[Frontiers in Bioscience, Landmark, 24, 628-647, March 1, 2019]

One-step strategy for chondral defect repair

Ying Zhang 1 , Hongbo Tan 2 , Guangxing Chen 1 , Weinan Zeng 1 , Junli Liu 1 , Liu Yang 1 , Fuyou Wang 1

1Center for Joint Surgery, Southwest Hospital, Third Military Medical University, No. 29 Gaotanyan Street, Shapingba District, Chongqing 400038, P.R. China, 2Department of Orthopaedics, Kunming General Hospital, No. 212 Daguan Road, Xishan District, Kunming, Yunnan 650228, P.R. China

TABLE OF CONTENTS

1. Abstract
2. Introduction
3. Materials and methods
    3.1. Animals
    3.2. Col-II isolation and purification
    3.3. Sol-gel transition of Col-II
    3.4. BMDC isolation and cell-sol mixture preparation
    3.5. Chondral defect creation and gel implantation
    3.6. MRI and macroscopic, and histological evaluation
    3.7. Cell imaging
    3.8. Statistical analysis
4. Results
    4.1. MRI observation
    4.2. Macroscopic and microscopic observation
    4.3. Histological results
    4.4. Cell imaging
5. Discussion
6. Acknowledgements
7. References

1. ABSTRACT

One-step clinical therapies of articular cartilage defects remains a challenge. In this study, a strategy was proposed to utilize type II collagen (Col-II) gels with autologous bone marrow-derived cells (BMDCs) embedded to repair full-thickness chondral defects. Its feasibility and efficacy were further assessed in a minipig model. An 8-mm full-thickness chondral defect was created on the femoral trochlea of two knees in Guizhou minipigs. One knee received Col-II gels with BMDCs implantation versus the untreated one as control. After 1, 3, and 6 months operation, the animals were sacrificed, and the repair outcomes of chondral defects were evaluated using Magnetic Resonance Imaging (MRI), macro- and microscopic observation, and histological analysis. The treatment group showed significantly better repair outcomes of chondral defects than that in the control group at each time point (P < 0.01). Furthermore, the image showed that the repaired tissue in the treatment group was more similar to the surrounding healthy cartilage tissue. Based on the hyaline-like tissue regenerated ability, this one-step strategy provides a promising therapeutic potential for clinical application.

7. REFERENCES

1. A. R. Poole, T. Kojima, T. Yasuda, F. Mwale, M. Kobayashi and S. Laverty: Composition and structure of articular cartilage: a template for tissue repair. Clin Orthop Relat Res, 391(391 Suppl), S26 (2001)
DOI: 10.1097/00003086-200110001-00004

2. J. A. Buckwalter and H. J. Mankin: Articular cartilage: tissue design and chondrocyte-matrix interactions. Instr Course Lect, 47(4), 477 (1998)

3. A. Mor, M. Grijota, M. Nørgaard, J. Munthe, M. Lind, A. Déruaz and A. B. Pedersen: Trends in arthroscopy-documented cartilage injuries of the knee and repair procedures among 15–60-year-old patients. Scand J Med Sci Sports, 25(4), e400-e407 (2015)
DOI: 10.1111/sms.12330

4. W. Widuchowski, J. Widuchowski and T. Trzaska: Articular cartilage defects: study of 25,124 knee arthroscopies. Knee, 14(3), 177-182 (2007)
DOI: 10.1016/j.knee.2007.02.001

5. J. A. Buckwalter and H. J. Mankin: Articular cartilage: degeneration and osteoarthritis, repair, regeneration, and transplantation. Instr Course Lect, 47, 487 (1998)

6. T. Minas: Autologous chondrocyte implantation for focal chondral defects of the knee. Clin Orthop Relat Res, 391(391 Suppl), S349 (2001)
DOI: 10.1097/00003086-200110001-00032

7. T. Minas and S. Nehrer: Current concepts in the treatment of articular cartilage defects. Orthopedics, 20(6), 525 (1997)

8. J. D. Harris, R. A. Siston, X. Pan and D. C. Flanigan: Autologous chondrocyte implantation: a systematic review. J Bone Joint Surg Am, 92(12), 2220 (2010)
DOI: 10.2106/JBJS.J.00049

9. A. Lahav and M. Eizenberg: Treatment of deep cartilage defects in the knee with autologous chondrocyte transplantation. N Engl J Med, 331(14), 889-95 (1994)
DOI: 10.1056/NEJM199410063311401

10. L. Peterson, H. S. Vasiliadis, M. Brittberg and A. Lindahl: Autologous chondrocyte implantation: a long-term follow-up. Am J Sports Med, 38(6), 1117 (2010)
DOI: 10.1177/0363546509357915

