Abstract
Adhesion formation after digital flexor tendon injury greatly affects gliding function of the tendon, which is a major clinical complication after hand surgery. Transforming growth factor beta 1 (TGF-β1) has a critical role in adhesion formation during tendon healing. Persistent regulation of TGF-β1 through application of microRNA (miRNA) specifically inhibiting the function of TGF-β1 (TGF-β1-miRNA) holds promise for treatment of such a complication. Adeno-associated virus (AAV) was used to transfer TGF-β1-miRNA to the chicken digital flexor tendons, which had been injured and surgically repaired. Four doses of AAV2-TGF-β1-miRNA (2 × 1011, 2 × 1010, 2 × 109 and 2 × 108 vector genomes (vg)) were used to determine the transfection efficiency. At postoperative 3 weeks, we found a positive correlation between the administered AAV2-TGF-β1-miRNA doses and transfection efficiency. The transfection rate ranged from 10% to 77% as the doses increased. Production of TGF-β1 protein in the tendons decreased on increasing vector dosage. When 2 × 1011 and 2 × 1010 vg were injected into the tendon, gliding excursion of the repaired tendon and work of flexion of chicken toes were significantly increased and adhesion score decreased 6 and 8 weeks later, indicating the improvement of tendon gliding and decreases in adhesion formations. However, the ultimate strength of the tendons transfected at the dose of 2 × 1010 vg was 12–24% lower than that of the control tendons. The results of this study demonstrate that application of TGF-β1-miRNA had a mixed impact on tendon healing: adhesion around the tendon is reduced but strength of the tendon healing is adversely affected. Future studies should aim at maintaining the beneficial effects of reducing tendon adhesions, while eliminating the adverse effects of decreasing the healing strength.
This is a preview of subscription content, access via your institution
Access options
Subscribe to this journal
Receive 12 print issues and online access
$259.00 per year
only $21.58 per issue
Buy this article
- Purchase on Springer Link
- Instant access to full article PDF
Prices may be subject to local taxes which are calculated during checkout
Similar content being viewed by others
References
Tang JB . Clinical outcomes associated with flexor tendon repair. Hand Clin 2005; 21: 199–210.
Wu YF, Tang JB . Recent developments in flexor tendon repair techniques and factors influencing strength of the tendon repair. J Hand Surg Eur Vol 2014; 39: 6–19.
Savage R . The search for the ideal tendon repair in zone 2: strand number, anchor points and suture thickness. J Hand Surg Eur Vol 2014; 39: 20–29.
Tang JB, Chang J, Elliot D, Lalonde DH, Sandow M, Vögelin E . IFSSH Flexor Tendon Committee report 2014: from the IFSSH Flexor Tendon Committee (Chairman: Jin Bo Tang). J Hand Surg Eur Vol 2014; 39: 107–115.
Bates SJ, Morrow E, Zhang AY, Pham H, Longaker MT, Chang J . Mannose-6-phosphate, an inhibitor of transforming growth factor-beta, improves range of motion after flexor tendon repair. J Bone Joint Surg Am 2006; 88: 2465–2472.
Tan V, Nourbakhsh A, Capo J, Cottrell JA, Meyenhofer M, O’Connor JP . Effects of nonsteroidal anti-inflammatory drugs on flexor tendon adhesion. J Hand Surg Am 2010; 35: 941–947.
Hung LK, Fu SC, Lee YW, Mok TY, Chan KM . Local vitamin-C injection reduced tendon adhesion in a chicken model of flexor digitorum profundus tendon injury. J Bone Joint Surg Am 2013; 95: e41.
Golash A, Kay A, Warner JG, Peck F, Watson JS, Lees VC . Efficacy of ADCON-T/N after primary flexor tendon repair in Zone II: a controlled clinical trial. J Hand Surg Br 2003; 28: 113–115.
Bhavsar D, Shettko D, Tenenhaus M . Encircling the tendon repair site with collagen-GAG reduces the formation of postoperative tendon adhesions in a chicken flexor tendon model. J Surg Res 2010; 159: 765–771.
Taguchi M, Zhao C, Sun YL, Jay GD, An KN, Amadio PC . The effect of surface treatment using hyaluronic acid and lubricin on the gliding resistance of human extrasynovial tendons in vitro. J Hand Surg Am 2009; 34: 1276–1281.
Håkansson J, Mahlapuu M, Ekström L, Olmarker K, Wiig M . Effect of lactoferrin peptide (PXL01) on rabbit digit mobility after flexor tendon repair. J Hand Surg Am 2012; 37: 2519–2525.
