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Pharmaceutical Research

, Volume 31, Issue 12, pp 3335–3347 | Cite as

Controlled Delivery of Fibroblast Growth Factor-9 from Biodegradable Poly(ester amide) Fibers for Building Functional Neovasculature

  • Somiraa S. Said
  • J. Geoffrey Pickering
  • Kibret Mequanint
Research Paper

Abstract

Purpose

For building functional vasculature, controlled delivery of fibroblast growth factor-9 (FGF9) from electrospun fibers is an appealing strategy to overcome challenges associated with its short half-life. FGF9 sustained delivery could potentially drive muscularization of angiogenic sprouts and help regenerate stable functional neovasculature in ischemic vascular disease patients.

Methods

Electrospinning parameters of FGF9-loaded poly(ester amide) (PEA) fibers have been optimized, using blend and emulsion electrospinning techniques. In vitro PEA matrix degradation, biocompatibility, FGF9 release kinetics, and bioactivity of the released FGF9 were evaluated. qPCR was employed to evaluate platelet-derived growth factor receptor-β (PDGFRβ) gene expression in NIH-3T3 fibroblasts, 10T1/2 cells, and human coronary artery smooth muscle cells cultured on PEA fibers at different FGF9 concentrations.

Results

Loaded PEA fibers exhibited controlled release of FGF9 over 28 days with limited burst effect while preserving FGF9 bioactivity. FGF9-loaded and unloaded electrospun fibers were found to support the proliferation of fibroblasts for five days even in serum-depleted conditions. Cells cultured on FGF9-supplemented PEA mats resulted in upregulation of PDGFRβ in concentration and cell type-dependent manner.

Conclusion

This study supports the premise of controlled delivery of FGF9 from PEA electrospun fibers for potential therapeutic angiogenesis applications.

Key Words

Electrospinning Fibroblast growth factor-9 Poly(ester amide)s Therapeutic angiogenesis 

Notes

Acknowledgments and Disclosures

The Authors acknowledge the financial support from The Heart and Stroke Foundation of Canada (T-7262), the Natural Sciences and Engineering Research Council of Canada, the Canadian Institutes for Health Research (FRN 11715), and the Canadian Cancer Society (Grant #701080). S. S. Said held a CIHR Strategic Training Fellowship in Vascular Research (2011–2013). J. G. Pickering holds the Heart and Stroke Foundation of Ontario/Barnett-Ivey Chair. Thanks to D. K. Knight for his assistance in the synthesis of PEA and performing the GPC molecular weight analysis.

