Abstract
To understand the role of tendon fibroblast contraction in tendon healing, we investigated the contraction of human patellar tendon fibroblasts (HPTFs) and its regulation by transforming growth factor-β1 (TGF-β1), TGF-β3, and prostaglandin E2 (PGE2). HPTFs were found to wrinkle the underlying thin silicone membranes, demonstrating that these tendon fibroblasts are contractile. Using fibroblast populated collagen gels (FPCGs), exogenous addition of TGF-β1 or TGF-β3 was found to increase fibroblast contraction compared to non-treated fibroblasts in serum-free medium, whereas PGE2 was found to decrease the tendon fibroblast contraction. Moreover, the tendon fibroblasts in collagen gels treated with TGF-β1 contracted to a greater degree than those treated with TGF-β3. Since the extent of fibroblast contraction is related to scar tissue formation, this differential effect of TGF-β1 and TGF-β3 on HPTF contraction supports the previous finding that TGF-β1 induces scar tissue formation, whereas TGF-β3 reduces its formation. Further, the reduced tendon fibroblast contraction by PGE2 suggests that excessive presence of this inflammatory mediator in the wound site might retard tendon healing. Taken together, the results of this study suggest that regulation of human tendon fibroblast contraction may reduce scar tissue formation and therefore improve the mechanical properties of healing tendons.
Similar content being viewed by others
References
Adzick NS, Lorenz HP (1994) Cells, matrix, growth factors, and the surgeon. The biology of scarless fetal wound repair. Ann Surg 220:10–18
Almekinders LC, Baynes AJ, Bracey LW (1995) An in vitro investigation into the effects of repetitive motion and nonsteroidal antiinflammatory medication on human tendon fibroblasts. Am J Sports Med 23:119–123
Bitar KN (2003) Function of gastrointestinal smooth muscle: from signaling to contractile proteins. Am J Med 115:15–23
Brown RA, Sethi KK, Gwanmesia I, Raemdonck D, Eastwood M, Mudera V (2002) Enhanced fibroblast contraction of 3D collagen lattices and integrin expression by TGF-beta1 and -beta3: mechanoregulatory growth factors? Exp Cell Res 274:310–322
Campbell BH, Clark WW, Wang JH (2003) A multi-station culture force monitor system to study cellular contractility. J Biomech 36:137–140
Carlstedt CA, Madsen K, Wredmark T (1986) Biomechanical and biochemical studies of tendon healing after conservative and surgical treatment. Arch Orthop Trauma Surg 105:211–215
Chrzanowska-Wodnicka M, Burridge K (1996) Rho-stimulated contractility drives the formation of stress fibers and focal adhesions. J Cell Biol 133:1403–1415
Coleman C, Tuan TL, Buckley S, Anderson KD, Warburton D (1998) Contractility, transforming growth factor-beta, and plasmin in fetal skin fibroblasts: role in scarless wound healing. Pediatr Res 43:403–409
Coulomb B, Dubertret L, Bell E, Touraine R (1984) The contractility of fibroblasts in a collagen lattice is reduced by corticosteroids. J Invest Dermatol 82:341–344
Cowin AJ, Holmes TM, Brosnan P, Ferguson MW (2001) Expression of TGF-beta and its receptors in murine fetal and adult dermal wounds. Eur J Dermatol 11:424–431
Cox DA (1995) Transforming growth factor-beta 3. Cell Biol Int 19:357–371
Eastwood M, Porter R, Khan U, McGrouther G, Brown R (1996) Quantitative analysis of collagen gel contractile forces generated by dermal fibroblasts and the relationship to cell morphology. J Cell Physiol 166:33–42
Ehrlich HP, Wyler DJ (1983) Fibroblast contraction of collagen lattices in vitro: inhibition by chronic inflammatory cell mediators. J Cell Physiol 116:345–351
Faryniarz DA, Chaponnier C, Gabbiani G, Yannas IV, Spector M (1996) Myofibroblasts in the healing lapine medial collateral ligament: possible mechanisms of contraction. J Orthop Res 14:228–237
Forslund C, Aspenberg P (2003) Improved healing of transected rabbit Achilles tendon after a single injection of cartilage-derived morphogenetic protein-2. Am J Sports Med 31:555–559
Fu SC, Wang W, Pau HM, Wong YP, Chan KM, Rolf CG (2002) Increased expression of transforming growth factor-beta1 in patellar tendinosis. Clin Orthop 174–183
