Skip to main content

Advertisement

SpringerLink
Log in

Fracture Healing in Mice Deficient in Plasminogen Activator Inhibitor-1

  • Published:
Calcified Tissue International Aims and scope Submit manuscript

Abstract

To evaluate the role of plasminogen activator inhibitor (PAI)-1, a key negative regulator of the plasmin system of extracellular matrix proteases in developmental bone growth and fracture repair, the bone phenotype of male adult PAI-1-deficient mice was determined and femoral fracture healing was compared with that of age- and sex-matched wild-type C57BL/6J control mice. Regarding bone phenotype, the length and size (but not cortical thickness) of the femur of male PAI-1-deficient mice were smaller than those of wild-type controls. Although the total bone mineral content of PAI-1-deficient mice was not significantly different from that of wild-type mice, the total bone area in PAI-1-deficient mice was smaller, leading to an increase in total bone mineral density. With respect to fracture healing, PAI-1-deficient mice developed fracture calluses that were larger and more mineralized than those of wild-type mice but only at 14 days postfracture. These changes were even greater given the smaller size of the normal femur in PAI-1-deficient mice. Surprisingly, the larger fracture callus remodeled rapidly to normal size and mineral content by 21 days postfracture. Examination of fracture histology revealed that these changes were associated with a dramatic increase followed by a rapid remodeling of the fracture callus cartilage. The remodeling of fracture callus cartilage in PAI-1-deficient mice also displayed an abnormal pattern. These findings demonstrate for the first time that PAI-1 (and potentially the plasminogen extracellular matrix protease system) is an important regulator of bone size during developmental growth and plays a regulatory role in the determination of fracture callus size, cartilage formation, and resorption during bone fracture repair.

This is a preview of subscription content, log in via an institution to check access.

Access this article

Log in via an institution

Price excludes VAT (USA)
Tax calculation will be finalised during checkout.

Instant access to the full article PDF.

Institutional subscriptions

Fig. 1
Fig. 2
Fig. 3
Fig. 4
Fig. 5

Similar content being viewed by others

References

  1. Romer J, Lund LR, Eriksen J, Ralfkiaer E, Zeheb R, Gelehrter TD, Dano K, Kristensen P (1991) Differential expression of urokinase-type plasminogen activator and its type-1 inhibitor during healing of mouse skin wounds. J Invest Dermatol 97:803–811

    Article  PubMed  CAS  Google Scholar 

  2. Chan JC, Duszcczyszyn DA, Castellino FJ, Ploplis VA (2001) Accelerated skin wound healing in plasminogen activator inhibitor-1-deficient mice. Am J Pathol 159:1681–1688

    PubMed  CAS  Google Scholar 

  3. Koh TJ, Bryer SC, Pucci AM, Sisson TH (2005) Mice deficient in plasminogen activator inhibitor-1 have improved skeletal muscle regeneration. Am J Physiol Cell Physiol 289:217–223

    Article  CAS  Google Scholar 

  4. Zhou A, Huntington JA, Pannu NS, Carrell RW, Read RJ (2003) How vitronectin binds to PAI-1 to modulate fibrinolysis and cell migration. Nat Struct Biol 10:541–544

    Article  PubMed  CAS  Google Scholar 

  5. Lazar MH, Christensen PJ, Du M, Yu B, Subbotina NM, Hanson KE, Hansen JM, White ES, Simon RH, Sisson TH (2004) Plasminogen activator inhibitor-1 impairs alveolar epithelial repair by binding to vitronectin. Am J Respir Cell Mol Biol 31:672–678

    Article  PubMed  CAS  Google Scholar 

  6. Carmeliet P, Moons L, Lijnen R, Janssens S, Lupu F, Collen D, Gerard RD (1997) Inhibitory role of plasminogen activator inhibitor-1 in arterial wound healing and neointima formation: a gene targeting and transfer study in mice. Circulation 96:3180–3191

