Tissue engineering approaches to tendon repair studies on the use of bone marrow stromal cells and knitted poly (d, l lactide co glycolide scaffold

TISSUE ENGINEERING APPROACHES TO TENDON REPAIR: STUDIES ON THE USE OF BONE MARROW STROMAL CELLS AND KNITTED POLY (D, L-LACTIDE-CO-GLYCOLIDE) SCAFFOLD OUYANG HONGWEI (Bachelor of Medicine/Bachelor of Surgery) A THESIS SUBMITED FOR THE DEGREE OF DOCTOR OF PHILOSOPHY DEPARTMENT OF ORTHOPAEDIC SURGERY NATIONAL UNIVERSITY OF SINGAPORE 2003 Acknowledgements i Acknowledgements I wish to express my deepest gratitude and heartfelt thanks to my supervisors: Professor Lee Eng Hin, Dean, Faculty of medicine, National University of Singapore, and Associate Professor James Goh Director of research, Department of Orthopedic Surgery, National University of Singapore, for their constant encouragement, invaluable guidance and infinite patience throughout the course of this study. I would like to express my sincere thanks to Professor K Satkunanantham Head, Department of Orthopedic Surgery, National University of Singapore, for his support. Without the excellent facilities, this work would not have been possible. I owe my thanks to Professor Teoh Swee Hin, Assistant Professor Dietmar Hutmacher, Dr Mo Xui-Mei, Division of Bioengineering, NUS for their assistance in the provision of biomaterials and reagents, as well as technique assistance in manufacturing of scaffolds. I would also like to express my appreciation to the following staff members Ms Chong Sue-Wee, Ms Julee Chan, Mr Ashvin Thambyah, Ms Grace Lee, Mr Barry P Pereira, Ms Jessie Tan, Mr Yong Soon Chiong, Mr Dominic Tey, Dr Li Li, Dr Ge Zi-Gang, and Dr Shao Xin-Xin for their kind help. I will always remember my friends in Singapore for their constant encouragement and kind help. This work was support by a grant from NMRC, Singapore. Acknowledgements ii Last but not least, I am grateful to my families, my parents, my wife Zou Xiao-Hui and my baby OuYang Xin-yi for their understanding and love during the years of my Ph.D. pursuit. . Table of Content Table of Content I. Acknowledgement…………………………………………………………………i II. Table of content………………………………………………………………… iii III. Summary………………………………………………………………………….ix IV. Publications………………………………………………………………………xii Chapter I: Introduction and Literature Review……………………………………………1 Chapter II Literature Review…………….……………………………………………… 2.0. Introduction to Literature Review……………………………………….… … .6 2.1 Tendon Anatomy, Physiology And Injury…………………………………… …6 2.1.1 Tendon Anatomy……………………………………………………………… .6 2.1.2 Tendon Composite…………… …………………………………….… ………8 2.1.3 Tendon to Bone Insertion…… …………………………………… ….….……13 2.1.4 Tendon Biomechanics…… …………………………………………… ……14 2.1.4.1 Tendon Mechanical Properties………… .…………… .…… .14 2.1.4.2 Effect of Biomechanical Load on Tendon…………………………………….16 2.2 Tendon Injury Healing and Current Therapy………………………………… 18 2.2.1 Tendon Injury Healing ……………………………………………………… 18 2.2.2 Several Concerns about Tendon Healing…………………………………… .20 2.2.2.1 The Amount of Reparative Cells………………………………………………20 2.2.2.2 The Mobilization of Reparative Tendon…………………………………….…22 iii . Table of Content 2.2.2.3 The Evaluation of Tendon Healing……………………………………………22 2.2.3. Current Therapy……………………………………………………………… 23 2.3. Tissue Engineering Approaches To Improve Tendon Healing……………… .24 2.3.1. Tissue Engineering Principles………………………………………………….24 2.3.2. Cell Source…………………………………………………………………… 25 2.3.3. Biomaterials and Scaffold……………………………………………… .29 2.3.4. Biomolecules………………………………………………………………… .33 2.3.5. Animal Model………………………………………………………………….36 2.3.6. Tissue Engineering Techniques for Tendon Insertion Healing……………… 39 2.4 Hypotheses and Objective of This study……………………… .…….…… 40 Chapter III: Materials and Methods…………………………………………….… ……42 3.0 Introduction to Material and Methodology…………………………….………43 3.1 Stage1. bMSCs Differentiation Study……………………………….…………43 3.1.1 Isolation And Culture Of Bone Marrow Stromal Cells……………………… 43 3.1.2 Osteogenesis Induction and Detection……………………………….……… .44 3.1.2.1 In vitro Osteogenesis Induction ……………………… …………….……… 44 3.1.2.2 Vonkossa Staining…………………………………………………………… 45 3.1.3 Chondrogensis Induction And Detection………………………………………46 3.1.3.1 In Vitro Chondrogenesis Induction ……………… ………………….…….…46 3.1.3.2 Collagen Type II Immunoassaying………………………………………….