ligament in the skeletally immature athlete: a review of current concepts: AAOS exhibit selection. , 2013, The Journal of bone and joint surgery. American volume.

[16]  Y. Morita,et al.  Osteogenic Matrix Cell Sheet Transplantation Enhances Early Tendon Graft to Bone Tunnel Healing in Rabbits , 2013, BioMed research international.

[17]  J. Ralphs,et al.  Where tendons and ligaments meet bone: attachment sites (‘entheses’) in relation to exercise and/or mechanical load , 2006, Journal of anatomy.

[18]  J. F. Suh,et al.  Periosteal Augmentation of a Tendon Graft Improves Tendon Healing in the Bone Tunnel , 2004, Clinical orthopaedics and related research.

[19]  Francis Berenbaum,et al.  Mesenchymal Stem Cell Therapy Regenerates the Native Bone-Tendon Junction after Surgical Repair in a Degenerative Rat Model , 2010, PloS one.

[20]  A. Weiler,et al.  The Influence of Locally Applied Platelet-Derived Growth Factor-BB on Free Tendon Graft Remodeling after Anterior Cruciate Ligament Reconstruction , 2004, The American journal of sports medicine.

[21]  Valerie A Longo,et al.  Synthesis of a tissue-engineered periosteum with acellular dermal matrix and cultured mesenchymal stem cells. , 2009, Tissue engineering. Part A.

[22]  S. Rodeo,et al.  The role of macrophages in early healing of a tendon graft in a bone tunnel. , 2008, The Journal of bone and joint surgery. American volume.

[23]  H. Uhthoff,et al.  Early reactions after reimplantation of the tendon of supraspinatus into bone. A study in rabbits. , 2000, The Journal of bone and joint surgery. British volume.

[24]  R. Takeuchi,et al.  Effects of collagen gel mixed with hydroxyapatite powder on interface between newly formed bone and grafted achilles tendon in rabbit femoral bone tunnel. , 2001, Biomaterials.

[25]  Young-Jin Ju,et al.  Synovial mesenchymal stem cells accelerate early remodeling of tendon-bone healing , 2008, Cell and Tissue Research.

[26]  Y. Toyama,et al.  Effects of Local Administration of Vascular Endothelial Growth Factor on Mechanical Characteristics of the Semitendinosus Tendon Graft after Anterior Cruciate Ligament Reconstruction in Sheep , 2006, The American journal of sports medicine.

[27]  Kai-Ming Chan,et al.  Healing of Bone-Tendon Junction in a Bone Trough: A Goat Partial Patellectomy Model , 2003, Clinical orthopaedics and related research.

[28]  H. Potter,et al.  Augmentation of Tendon Healing in an Intraarticular Bone Tunnel with Use of a Bone Growth Factor * , 2001, The American journal of sports medicine.

[29]  W. Grana,et al.  An Analysis of Autograft Fixation After Anterior Cruciate Ligament Reconstruction in a Rabbit Model , 1994, The American journal of sports medicine.

[30]  P. Chambat,et al.  The evolution of ACL reconstruction over the last fifty years , 2013, International Orthopaedics.

[31]  W. Nebelung,et al.  Histological findings of tendon-bone healing following anterior cruciate ligament reconstruction with hamstring grafts , 2003, Archives of Orthopaedic and Trauma Surgery.

[32]  Norbert P Südkamp,et al.  Tendon healing in a bone tunnel. Part II: Histologic analysis after biodegradable interference fit fixation in a model of anterior cruciate ligament reconstruction in sheep. , 2002, Arthroscopy : the journal of arthroscopic & related surgery : official publication of the Arthroscopy Association of North America and the International Arthroscopy Association.

[33]  Constance R. Chu,et al.  The Clinical Use of Human Culture–Expanded Autologous Bone Marrow Mesenchymal Stem Cells Transplanted on Platelet-Rich Fibrin Glue in the Treatment of Articular Cartilage Defects , 2010, Cartilage.

[34]  Freddie H. Fu,et al.  HGF Mediates the Anti-inflammatory Effects of PRP on Injured Tendons , 2013, PloS one.

[35]  L. Qin,et al.  Enlargement of remaining patella after partial patellectomy in rabbits. , 1999, Medicine and science in sports and exercise.

[36]  H. Uhthoff,et al.  Early reactions after reimplantation of the tendon of supraspinatus into bone , 2000 .

[37]  Freddie H. Fu,et al.  Enhancement of Tendon-Bone Integration of Anterior Cruciate Ligament Grafts with Bone Morphogenetic Protein-2 Gene Transfer: A Histological and Biomechanical Study , 2002, The Journal of bone and joint surgery. American volume.

[38]  M. Raspanti,et al.  Structure and ultrastructure of the bone/ligament junction. , 1996, Italian journal of anatomy and embryology = Archivio italiano di anatomia ed embriologia.

[39]  Matthew J. Silva,et al.  Characteristics of the rat supraspinatus tendon during tendon‐to‐bone healing after acute injury , 2006, Journal of orthopaedic research : official publication of the Orthopaedic Research Society.

[40]  K. Johnson A stem cell-based approach to cartilage repair , 2013 .

[41]  J. Voncken,et al.  A novel in vivo model to study endochondral bone formation; HIF-1alpha activation and BMP expression. , 2007, Bone.

[42]  Shinichi Yoshiya,et al.  Graft Healing in the Bone Tunnel in Anterior Cruciate Ligament Reconstruction , 2000, Clinical orthopaedics and related research.

[43]  E. Nauman,et al.  In vitro and in vivo evaluation of orthopedic interface repair using a tissue scaffold with a continuous hard tissue-soft tissue transition , 2013, Journal of Orthopaedic Surgery and Research.

[44]  L. Qin,et al.  A comparative study of bone to bone repair and bone to tendon healing in patella-patellar tendon complex in rabbits. , 2002, Clinical biomechanics.

[45]  M. Gnecchi,et al.  Bone marrow-derived mesenchymal stem cells: isolation, expansion, characterization, viral transduction, and production of conditioned medium. , 2009, Methods in molecular biology.

[46]  W. Grana,et al.  Tendon-to-bone healing of a semitendinosus tendon autograft used for ACL reconstruction in a sheep model. , 2000, The American journal of knee surgery.

[47]  J. Wang,et al.  Characterization of differential properties of rabbit tendon stem cells and tenocytes , 2010, BMC musculoskeletal disorders.