11. A. E. Beris, M. G. Lykissas, I. Kostasagnantis and G. N. Manoudis: Treatment of full-thickness chondral defects of the knee with autologous chondrocyte implantation: a functional evaluation with long-term follow-up. Am J Sports Med, 40(3), 562-7 (2012)
DOI: 10.1177/0363546511428778

12. B. L. Clair, A. R. Johnson and T. Howard: Cartilage repair: current and emerging options in treatment. Foot Ankle Spec, 2(4), 179 (2009)
DOI: 10.1177/1938640009342272

13. A. Gigante, C. Bevilacqua, C. Zara, M. Travasi and C. Chillemi: Autologous chondrocyte implantation: cells phenotype and proliferation analysis. Knee Surg Sports Traumatol Arthrosc, 9(4), 254-258 (2001)
DOI: 10.1007/s001670100200

14. M. Bhattacharjee, J. Coburn, M. Centola, S. Murab, A. Barbero, D. L. Kaplan, I. Martin and S. Ghosh: Tissue engineering strategies to study cartilage development, degeneration and regeneration. Adv Drug Deliv Rev, 84, 107 (2015)
DOI: 10.1016/j.addr.2014.08.010

15. A. Gigante, S. Calcagno, S. Cecconi, D. Ramazzotti, S. Manzotti and D. Enea: Use of collagen scaffold and autologous bone marrow concentrate as a one-step cartilage repair in the knee: histological results of second-look biopsies at 1 year follow-up. Int J Immunopathol Pharmacol, 24(1 Suppl 2), 69 (2011)
DOI: 10.1177/03946320110241S213

16. A. Gobbi, G. Karnatzikos, C. Scotti, V. Mahajan, L. Mazzucco and B. Grigolo: One-Step Cartilage Repair with Bone Marrow Aspirate Concentrated Cells and Collagen Matrix in Full-Thickness Knee Cartilage Lesions: Results at 2-Year Follow-up. Cartilage, 2(3), 286 (2011)
DOI: 10.1177/1947603510392023

17. J. S. Pieper, V. D. K. Pm, T. Hafmans, J. Kamp, P. Buma, J. L. van Susante, V. D. B. Wb, J. H. Veerkamp and T. H. van Kuppevelt: Crosslinked type II collagen matrices: preparation, characterization, and potential for cartilage engineering. Biomaterials, 23(15), 3183-3192 (2002)
DOI: 10.1016/S0142-9612(02)00067-4

18. S. Li, C. Ye, H. Zou, Y. Peng and X. Yang: Preparation and analysis of pure type II collagen from porcine articular cartilage. Sheng Wu Yi Xue Gong Cheng Xue Za Zhi, 18(4), 592-594 (2001)

19. S. Marlovits, P. Singer, P. Zeller, I. Mandl, J. Haller and S. Trattnig: Magnetic resonance observation of cartilage repair tissue (MOCART) for the evaluation of autologous chondrocyte transplantation: Determination of interobserver variability and correlation to clinical outcome after 2 years. Eur J Radiol, 57(1), 16-23 (2006)
DOI: 10.1016/j.ejrad.2005.08.007

20. H. Nejadnik, T. D. Henning, T. Do, E. J. Sutton, F. Baehner, A. Horvai, B. Sennino, D. Mcdonald, R. Meier and B. Misselwitz: MR imaging features of gadofluorine-labeled matrix-associated stem cell implants in cartilage defects. Plos One, 7(12), e49971 (2012)
DOI: 10.1371/journal.pone.0049971

21. S. Quirbach, S. Trattnig, S. Marlovits, V. Zimmermann, S. Domayer, R. Dorotka, T. C. Mamisch, K. Bohndorf and G. H. Welsch: Initial results of in vivo high-resolution morphological and biochemical cartilage imaging of patients after matrix-associated autologous chondrocyte transplantation (MACT) of the ankle. Skeletal Radiol, 38(8), 751-760 (2009)
DOI: 10.1007/s00256-009-0682-1

22. M. D. B. Van, N. J. Raijmakers, J. Vanlauwe, J. Victor, S. J. De, J. Bellemans and D. B. Saris: International Cartilage Repair Society (ICRS) and Oswestry macroscopic cartilage evaluation scores validated for use in Autologous Chondrocyte Implantation (ACI) and microfracture. Osteoarthritis Cartilage, 15(12), 1397 (2007)
DOI: 10.1016/j.joca.2007.05.005

23. Y. Hagiwara, A. Ando, E. Chimoto, M. Tsuchiya, I. Takahashi, Y. Sasano, Y. Onoda, H. Suda and E. Itoi: Expression of collagen types I and II on articular cartilage in a rat knee contracture model. Connect Tissue Res, 51(1), 22-30 (2010)
DOI: 10.3109/03008200902859406