…47 3.1.4 Adipogenesis Induction And Detection……………………………………… 48 3.1.4.1 In Vitro Adipogenesis Induction …………………………………… ……….49 iv . Table of Content v 3.1.4.2 Oil Red Staining……………………………………………………………….50 3.2 Stage II. Trace Study On The Fate of bMSCs After Implantation…………….51 3.2.1. Animal Model ……………………………………………………………… .51 3.2.2. Cells Labeling And Detection………………………………………………….52 3.2.2.1 CFDA Labeling And Detection……………………………………………… 52 3.2.2.2 GFP Gene Transfection And Detection……………………………………….53 3.2.3. Tissue Preparation For Cryostat Section……………………………………….54 3.3 Stage 3. Study On The behavior of bMSCs On Various Polymer Films …… .55 3.3.1. Materials……………………………………………………………………….55 3.3.2. Polymer Film Manufacture…………………………………………………….56 3.3.3. Polymer film Sterilization and Prewetting…………………………….……….56 3.3.4. Water Contact Angle Test…………………………………………………… .57 3.3.5. bMSCs Adhesion Assay ………………………………………………………57 3.3.6. bMSCs Proliferation Assay…………………………………………………….57 3.3.7. MTS Assay For Cell Proliferation…………………………………………… 58 3.3.8. Statistic Analysis Method…………………………………………………… .58 3.4 Stage 4. Study On The Effect Of bMSCs Seeded knitted PLGA For Achilles Tendon Repair……………………………………………………………………………….……59 3.4.1 Animal Surgery……………………………………………………………… .59 3.4.2 Knitting PLGA Fiber Scaffold…………………………………………………61 3.4.3 Tissue Preparation For Paraffin Section……………………………………….61 3.4.4 H&E Staining…………………………………………………………… …….62 3.4.5 Immunohistochemical Staining……………………………………………… 63 . Table of Content vi 3.4.6 Transmission Electronic Microscopy……………………………………… ….64 3.4.7 Biomechanical Test………………………………………………………….…64 3.5. Stage 5. Study on the Effect Of bMSCs On the Tendon Insertion Healing… … 66 3.5.1. Animal Surgery………………………… …………………………….……… .66 3.5.2. Hard Tissue preparation For Paraffin Section……… ………………….………67 3.5.3. H&E & Immunohistochemical Staining ……………………………………… 67 Chapter IV: Results………………………………………………………………………68 4.1 Stage 1.The Differentiation Of bMSCs………… ………………………….….69 4.1.1. bMSCs Isolation……………………………………………… …69 4.1.2. Osteo-lineage Differentiation………………………………………………… 70 4.1.3. Chondro-lineage Differentiation ………………………………………… … .71 4.1.4. Adipo-lineage Differentiation………………………………………… … ……72 4.2 Stage 2. The Fate of bMSCs After Implantation…… …………………… ……73 4.3 Stage3. The Adhesion and Proliferation of bMSCs on Various Polymer Films…………………………………………………………………………………… 76 4.3.1. Polymer Films……………………………………………………………………76 4.3.2. Cell Adhesion…………………………………………………………………….77 4.3.3. Cell Proliferation ………………………………… …………………….………79 4.3.4. Cell Morphology………………………………………… …………….……….81 4.4 Stage4. The Efficacy Of Allogeneic bMSCs Seeded Knitted PLGA Scaffold For Achilles Tendon Repair…………………………………………………………… ….82 4.4.1. The Histology of Tendon Repair………………………………………… ……82 . Table of Content vii 4.4.2. The Immunohistology of Tendon Repair……………………………… ………90 4.4.3. The Biomechanics of Tendon Repair…………………………… .…… … … 91 4.5 Stage5. The Effect of bMSCs on Tendon Insertion Healing………… …….….94 5.5.1. The Natural Healing of Tendon Insertion……………………………… ….… 94 5.5.2. The Effect of bMSCs on Tendon Insertion Healing …………………… … …96 Chapter V: Discussion…………………………………………………………………99 5.1. The Isolation and Differentiation of bMSCs…………………….…………… 100 5.1.1. The isolation of bMSCs………………………………………………….…… 100 5.1.2. The Multipotential of bMSCs…………………………………………….…….101 5.1.3. Bone Marrow Stromal Cells as Tendon progenitor Cells………………….… 102 5.2. The Fate of bMSCs After Implantation………………………………… … .103 5.2.1. The Methods of Cell Trace Study………………………………………… … 103 5.2.2. The Fate of bMSCs after Implantation into Tendon Wound Site….……… .…103 5.2.3. The Possibility of Allogeneic bMSCs for Tissue Engineering…………….… .105 5.2.4. The Potential of bMSCs For Gene Delivery……………………… …….…….107 5.3. The Behavior of bMSCs on Various Polymer Films……………… ……… …108 5.3.1. Method for Characterizing Cell-Polymer Interaction……………………… ….108 5.3.2. The Effect of Cell Source on The Cell-Polymer Interaction………………… 109 5.3.3. The Effect of Substrate on The bMSCs Adhesion And Proliferation……… 110 5.4. The Efficacy of bMSCs and Knitted PLGA scaffold for Achilles Tendon Repair……………………………………………………………………………… … 113 5.4.1. The Effect of Knitted PLGA on Tendon Repair…………………………… ….113 . Table of Content 5.4.2. The Effect of bMSCs on Tendon Repair…………………………………… 116 5.4.3. Tendon Repair versus Tendon Regeneration……………………………… …118 5.5. The Effect of bMSCs on Tendon Insertion Healing ……………………… …120 5.5.1. The Natural Healing of Tendon Insertion………………………………… ….120 5.5.2. The Effect of bMSCs on Tendon Insertion Healing……………………… ….121 Chapter VI Conclusion and Recommended Future Study……………………… ….