    PubMed  CAS  Google Scholar 

  7. Carmeliet P, Kieckens L, Schoonjans L, Ream B, van Nuffelen A, Prendergast G, Cole M, Bronson F, Collen D, Mulligan RC (1993) Plasminogen activator inhibitor-1 gene-deficient mice. I. Generation by homologous recombination and characterization. J Clin Invest 92:2746–2755

    Article  PubMed  CAS  Google Scholar 

  8. Wergedal JE, Sheng MH, Ackert-Bicknell CL, Beamer WG, Baylink DJ (2005) Genetic variation in femur extrinsic strength in 29 different inbred strains of mice is dependent on variations in femur cross-sectional geometry and bone density. Bone 36:111–122

    Article  PubMed  Google Scholar 

  9. Bonnarens F, Einhorn TA (1984) Production of a standard closed fracture in laboratory animal bone. J Orthop Res 2:97–101

    Article  PubMed  CAS  Google Scholar 

  10. Bancroft JD (2002) Enzyme histochemistry and its diagnostic applications. In: Bancroft JD, Gamble M (eds) Theory and practice of histological techniques. Harcourt, New York, pp 593–620

    Google Scholar 

  11. Carmeliet P, Stassen JM, Schoonjans L, Ream B, van den Oord JJ, De Mol M, Mulligan RC, Collen D (1993) Plasminogen activator inhibitor-1 gene-deficient mice. II. Effects on hemostasis, thrombosis, and thrombolysis. J Clin Invest 92:2756–2760

    Article  PubMed  CAS  Google Scholar 

  12. Daci E, Verstuyf A, Moermans K, Bouillon R, Carmeliet G (2000) Mice lacking the plasminogen activator inhibitor 1 are protected from trabecular bone loss induced by estrogen deficiency. J Bone Miner Res 8:1510–1516

    Article  Google Scholar 

  13. Nordstrom SM, Carleton SM, Carson WL, Eren M, Phillips CL, Vaughan DE (2007) Transgenic over-expression of plasminogen activator inhibitor-1 results in age-dependent and gender-specific increases in bone strength and mineralization. Bone 41:995–1004

    Article  PubMed  CAS  Google Scholar 

  14. Daci E, Everts V, Torrekens S, van Herck E, Tigchelaar-Gutterr W, Bouillon R, Carmeliet G (2003) Increased bone formation in mice lacking plasminogen activators. J Bone Miner Res 18:1167–1176

    Article  PubMed  CAS  Google Scholar 

  15. Pedrozo HA, Schwartz Z, Robinson M, Gomez R, Dean DD, Bonewald LF, Boyan BD (1999) Potential mechanisms for the plasmin-mediated release and activation of latent transforming growth factor-β1 from the extracellular matrix of growth plate chondrocytes. Endocrinol 140:5806–5816

    Article  CAS  Google Scholar 

  16. Saksela O, Moscatelli D, Sommer A, Rifkin DB (1988) Endothelial cell–derived heparin sulfate binds basic fibroblast growth factor and protects it from proteolytic degradation. J Cell Biol 107:743–751

    Article  PubMed  CAS  Google Scholar 

  17. Campbell PG, Novak JF, Yanosick TB, McMaster JH (1992) Involvement of the plasmin system in dissociation of the insulin-like growth factor-binding protein complex. Endocrinology 130:1401–1412

    Article  PubMed  CAS  Google Scholar 

  18. Lalou C, Silve C, Rosato R, Segovia B, Binoux M (1994) Interactions between insulin-like growth factor-I (IGF-I) and the system of plasminogen activators and their inhibitors in the control of IGF-binding protein-3 production and proteolysis in human osteosarcoma cells. Endocrinology 135:2318–2326

    Article  PubMed  CAS  Google Scholar 

  19. Hughes DE, Salter DM, Dedhar S, Simpson R (1993) Integrin expression in human bone. J Bone Miner Res 8:527–533

    PubMed  CAS  Google Scholar 

  20. Daci E, Udagawa N, Martin TJ, Bouillon R, Carmeliet G (1999) The role of the plasminogen system in bone resorption in vitro. J Bone Miner Res 14:946–952

