Abstract
Background
Extracapsular ligament injuries of the knee and ankle are common injuries. Ligaments heal slowly, usually over months or longer by scar formation rather than by tissue regeneration. This study was performed to evaluate the therapeutic effect of locally delivered recombinant hepatocyte growth factor (HGF) on the early healing of ligaments in a rabbit model.
Methods
Japanese white rabbits were subjected to a standardized gap injury in the medial collateral ligaments (MCLs) of both knees. Each rabbit underwent bilateral transection of the midsubstance of the MCL, which was not repaired. During postoperative days 0–6, the rabbits were injected with 10 μg human recombinant HGF into the right MCL, while the left MCL was injected with saline alone. One, 3, 6, and 12 weeks after surgery, experimental rabbits were sacrificed. The structural properties of the femur-MCL-tibia complex were then assessed and the tissue was subjected to histological evaluation. To see the distribution of cells that express c-Met receptor, the tissue was subjected to immunohistochemistry.
Results
Immunohistochemical evaluation revealed that c-Met expression was observed particularly at opposing ligament ends in the HGF-treated limbs 1 week after surgery. Histological evaluation revealed earlier neovascularization and more aligned collagen fibers in the MCLs of the HGF-treated group than the control group. In mechanical evaluations, similar ligament failure modes were noted in the two groups. After 3 weeks, HGF-treated limbs had significantly improved structural properties than the paired control limbs.
Conclusions
Our findings indicate local administration of recombinant HGF promotes early steps in ligament healing and the repair of structural properties in a rabbit model. Local administration of HGF may represent a new therapeutic approach to accelerating healing and rehabilitation after ligament injury.
Similar content being viewed by others
References
Miyamoto RG, Bosco JA, Sherman OH. Treatment of medial collateral ligament injuries. J Am Acad Orthop Surg. 2009;17:152–61.
Frank C, Schachar N, Dittrich D. Natural history of healing in the repaired medial collateral ligament. J Orthop Res. 1983;1:179–88.
Niyibizi C, Kavalkovich K, Yamaji T, Woo SL-Y. Type V collagen is increased during rabbit medial collateral ligament healing. Knee Surg Sports Traumatol Arthrosc. 2000;8:281–5.
Woo SL-Y, Abramowitch SD, Kilger R, Liang R. Biomechanics of knee ligaments: injury, healing, and repair. J Biomech. 2006;39:1–20.
Takeyama N, Sakai H, Ohtake H, Mashitori H, Tamai K, Saotome K. Effects of hyperbaric oxygen on gene expressions of procollagen, matrix metalloproteinase and tissue inhibitor of metalloproteinase in injured medial collateral ligament and anterior cruciate ligament. Knee Surg Sports Traumatol Arthrosc. 2007;15:443–52.
Delbari A, Bayat M, Bayat M. Effect of low-level laser therapy on healing of medial collateral ligament injuries in rats: an ultrastructural study. Photomed Laser Surg. 2007;25:191–6.
Nakamura T, Nawa K, Ichihara A. Partial purification and characterization of hepatocyte growth factor from serum of hepatectomized rats. Biochem Biophys Res Commun. 1984;122:1450–9.
Nakamura T, Nishizawa T, Hagiya M. Molecular cloning and expression of human hepatocyte growth factor. Nature. 1989;342:440–3.
Matsubara H, Tsuchiya H, Watanabe K, Takeuchi A, Tomita K. Percutaneous nonviral delivery of hepatocyte growth factor in an osteotomy gap promotes bone repair in rabbits: a preliminary study. Clin Orthop Relat Res. 2008;466:2962–72.
Nakase J, Kitaoka K, Matsumoto K, Tomita K. Facilitated tendon-bone healing by local delivery of recombinant hepatocyte growth factor in rabbits. Arthroscopy. 2010;26:84–90.
Ono I, Yamashita T, Hida T, Jin HY, Ito Y, Hamada H, Akasaka Y, Ishii T, Jimbow K. Local administration of hepatocyte growth factor gene enhances the regeneration of dermis in acute incisional wounds. J Surg Res. 2004;120:47–55.
Jiang D, Jiang Z, Han F, Zhang Y, Li Z. HGF suppresses the production of collagen type III and α-SMA induced by TGF-β1 in healing fibroblasts. Eur J Appl Physiol. 2008;103:489–93.