…124 ChapterVII References………………………………………………………… …… 128 viii Summary ix Summary Background: Unlike bone that is able to heal by regenerating normal bone in most cases, tendons often heal by forming scar tissue. The limited capacity for injured tendon to regenerate poses a great challenge and creates an opportunity for engineering new tendons. To date, less work has been done on tendon tissue engineering compared to the extensive work on the bone and cartilage tissue engineering. Several studies have investigated the use of gel and braided scaffold with or without cells for tendon repair; however, the inferior mechanical strength of gel carrier and the poor tissue ingrowths associated with the braided fiber scaffold has limited the success. Many other problems have yet to be addressed: the fate of implanted bone marrow stromal cells (bMSCs) at the tendon site has not yet been studied; no general principles have been established to select material for bMSCs delivery; the role of bMSCs in tendon repair has been arguable due to the lack of appropriate controls in previous studies, and limited attention has been placed on the tendon-to-bone healing when engineering tendon graft for tendon repair. Hypotheses: The main hypothesis is that tissue engineered graft composed of bMSCs and knitted PLGA can improve tendon repair. To support this hypothesis: (a) bMSCs should have multipotential and be good candidates of cell source for tendon repair. (b) The knitted PLGA should be promising for bMSCs delivery and tendon repair. Chapter VII References 123. 145 Kadiyala S, Young RG, Thiede MA, Bruder SP. Culture expanded canine mesenchymal stem cells possess osteochondrogenic potential in vivo and in vitro.Cell Transplant 6(2), 125, 1997. 124. Kang HJ, Kang ES. Ideal concentration of growth factors in rabbit's flexor tendon culture. Yonsei Med J 40(1): 26, 1999. 125. Kaigler D, Krebsbach PH, Polverini PJ, Mooney DJ. Role of vascular endothelial growth factor in bone marrow stromal cell modulation of endothelial cells.Tissue EngFeb; 9(1):95-103, 2003. 126. Kainberger FM, Engel A, Barton P, Huebsch P, Neuhold A, Salomonowitz E. Injury of the Achilles tendon: diagnosis with sonography. AJR Am J Roentgenol. Nov; 155(5):1031-6. 1990. 127. Karrer FM, Reitz BL, Hao L, Lafferty KJ. Fluorescein labeling of murine hepatocytes for identification after intrahepatic transplantation.Transplant Proc. Dec; 24(6):2820-1, 1992. 128. Karpakka J, Virtanen P, Vaananen K, Orava S, Takala TE. Collagen synthesis in rat skeletal muscle during immobilization and remobilization. J Appl Physiol. Apr; 70(4):1775-80, 1991. 129. Kastelic J, Galeski A, Baer E. The multicomposite structure of tendon. Connect Tissue Res. 6(1):11-23, 1978. 130. Kato YP, Dunn MG, Zawadsky JP, Tria AJ, Silver FH. Regeneration of Achilles tendon with a collagen tendon prosthesis. Results of a one-year implantation study. J Bone Joint Surg Am. Apr; 73(4):561-74. 1991. Chapter VII References 131. 146 Kawaja MD, Fagan AM, Firestein BL, Gage FH. Intracerebral grafting of cultured autologous skin fibroblasts into the rat striatum: an assessment of graft size and ultrastructure.J Comp Neurol. May 22;307(4):695-706, 1991. 132. Kessler FB, Epstein MJ, Lannik D, Maher D, Pappu S. Fascia patch graft for a digital flexor sheath defect over primary tendon repair in the chicken.J Hand Surg [Am]. Mar; 11(2):241-5. 1986. 133. Ketchum LD. Tendon healing. In Fundamentals of Wound Management in Surgery: Selected Tissue. Chirgecom, New Jersy, pp 122-153. 1977. 134. Klebe RJ, Caldwell H, Milam S. Cells transmit spatial information by orienting collagen fibers. Matrix.9 (6):451-8, 1989. 135. Kobayashi D, Kurosaka M, Yoshiya S, Mizuno K. Effect of basic fibroblast growth factor on the healing of defects in the canine anterior cruciate ligament. Knee Surg Sports Traumatol Arthrosc 5(3): 189, 1997. 136. Kurtz CA, Loebig TG, Anderson DD, DeMeo PJ, Campbell PG. Insulin-like growth factor I accelerates functional recovery from Achilles tendon injury in a rat model. Am J Sports Med 27(3): 363, 1999. 137. Kvist M, Jozsa L, Jarvinen M, Kvist H. Fine structural alterations in chronic Achilles paratenonitis in athletes. Pathol Res Pract. Oct; 180(4):416-23. 1985. 138. Kvist M, Jozsa L, Jarvinen M. Vascular changes in the ruptured Achilles tendon and paratenon. Int Orthop.16 (4):377-82, 1992. 139. Laitinen O, Tormala P, Taurio R, Skutnabb K, Saarelainen K, Iivonen T, Vainionpaa S. Mechanical properties of biodegradable ligament augmentation Chapter VII References 147 device of poly(L-lactide) in vitro and in vivo. Biomaterials 13(14): 1012, 1992. 140. Lampin M, Warocquier-Clerout, Legris C, Degrange M, Sigot-Luizard MF. Correlation between substratum roughness and wettability, cell adhesion, and cell migration.J Biomed Mater Res. Jul;36(1):99-108, 1997. 141. Langer R, Vancanti JP. Tissue engineering. Science, 260: 920-926. 