Tissue engineering of tendons and ligaments by human bone marrow stromal cells in a liquid fibrin matrix in immunodeficient rats: Results of a histologic study

IntroductionThe original complex structure and mechanical properties are not fully restored after ligament and tendon injuries. Due to their high proliferation rate and differentiation potential, Bone Marrow Stromal Cells (BMSC) are considered to be an ideal cell source for tissue engineering to optimize the healing process. Ideal matrices for tissue engineering of ligaments and tendons should allow for homogenous cell seeding and offer sufficient stability.Material and methodsA mixture of human BMSC and liquid fibrin glue was injected into a standardized full-thickness window defect of the patellar tendon of immunodeficient rats (BMSC group). The histology of the tissue was analysed 10 and 20 days postoperatively and compared to four control groups. These groups consisted of a cohort with a mixture of human fibroblasts and fibrin glue, fibrin glue without cells, a defect group without treatment, and a group with uninjured patellar tendon tissue.ResultsTendon defects in the BMSC group revealed dense collagen fibres and spindle-shaped cells, which were mainly orientated along the loading axis. Histologic sections of the control groups, especially of untreated defects and of defects filled with fibrin glue only, showed irregular patterns of cell distribution, irregular formed cell nucleoli and less tissue maturation. Compared to healthy tendon tissue, higher numbers of cells and less intense matrix staining was observed in the BMSC group. No ectopic bone or cartilage formation was observed in any specimen.ConclusionsInjection of human BMSC in a fibrin glue matrix appears to lead to more mature tissue formation with more regular patterns of cell distribution. Advantages of this “in-vivo” tissue engineering approach are a homogenous cell-matrix mixture in a well-known and approved biological matrix, and simple, minimally-invasive application by injection.

[1]  D L Butler,et al.  Comparison of material properties in fascicle-bone units from human patellar tendon and knee ligaments. , 1986, Journal of biomechanics.

[2]  R. Morgan-Jones,et al.  The intercruciate band of the human knee , 1999 .

[3]  E. Hunziker,et al.  Ultrastructural Determinants of Murine Achilles Tendon Strength During Healing , 2003, Connective tissue research.

[4]  Igor Tudorache,et al.  Biological vascularized matrix for bladder tissue engineering: matrix preparation, reseeding technique and short-term implantation in a porcine model. , 2005, The Journal of urology.

[5]  Mark J. Sussman Cardiovascular biology: Hearts and bones , 2001, Nature.

[6]  Albert J Banes,et al.  Novel system for engineering bioartificial tendons and application of mechanical load. , 2003, Tissue engineering.

[7]  S L Woo,et al.  The use of a laser micrometer system to determine the cross-sectional shape and area of ligaments: a comparative study with two existing methods. , 1990, Journal of biomechanical engineering.

[8]  Cain Je,et al.  The effect of fibrin sealant on the strength of tendon repair of full thickness tendon lacerations in the rabbit Achilles tendon. , 1994 .

[9]  M. van Griensven,et al.  A modified method to culture human osteoblasts from bone tissue specimens using fibrin glue. , 2002, Experimental and toxicologic pathology : official journal of the Gesellschaft fur Toxikologische Pathologie.

[10]  David L Butler,et al.  Functional efficacy of tendon repair processes. , 2004, Annual review of biomedical engineering.

[11]  Ivan Martin,et al.  The FASEB Journal express article 10.1096/fj.01-0656fje. Published online December 28, 2001. Cell differentiation by mechanical stress , 2022 .

[12]  K. Brandt,et al.  Osteoarthritic changes in canine articular cartilage, subchondral bone, and synovium fifty-four months after transection of the anterior cruciate ligament. , 2010, Arthritis and rheumatism.

[13]  L. Soslowsky,et al.  Biomechanics of tendon injury and repair. , 2004, Journal of biomechanics.

[14]  H. Ouyang,et al.  Viability of Allogeneic Bone Marrow Stromal Cells following Local Delivery into Patella Tendon in Rabbit Model , 2004, Cell transplantation.

[15]  A I Caplan,et al.  The mesengenic process. , 1994, Clinics in plastic surgery.

[16]  W. Plitz,et al.  Achilles tendon rupture: experimental results on spontaneous repair in a sheep-model , 2000, Knee Surgery, Sports Traumatology, Arthroscopy.

[17]  Z. Estrov,et al.  Adult stem cells for tissue repair - a new therapeutic concept? , 2003, The New England journal of medicine.

[18]  T. Best,et al.  Achilles Tendon Injuries: A Comparison of Surgical Repair Versus No Repair in a Rat Model ∗ , 1993, Foot & ankle.

[19]  T. Simon,et al.  Characterization of the Repair Tissue after Removal of the Central One-Third of the Patellar Ligament. An Experimental Study in a Goat Model* , 1997, The Journal of bone and joint surgery. American volume.

