Generation of tendon‐to‐bone interface “enthesis” with use of recombinant BMP‐2 in a rabbit model

The anatomical structure at bone‐tendon and bone‐ligament interfaces is called the enthesis. Histologically, the enthesis is characterized by a transitional series of tissue layers from the end of the tendon to bone, including tendon, fibrocartilage, calcified fibrocartilage, and bone. This arrangement yields stronger direct connection of the soft tissues to bone. In surgical repair, the enthesis has proven difficult to reproduce, and the success of ligament‐bone bonding has depended on the fibrous attachment that forms after any ligament reconstructions. In this study, we attempted to generate a direct‐insertion enthesis in two stages. First, recombinant human bone morphogenetic protein‐2 (rhBMP‐2) was injected into the flexor digitorum communis tendon in the rabbit hind limb to induce ectopic ossicle formation. In a second step, the resultant tendon/ossicle complex was then surgically transferred onto the surface of the rabbit tibia to generate a stable tendon‐bone junction. One month following surgery, histomorphological examination confirmed direct insertion of tendon‐bone structures in the proximal tibia of the rabbit. Ultimate failure loads of the BMP‐2‐generated tendon‐bone junction were significantly higher than in the control group (p < 0.01). These findings suggest that it is possible to successfully regenerate a direct tendon‐to‐bone enthesis. Use of this approach may enable successful reconstruction of joints rendered unstable after ligamentous rupture or laxity after anterior cruciate ligament injury. © 2007 Orthopaedic Research Society. Published by Wiley Periodicals, Inc. J Orthop Res 25:1415–1424, 2007

[1]  R. Rutherford,et al.  Bone morphogenetic protein-transduced human fibroblasts convert to osteoblasts and form bone in vivo. , 2002, Tissue engineering.

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

[3]  J Kumagai,et al.  Immunohistochemical distribution of type I, II and III collagens in the rabbit supraspinatus tendon insertion. , 1994, Journal of anatomy.

[4]  K. Takaoka,et al.  Purification and characterization of a bone-inducing protein from a murine osteosarcoma (Dunn type). , 1993, Clinical orthopaedics and related research.

[5]  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.

[6]  K. Miyazono,et al.  Bone morphogenetic protein receptors and activin receptors are highly expressed in ossified ligament tissues of patients with ossification of the posterior longitudinal ligament. , 1997, The American journal of pathology.

[7]  P. Annear,et al.  Tunnel widening in anterior cruciate ligament reconstruction: a prospective evaluation of hamstring and patella tendon grafts , 1999, Knee Surgery, Sports Traumatology, Arthroscopy.

[8]  J. Tanaka,et al.  Calcium-phosphate-hybridized tendon directly promotes regeneration of tendon-bone insertion. , 2004, Journal of biomedical materials research. Part A.

[9]  J. Ralphs,et al.  The skeletal attachment of tendons--tendon "entheses". , 2002, Comparative biochemistry and physiology. Part A, Molecular & integrative physiology.

[10]  C B Frank,et al.  Tensile properties of the medial collateral ligament as a function of age , 1986, Journal of orthopaedic research : official publication of the Orthopaedic Research Society.

[11]  W. Grana,et al.  Analysis of a Semitendinosus Autograft in a Rabbit Model , 1997, The American journal of sports medicine.

[12]  H. Fujioka,et al.  Comparison of mechanical and histological properties between the immature and mature tendon attachment , 2002, International Orthopaedics.

[13]  H. Terai,et al.  Repair of long intercalated rib defects using porous beta-tricalcium phosphate cylinders containing recombinant human bone morphogenetic protein-2 in dogs. , 2006, Biomaterials.

[14]  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.

[15]  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.

[16]  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.

[17]  R. Warren,et al.  Tendon-healing in a bone tunnel. A biomechanical and histological study in the dog. , 1993, The Journal of bone and joint surgery. American volume.

[18]  T. Wredmark,et al.  There are differences in early morbidity after ACL reconstruction when comparing patellar tendon and semitendinosus tendon graft , 2001, Scandinavian journal of medicine & science in sports.

[19]  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.

[20]  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.

[21]  Kazutoshi Nozaki,et al.  A biodegradable polymer as a cytokine delivery system for inducing bone formation , 2001, Nature Biotechnology.

