Effect of scaffold dilution on migration of mesenchymal stem cells from fibrin hydrogels

Benjamin W. Hale Orthopaedic Research Center, Department of Clinical Science, College of Veterinary Medicine and Biomedical Sciences, Colorado State University, Fort Collins, CO 80523.

Search for other papers by Benjamin W. Hale in
Current site
Google Scholar
PubMed Close
 MS
,
Laurie R. Goodrich Orthopaedic Research Center, Department of Clinical Science, College of Veterinary Medicine and Biomedical Sciences, Colorado State University, Fort Collins, CO 80523.

Search for other papers by Laurie R. Goodrich in
Current site
Google Scholar
PubMed Close
 DVM, PhD
,
David D. Frisbie Orthopaedic Research Center, Department of Clinical Science, College of Veterinary Medicine and Biomedical Sciences, Colorado State University, Fort Collins, CO 80523.

Search for other papers by David D. Frisbie in
Current site
Google Scholar
PubMed Close
 DVM, PhD
,
C. Wayne McIlwraith Orthopaedic Research Center, Department of Clinical Science, College of Veterinary Medicine and Biomedical Sciences, Colorado State University, Fort Collins, CO 80523.

Search for other papers by C. Wayne McIlwraith in
Current site
Google Scholar
PubMed Close
 DVM, PhD, DSc
, and
John D. Kisiday Orthopaedic Research Center, Department of Clinical Science, College of Veterinary Medicine and Biomedical Sciences, Colorado State University, Fort Collins, CO 80523.

Search for other papers by John D. Kisiday in
Current site
Google Scholar
PubMed Close
 PhD
DOI:
https://doi.org/10.2460/ajvr.73.2.313
Volume/Issue:
Volume 73: Issue 2
Online Publication Date:
01 Feb 2012
Restricted access
Sign In Reprints and Permissions Purchase Article

Abstract

Objective—To evaluate the effect of fibrin concentrations on mesenchymal stem cell (MSC) migration out of autologous and commercial fibrin hydrogels.

Sample—Blood and bone marrow from six 2- to 4-year-old horses.

Procedures—Autologous fibrinogen was precipitated from plasma and solubilized into a concentrated solution. Mesenchymal stem cells were resuspended in fibrinogen solutions containing 100%, 75%, 50%, and 25% of the fibrinogen precipitate solution. Fibrin hydrogels were created by mixing the fibrinogen solutions with MSCs and thrombin on tissue culture plates. After incubation for 24 hours in cell culture medium, the MSCs that had migrated onto the tissue culture surface and beyond the boundary of the hydrogels were counted. This procedure was repeated with a commercial fibrin sealant.

Results—Hydrogel-to-surface MSC migration was detected for all fibrin hydrogels. Migration from the 25% autologous hydrogels was 7.3-, 5.2-, and 4.6-fold higher than migration from 100%, 75%, and 50% autologous hydrogels, respectively. The number of migrating cells from 100%, 75%, and 50% autologous hydrogels did not differ significantly. With commercial fibrin sealant, the highest magnitude of migration was from the 25% hydrogels, and it was 26-fold higher than migration from 100% hydrogels. The 75% and 50% hydrogels resulted in migration that was 9.5- and 4.2-fold higher than migration from the 100% hydrogels, respectively.

Conclusions and Clinical Relevance—MSC migration from fibrin hydrogels increased with dilution of the fibrinogen component for both autologous and commercial sources. These data supported the feasibility of using diluted fibrin hydrogels for rapid delivery of MSCs to the surface of damaged tissues.

Abstract

Objective—To evaluate the effect of fibrin concentrations on mesenchymal stem cell (MSC) migration out of autologous and commercial fibrin hydrogels.

Sample—Blood and bone marrow from six 2- to 4-year-old horses.

Procedures—Autologous fibrinogen was precipitated from plasma and solubilized into a concentrated solution. Mesenchymal stem cells were resuspended in fibrinogen solutions containing 100%, 75%, 50%, and 25% of the fibrinogen precipitate solution. Fibrin hydrogels were created by mixing the fibrinogen solutions with MSCs and thrombin on tissue culture plates. After incubation for 24 hours in cell culture medium, the MSCs that had migrated onto the tissue culture surface and beyond the boundary of the hydrogels were counted. This procedure was repeated with a commercial fibrin sealant.

Results—Hydrogel-to-surface MSC migration was detected for all fibrin hydrogels. Migration from the 25% autologous hydrogels was 7.3-, 5.2-, and 4.6-fold higher than migration from 100%, 75%, and 50% autologous hydrogels, respectively. The number of migrating cells from 100%, 75%, and 50% autologous hydrogels did not differ significantly. With commercial fibrin sealant, the highest magnitude of migration was from the 25% hydrogels, and it was 26-fold higher than migration from 100% hydrogels. The 75% and 50% hydrogels resulted in migration that was 9.5- and 4.2-fold higher than migration from the 100% hydrogels, respectively.

