A comparison of tenocytes and mesenchymal stem cells for use in flexor tendon tissue engineering.

PURPOSE Tissue-engineered tendon grafts will meet an important clinical need. To engineer tendons, we used acellularized allogeneic tendon as scaffold material. To determine the ideal cell type to seed the scaffolds, we studied in vitro characteristics of epitenon tenocytes, tendon sheath fibroblasts, bone marrow-derived mesenchymal stem cells (BMSCs), and adipoderived mesenchymal stem cells (ASCs). Subsequently, we implanted reseeded acellularized tendons in vivo as flexor tendon grafts. METHODS Tenocytes, sheath fibroblasts, BMSCs, and ASCs were obtained from adult rabbits. For all cell lines, collagen 1, 2, and 3 immunocytochemistry was performed, and proliferation was assessed by hemacytometry and senescence by beta-galactosidase staining. Flexor tendons were acellularized after harvest. Tendons were assessed by histology after in vitro reseeding with each of the cell types after 1, 4, and 8 weeks. Finally, reseeded tendons and controls were implanted in a flexor profundus tendon defect. After 6 weeks, the reseeded tendons were harvested and assessed by histology. Statistical analysis for cell proliferation was performed using analysis of variance and t-tests with Bonferroni correction. RESULTS All cell types had similar collagen expression. Cell proliferation was higher in ASCs in late passage compared with early passage and in ASCs compared with epitenon tenocytes at late passage. The other cell types were similar in growth characteristics. No senescence was detected. In vitro assessment of reseeded constructs showed the presence of cells on the construct surface. In vivo assessment after implantation showed viable cells seen within the tendon architecture in all cell types. CONCLUSIONS This study suggests that the four cell types may be successfully used to engineer tendons. Adipoderived mesenchymal stem cells proliferate faster in cell culture, but the cell types were similar in other respects. All could be used to successfully repopulate acellularized tendon in vivo as flexor tendon grafts.

[1]  H. Lorenz,et al.  Multilineage cells from human adipose tissue: implications for cell-based therapies. , 2001, Tissue engineering.

[2]  C. Hecquet,et al.  Culture and characterization of juvenile rabbit tenocytes , 1997, Cell Biology and Toxicology.

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

[4]  J. Goh,et al.  Selection of Cell Source for Ligament Tissue Engineering , 2005, Cell transplantation.

[5]  Min Zhu,et al.  Comparison of Multi-Lineage Cells from Human Adipose Tissue and Bone Marrow , 2003, Cells Tissues Organs.

[6]  W. L. White.  Tendon grafts: a consideration of their source, procurement and suitability. , 1960, The Surgical clinics of North America.

[7]  Jennifer H. Elisseeff,et al.  Engineering Structurally Organized Cartilage and Bone Tissues , 2004, Annals of Biomedical Engineering.

[8]  S. Woo,et al.  The Marshall R. Urist Young Investigator Award , 1997 .

[9]  D J Prockop,et al.  Rapid expansion of recycling stem cells in cultures of plastic-adherent cells from human bone marrow. , 2000, Proceedings of the National Academy of Sciences of the United States of America.

[10]  Yilin Cao,et al.  Bridging tendon defects using autologous tenocyte engineered tendon in a hen model. , 2002, Plastic and reconstructive surgery.

[11]  Frank P Barry,et al.  Mesenchymal stem cells avoid allogeneic rejection , 2005, Journal of Inflammation.

[12]  Beng Hai Lim,et al.  Bone marrow-derived mesenchymal stem cells influence early tendon-healing in a rabbit achilles tendon model. , 2007, The Journal of bone and joint surgery. American volume.

[13]  Swee Hin Teoh,et al.  The Efficacy of Bone Marrow Stromal Cell‐Seeded Knitted PLGA Fiber Scaffold for Achilles Tendon Repair , 2002, Annals of the New York Academy of Sciences.

[14]  N. Occleston,et al.  Differences in proliferative rate and collagen lattice contraction between endotenon and synovial fibroblasts. , 1998, The Journal of hand surgery.

[15]  M. Kassem,et al.  Mesenchymal stem cells: cell biology and potential use in therapy. , 2004, Basic & clinical pharmacology & toxicology.

[16]  M. Klein,et al.  Flexor tendon wound healing in vitro: the effect of lactate on tendon cell proliferation and collagen production. , 2001, Journal of Hand Surgery-American Volume.

[17]  C Roskelley,et al.  A biomarker that identifies senescent human cells in culture and in aging skin in vivo. , 1995, Proceedings of the National Academy of Sciences of the United States of America.

[18]  James Chang,et al.  Tissue engineering of flexor tendons. , 2003, Clinics in plastic surgery.

[19]  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.

[20]  A Haverich,et al.  Tissue engineering of heart valves--human endothelial cell seeding of detergent acellularized porcine valves. , 1998, European journal of cardio-thoracic surgery : official journal of the European Association for Cardio-thoracic Surgery.

[21]  S. Woo,et al.  Autogenous flexor tendon grafts: Biologic mechanisms for incorporation , 1997 .

[22]  A. Chong,et al.  Tissue engineering for the hand surgeon: a clinical perspective. , 2006, The Journal of hand surgery.

[23]  R. Gelberman,et al.  Flexor tendon grafting to the hand: an assessment of the intrasynovial donor tendon-A preliminary single-cohort study. , 2000, The Journal of hand surgery.

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

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