[20]  S. Badylak,et al.  The use of xenogeneic small intestinal submucosa as a biomaterial for Achilles tendon repair in a dog model. , 1995, Journal of biomedical materials research.

[21]  J. Zuckerman,et al.  Long-term functional outcome of repair of large and massive chronic tears of the rotator cuff. , 1999, The Journal of bone and joint surgery. American volume.

[22]  D. A. Lusardi,et al.  The effect of fibrin sealant on the strength of tendon repair of full thickness tendon lacerations in the rabbit Achilles tendon. , 1994, The Journal of foot and ankle surgery : official publication of the American College of Foot and Ankle Surgeons.

[23]  Randall J Lee,et al.  Fibrin glue alone and skeletal myoblasts in a fibrin scaffold preserve cardiac function after myocardial infarction. , 2004, Tissue engineering.

[24]  U Bosch,et al.  The Proliferative Response of Isolated Human Tendon Fibroblasts to Cyclic Biaxial Mechanical Strain * , 2000, The American journal of sports medicine.

[25]  D L Butler,et al.  Effects of age on the repair ability of mesenchymal stem cells in rabbit tendon , 2005, Journal of orthopaedic research : official publication of the Orthopaedic Research Society.

[26]  Freddie H Fu,et al.  Gene therapy and tissue engineering in orthopaedic surgery. , 2002, The Journal of the American Academy of Orthopaedic Surgeons.

[27]  D. Butler,et al.  Use of mesenchymal stem cells in a collagen matrix for achilles tendon repair , 1998, Journal of orthopaedic research : official publication of the Orthopaedic Research Society.

[28]  G Schlag,et al.  Fibrin sealant in orthopedic surgery. , 1988, Clinical orthopaedics and related research.

[29]  A. Caplan,et al.  Myogenic Expression of Mesenchymal Stem Cells within Myotubes of mdx Mice in Vitro and in Vivo. , 1995, Tissue engineering.

[30]  S. Grässel,et al.  Alteration of fracture stability influences chondrogenesis, osteogenesis and immigration of macrophages , 2001, Journal of orthopaedic research : official publication of the Orthopaedic Research Society.

[31]  Christian Krettek,et al.  Modulation of proliferation and differentiation of human bone marrow stromal cells by fibroblast growth factor 2: potential implications for tissue engineering of tendons and ligaments. , 2005, Tissue engineering.

[32]  M. Pittenger,et al.  Multilineage potential of adult human mesenchymal stem cells. , 1999, Science.

[33]  R E Horch,et al.  Single-cell suspensions of cultured human keratinocytes in fibrin-glue reconstitute the epidermis. , 1998, Cell transplantation.

[34]  R. Hunter,et al.  Current Concepts in Anterior Cruciate Ligament Reconstruction , 2001 .

[35]  M. Dunn,et al.  Development of cell-seeded patellar tendon allografts for anterior cruciate ligament reconstruction. , 2004, Tissue engineering.

[36]  C. Ohlsson,et al.  Treatment of deep cartilage defects in the knee with autologous chondrocyte transplantation. , 1994, The New England journal of medicine.

[37]  Y. Tabata,et al.  Homogeneous seeding of mesenchymal stem cells into nonwoven fabric for tissue engineering. , 2003, Tissue engineering.

[38]  C. Frank,et al.  Molecular biology and biomechanics of normal and healing ligaments--a review. , 1999, Osteoarthritis and cartilage.

[39]  L. A. Dahlgren,et al.  Temporal expression of growth factors and matrix molecules in healing tendon lesions , 2005, Journal of orthopaedic research : official publication of the Orthopaedic Research Society.

[40]  D. Butler,et al.  Repair of patellar tendon injuries using a cell–collagen composite , 2003, Journal of orthopaedic research : official publication of the Orthopaedic Research Society.

[41]  W. Domschke,et al.  Biological in vitro effects of fibrin glue: fibroblast proliferation, expression and binding of growth factors , 2004, Scandinavian journal of gastroenterology.

[42]  R Cancedda,et al.  Repair of large bone defects with the use of autologous bone marrow stromal cells. , 2001, The New England journal of medicine.

[43]  David M. Bodine,et al.  Bone marrow cells regenerate infarcted myocardium , 2001, Nature.

[44]  Eric A Nauman,et al.  Mechanical characterization of collagen fibers and scaffolds for tissue engineering. , 2003, Biomaterials.

[45]  R Cancedda,et al.  Clonal mesenchymal progenitors from human bone marrow differentiate in vitro according to a hierarchical model. , 2000, Journal of cell science.

[46]  R. Vanderby,et al.  Microstructural Morphology in the Transition Region Between Scar and Intact Residual Segments of a Healing Rat Medial Collateral Ligament , 2001, Connective tissue research.