1993. 142. Lappalainen R, Knuuttila M, Lammi S, Alhava EM, Olkkonen H. Zn and Cu content in human cancellous bone. Acta Orthop Scand. Feb; 53(1):51-5, 1982. 143. Laurencin C, Attawia M, Botchwey E, Warren R, Attia E. Cell-material systems for anterior cruciate ligament regeneration. In Vitro Cell Dev Biol Anim 34(2), 90, 1998. 144. Lazarus HM, Haynesworth SE, Gerson SL, Rosenthal NS, Caplan AI. Ex vivo expansion and subsequent infusion of human bone marrow-derived stromal progenitor cells (mesenchymal progenitor cells): implications for therapeutic use.Bone Marrow Transplant. Oct; 16(4):557-64. 1995. 145. Lawrence MB, McIntire LV, Eskin SG. Effect of flow on polymorphonuclear leukocyte/endothelial cell adhesion.Blood. Nov;70(5):1284-90, 1987. 146. Leadbetter WB, Mooar PA, Lane GJ, Lee SJ. The surgical treatment of tendinitis. Clinical rationale and biologic basis. Clin Sports Med. Oct;11(4):679-712. 1992. 147. Letson AK, Dahners LE. The effect of combinations of growth factors on ligament healing. Clin Orthop 308, 207, 1994. Chapter VII References 148. 148 Lodie TA, Blickarz CE, Devarakonda TJ, He C, Dash AB, Clarke J, Gleneck K, Shihabuddin L, Tubo R.Systematic analysis of reportedly distinct populations of multipotent bone marrow-derived stem cells reveals a lack of distinction.Tissue Eng. Oct;8(5):739-5, 2002. 149. Levine SN. Survey of biomedical materials and some relevant problems.Ann N Y Acad Sci. Jan; 146(1):3-10. 1968. 150. Liechty KW, MacKenzie TC, Shaaban AF, Radu A, Moseley AM, Deans R, Marshak DR, Flake AW. Human mesenchymal stem cells engraft and demonstrate site-specific differentiation after in utero transplantation in sheep. Nature Medicine, 6(11), 1282, 2000. 151. Lin VS, Lee MC, O'Neal S, McKean J, Sung KL. Ligament tissue engineering using synthetic biodegradable fiber scaffolds. Tissue Eng 5(5), 443, 1999. 152. Liu SH, Yang RS, al-Shaikh R, Lane JM. Collagen in tendon, ligament, and bone healing. A current review.Clin Orthop. Sep;(318):265-78, 1995. 153. Liu SH, Panossian V, al-Shaikh R, Tomin E, Shepherd E, Finerman GA, Lane JM. Morphology and matrix composition during early tendon to bone healing. Clin Orthop 339: 253-260, 1997. 154. Liu YT, Zhang ZF, Wu JJ, Shang QX, Cao YL. A experiment study on repair of tendon defect using tissue-engineered tendon. Tissue Eng. 6, 677, 2000. 155. Lou J, Manske PR, Aoki M, Joyce ME. Adenovirus-mediated gene transfer into tendon and tendon sheath. J Orthop Res. Jul; 14(4):513-7. 1996. 156. Lou J, Kubota H, Hotokezaka S, Ludwig FJ, Manske PR. In vivo gene transfer and overexpression of focal adhesion kinase (pp125 FAK) mediated by Chapter VII References 149 recombinant adenovirus-induced tendon adhesion formation and epitenon cell change. J Orthop Res. Nov; 15 (6):911-8. 1997. 157. Louie LK, Yannas IV, Hsu HP, Spector M Healing of tendon defects implanted with a porous collagen-GAG matrix: Histological evaluation. Tissue Eng (2): 187-195 SUM 1997. 158. Lu L, Peter SJ, Lyman MD, Lai HL, Leite SM, Tamada JA, Uyama, S, Vacanti, JP, Langer, R, Mikos AG. In vitro and in vivo degradation of porous poly(DL-lactic-co-glycolic acid) foams, Biomaterials, Volume 21, Issue 18: 1837, 2000. 159. Lucas PA, Warejcka DJ, Zhang LM, Newman WH, Young HE. Effect of rat mesenchymal stem cells on development of abdominal adhesions after surgery. J Surg Res 62(2), 229, 1996. 160. Mackay AM, Beck SC, Murphy JM, Barry FP, Chichester CO, Pittenger MF. Chondrogenic differentiation of cultured human mesenchymal stem cells from marrow. Tissue Eng. Winter; 4(4):415-28, 1998. 161. Maffulli N, King JB. Effects of physical activity on some components of the skeletal system. Sports Med. Jun; 13 (6):393-407. 1992. 162. Manske PR, Lesker PA. Histologic evidence of intrinsic flexor tendon repair in various experimental animals. An in vitro study. Clin Orthop. JanFeb;(182):297-304, 1984. 163. Marui T, Niyibizi C, Georgescu HI, Cao M, Kavalkovich KW, Levine RE, Woo SLY. Effect of growth factors on matrix synthesis by ligament fibroblasts. J Orthop Res 15(1): 18, 1997. Chapter VII References 164. 150 Mason ML. Allen HS. The rate of healing of tendons. An experiment study of tensile strength. Ann Surg. 113. 424-459. 1941. 165. Mass DP, Tuel RJ. Participation of human superficialis flexor tendon segments in repair in vitro.J Orthop Res. Jan; 8(1):21-34. 1990. 166. Matyas JR, Frank CB. Development of a periosteal ligament insertion : The origin of Sharpey’s fibers. Trans Orthop Res Soc 14: 185, 1989. 167. McGaw WT. The effect of tension on collagen remodelling by fibroblasts: a stereological ultrastructural study. Connect Tissue Res. 14(3):229-35, 1986. 