Temporal and Spatial Expression of TGF-b1 in the Early Phase of Patellar Tendon Healing after Application of Platelet Rich Plasma

Document Type: RESEARCH PAPER

Authors

1 Democritus University of Thrace, Orthopaedic Clinic, Dragana, Alexandroupolis, Greece

2 Kapodistrian University of Athens, Department of Pathology, Dragana, Alexandroupolis, Greece

3 Democritus University of Thrace, Laboratory of Experimental Surgery and Surgical Research, Dragana, Alexandroupolis, Greece

Abstract

 
 Background:The aim of this study is to find out the spatial and temporal expression of TGF-b1 during the tendon healing, after application of Platelet Rich Plasma (PRP).
Methods: A patellar tendon defect model in rabbits was used for this purpose. 48 skeletally mature New Zealand White rabbits, weighing 3.5 kg, were used for this study. Equal numbers of animals from both groups were sacrificed at 4 different time points (1st, 2nd, 3rd, and 4th week). A full thickness patellar tendon substance in the right limb of each animal was excised from its central portion during the operation. PRP with a gel form was applied and filled the tendon defect in PRP group. No PRP was applied in the tendon defect of controls. Histological sections with hematoxylin-eosin and immunohistochemical sections with an anti-TGF-b1 primary antibody were made for the evaluation of the results.
Results: A differentiation of the healing process was observed in the PRP group in comparison with the control group. TGF-b1 expression was detected in various cell populations (inflammatory cells, endothelial cells, macrophages, and tenocytes). Both cytoplasmic and nuclear expressions were present. The larger amounts of immunoexpression were localized in epitenon and in the repair site. PRP group showed stronger and more extensive staining at 1st and 2nd week (P<0.0001), whereas control group showed more extensive staining at the 3rd and 4th week (P<0.0001).
Conclusions: Our study demonstrates that locally application of PRP result in an alteration of TGF-b1 expression during the healing of a patellar tendon defect.

Keywords


  1. Marx RE. Platelet-rich plasma (PRP): what is PRP and what is not PRP? Implant Dent. 2001; 10(4):225–8.
  2. Kajikawa Y, Morihara T, Sakamoto H, Matsuda K, Oshima Y, Yoshida A, et al. Platelet-rich plasma enhances the initial mobilization of circulation-derived cells for tendon healing. J Cell Physiol. 2008; 215(3):837-45.
  3. Aspenberg P, Virchenko O. Platelet concentrate injection improves Achilles tendon repair in rats. Acta Orthop Scand. 2004; 75(1):93–9.
  4. Murray MM, Spindler KP, Devin C, Snyder BS, Muller J, Takahashi M, et al. Use of a collagen-platelet rich plasma scaffold to stimulate healing of a central defect in the canine ACL. J Orthop Res. 2006; 24(4):820–30.
  5. Murray MM, Spindler KP, Abreu E, Muller JA, Nedder A, Kelly M, et al. Collagen-platelet rich plasma hydrogel enhances primary repair of the porcine anterior cruciate ligament. J Orthop Res. 2007; 25(1):81–91.
    1. Lyras D, Kazakos K, Verettas D, Polychronidis A, Simopoulos C, Botaitis S, et al. Immunohistochemical study of angiogenesis after local administration of platelet-rich plasma in a patellar tendon defect. Int Orthop. 2010; 34(1):143-8.
    2. Lyras DN, Kazakos K, Verettas D, Polychronidis A, Tryfonidis M, Botaitis S, et al. The influence of platelet-rich plasma on angiogenesis during the early phase of tendon healing. Foot Ankle Int. 2009; 30(11):1101-6.
    3. Lyras DN, Kazakos K, Agrogiannis G, Verettas D, Kokka A, Kiziridis G, et al. Experimental study of tendon healing early phase: Is IGF-1 expression influenced by platelet rich plasma gel? Orthop Traumatol Surg Res. 2010; 96(4):381-7.
    4. Lyras DN, Kazakos K, Verettas D, Botaitis S, Agrogiannis G, Kokka A, et al. The effect of platelet-rich plasma gel in the early phase of patellar tendon healing. Arch Orthop Trauma Surg. 2009; 129(11):1577-82.
    5. Anaguchi Y, Yasuda K, Majima T, Tohyama H, Minami A, Hayasi K, et al. The effect of transforming growth factor-beta on mechanical properties of the fibrous tissue regenerated in the patellar tendon after resecting the central portion. Clin Biomech (Bristol, Avon). 2005; 20(9):959-65.
    6. Zafirellis K, Agrogiannis G, Zachaki A, Gravani K, Karameris A, Kombouras C. Prognostic significance of VEGF expression evaluated by quantitative immunohistochemical analysis in colorectal cancer. J Surg Res. 2008; 147(1):99-107.
    7. 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. 1997; 15(6):837-43.
    8. Chan KM, Fu SC, Wong YP, Hui WC, Cheuk YC, Wong MW. Expression of transforming growth factor beta isoforms and their roles in tendon healing. Wound Repair Regen. 2008; 16(3):399–407.
    9. Yamazaki S, Yasuda K, Tomita F, Tohyama H, Minami A. The effect of transforming growth factor-beta1 on intraosseous healing of flexor tendon autograft replacement of anterior cruciate ligament in dogs. Arthroscopy. 2005; 21(9):1034-41.
    10. Anitua E, Sanchez M, Nurden AT, Zalduendo M, de la Fuente M, Orive G, et al. Autologous fibrin matrices: a potential source of biological mediators that modulate tendon cell activities. J Biomed Mater Res A. 2006; 77(2):285-93.
    11. Wakefield LM, Smith DM, Flanders KC, Sporn MB. Latent transforming growth factor-beta from human platelets. A high molecular weight complex containing precursor sequences. J Biol Chem. 1988; 263(16):7646-54.
    12. Assoian RK, Fleurdelys BE, Stevenson HC, Miller PJ, Madtes DK, Raines EW, et al. Expression and secretion of type beta transforming growth factor by activated human macrophages. Proc Natl Acad Sci U S A. 1987; 84(17):6020-4.
    13. 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(1):148-55.
    14. Klein MB, Yalamanchi N, Pham H, Longaker MT, Chang J. Flexor tendon healing in vitro: effects of TGF-beta on tendon cell collagen production. J Hand Surg Am. 2002; 27(4):615-20.