Development of cell-seeded patellar tendon allografts for anterior cruciate ligament reconstruction.

Patellar tendon (PT) allografts for anterior cruciate ligament (ACL) reconstruction are potentially immunogenic and incorporate slowly compared with autografts. Our tissue-engineering approach to improve allograft efficacy is to (1) remove intrinsic cells from the graft to reduce antigenicity and then (2) seed the graft with extrinsic cells to improve ligamentization. To remove cells, tendons were soaked in 1% extraction solutions of tri(n-butyl)phosphate (TBP) or sodium dodecyl sulfate (SDS) for various time periods (24-72 h) and rinsed exhaustively. After treatment, we measured tendon cellularity, crimp structure, and mechanical properties. Treatment with either SDS or TBP removed approximately 70-90% of the intrinsic PT cells. Mechanical properties of treated PTs were similar to those of controls, despite changes in appearance. TBP- and SDS-treated PTs were then seeded with fibroblasts and cultured for up to 2 weeks in vitro. Fibroblast proliferation was retarded on SDS-treated PTs; in contrast, TBP-treated PTs supported cell proliferation similar to that of untreated controls. Extrinsic fibroblasts were successfully cultured on the TBP-treated PTs in vitro, creating viable tissue-engineered grafts potentially useful for ACL reconstruction. These modified allografts have the potential to be developed into mechanically functional delivery vehicles for cells, gene therapy vectors, or other biological agents.

[1]  D. Acosta,et al.  Prolonged adverse effects of benzalkonium chloride and sodium dodecyl sulfate in a primary culture system of rabbit corneal epithelial cells. , 1996, Fundamental and applied toxicology : official journal of the Society of Toxicology.

[2]  K. Miyasaka.  The incidece of knee ligament injuries in the general population , 1991 .

[3]  Savio Lau-Yuen Woo,et al.  Biomechanics of diarthrodial joints , 1990 .

[4]  Dale M. Daniel,et al.  Knee Ligaments: Structure, Function, Injury, and Repair , 1991 .

[5]  M. Weiner,et al.  A dietary toxicity/oncogenicity study of tributyl phosphate in the rat. , 1998, Toxicology.

[6]  S. Dye.  The future of anterior cruciate ligament restoration. , 1996, Clinical orthopaedics and related research.

[7]  D. Butler,et al.  Dose‐dependent response of gamma irradiation on mechanical properties and related biochemical composition of goat bone‐patellar tendon‐bone allografts , 1995, Journal of orthopaedic research : official publication of the Orthopaedic Research Society.

[8]  D. Herndon,et al.  Transplanted acellular allograft dermal matrix. Potential as a template for the reconstruction of viable dermis. , 1995, Transplantation.

[9]  D. Buttle,et al.  Improved quantitation and discrimination of sulphated glycosaminoglycans by use of dimethylmethylene blue. , 1986, Biochimica et biophysica acta.

[10]  A. Minami,et al.  Effect of the Immunological Antigenicity of the Allogeneic Tendons on Tendon Grafting , 1982, The Hand.

[11]  B. Romanus,et al.  Morphological and biochemical effects of sodium morrhuate on tendons , 1985, Journal of orthopaedic research : official publication of the Orthopaedic Research Society.

[12]  L. Coiffard,et al.  Comparison of cytotoxicity of various surfactants tested on normal human fibroblast cultures using the neutral red test, MTT assay and LDH release , 1999, Journal of applied toxicology : JAT.

[13]  J. Michel,et al.  Cell-free arterial grafts: morphologic characteristics of aortic isografts, allografts, and xenografts in rats. , 1994, Journal of vascular surgery.

[14]  L. Paulos,et al.  Why Grafts Fail , 1996, Clinical orthopaedics and related research.

[15]  S. Morita,et al.  Effects of TRI-n-butyl phosphate on pregnancy in rats. , 1994, Food and chemical toxicology : an international journal published for the British Industrial Biological Research Association.

[16]  G. Gerosa,et al.  Thermal analysis characterization of aortic tissues for cardiac valve bioprostheses. , 1999, Journal of biomedical materials research.

[17]  A. Prince,et al.  Solvent/detergent-treated plasma: a virus-inactivated substitute for fresh frozen plasma. , 1992, Blood.

[18]  L. A. Kotkoskie,et al.  A dietary oncogenicity study of tributyl phosphate in the CD-1 mouse. , 1998, Toxicology.

[19]  C. Vangsness,et al.  Gamma Irradiation: Effects on Biomechanical Properties of Human Bone-Patellar Tendon-Bone Allografts , 1995, The American journal of sports medicine.

[20]  J. B. Quattlebaum,et al.  Antigenicity of freeze-dried cortical bone allograft in human periodontal osseous defects. , 1988, Journal of periodontology.

[21]  P. Indelicato,et al.  Bone Tunnel Enlargement After Anterior Cruciate Ligament Replacement , 1994, The American journal of sports medicine.

[22]  J. B. Liesch,et al.  Development of fibroblast-seeded ligament analogs for ACL reconstruction. , 1995, Journal of biomedical materials research.

[23]  N. Sharkey,et al.  Replacement of the anterior cruciate ligament. A comparison of autografts and allografts in dogs. , 1989, Acta orthopaedica Scandinavica.

[24]  G. Fields,et al.  Melanoma cell adhesion and spreading activities of a synthetic 124-residue triple-helical "mini-collagen". , 1993, The Journal of biological chemistry.

[25]  R J Johnson,et al.  The treatment of injuries of the anterior cruciate ligament. , 1992, The Journal of bone and joint surgery. American volume.

[26]  Cyril B. Frank,et al.  Current Concepts Review - The Science of Reconstruction of the Anterior Cruciate Ligament* , 1997 .

[27]  J. Dayer,et al.  Stimulation of collagenase production by collagen in mammalian cell cultures , 1979, Cell.

[28]  N. Sharkey,et al.  Immune Response to Nonspecific and Altered Tissue Antigens in Soft Tissue Allografts , 1996, Clinical orthopaedics and related research.

[29]  D. W. Jackson,et al.  Biologic and synthetic implants to replace the anterior cruciate ligament. , 1994, Arthroscopy : the journal of arthroscopic & related surgery : official publication of the Arthroscopy Association of North America and the International Arthroscopy Association.

[30]  M. Dunn,et al.  Effect of chemical treatments on tendon cellularity and mechanical properties. , 2000, Journal of biomedical materials research.

[31]  J. Minei,et al.  Replacement of the anterior cruciate ligament using a synthetic prosthesis , 1986, The American journal of sports medicine.

[32]  J. Pinkowski,et al.  Human lymphocyte reaction to freeze-dried allograft and xenograft ligamentous tissue , 1989, The American journal of sports medicine.

[33]  C. Frank,et al.  The Biology of Tendons and Ligaments , 1990 .

[34]  D McComb,et al.  Development of a pericardial acellular matrix biomaterial: biochemical and mechanical effects of cell extraction. , 1994, Journal of biomedical materials research.

[35]  Sheldon R. Simon,et al.  Orthopaedic basic science : biology and biomechanics of the musculoskeletal system , 2000 .