Conclusions and Clinical Relevance—MSC migration from fibrin hydrogels increased with dilution of the fibrinogen component for both autologous and commercial sources. These data supported the feasibility of using diluted fibrin hydrogels for rapid delivery of MSCs to the surface of damaged tissues.

Contributor Notes

Mr. Hale is presently a student at the Department of Science, School of Medicine, University of Colorado, Denver, CO 80217.

Supported by the Orthopaedic Research Center.

Address correspondence to Dr. Kisiday (john.kisiday@colostate.edu).
Cover American Journal of Veterinary Research
American Journal of Veterinary Research
Publication Date:
01 Feb 2012
  • 1.

    Caplan AI. Adult mesenchymal stem cells for tissue engineering versus regenerative medicine. J Cell Physiol 2007; 213:341–347.

  • 2.

    Caplan AI, Dennis JE. Mesenchymal stem cells as trophic mediators. J Cell Biochem 2006; 98:1076–1084.

  • 3.

    Frisbie DD, Smith RK. Clinical update on the use of mesenchymal stem cells in equine orthopaedics. Equine Vet J 2010; 42:86–89.

  • 4.

    Ferris DJ, Frisbie DD, Kisiday JD, et al. Clinical follow-up of horses treated with bone marrow–derived mesenchymal stem cells for musculoskeletal lesions, in Proceedings. 55th Annu Meet Am Assoc Equine Pract 2009;59–60.

    • Search Google Scholar
    • Export Citation
  • 5.

    Fortier LA, Smith RK. Regenerative medicine for tendinous and ligamentous injuries of sport horses. Vet Clin North Am Equine Pract 2008; 24:191–201.

    • Crossref
    • Search Google Scholar
    • Export Citation
  • 6.

    Pacini S, Spinabella S, Trombi L, et al. Suspension of bone marrow–derived undifferentiated mesenchymal stromal cells for repair of superficial digital flexor tendon in race horses. Tissue Eng 2007; 13:2949–2955.

    • Crossref
    • Search Google Scholar
    • Export Citation
  • 7.

    Schnabel LV, Lynch ME, van der Meulen MC, et al. Mesenchymal stem cells and insulin-like growth factor-I gene-enhanced mesenchymal stem cells improve structural aspects of healing in equine flexor digitorum superficialis tendons. J Orthop Res 2009; 27:1392–1398.

    • Crossref
    • Search Google Scholar
    • Export Citation
  • 8.

    Smith RK. Mesenchymal stem cell therapy for equine tendinopathy. Disabil Rehabil 2008; 30:1752–1758.

  • 9.

    Wilke MM, Nydam DV, Nixon AJ. Enhanced early chondrogenesis in articular defects following arthroscopic mesenchymal stem cell implantation in an equine model. J Orthop Res 2007; 25:913–925.

    • Crossref
    • Search Google Scholar
    • Export Citation
  • 10.

    Frisbie DD, Kisiday JD, Kawcak CE, et al. Evaluation of adipose-derived stromal vascular fraction or bone marrow–derived mesenchymal stem cells for treatment of osteoarthritis. J Orthop Res 2009; 27:1675–1680.

    • Crossref
    • Search Google Scholar
    • Export Citation
  • 11.

    Silver FH, Wang MC, Pins GD. Preparation of fibrin glue: a study of chemical and physical methods. J Appl Biomater 1995; 6:175–183.

  • 12.

    Silver FH, Wang MC, Pins GD. Preparation and use of fibrin glue in surgery. Biomaterials 1995; 16:891–903.

  • 13.

    Brittberg M, Sjogren-Jansson E, Lindahl A, et al. Influence of fibrin sealant (Tisseel) on osteochondral defect repair in the rabbit knee. Biomaterials 1997; 18:235–242.

    • Crossref
    • Search Google Scholar
    • Export Citation
  • 14.

    Lechner S, Huss R. Bone engineering: combining smart biomaterials and the application of stem cells. Artif Organs 2006; 30:770–774.

  • 15.

    Soon MY, Hassan A, Hui JH, et al. An analysis of soft tissue allograft anterior cruciate ligament reconstruction in a rabbit model: a short-term study of the use of mesenchymal stem cells to enhance tendon osteointegration. Am J Sports Med 2007; 35:962–971.

    • Crossref
    • Search Google Scholar
    • Export Citation
  • 16.

    Chong AK, Ang AD, Goh JC, et al. Bone marrow-derived mesenchymal stem cells influence early tendon-healing in a rabbit achilles tendon model. J Bone Joint Surg Am 2007; 89:74–81.

    • Crossref
    • Search Google Scholar
    • Export Citation
  • 17.