168. McNeilly CM, Banes AJ, Benjamin M, Ralphs JR. Tendon cells in vivo form a three dimensional network of cell processes linked by gap junctions. J Anat 189(3), 593, 1996. 169. Michna H. Morphometric analysis of loading-induced changes in collagenfibril populations in young tendons. Cell Tissue Res. 1984;236(2):465-70. 170. Michna H, Hartmann G. Adaptation of tendon collagen to exercise. Int Orthop. 13(3):161-5, 1989. 171. Murphy PG, Loitz BJ, Frank CB, Hart DA. Influence of exogenous growth factors on the synthesis and secretion of collagen types I and III by explants of normal and healing rabbit ligaments. Biochem Cell Biol 72(9-10): 403, 1994. 172. Murrell GA, Lilly EG 3rd, Goldner RD, Seaber AV, Best TM. Effects of immobilization on Achilles tendon healing in a rat model.J Orthop Res. Jul;12(4):582-91. 1994. 173. Muschler GF, Midura RJ. Connective tissue progenitors: practical concepts for clinical applications. Clin Orthop. Feb;(395):66-80. 2002. Chapter VII References 174. 151 Nakamura N, Horibe S, Matsumoto N, Tomita T, Natsuume T, Kaneda Y, Shino K, Ochi T. Transient introduction of a foreign gene into healing rat patellar ligament.J Clin Invest. Jan 1;97(1):226-31. 1996. 175. Nakamura N, Shino K, Natsuume T, Horibe S, Matsumoto N, Kaneda Y, Ochi T. Early biological effect of in vivo gene transfer of platelet-derived growth factor (PDGF)-B into healing patellar ligament. Gene Ther. Sep;5(9):1165-70. 1998. 176. Natsu-ume T, Nakamura N, Shino K, Toritsuka Y, Horibe S, Ochi T. Temporal and spatial expression of transforming growth factor-beta in the healing patellar ligament of the rat. J Orthop Res. Nov;15(6):837-43. 1997. 177. O'Brien M. Functional anatomy and physiology of tendons. Clin Sports Med. Jul;11(3):505-20. 1992. 178. Oguma H, Murakami G, Takahashi-Iwanaga H, Aoki M, Ishii S. 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. J Orthop Res. 19(5): 873-80, 2001. 179. Ohtera K, Yamada Y, Aoki M, Sasaki T, Yamakoshi K. Effects of periosteum wrapped around tendon in a bone tunnel: A biomechanical and histological study in rabbits. Crit Rev Biomed Eng 28(1&2):115, 2000. 180. Olivieri MP, Baier RE, Loomis RE. Surface properties of mussel adhesive protein component films. Biomaterials.13(14):1000-8, 1992. Chapter VII References 181. 152 Petersen W, Laprell H: Insertion of autologous tendon graft to the bone: a histological and immunohistochemical study of hamstring and patellar tendon grafts. Knee Surg Sports Traumatol Arthrosc 8(1): 26-31, 2000. 182. Pittenger MF, Mackay AM, Beck SC, Jaiswal RK, Douglas R, Mosca JD, Moorman MA, Simonetti DW, Craig S, Marshak DR. Multilineage potential of adult human mesenchymal stem cells. Science 284(5411), 143, 1999. 183. Pittenger MF, Mosca JD, McIntosh KR. Human mesenchymal stem cells: progenitor cells for cartilage, bone, fat and stroma. Curr Top Microbiol Immunol 251, 3, 2000. 184. Pneumaticos SG, Phd PCN, McGarvey WC, Mody DR, Trevino SG. The effects of early mobilization in the healing of achilles tendon repair.Foot Ankle Int. Jul;21 (7):551-7, 2000. 185. Prockop DJ. Marrow stromal cells as stem cells for nonhematopoietic tissues.Science. Apr 4; 276(5309):71-4. 1997. 186. Proctor CS, Jackson DW, Simon TM. Characterization of the repair tissue after removal of the central one-third of the patellar ligament. An experimental study in a goat model. J Bone Joint Surg Am. Jul;79(7):9971006. 1997. 187. Postacchini F, Accinni L, Natali PG, Ippolito E, DeMartino C. Regeneration of rabbit calcaneal tendon: a morphological and immunochemical study. Cell Tissue Res. Dec 14;195(1):81-97, 1978. Chapter VII References 188. 153 Potenza AD. Tendon healing within the flexor digital sheath of the dog. J Bone J Surg , 44A, 49-64. 1962. 189. Potenza AD. Concepts of tendon healing and repair. In symposium on tendon surgery in the hand. American Academy of Orthopaedic Surgeons. C.V. Mosby Co. St Louis pp18-47. 1975. 190. Proctor CS, Jackson DW, Simon TM. Characterization of the repair tissue after removal of the central one-third of the patellar ligament. An experimental study in a goat model.J Bone Joint Surg Am. Jul;79(7):9971006. 1997. 191. Ralphs JR, Waggett AD, Benjamin M. Actin stress fibres and cell-cell adhesion molecules in tendons: organisation in vivo and response to mechanical loading of tendon cells in vitro.Matrix Biol.21(1):67, 2002. 192. Rodeo SA, Arnoczky SP, Torzilli PA, Tendon-healing in a bone tunnel. A biomechanical and histological study in the dog. J Bone Joint Surg 75A: 1795-1803, 1993. 