24. S. W. O’Driscoll, F. W. Keeley and R. B. Salter: Durability of regenerated articular cartilage produced by free autogenous periosteal grafts in major full-thickness defects in joint surfaces under the influence of continuous passive motion. A follow-up report at one year. J Bone Joint Surg Am, 70(4), 595 (1988)
DOI: 10.2106/00004623-198870040-00017

25. X. Pan, K. Xu, X. Qiu, W. Zhao, D. Wang and L. Yang: The Extract of Fructus Psoraleae Promotes Viability and Cartilaginous Formation of Rat Chondrocytes In vitro. Evid Based Complement Alternat Med, 2016, 2057631 (2016)
DOI: 10.1155/2016/2057631

26. E. A. Makris, A. H. Gomoll, K. N. Malizos, J. C. Hu and K. A. Athanasiou: Repair and tissue engineering techniques for articular cartilage. Nat Rev Rheumatol, 11(1), 21 (2015)
DOI: 10.1038/nrrheum.2014.157

27. G. Knutsen, J. O. Drogset, L. Engebretsen, T. Grøntvedt, T. C. Ludvigsen, S. Løken, E. Solheim, T. Strand and O. Johansen: A Randomized Multicenter Trial Comparing Autologous Chondrocyte Implantation with Microfracture: Long-Term Follow-up at 14 to 15 Years. J Bone Joint Surg Am, 98(16), 1332 (2016)
DOI: 10.2106/JBJS.15.01208

28. Z. Li, T. Zhu and W. Fan: Osteochondral autograft transplantation or autologous chondrocyte implantation for large cartilage defects of the knee: a meta-analysis. Cell Tissue Bank, 17(1), 1-9 (2016)
DOI: 10.1007/s10561-015-9515-8

29. C. R. Chu, M. Szczodry and S. Bruno: Animal Models for Cartilage Regeneration and Repair. Tissue Eng Part B Rev, 16(1), 105 (2010)
DOI: 10.1089/ten.teb.2009.0452

30. G. Schulzetanzil, R. D. Müller, B. Kohl, N. Schneider, W. Ertel, K. Ipaktchi, H. Hünigen, O. Gemeinhardt, R. Stark and T. John: Differing in vitro biology of equine, ovine, porcine and human articular chondrocytes derived from the knee joint: an immunomorphological study. Histochem Cell Biol, 131(2), 219-229 (2009)
DOI: 10.1007/s00418-008-0516-6

31. E. J. Caterson, L. J. Nesti, W. J. Li, K. G. Danielson, T. J. Albert, A. R. Vaccaro and R. S. Tuan: Three-dimensional cartilage formation by bone marrow-derived cells seeded in polylactide/alginate amalgam. J Biomed Mater Res, 57(3), 394 (2001)
DOI: 10.1002/1097-4636(20011205)57:3 <394::AID-JBM1182>3.0.CO;2-9

32. H. Park, J. S. Temenoff, Y. Tabata, A. I. Caplan and A. G. Mikos: Injectable biodegradable hydrogel composites for rabbit marrow mesenchymal stem cell and growth factor delivery for cartilage tissue engineering. Biomaterials, 28(21), 3217-3227 (2007)
DOI: 10.1016/j.biomaterials.2007.03.030

33. H. Fan, Y. Hu, C. Zhang, X. Li, R. Lv, L. Qin and R. Zhu: Cartilage regeneration using mesenchymal stem cells and a PLGA-gelatin/chondroitin/hyaluronate hybrid scaffold. Biomaterials, 27(26), 4573 (2006)
DOI: 10.1016/j.biomaterials.2006.04.013

34. S. Wakitani, K. Imoto, T. Yamamoto, M. Saito, N. Murata and M. Yoneda: Human autologous culture expanded bone marrow mesenchymal cell transplantation for repair of cartilage defects in osteoarthritic knees. Osteoarthritis Cartilage, 10(3), 199 (2002)
DOI: 10.1053/joca.2001.0504

35. A. Gobbi, G. Karnatzikos and S. R. Sankineani: One-step surgery with multipotent stem cells for the treatment of large full-thickness chondral defects of the knee. Am J Sports Med, 42(3), 648 (2014)
DOI: 10.1177/0363546513518007

36. A. I. Caplan: Review: mesenchymal stem cells: cell-based reconstructive therapy in orthopedics. Tissue Eng, 11(7-8), 1198 (2005)
DOI: 10.1089/ten.2005.11.1198

37. K. Gopal, H. A. Amirhamed and T. Kamarul: Advances of human bone marrow-derived mesenchymal stem cells in the treatment of cartilage defects: a systematic review. Exp Biol Med, 239(6), 663-669 (2014)
DOI: 10.1177/1535370214530364