    Catelas I, Sese N, Wu BM, et al. Human mesenchymal stem cell proliferation and osteogenic differentiation in fibrin gels in vitro. Tissue Eng 2006; 12:2385–2396.

    • Crossref
    • Search Google Scholar
    • Export Citation
  • 18.

    Ho W, Tawil B, Dunn JC, et al. The behavior of human mesenchymal stem cells in 3D fibrin clots: dependence on fibrinogen concentration and clot structure. Tissue Eng 2006; 12:1587–1595.

    • Crossref
    • Search Google Scholar
    • Export Citation
  • 19.

    Huang CY, Deitzer MA, Cheung HS. Effects of fibrinolytic inhibitors on chondrogenesis of bone-marrow derived mesenchymal stem cells in fibrin gels. Biomech Model Mechanobiol 2007; 6:5–11.

    • Crossref
    • Search Google Scholar
    • Export Citation
  • 20.

    Dickhut A, Gottwald E, Steck E, et al. Chondrogenesis of mesenchymal stem cells in gel-like biomaterials in vitro and in vivo. Front Biosci 2008; 13:4517–4528.

    • Search Google Scholar
    • Export Citation
  • 21.

    Weinand C, Gupta R, Huang AY, et al. Comparison of hydrogels in the in vivo formation of tissue-engineered bone using mesenchymal stem cells and beta-tricalcium phosphate. Tissue Eng 2007; 13:757–765.

    • Crossref
    • Search Google Scholar
    • Export Citation
  • 22.

    Bensaid W, Triffitt JT, Blanchat C, et al. A biodegradable fibrin scaffold for mesenchymal stem cell transplantation. Biomaterials 2003; 24:2497–2502.

    • Crossref
    • Search Google Scholar
    • Export Citation
  • 23.

    Gregory CA, Reyes E, Whitney MJ, et al. Enhanced engraftment of mesenchymal stem cells in a cutaneous wound model by culture in allogenic species-specific serum and administration in fibrin constructs. Stem Cells 2006; 24:2232–2243.

    • Crossref
    • Search Google Scholar
    • Export Citation
  • 24.

    Falanga V, Iwamoto S, Chartier M, et al. Autologous bone marrow–derived cultured mesenchymal stem cells delivered in a fibrin spray accelerate healing in murine and human cutaneous wounds. Tissue Eng 2007; 13:1299–1312.

    • Crossref
    • Search Google Scholar
    • Export Citation
  • 25.

    Qiu LL, Levinson SS, Keeling KL, et al. Convenient and effective method for removing fibrinogen from serum specimens before protein electrophoresis. Clin Chem 2003; 49:868–872.

    • Crossref
    • Search Google Scholar
    • Export Citation
  • 26.

    Kisiday JD, Kopesky PW, Evans CH, et al. Evaluation of adult equine bone marrow- and adipose-derived progenitor cell chondrogenesis in hydrogel cultures. J Orthop Res 2008; 26:322–331.

    • Crossref
    • Search Google Scholar
    • Export Citation
  • 27.

    Yoshida H, Hirozane K, Kamiya A. Comparative study of autologous fibrin glues prepared by cryo-centrifugation, cryo-filtration, and ethanol precipitation methods. Biol Pharm Bull 1999; 22:1222–1225.

    • Crossref
    • Search Google Scholar
    • Export Citation
  • 28.

    Siddappa R, Licht R, van Blitterswijk C, et al. Donor variation and loss of multipotency during in vitro expansion of human mesenchymal stem cells for bone tissue engineering. J Orthop Res 2007; 25:1029–1041.

    • Crossref
    • Search Google Scholar
    • Export Citation
  • 29.

    Solchaga LA, Penick K, Porter JD, et al. FGF-2 enhances the mitotic and chondrogenic potentials of human adult bone marrow–derived mesenchymal stem cells. J Cell Physiol 2005; 203:398–409.

    • Crossref
    • Search Google Scholar
    • Export Citation
  • 30.

    Glidden PF, Malaska C, Herring SW. Thromboelastograph assay for measuring the mechanical strength of fibrin sealant clots. Clin Appl Thromb Hemost 2000; 6:226–233.

    • Crossref
    • Search Google Scholar
    • Export Citation
  • 31.

    Koga H, Shimaya M, Muneta T, et al. Local adherent technique for transplanting mesenchymal stem cells as a potential treatment of cartilage defect. Arthritis Res Ther 2008; 10:R84.

    • Crossref
    • Search Google Scholar
    • Export Citation
  • 32.

    Kalbermatten DF, Kingham PJ, Mahay D, et al. Fibrin matrix for suspension of regenerative cells in an artificial nerve conduit. J Plast Reconstr Aesthet Surg 2008; 61:669–675.

    • Crossref
    • Search Google Scholar
    • Export Citation

Advertisement