193. Rodeo SA, Suzuki K, Deng XH, Wozney J, Warren RF,Use of recombinant human bone morphogenetic protein-2 to enhance tendon healing in a bone tunnel. Am J Sports Med. Jul-Aug;27(4):476-88, 1999. 194. Reynolds NL, Worrell TW. Chronic Achilles peritenditis: Etiology, pathophysiology, and treatment. J Orthop Sports Phys Ther 13, 171-176. 1991. Chapter VII References 195. 154 Rodkey WG, Cabaud HE, Feagin JA, Perlik PC. A partially biodegradable material device for repair and reconstruction of injured tendons:experimental studies. Am J Sports Med 13: 242, 1985. 196. Rowe RWD. The structure of rat tail tendon fascicles. Connect Tissue Res.14(1):21-30,1985. 197. Russell JE, Manske PR. Collagen synthesis during primate flexor tendon repair in vitro.J Orthop Res. Jan;8(1):13-20. 1990. 198. Salamon A, Hamori J, deak, GY, Mayer F Submicroscopic investigation of autologous tendon grafts. Acta morph Hung 18, 23-40.1970. 199. Saltzman WM. Cell interactions with polymers. In: Lanza RP, Langer R, Vancanti J eds. Principles of Tissue Engineering. 2nd Edition, Academic Press, California, pp: 221-237, 2000. 200. Sato M, Maeda M, Kurosawa H, Inoue Y, Yamauchi Y, Iwase H. Reconstruction of rabbit Achilles tendon with three bioabsorbable materials: histological and biomechanical studies.J Orthop Sci.5(3):256-67. 2000. 201. Schemidt-Rhfing B, Graf J, Schneider U, Niethard FU. The blood supply of the Achilles tendon. Int Orthop 16, 29-31.1992. 202. Scherping SC Jr, Schmidt CC, Georgescu HI, Kwoh CK, Evans CH, Woo SL. Effect of growth factors on the proliferation of ligament fibroblasts from skeletally mature rabbits. Connect Tissue Res 36(1): 1, 1997. 203. Schiavone PA, Fabbriciani C, Delcogliano A, Franzese S. Bone-ligament interaction in patellar tendon reconstruction of the ACL. Knee Surg Sports Traumatol Arthrosc.1 (1):4-8, 1993. Chapter VII References 204. 155 Shino K, Kawasaki T, Hirose H, Gotoh I, Inoue M, Ono K. Replacement of the anterior cruciate ligament by an allogeneic tendon graft. An experimental study in the dog. J Bone Joint Surg Br. Nov;66(5):672-81, 1984. 205. Skalak R, Fox CF. Tissue engineering. Granlibakken, Lake Tahoe: Proc workshop; new York: Liss p26-29,1988. 206. Spadaro JA, Becker RO, Bachman CH. The distribution of trace metal ions in bone and tendon. Calcif Tissue Res. 6(1):49-54, 1970. 207. Spindler KP, Imro AK, Mayes CE, Davidson JM. Patellar tendon and anterior cruciate ligament have different mitogenic responses to platelet-derived growth factor and transforming growth factor beta. J Orthop Res 14(4): 542, 1996. 208. Steenbrugge F, Vrdonk R, Vorlat P, Mortier F, Verstraete K. Repair of chronic ruptures of the anterior cruciate ligament using allograft reconstruction and a ligament augmentation device. Acta Orthop Belg 67(3), 252, 2001. 209. Stein SR, Luekens CA. Methods and rationale for closed treatment of Achilles tendon ruptures.Am J Sports Med. Jul-Aug;4(4):162-9. 1976. 210. Sato M, Maeda M, Kurosawa H, Inoue Y, Yamauchi Y, Iwase H. Reconstruction of rabbit Achilles tendon with three bioabsorbable materials: histological and biomechanical studies. J Orthop Sci 5(3): 256, 2000. 211. Strehlow CD, Kneip TJ. The distribution of lead and zinc in the human skeleton. Am Ind Hyg Assoc J. Jul-Aug;30(4):372-8. 1969. Chapter VII References 212. 156 Streit M, Braathen LR. Apligraf - a living human skin equivalent for the treatment of chronic wounds. Int J Artif Organs 23(12), 831, 2000. 213. Sutker BD, lester GGE, Beanes AJ. Cyclic strain stimulates DNA and collagen synthesis in fibroblasts cultured from rat medial collateral ligaments. Trans Orthop Res 15, 130, 1990. 214. Tamada Y, Ikata Y. Fibroblast growth on polymer surfaces and biosynthesis of collagen. J Biomed Mat Res. 28:783-189, 1994. 215. Takai S, Woo SL, Horibe S, Tung DK, Gelberman RH. The effects of frequency and duration of controlled passive mobilization on tendon healing.J Orthop Res. Sep;9(5):705-13, 1991. 216. Tauro JC, Parsons JR, Ricci J, Alexander H. Comparison of bovine collagen xenografts to autografts in the rabbit. Clin Orthop May;(266):271-84. 1991. 217. Thambyah, A., Thiagarajan, P, and Goh, J.C.study on the effect of twisted human patellar tendon.Clin. Biomech. 15, 756, 2000. 218. Thomopoulos S, Hattersley G, Rosen V, Mertens M, Galatz L, Williams GR, Soslowsky LJ. The localized expression of extracellular matrix components in healing tendon insertion sites: an in situ hybridization study. J Orthop Res. 20(3): 454-63, 2002. 219. Tozeren A. Adhesion induced by mobile cross-bridges: steady state peeling of conjugated cell pairs. J Theor Biol. Sep 11;140(1):1-17, 1989. 