Matsumoto K, Nakamura T. Hepatocyte growth factor: renotropic role and potential therapeutics for renal diseases. Kidney Int. 2001;59:2023–38.
Yoshida S, Matsumoto K, Tomioka D, Bessho K, Itami S, Yoshikawa K, Nakamura T. Recombinant hepatocyte growth factor accelerates cutaneous wound healing in a diabetic mouse model. Growth Factors. 2004;22:111–9.
Woo SL-Y, Orlando CA, Camp JF, Akeson WH. Effects of postmortem storage by freezing on ligament tensile behavior. J Biomech. 1986;19:399–404.
Shimode K, Iwasaki N, Majima T, Funakoshi T, Sawaguchi N, Onodera T, Sawaguchi N, Onodera T, Minami A. Local upregulation of stromal cell-derived factor-1 after ligament injuries enhances homing rate of bone marrow stromal cells in rats. Tissue Eng Part A. 2009;15:2277–84.
Ishizawa K, Kubo H, Yamada M, Kobayashi S, Suzuki T, Mizuno S, Nakamura T, Sasaki H. Hepatocyte growth factor induces angiogenesis in injured lungs through mobilizing endothelial progenitor cells. Biochem Biophys Res Commun. 2004;324:276–80.
Bussolino F, DiRenzo MF, Ziche M, Bocchietto E, Olivero M, Naidini L, Gaudino G, Tamagnone L, Coffer A, Comoglio PM. Hepatocyte growth factor is a potent angiogenic factor which stimulates endothelial cell motility and growth. J Cell Biol. 1992;119:629–41.
Van Belle E, Witzenbichler B, Chen D, Silver M, Chang L, Schwall R, Jeffrey M. Potential angiogenic effect of scatter factor/hepatocyte growth factor via induction of vascular endothelial growth factor: the case for paracrine amplification of angiogenesis. Circulation. 1998;97:381–90.
Morishita R, Nakamura S, Hayashi S, Taniyama Y, Moriguchi A, Nagano T, Taiji M, Noguchi H, Takeshita S, Matsumoto K, Nakamura T, Higaki J, Ogihara T. Therapeutic angiogenesis induced by human recombinant hepatocyte growth factor in rabbit hind limb ischemia model as cytokine supplement therapy. Hypertension. 1999;33:1379–84.
Molloy T, Wang Y, Murrell G. The roles of growth factors in tendon and ligament healing. Sports Med. 2003;33:381–94.
Bray RC, Leonard CA, Salo PT. Correlation of healing capacity with vascular response in the anterior cruciate and medial collateral ligaments of the rabbit. J Orthop Res. 2003;21:1118–23.
Mammoto T, Seerattan RA, Paulson KD, Leonard CA, Bary RC, Salo PT. Nerve growth factor improves ligament healing. J Orthop Res. 2008;26:957–64.
Spindler KP, Murray MM, Detwiler KB, Tarter JT, Dawson JM, Nanney LB, Davidson JM. The biomechanical response to dose of TGF-β2 in the healing rabbit medial collateral ligament. J Orthop Res. 2003;21:245–9.
Virchenko O, Aspenberg P. How can one platelet injection after tendon injury lead to a stronger tendon after 4 weeks? Interplay between early regeneration and mechanical stimulation. Acta Orthop. 2006;77:806–12.
Hannafin JA, Attia ET, Warren RF, Bhargava MM. Characterization of chemotactic migration and growth kinetics of canine knee ligament fibroblasts. J Orthop Res. 1999;17:398–404.
Acknowledgments
The authors thank Ms. Yoko Kasai for her skillful technical assistance.
Conflict of interest
Dr. Matsumoto is a co-founder and shareholder of Kringle Pharma (Toyonaka, Japan). He has served as a Chief Scientific Officer of Kringle Pharma and received payment for this role. The other authors did not receive and will not receive any benefits and funding from any commercial party related directly or indirectly to the subject of this article.
Author information
Authors and Affiliations
Corresponding author
About this article
Cite this article
Ueshima, K., Kitaoka, K., Nakase, J. et al. Promotion of rabbit ligament healing by local delivery of hepatocyte growth factor. J Orthop Sci 16, 451–457 (2011). https://doi.org/10.1007/s00776-011-0101-z
Received:
Accepted:
Published:
Issue Date:
DOI: https://doi.org/10.1007/s00776-011-0101-z