[22]  Y. Hashizume,et al.  Immunohistochemistry of symptomatic hypertrophy of the posterior longitudinal ligament with special reference to ligamentous ossification , 2006, Spinal Cord.

[23]  M. Röpke,et al.  Bone tunnel enlargement after anterior cruciate ligament reconstruction with semitendinosus tendon using Endobutton fixation on the femoral side. , 1998, Arthroscopy : the journal of arthroscopic & related surgery : official publication of the Arthroscopy Association of North America and the International Arthroscopy Association.

[24]  K. Messner Postnatal development of the cruciate ligament insertions in the rat knee. morphological evaluation and immunohistochemical study of collagens types I and II. , 1997, Acta anatomica.

[25]  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.

[26]  F R Noyes,et al.  Biomechanics of ligament failure. II. An analysis of immobilization, exercise, and reconditioning effects in primates. , 1974, The Journal of bone and joint surgery. American volume.

[27]  D Amiel,et al.  Tendons and ligaments: A morphological and biochemical comparison , 1984, Journal of orthopaedic research : official publication of the Orthopaedic Research Society.

[28]  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.

[29]  Torsten Wredmark,et al.  There are differences in early morbidity after ACL reconstruction when comparing patellar tendon and semitendinosus tendon graft. A prospective randomized study of 107 patients , 2001 .

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

[31]  C. Fabbriciani,et al.  Graft healing after anterior cruciate ligament reconstruction in rabbits. , 1997, Clinical orthopaedics and related research.

[32]  L. Galatz,et al.  The localized expression of extracellular matrix components in healing tendon insertion sites: an in situ hybridization study , 2002, Journal of orthopaedic research : official publication of the Orthopaedic Research Society.

[33]  M. Obinata,et al.  Establishment of tendon-derived cell lines exhibiting pluripotent mesenchymal stem cell-like property. , 2003, Experimental cell research.

[34]  H. Terai,et al.  Experimental Spinal Fusion With Recombinant Human Bone Morphogenetic Protein-2 Delivered by a Synthetic Polymer and &bgr;-Tricalcium Phosphate in a Rabbit Model , 2005, Spine.

[35]  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.

[36]  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.

[37]  D. Kohn,et al.  Donor-site morbidity after harvest of a bone-tendon-bone patellar tendon autograft , 2005, Knee Surgery, Sports Traumatology, Arthroscopy.

[38]  H. Yoshikawa,et al.  Graft healing in a bone tunnel: bone-attached graft with screw fixation versus bone-free graft with extra-articular suture fixation , 2004, Knee Surgery, Sports Traumatology, Arthroscopy.

[39]  J. Ralphs,et al.  Characterization of collagens and proteoglycans at the insertion of the human Achilles tendon. , 1998, Matrix biology : journal of the International Society for Matrix Biology.

[40]  H. Uhthoff,et al.  The collagen types in the attachment zone of rotator cuff tendons in the elderly: an immunohistochemical study. , 1994, The Journal of rheumatology.

[41]  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.

[42]  K. Nawata,et al.  Development of the attachment zones in the rat anterior cruciate ligament: changes in the distributions of proliferating cells and fibrillar collagens during postnatal growth , 2002, Journal of orthopaedic research : official publication of the Orthopaedic Research Society.

[43]  Braden C. Fleming,et al.  Anterior Cruciate Ligament Replacement: Comparison of Bone-Patellar Tendon-Bone Grafts with Two-Strand Hamstring Grafts A Prospective, Randomized Study , 2002, The Journal of bone and joint surgery. American volume.

[44]  H. Tohyama,et al.  Comparisons of intraosseous graft healing between the doubled flexor tendon graft and the bone-patellar tendon-bone graft in anterior cruciate ligament reconstruction. , 2001, Arthroscopy : the journal of arthroscopic & related surgery : official publication of the Arthroscopy Association of North America and the International Arthroscopy Association.

[45]  J. Ralphs,et al.  Development and ageing of phenotypically distinct fibrocartilages associated with the rat Achilles tendon , 1992, Anatomy and Embryology.

[46]  G. Finerman,et al.  Morphology and Matrix Composition During Early Tendon to Bone Healing , 1997, Clinical orthopaedics and related research.