References

  1. 1.
    Dragneva G, Korpisalo P, Yla-Herttuala S. Promoting Blood Vessel Growth in Ischemic Diseases: Challenges in Translating Preclinical Potential into Clinical Success. Dis Model Mech. 2013;6(2):312–22.PubMedCentralPubMedCrossRefGoogle Scholar
  2. 2.
    Lusis AJ. Atherosclerosis. Nature. 2000;407(6801):233–41.PubMedCentralPubMedCrossRefGoogle Scholar
  3. 3.
    Lee KY, Peters MC, Mooney DJ. Comparison of Vascular Endothelial Growth Factor and Basic Fibroblast Growth Factor on Angiogenesis in SCID Mice. J Control Release. 2003;87(1–3):49–56.PubMedCrossRefGoogle Scholar
  4. 4.
    Said SS, Pickering JG, Mequanint K. Advances in Growth Factor Delivery for Therapeutic Angiogenesis. J Vasc Res. 2013;50(1):35–51.PubMedCrossRefGoogle Scholar
  5. 5.
    Sahoo S, Ang LT, Goh JC-H, Toh SL. Growth Factor Delivery Through Electrospun Nanofibers in Scaffolds for Tissue Engineering Applications. J Biomed Mater Res. 2010;93A:1539–50.Google Scholar
  6. 6.
    Yang Y, Xia T, Zhi W, Wei L, Weng J, Zhang C, et al. Promotion of Skin Regeneration in Diabetic Rats by Electrospun Core-Sheath Fibers Loaded With Basic Fibroblast Growth Factor. Biomaterials. 2011;32(18):4243–54.PubMedCrossRefGoogle Scholar
  7. 7.
    Seyednejad H, Ji W, Yang F, van Nostrum CF, Vermonden T, van den Beucken JJ, et al. Coaxially Electrospun Scaffolds Based on Hydroxyl-Functionalized Poly (Epsilon-Caprolactone) and Loaded With VEGF for Tissue Engineering Applications. Biomacromolecules. 2012;13(11):3650–60.PubMedCrossRefGoogle Scholar
  8. 8.
    Roy RS, Roy B, Sengupta S. Emerging Technologies for Enabling Proangiogenic Therapy. Nanotechnology. 2011;22(49):494004.PubMedCrossRefGoogle Scholar
  9. 9.
    Rubanyi GM. Angiogenesis in health and disease: Basic mechanisms and clinical applications. New york: Marcel Dekker Inc.; 2000.Google Scholar
  10. 10.
    Agrotis A, Kanellakis P, Kostolias G, Di Vitto G, Wei C, Hannan R, et al. Proliferation of Neointimal Smooth Muscle Cells After Arterial Injury. Dependence on Interactions Between Fibroblast Growth Factor Receptor-2 and Fibroblast Growth Factor-9. J Biol Chem. 2004;279(40):42221–9.PubMedCrossRefGoogle Scholar
  11. 11.
    Spicer D. FGF9 on the Move. Nat Genet. 2009;41(3):272–3.PubMedCrossRefGoogle Scholar
  12. 12.
    Frontini MJ, Nong Z, Gros R, Drangova M, O’Neil C, Rahman MN, et al. Fibroblast Growth Factor 9 Delivery During Angiogenesis Produces Durable, Vasoresponsive Microvessels Wrapped by Smooth Muscle Cells. Nat Biotechnol. 2011;29(5):421–7.PubMedCrossRefGoogle Scholar
  13. 13.
    Richardson TP, Peters MC, Ennett AB, Mooney DJ. Polymeric System for Dual Growth Factor Delivery. Nat Biotechnol. 2001;19(11):1029–34.PubMedCrossRefGoogle Scholar
  14. 14.
    Knight DK, Gillies ER, Mequanint K. Strategies in Functional Poly (ester amide) Syntheses to Study Human Coronary Artery Smooth Muscle Cell Interactions. Biomacromolecules. 2011;12(7):2475–87.PubMedCrossRefGoogle Scholar
  15. 15.
    Vert M, Li S, Garreau H. New Insights on the Degradation of Bioresorbable Polymeric Devices Based on Lactic and Glycolic Acids. Clin Mater. 1992;10(1–2):3–8.PubMedCrossRefGoogle Scholar
  16. 16.
    Szentivanyi A, Chakradeo T, Zernetsch H, Glasmacher B. Electrospun Cellular Microenvironments: Understanding Controlled Release and Scaffold Structure. Adv Drug Delivery Rev. 2011;63(4–5):209–20.CrossRefGoogle Scholar