Lees VC, Warwick D, Gillespie P, Brown A, Akhavani M, Dewer D et al. A multicentre, randomized, double-blind trial of the safety and efficacy of mannose-6-phosphate in patients having Zone II flexor tendon repairs. J Hand Surg Eur Vol 2014; 40: 682–694.
Tsubone T, Moran SL, Amadio PC, Zhao C, An KN . Expression of growth factors in canine flexor tendon after laceration in vivo. Ann Plast Surg 2004; 53: 393–397.
Berglund M, Reno C, Hart DA, Wiig M . Patterns of mRNA expression for matrix molecules and growth factors in flexor tendon injury: differences in the regulation between tendon and tendon sheath. J Hand Surg Am 2006; 31: 1279–1287.
Chang J, Thunder R, Most D, Longaker MT, Lineaweaver WC . Studies in flexor tendon wound healing: neutralizing antibody to TGF-beta1 increases postoperative range of motion. Plast Reconstr Surg 2000; 105: 148–155.
Xia C, Yang X, Wang YZ, Sun K, Ji L, Tian S . Tendon healing in vivo and in vitro: neutralizing antibody to TGF-β improves range of motion after flexor tendon repair. Orthopedics 2010; 33: 809.
Chen CH, Zhou YL, Wu YF, Cao Y, Gao JS, Tang JB . Effectiveness of microRNA in down-regulation of TGF-beta gene expression in digital flexor tendons of chickens: in vitro and in vivo study. J Hand Surg Am 2009; 34: 1777–1784.
Wang XT, Liu PY, Tang JB, Mizukami H, Xin KQ, Ozawa K et al. Tendon healing in vitro: adeno-associated virus-2 effectively transduces intrasynovial tenocytes with persistent expression of the transgene, but other serotypes do not. Plast Reconstr Surg 2007; 119: 227–234.
Mason MR, Ehlert EM, Eggers R, Pool CW, Hermening S, Huseinovic A et al. Comparison of AAV serotypes for gene delivery to dorsal root ganglion neurons. Mol Ther 2010; 18: 715–724.
Markakis EA, Vives KP, Bober J, Leichtle S, Leranth C, Beecham J et al. Comparative transduction efficiency of AAV vector serotypes 1-6 in the substantia nigra and striatum of the primate brain. Mol Ther 2010; 18: 588–593.
Hasslund S, Dadali T, Ulrich-Vinther M, Søballe K, Schwarz EM, Awad HA . Freeze-dried allograft-mediated gene or protein delivery of growth and differentiation factor 5 reduces reconstructed murine flexor tendon adhesions. J Tissue Eng 2014; 19: 5.
Basile P, Dadali T, Jacobson J, Hasslund S, Ulrich-Vinther M, Søballe K et al. Freeze-dried tendon allografts as tissue-engineering scaffolds for Gdf5 gene delivery. Mol Ther 2008; 16: 466–473.
Wang XT, Liu PY, Xin KQ, Tang JB . Tendon healing in vitro: bFGF gene transfer to tenocytes by adeno-associated viral vectors promotes expression of collagen genes. J Hand Surg Am 2005; 30: 1255–1261.
Zhu B, Cao Y, Xin KQ, Wang XT, Summerhayes IC, Liu PY et al. Tissue reactions of adenoviral, adeno-associated viral, and liposome-plasmid vectors in tendons and comparison with early-stage healing responses of injured flexor tendons. J Hand Surg Am 2006; 31: 1652–1660.
Kaufman HL, Kim DW, Kim-Schulze S, DeRaffele G, Jagoda MC, Broucek JR . Results of a randomized phase I gene therapy clinical trial of nononcolytic fowlpox viruses encoding T cell costimulatory molecules. Hum Gene Ther 2014; 25: 452–460.
MacLaren RE, Groppe M, Barnard AR, Cottriall CL, Tolmachova T, Seymour L et al. Retinal gene therapy in patients with choroideremia: initial findings from a phase 1/2 clinical trial. Lancet 2014; 383: 1129–1137.
Foust KD, Nurre E, Montgomery CL, Hernandez A, Chan CM, Kaspar BK . Intravascular AAV9 preferentially targets neonatal neurons and adult astrocytes. Nat Biotechnol 2009; 27: 59–65.
Wang D, Zhong L, Nahid MA, Gao G . The potential of adeno-associated viral vectors for gene delivery to muscle tissue. Expert Opin Drug Deliv 2014; 11: 345–364.