220. Viidik A. Biomechanics and functional adaptation of tendons and joint ligaments. In: Evans GF eds. Studies on the anatomy and function of bone and joints. New York: Springer, pp.17-39. 1966. Chapter VII References 221. 157 Viidik A. Functional properties of collagenous tissues. Int Rev conn Tissue res6. 127-215.1973. 222. Wada A, Kubota H, Miyanishi K, Hatanaka H, Miura H, Iwamoto Y. Comparison of postoperative early active mobilization and immobilization in vivo utilising a four-strand flexor tendon repair. J Hand Surg [Br]. Aug;26(4):301-6. 2001. 223. Wagers AJ, Sherwood RI, Christensen JL, Weissman IL. Little evidence for developmental plasticity of adult hematopoietic stem cells.Science. Sep 27;297(5590):2256-9, 2002. 224. Wasserman AJ, Kato YP, Christiansen D, Dunn MG, Silver FH. Achilles tendon replacement by collagen fiber prosthesis: morphological evaluation of neotendon formation. Scanning Microsc 3(4):1183, 1989. 225. Watanabe N, Taskai S, Morita N, Kawata T, Hirasawa Y. New method of distinguishing between intrinsic cells in situ and extrinsic cells supplied by the atuogeneic transplantation, employing transgenic rats. Trans Orthop Res Soc 23: 1035, 1998. 226. Whitby DJ, Ferguson MW. The extracellular matrix of lip wounds in fetal, neonatal and adult mice. Development. Jun;112(2):651-68, 1991. 227. Whittaker P, Canham PB. Demonstration of quantitative fabric analysis of tendon collagen using two-dimensional polarized light microscopy. Matrix. Feb;11(1):56-62, 1991. Chapter VII References 228. 158 Williams JG. Achilles tendon lesions in sport. Sports Med. Sep;16(3):216-20, 1993. 229. Williams SK, Kleinert LB, Rose D, McKenney S. Origin of endothelial cells that line expanded polytetrafluorethylene vascular grafts sodded with cells from microvascularized fat. J Vasc Surg. Apr;19(4):594-604, 1994. 230. Wolfman NM, Hattersley G, Cox K, Celeste AJ, Nelson R, Yamaji N, Dube JL, DiBlasio-Smith E, Nove J, Song JJ, Wozney JM, Rosen V. Ectopic induction of tendon and ligament in rats by growth and differentiation factors 5, 6, and 7, members of the TGF-beta gene family. J Clin Invest 100(2): 321, 1997. 231. Woo SL, Ritter MA, Amiel D, Sanders TM, Gomez MA, Kuei SC, Garfin SR, Akeson WH. The biomechanical and biochemical properties of swine tendons--long term effects of exercise on the digital extensors.Connect Tissue Res.7(3):177, 1980. 232. Woo SL, Gomez MA, Amiel D, Ritter MA, Gelberman RH, Akeson WH. The effects of exercise on the biomechanical and biochemical properties of swine digital flexor tendons.J Biomech Eng. Feb;103(1):51, 1981. 233. Woo SL, Gomez MA, Woo YK, Akeson WH Mechanical properties of tendons and ligaments. II. The relationships of immobilization and exercise on tissue remodeling. Biorheology. 19(3):397,1982. 234. Woo SL, An KN, Arnoczky SP, Wayne WH. Anatomy, biology and biomechanics of tendon, ligament, and menicus. In Simon SR eds. Chapter VII References 159 Orthopaedic Basic Science. American Academy of Orthopaedic Surgeons, Park Ridge, IL, pp45-87, 1994. 235. Yamazaki S, Yasuda K, Tohyama H, Tomita F, Minami A. The effect of transforming growth factor- beta1 on intraosseous healing of the flexor tendon autograft in anterior cruciate ligament reconstruction. 48th Annual Meeting of the Orthopaedic Research Society, Poster 0627, 2001. 236. Yoshikawa Y, Abrahamsson SO. Dose-related cellular effects of plateletderived growth factor-BB differ in various types of rabbit tendons in vitro. Acta Orthop Scand 72(3): 287, 2001. 237. Young HE, Steele TA, Bray RA, et al. Human reserve pluripotent mesenchmal stem cells are present in the connective tissue of skeletal muscle and dermis derived from fetal, adult, and geriatric donors. The Anat Rec 264, 51, 2001. 238. Young RG, Butler DL, Weber W, Caplan AI, Gordon SL, Fink DJ. Use of mesenchymal stem cells in a collagen matrix for Achilles tendon repair. J Orthop Res. Jul;16(4), 406,1998. 239. Zange R, Kissel T. Comparative in vitro biocompatibility testing of polycyanoacrylates and poly(D,L-lactide-co-glycolide) using different mouse fibroblast (L929) biocompatibility test models. Europ J Pharma Biopharma .44(2): 149,1997. 240. Zdrahala RJ. Small caliber vascular grafts. Part I: state of the art. J Biomater Appl 10 (4):309, 1996. Chapter VII References 241. 160 Zernicke RF, Loitz BJ. Exercise-related adaptations in connective tissue. In Komi PV eds. Strength and power in sport. Oxford: Blackwell, pp77-95. 1992. 242. Zhang XY, La Russa VF, Bao L, Kolls J, Schwarzenberger P, Reiser J. Lentiviral vectors for sustained transgene expression in human bone marrowderived stromal cells. Mol Ther. May;5(5 Pt 1):555-65. 2002. [...]