Gigante A, Specchia N, Rapali S, Ventura A, de Palma L (1996) Fibrillogenesis in tendon healing: an experimental study. Boll Soc Ital Biol Sper 72:203–210
Greco RM, Iocono JA, Ehrlich HP (1998) Hyaluronic acid stimulates human fibroblast proliferation within a collagen matrix. J Cell Physiol 177:465–473
Grinnell F, Ho CH (2002) Transforming growth factor beta stimulates fibroblast-collagen matrix contraction by different mechanisms in mechanically loaded and unloaded matrices. Exp Cell Res 273:248–255
Harris AK, Wild P, Stopak D (1980) Silicone rubber substrata: a new wrinkle in the study of cell locomotion. Science 208:177–179
Khan KM, Maffuli N, Coleman BD, Cook JL, Taunton JE (2000) Patellar tendinopathy: some aspects of basic science and clinical management. Br J Sports Med 32:346–355
Khan U, Occleston NL, Khaw PT, McGrouther DA (1997) Single exposures to 5-fluorouracil: a possible mode of targeted therapy to reduce contractile scarring in the injured tendon. Plast Reconstr Surg 99:465–471
Khan U, Occleston NL, Khaw PT, McGrouther DA (1998) Differences in proliferative rate and collagen lattice contraction between endotenon and synovial fibroblasts. J Hand Surg [Am] 23:266–273
Milano G, Gigante A, Panni AS, Mulas PD, Fabbriciani C (2001) Patellar tendon healing after removal of its central third. A morphologic evaluation in rabbits. Knee Surg Sports Traumatol Arthrosc 9:92–101
Murata H, Zhou L, Ochoa S, Hasan A, Badiavas E, Falanga V (1997) TGF-beta3 stimulates and regulates collagen synthesis through TGF-beta1- dependent and independent mechanisms. J Invest Dermatol 108:258–262
Nedelec B, Ghahary A, Scott PG, Tredget EE (2000) Control of wound contraction. Basic and clinical features. Hand Clin 16:289–302
Reddy GK, Stehno-Bittel L, Enwemeka CS (1999) Matrix remodeling in healing rabbit Achilles tendon. Wound Repair Regen 7:518–527
Sears CE, Bryant SM, Ashley EA, Lygate CA, Rakovic S, Wallis HL, Neubauer S, Terrar DA, Casadei B (2003) Cardiac neuronal nitric oxide synthase isoform regulates myocardial contraction and calcium handling. Circ Res 92:e52–59
Shah M, Foreman DM, Ferguson MW (1995) Neutralisation of TGF-beta 1 and TGF-beta 2 or exogenous addition of TGF-beta 3 to cutaneous rat wounds reduces scarring. J Cell Sci 108:985–1002
Skold CM, Liu XD, Zhu YK, Umino T, Takigawa K, Ohkuni Y, Ertl RF, Spurzem JR, Romberger DJ, Brattsand R, Rennard SI (1999) Glucocorticoids augment fibroblast-mediated contraction of collagen gels by inhibition of endogenous PGE production. Proc Assoc Am Physicians 111:249–258
Solito E, Parente L (1989) Modulation of phospholipase A2 activity in human fibroblasts. Br J Pharmacol 96:656–660
Torres DS, Freyman TM, Yannas IV, Spector M (2000) Tendon cell contraction of collagen-GAG matrices in vitro: effect of cross-linking. Biomaterials 21:1607–1619
Woo SL, Hildebrand K, Watanabe N, Fenwick JA, Papageorgiou CD, Wang JH (1999) Tissue engineering of ligament and tendon healing. Clin Orthop S312–S323
Younai S, Venters G, Vu S, Nichter L, Nimni ME, Tuan TL (1996) Role of growth factors in scar contraction: an in vitro analysis. Ann Plast Surg 36:495–501
Acknowledgements
BHC was supported in part by the Wellington C. Carl Scholarship from The Pittsburgh Foundation. This work was supported by the Arthritis Investigator Award, Whitaker Biomedical Engineering Grant, NIH grant AR049921, and CMRF from the University of Pittsburgh Medical Center (JHW). We thank Drs. Savio L-Y. Woo and Patricia Hebda for their helpful discussion during this study. We also thank Philip Magcalas for his technical assistance in this study.
Author information
Authors and Affiliations
Corresponding author
Rights and permissions
About this article
Cite this article
Campbell, B.H., Agarwal, C. & Wang, J.HC. TGF-β1, TGF-β3, and PGE2 regulate contraction of human patellar tendon fibroblasts. Biomech Model Mechanobiol 2, 239–245 (2004). https://doi.org/10.1007/s10237-004-0041-z
Received:
Accepted:
Published:
Issue Date:
DOI: https://doi.org/10.1007/s10237-004-0041-z