Ferreira V, Twisk J, Kwikkers K, Aronica E, Brisson D, Methot J et al. Immune responses to intramuscular administration of alipogene tiparvovec (AAV1-LPL(S447X)) in a phase II clinical trial of lipoprotein lipase deficiency gene therapy. Hum Gene Ther 2014; 25: 180–188.
Smith BK, Collins SW, Conlon TJ, Mah CS, Lawson LA, Martin AD et al. Phase I/II trial of adeno-associated virus-mediated alpha-glucosidase gene therapy to the diaphragm for chronic respiratory failure in Pompe disease: initial safety and ventilatory outcomes. Hum Gene Ther 2013; 24: 630–640.
Tang JB, Cao Y, Zhu B, Xin KQ, Wang XT, Liu PY . Adeno-associated virus-2-mediated bFGF gene transfer to digital flexor tendons significantly increases healing strength. an in vivo study. J Bone Joint Surg Am 2008; 90: 1078–1089.
Wong JK, Lui YH, Kapacee Z, Kadler KE, Ferguson MW, McGrouther DA . The cellular biology of flexor tendon adhesion formation: an old problem in a new paradigm. Am J Pathol 2009; 175: 1938–1951.
Wu YF, Tang JB . Apoptosis in adhesions and the adhesion-tendon gliding interface: relationship to adhesion-tendon gliding mechanics. J Hand Surg Am 2013; 38: 1071–1078.
Margadant C, Sonnenberg A . Integrin-TGF-beta crosstalk in fibrosis, cancer and wound healing. EMBO Rep 2010; 11: 97–105.
Wu YF, Tang JB . Tendon healing, edema, and resistance to flexor tendon gliding: clinical implications. Hand Clin 2013; 29: 167–178.
Chen CH, Cao Y, Wu YF, Bais AJ, Gao JS, Tang JB . Tendon healing in vivo: gene expression and production of multiple growth factors in early tendon healing period. J Hand Surg Am 2008; 33: 1834–1842.
Katzel EB, Wolenski M, Loiselle AE, Basile P, Flick LM, Langstein HN et al. Impact of Smad3 loss of function on scarring and adhesion formation during tendon healing. J Orthop Res 2011; 29: 684–693.
Zhang AY, Pham H, Ho F, Teng K, Longaker MT, Chang J . Inhibition of TGF-beta-induced collagen production in rabbit flexor tendons. J Hand Surg Am 2004; 29: 230–235.
Branford OA, Klass BR, Grobbelaar AO, Rolfe KJ . The growth factors involved in flexor tendon repair and adhesion formation. J Hand Surg Eur Vol 2014; 39: 60–70.
Alam N, McGrouther DA, Wong JK . The cellular biology of tendon grafting. J Hand Surg Eur Vol 2014; 39: 79–92.
Wiig ME, Dahlin LB, Fridén J, Hagberg L, Larsen SE, Wiklund K et al. PXL01 in sodium hyaluronate for improvement of hand recovery after flexor tendon repair surgery: randomized controlled trial. PLoS One 2014; 9: e110735.
Wu YF, Zhou YL, Tang JB . Relative contribution of tissue oedema and the presence of an A2 pulley to resistance to flexor tendon movement: an in vitro and in vivo study. J Hand Surg Eur Vol 2012; 37: 310–315.
Cao Y, Tang JB . Strength of tendon repair decreases in the presence of an intact A2 pulley: biomechanical study in a chicken model. J Hand Surg Am 2009; 34: 1763–1770.
Acknowledgements
This work was supported by grants from the Natural Science Foundation of China (Nos. 81271985, 81030035), the Priority of Academic Program Development of Jiangsu Higher Education Institutions.
Author information
Authors and Affiliations
Corresponding author
Ethics declarations
Competing interests
The authors declare no conflict of interest.
Rights and permissions
About this article
Cite this article
Wu, Y., Mao, W., Zhou, Y. et al. Adeno-associated virus-2-mediated TGF-β1 microRNA transfection inhibits adhesion formation after digital flexor tendon injury. Gene Ther 23, 167–175 (2016). https://doi.org/10.1038/gt.2015.97
Received:
Revised:
Accepted:
Published:
Issue Date:
DOI: https://doi.org/10.1038/gt.2015.97
This article is cited by
-
Battling adhesions: from understanding to prevention
BMC Biomedical Engineering (2019)
-
Modulation of digital flexor tendon healing by vascular endothelial growth factor gene transfection in a chicken model
Gene Therapy (2017)