... successful will depend on the degree to which the normal composition and architecture of the extracellular matrix is restored Apart from the extensive work on the biology of the extracellullar components of tendons, there are also many studies that focused on understanding the biology of tendon cells Tendons have a variety of cell types, including fibroblasts, fibrocartilage cells and occasional fat cells. .. for skeletal locomotion; for example, the Achilles tendon, one of the largest in the body, links the triceps surae muscle, the grouping of gastrocnemius, the soleus, and the plantaris muscle to the calcaneus bone As organs, tendons consist of three parts: the muscle attachment region, the substance of the tendon itself, and tendon to bone insertion region These three parts vary in their cellular Chapter... tendon junction Vascular basement membrane, Muscle tendon junction Vascular walls, Muscle tendon junction Type I collagen has predilection to form parallel fibers One collagen type I molecule consists of three helical polypeptide alpha-chains that wind around each other to form a right-hand triple helix that extends collinear through out the length of the tropocollagen molecule or microfibril (Ghadially... Fibroblasts are the most common and can be found in all regions of tendons and ligaments They are typically arranged in elongated rows within the parallel bundles of collagen fibers In longitudinal section, the cells are elongated and have spindle shape nuclei In addition to the responsibility of producing extracellular matrix, fibroblasts also mediate the tendons response’ to biomechanical load Chapter... appropriate controls in previous studies, and 4) limited attention has been placed on the tendon -to- bone healing when engineering tendon graft for tendon repair It is clear that the continued development of tendon tissue engineering will depend upon identification and characterization of appropriate sources of cells as well as the development of new scaffolds The identification of an optimal cell source... quality of tendon repair 2.2.2 Issues in Tendon Healing: 2.2.2.1 The Amount of Reparative Cells Many cell-mediated processes related to the generation of skeletal tissue depend on the number of cells involved, both in the rate and magnitude of the effect For example, in the in-vitro model of production of connective tissue, the rate of collagen gel contraction by fibroblasts embedded in the gel is dependent... healing in the stage 5 experiment Conclusion: In all, these sequential experiments proved that the bMSCs were able to be the seed cells for tendon repair; the knitted PLGA scaffolds possess optimal material and structural properties for bMSCs delivery and tendon tissue formation; and the composite of bMSCs and knitted PLGA could be an ideal substitute for tendon repair This work could be logically... extremely high tensile strength The extracellular tendon matrix is composed of collagen and elastin fibers, the ground substance, and the inorganic components Collagen constitutes approximately 90% of the total protein of the tendons or 65-75% of the dry mass of tendons Elastin accounts for only about 2 % of the dry mass of tendons (Hess, 1989, Jozsa 1989b) The ground substance, which surrounds the collagen,... HW, Lee EH Survivability and functionality of bone marrow stromal cells following implantation in tendon regeneration in rabbit model 49th annual meeting of orthopedic research society New Orleans, LA, USA Feb 2-5, 2003 2 Ouyang HW, Goh JCH, Lee EH Application of mesenchymal stem cells in the repair of tendon and ligament 11th international conference of biological and medical engineering 4th to 7th... those of non athletes It suggests that localized physiological adaptation to mechanical stress occurs in human tendons (Kainberger 1990, Maffulli 1992) In in-vitro studies, tendon cells have been shown to respond to mechanical loading The responses include release of intracellular calcium, alteration of their cytoplasmic filament organization and content, polymerization of actins and alteration of protein . TISSUE ENGINEERING APPROACHES TO TENDON REPAIR: STUDIES ON THE USE OF BONE MARROW STROMAL CELLS AND KNITTED POLY (D, L- LACTIDE- CO- GLYCOLIDE) SCAFFOLD OUYANG HONGWEI. dense connective tissue that links muscle to bone and allows for transmission of muscle contraction forces to the bone for skeletal locomotion; for example, the Achilles tendon, one of the largest. Understanding of the cellular and extracellular components of tendon is essential for determining the methodology required to successfully engineer tissue, and provide useful information in the