38. H. Nejadnik, J. H. Hui, C. E. Feng, B. C. Tai and E. H. Lee: Autologous bone marrow-derived mesenchymal stem cells versus autologous chondrocyte implantation: an observational cohort study. Am J Sports Med, 38(6), 1110-1116 (2010)
DOI: 10.1177/0363546509359067

39. M. Jung, B. Kaszap, A. Redöhl, E. Steck, S. Breusch, W. Richter and T. Gotterbarm: Enhanced early tissue regeneration after matrix-assisted autologous mesenchymal stem cell transplantation in full thickness chondral defects in a minipig model. Cell Transplant, 18(8), 923-32 (2009)
DOI: 10.3727/096368909X471297

40. C. H. Chang, T. F. Kuo, F. H. Lin, J. H. Wang, Y. M. Hsu, H. T. Huang, S. T. Loo, H. W. Fang, H. C. Liu and W. C. Wang: Tissue engineering-based cartilage repair with mesenchymal stem cells in a porcine model. J Orthop Res, 29(12), 1874-80 (2011)
DOI: 10.1002/jor.21461

41. K. B. L. Lee, J. H. P. H. M.B.B.S, I. C. Song, L. Ardany and E. H. Lee: Injectable Mesenchymal Stem Cell Therapy for Large Cartilage Defects—A Porcine Model. Stem Cells, 25(11), 2964 (2007)
DOI: 10.1634/stemcells.2006-0311

42. K. Y. Saw, P. Hussin, S. C. Loke, M. Azam, H. C. Chen, Y. G. Tay, S. Low, K. L. Wallin and K. Ragavanaidu: Articular cartilage regeneration with autologous marrow aspirate and hyaluronic Acid: an experimental study in a goat model. Arthroscopy, 25(12), 1391 (2009)
DOI: 10.1016/j.arthro.2009.07.011

43. D. E. Trentham, A. S. Townes and A. H. Kang: Autoimmunity to type II collagen an experimental model of arthritis. J Exp Med, 146(3), 857 (1977)
DOI: 10.1084/jem.146.3.857

44. D. Bosnakovski, M. Mizuno, G. Kim, S. Takagi, M. Okumura and T. Fujinaga: Chondrogenic differentiation of bovine bone marrow mesenchymal stem cells (MSCs) in different hydrogels: influence of collagen type II extracellular matrix on MSC chondrogenesis. Biotechnol Bioeng, 93(6), 1152 (2006)
DOI: 10.1002/bit.20828

45. P. Buma, J. S. Pieper, T. T. Van, J. L. van Susante, V. D. K. Pm, J. H. Veerkamp, V. D. B. Wb, R. P. Veth and T. H. van Kuppevelt: Cross-linked type I and type II collagenous matrices for the repair of full-thickness articular cartilage defects--a study in rabbits. Biomaterials, 24(19), 3255-3263 (2003)
DOI: 10.1016/S0142-9612(03)00143-1

46. C. R. Lee, A. J. Grodzinsky, H. P. Hsu and M. Spector: Effects of a cultured autologous chondrocyte-seeded type II collagen scaffold on the healing of a chondral defect in a canine model. J Orthop Res, 21(2), 272-281 (2003)
DOI: 10.1016/S0736-0266(02)00153-5

47. A. Funayama, Y. Niki, H. Matsumoto, S. Maeno, T. Yatabe, H. Morioka, S. Yanagimoto, T. Taguchi, J. Tanaka and Y. Toyama: Repair of full-thickness articular cartilage defects using injectable type II collagen gel embedded with cultured chondrocytes in a rabbit model. J Orthop Sci, 13(3), 225 (2008)
DOI: 10.1007/s00776-008-1220-z

48. J. H. Guettler, C. K. Demetropoulos, K. H. Yang and K. A. Jurist: Osteochondral defects in the human knee: influence of defect size on cartilage rim stress and load redistribution to surrounding cartilage. Am J Sports Med, 32(6), 1451 (2004)
DOI: 10.1177/0363546504263234

Abbreviations: MSCs: senchymal stem cells; Col-II: type II collagen; MRI: Magnetic Resonance Imaging; ACI: autologous chondrocyte implantation; MOCART: Magnetic resonance Observation of CArtilage Repair Tissue; ICRS: International Cartilage Repair Society; EDTA: Ethylene diamine tetraacetic acid; FISH: Fluorescence in situ hybridization; IHC: Immunohistochemistry

Key Words: Articular Cartilage Defect, BMDCs, Type II Collagen Gel, Regeneration, Repair

Send correspondence to: Fuyou Wang, Center for Joint Surgery, Southwest Hospital, Third Military Medical University, No. 29 Gaotanyan Street, Shapingba District, Chongqing 400038, P.R. China, Tel: 86-68765290, Fax: 86-68765293, E-mail: wfy731023@163.com