  17. 17.
    Li L, Chu CC. Nitroxyl Radical Incorporated Electrospun Biodegradable Poly (Ester Amide) Nanofiber Membranes. J Biomater Sci Polym Ed. 2009;20(3):341–61.PubMedCrossRefGoogle Scholar
  18. 18.
    Srinath D, Lin S, Knight DK, Rizkalla AS, Mequanint K. Fibrous Biodegradable L-alanine-based Scaffolds for Vascular Tissue Engineering. J Tissue Eng Regen Med. 2012.Google Scholar
  19. 19.
    Valle L, Roa M, Diaz A, Casas M, Puiggali J, Rodriguez-Galan A. Electrospun Nanofibers of a Degradable Poly (Ester amide). Scaffolds Loaded with Antimicrobial Agents. J Polym Res. 2012;19(2):1–13.Google Scholar
  20. 20.
    Morgan PW. Interfacial Polymerization. Encyclopedia of Polymer Science and Technology: John Wiley & Sons, Inc.; 2002.Google Scholar
  21. 21.
    Pham QP, Sharma U, Mikos AG. Electrospinning of Polymeric Nanofibers for Tissue Engineering Applications: A Review. Tissue Eng. 2006;12(5):1197–211.PubMedCrossRefGoogle Scholar
  22. 22.
    Zong X, Kim K, Fang D, Ran S, Hsiao BS, Chu B. Structure and Process Relationship of Electrospun Bioabsorbable Nanofiber Membranes. Polymer. 2002;43(16):4403–12.CrossRefGoogle Scholar
  23. 23.
    Jarusuwannapoom T, Hongrojjanawiwat W, Jitjaicham S, Wannatong L, Nithitanakul M, Pattamaprom C, et al. Effect of Solvents on Electro-Spinnability of Polystyrene Solutions and Morphological Appearance of Resulting Electrospun Polystyrene Fibers. Eur Polym J. 2005;41(3):409–21.CrossRefGoogle Scholar
  24. 24.
    Tsitlanadze G, Machaidze M, Kviria T, Djavakhishvili N, Chu CC, Katsarava R. Biodegradation of Amino-Acid-Based Poly (Ester Amide)s: In Vitro Weight Loss and Preliminary in Vivo Studies. J Biomater Sci Polym Ed. 2004;15(1):1–24.PubMedCrossRefGoogle Scholar
  25. 25.
    Zamani M, Morshed M, Varshosaz J, Jannesari M. Controlled Release of Metronidazole Benzoate from Poly Epsilon-Caprolactone Electrospun Nanofibers for Periodontal Diseases. Eur J Pharm Biopharm. 2010;75(2):179–85.PubMedCrossRefGoogle Scholar
  26. 26.
    Zeng J, Yang L, Liang Q, Zhang X, Guan H, Xu X, et al. Influence of the Drug Compatibility With Polymer Solution on the Release Kinetics of Electrospun Fiber Formulation. J Control Release. 2005;105(1–2):43–51.PubMedCrossRefGoogle Scholar
  27. 27.
    Maretschek S, Greiner A, Kissel T. Electrospun Biodegradable Nanofiber Nonwovens for Controlled Release of Proteins. J Control Release. 2008;127(2):180–7.PubMedCrossRefGoogle Scholar
  28. 28.
    Sy JC, Klemm AS, Shastri VP. Emulsion as a Means of Controlling Electrospinning of Polymers. Adv Mater. 2009;21(18):1814–9.CrossRefGoogle Scholar
  29. 29.
    Yang Y, Li X, He S, Cheng L, Chen F, Zhou S, et al. Biodegradable Ultrafine Fibers With Core–Sheath Structures for Protein Delivery and its Optimization. Polym Adv Technol. 2011;22(12):1842–50.CrossRefGoogle Scholar
  30. 30.
    Ritger PL, Peppas NA. A Simple Equation for Description of Solute Release I. Fickian and non-Fickian Release from non-Swellable Devices in the Form of Slabs, Spheres, Cylinders or Discs. J Control Release. 1986;5(1):23–36.CrossRefGoogle Scholar
  31. 31.
    Baker SC, Southgate J. Towards Control of Smooth Muscle Cell Differentiation in Synthetic 3D Scaffolds. Biomaterials. 2008;29(23):3357–66.PubMedCrossRefGoogle Scholar