    Article  PubMed  CAS  Google Scholar 

  21. Weisel JW (2005) Fibrinogen and fibrin. Adv Protein Chem 70:247–299

    Article  PubMed  CAS  Google Scholar 

  22. Colnot C, Thompson Z, Miclau T, Werb Z, Helms JA (2003) Altered fracture repair in the absence of MMP9. Development 130:4123–4133

    Article  PubMed  CAS  Google Scholar 

  23. Tumber A, Papaioannou S, Breckon J, Meikle MC, Reynolds JJ, Hill PA (2003) The effects of serine protease inhibitors on bone resorption in vitro. J Endocrinol 178:437–447

    Article  PubMed  CAS  Google Scholar 

  24. Leloup G, Lemoine P, Carmeliet P, Vaes G (1996) Bone resorption and response to calcium-regulating hormones in the absence of tissue or urokinase plasminogen activator or of their type 1 inhibitor. J Bone Miner Res 11:1146–1157

    Article  PubMed  CAS  Google Scholar 

  25. Furlan F, Galbiati C, Jorgensen NR, Jensen J-EB, Mrak E, Rubinacci A, Talotta F, Verde P, Blasi F (2007) Urokinase plasminogen activator receptor affects bone homeostasis by regulating osteoblast and osteoclast function. J Bone Miner Res 22:1387–1396

    Article  PubMed  CAS  Google Scholar 

Download references

Acknowledgement

This work was supported by the U.S. Army Medical Research Acquisition Activity Assistance Award (DAMD17-03-2-0021). The U.S. Army Medical Research Acquisition Activity (820 Chandler Street, Fort Detrick, MD 21702-5014) is the awarding and administering acquisition office. The information contained in this publication does not necessarily reflect the position or the policy of the government, and no official endorsement should be inferred. We are grateful to Emile Davis, Nancy Lowen, Ryan Porte, and Jann Smallwood for technical assistance. All work was performed in facilities provided by the Department of Veterans Affairs.

Author information

Authors and Affiliations

  1. Musculoskeletal Disease Center, Jerry L. Pettis Memorial Veterans Administration Medical Center, 11201 Benton Street, Loma Linda, CA, 92357, USA

    Charles H. Rundle, Xiaoguang Wang, Jon E. Wergedal, Subburaman Mohan & K.-H. William Lau

  2. Department of Medicine, Loma Linda University, Loma Linda, CA, 92350, USA

    Charles H. Rundle, Jon E. Wergedal, Subburaman Mohan & K.-H. William Lau

  3. Department of Biochemistry, Loma Linda University, Loma Linda, CA, 92350, USA

    Jon E. Wergedal, Subburaman Mohan & K.-H. William Lau

  4. Department of Physiology, Loma Linda University, Loma Linda, CA, 92350, USA

    Subburaman Mohan

Authors
  1. Charles H. Rundle
    View author publications

    You can also search for this author in PubMed Google Scholar

  2. Xiaoguang Wang
    View author publications

    You can also search for this author in PubMed Google Scholar

  3. Jon E. Wergedal
    View author publications

    You can also search for this author in PubMed Google Scholar

  4. Subburaman Mohan
    View author publications

    You can also search for this author in PubMed Google Scholar

  5. K.-H. William Lau
    View author publications

    You can also search for this author in PubMed Google Scholar

Corresponding author

Correspondence to K.-H. William Lau.

Rights and permissions

Reprints and permissions

About this article

Cite this article

Rundle, C.H., Wang, X., Wergedal, J.E. et al. Fracture Healing in Mice Deficient in Plasminogen Activator Inhibitor-1. Calcif Tissue Int 83, 276–284 (2008). https://doi.org/10.1007/s00223-008-9169-7

Download citation

  • Received:

  • Accepted:

  • Published:

  • Issue Date:

  • DOI: https://doi.org/10.1007/s00223-008-9169-7

Keywords

Search

Navigation