  32. 32.
    Naruo K, Seko C, Kuroshima K, Matsutani E, Sasada R, Kondo T, et al. Novel Secretory Heparin-Binding Factors from Human Glioma Cells (Glia-Activating Factors) Involved in Glial Cell Growth. Purification and Biological Properties. J Biol Chem. 1993;268(4):2857–64.PubMedGoogle Scholar
  33. 33.
    Rubin JS, Chan AM, Bottaro DP, Burgess WH, Taylor WG, Cech AC, et al. A Broad-Spectrum Human Lung Fibroblast-Derived Mitogen is a Variant of Hepatocyte Growth Factor. Proc Natl Acad Sci. 1991;88(2):415–9.PubMedCentralPubMedCrossRefGoogle Scholar
  34. 34.
    Richardson WJ, Wilson E, Moore Jr JE. Altered Phenotypic Gene Expression of 10T1/2 Mesenchymal Cells in Nonuniformly Stretched PEGDA Hydrogels. Am J Physiol Cell Physiol. 2013;305(1):C100–110.Google Scholar
  35. 35.
    Horwitz JA, Shum KM, Bodle JC, Deng M, Chu CC, Reinhart-King CA. Biological Performance of Biodegradable Amino Acid-Based Poly (Ester Amide)s: Endothelial Cell Adhesion and Inflammation In Vitro. J Biomed Mater Res A. 2010;95(2):371–80.PubMedCrossRefGoogle Scholar
  36. 36.
    Deng M, Wu J, Reinhart-King CA, Chu CC. Biodegradable Functional Poly (Ester Amide)s With Pendant Hydroxyl Functional Groups: Synthesis, Characterization, Fabrication and In Vitro Cellular Response. Acta Biomater. 2011;7(4):1504–15.PubMedCrossRefGoogle Scholar
  37. 37.
    Lin S, Sandig M, Mequanint K. Three-Dimensional Topography of Synthetic Scaffolds Induces Elastin Synthesis by Human Coronary Artery Smooth Muscle Cells. Tissue Eng Part A. 2011;17(11–12):1561–71.PubMedCrossRefGoogle Scholar
  38. 38.
    Carlson AL, Florek CA, Kim JJ, Neubauer T, Moore JC, Cohen RI, et al. Microfibrous Substrate Geometry as a Critical Trigger for Organization, Self-Renewal, and Differentiation of Human Embryonic Stem Cells Within Synthetic 3-Dimensional Microenvironments. Faseb J. 2012;26(8):3240–51.PubMedCentralPubMedCrossRefGoogle Scholar
  39. 39.
    Zhou L, Takayama Y, Boucher P, Tallquist MD, Herz J. LRP1 Regulates Architecture of the Vascular Wall by Controlling PDGFRÎ2-Dependent Phosphatidylinositol 3-Kinase Activation. PLoS One. 2009;4(9):e6922.PubMedCentralPubMedCrossRefGoogle Scholar
  40. 40.
    Hellstrom M, Kalen M, Lindahl P, Abramsson A, Betsholtz C. Role of PDGF-B and PDGFR-Beta in Recruitment of Vascular Smooth Muscle Cells and Pericytes During Embryonic Blood Vessel Formation in the Mouse. Development. 1999;126(14):3047–55.PubMedGoogle Scholar
  41. 41.
    Zhang H, Jia X, Han F, Zhao J, Zhao Y, Fan Y, et al. Dual-Delivery of VEGF and PDGF by Double-Layered Electrospun Membranes for Blood Vessel Regeneration. Biomaterials. 2012;34(9):2202–12.CrossRefGoogle Scholar

Copyright information

© Springer Science+Business Media New York 2014

Authors and Affiliations

  • Somiraa S. Said
    • 1
  • J. Geoffrey Pickering
    • 2
  • Kibret Mequanint
    • 3
  1. 1.Biomedical Engineering Graduate ProgramThe University of Western OntarioLondonCanada
  2. 2.Department of Medicine (Cardiology), Department of Biochemistry, and Department of Medical BiophysicsThe University of Western OntarioLondonCanada
  3. 3.Department of Chemical and Biochemical Engineering and Biomedical Engineering Graduate ProgramThe University of Western OntarioLondonCanada

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