OJRM  Vol.3 No.1 , March 2014
Electrospun Gelatin Constructs with Tunable Fiber Orientation Promote Directed Angiogenesis

The field of therapeutic angiogenesis has been predominantly concentrated in modalities that incorporate pro-angiogenic growth factors and/or cells within polymeric constructs that are implanted into the ischemic region. There is growing evidence that construct architecture can significantly affect growth factor activity, cellular viability and differentiation potential. Electrospinning is an attractive but simple scaffold fabrication technique that offers several advantages over traditional fabrication approaches to prepare highly organized structures for therapeutic angiogenesis applications. We recently described the fabrication of nanofibrous scaffolds with aligned fiber orientation that directed cell migration and orientation (i.e.human umbilical vein endothelial cells). Herein we demonstrate the ability of bFGF containing nanofibrous gelatin B scaffolds with controlled fiber orientation to promote capillary formation in vivo. Aligned scaffolds loaded with bFGF induced the highest levels of reperfusion (73% increased in LDPI ratios by day 21 post ischemia induction) in comparison to all other groups including scaffolds with random fiber orientation. Furthermore, the newly formed vasculature, assessed by confocal microscopy, had a parallel alignment along the axis of the scaffold’s fibers. In contrast, no vessel directionality was observed in the animals treated with scaffolds with random fiber orientation in the presence or absence of bFGF.

Cite this paper
Montero, R. , Vazquez-Padron, R. , Pham, S. , D’Ippolito, G. and Andreopoulos, F. (2014) Electrospun Gelatin Constructs with Tunable Fiber Orientation Promote Directed Angiogenesis. Open Journal of Regenerative Medicine, 3, 1-12. doi: 10.4236/ojrm.2014.31001.
[1]   Layman, H., Sacasa, M., Murphy, A.E., Murphy, A.M., Pham, S.M. and Andreopoulos, F.M. (2009) Co-Delivery of FGF-2 and G-CSF from Gelatin-Based Hydrogels as Angiogenic Therapy in a Murine Critical Limb Ischemic Model. Acta Biomater, 5, 230-239. http://dx.doi.org/10.1016/j.actbio.2008.07.024

[2]   Layman, H., Rahnemai-Azar, A.A., Pham, S.M., Tsechpenakis, G. and Andreopoulos, F.M. (2011) Synergistic Angiogenic Effect of Codelivering Fibroblast Growth Factor 2 and Granulocyte-Colony Stimulating Factor from Fibrin Scaffolds and Bone Marrow Transplantation in Critical Limb Ischemia. Tissue Engineering, Part A, 17, 243-254.

[3]   Ma, Z., He, W., Yong, T. and Ramakrishna, S. (2005) Grafting of Gelatin on Electrospun Poly(caprolactone) Nanofibers to Improve Endothelial Cell Spreading and Proliferation and to Control Cell Orientation. Tissue Engineering, 11, 1149-1158. http://dx.doi.org/10.1089/ten.2005.11.1149

[4]   Zisch, A.H., Lutolf, M.P. and Hubbell, J.A. (2003) Biopolymeric Delivery Matrices for Angiogenic Growth Factors. Cardiovascular Pathology, 12, 295-310. http://dx.doi.org/10.1016/S1054-8807(03)00089-9

[5]   Bouta, E.M., McCarthy, C.W., Keim, A., Wang, H.B., Gilbert, R.J. and Goldman, J. (2011) Biomaterial Guides for Lymphatic Endothelial Cell Alignment and Migration. Acta Biomater, 7, 1104-1113.

[6]   Hadjizadeh, A. and Doillon, C.J. (2010) Directional Migration of Endothelial Cells towards Angiogenesis Using Polymer Fibres in a 3D Co-Culture System. Journal of Tissue Engineering and Regenerative Medicine, 4, 524-531.

[7]   Heath, D.E., Lannutti, J.J. and Cooper, S.L. (2010) Electrospun Scaffold Topography Affects Endothelial Cell Proliferation, Metabolic Activity, and Morphology. Journal of Biomedical Materials Research Part A, 94A, 1195-1204.

[8]   Spadaccio, C., Chello, M., Trombetta, M., Rainer, A., Toyoda, Y. and Genovese, J.A. (2009) Drug Releasing Systems in Cardiovascular Tissue Engineering. Journal of Cellular and Molecular Medicine, 13, 422-439.

[9]   Eble, J.A. and Niland, S. (2009) The Extracellular Matrix of Blood Vessels. Current Pharmaceutical Design, 15, 1385-1400. http://dx.doi.org/10.2174/138161209787846757

[10]   Chien, K.R., Domian, I.J. and Parker, K.K. (2008) Cardiogenesis and the Complex Biology of Regenerative Cardiovascular Medicine. Science (Washington DC, U S), 322, 1494-1497. http://dx.doi.org/10.1126/science.1163267

[11]   Lutolf, M.P. and Hubbell, J.A. (2005) Synthetic Biomaterials as Instructive Extracellular Microenvironments for Morphogenesis in Tissue Engineering. Nature Biotechnology, 23, 47-55. http://dx.doi.org/10.1038/nbt1055

[12]   Bu, X., Yan, Y., Zhang, Z., et al. (2010) Properties of Extracellular Matrix-Like Scaffolds for the Growth and Differentiation of Endothelial Progenitor Cells. Journal of Surgical Research, 164, 50-57.

[13]   Hu, X., Shen, H., Yang, F., Bei, J. and Wang, S. (2008) Preparation and Cell Affinity of Microtubular Orientation-Structured PLGA(70/30) Blood Vessel Scaffold. Biomaterials, 29, 3128-3136.

[14]   Indolfi, L., Baker, A.B. and Edelman, E.R. (2012) The Role of Scaffold Microarchitecture in Engineering Endothelial Cell Immunomodulation. Biomaterials, 33, 7019-7027. http://dx.doi.org/10.1016/j.biomaterials.2012.06.052

[15]   Moroni, L., Licht, R., de Boer, J., de Wijn, J.R. and van Blitterswijk, C.A. (2006) Fiber Diameter and Texture of Electrospun PEOT/PBT Scaffolds Influence Human Mesenchymal Stem Cell Proliferation and Morphology, and the Release of Incorporated Compounds. Biomaterials, 27, 4911-4922. http://dx.doi.org/10.1016/j.biomaterials.2006.05.027

[16]   Ragetly, G.R., Griffon, D.J., Lee, H.-B., Fredericks, L.P., Gordon-Evans, W. and Chung, Y.S. (2010) Effect of Chitosan Scaffold Microstructure on Mesenchymal Stem Cell Chondrogenesis. Acta Biomaterialia, 6, 1430-1436.

[17]   Montero, R.B., Vial, X., Nguyen, D.T., et al. (2012) bFGF-Containing Electrospun Gelatin Scaffolds with Controlled Nano-Architectural Features for Directed Angiogenesis. Acta Biomaterialia, 8, 1778-1791.

[18]   Qian, T. and Wang, Y. (2010) Micro/Nano-Fabrication Technologies for Cell Biology. Medical & Biological Engineering & Computing, 48, 1023-1032. http://dx.doi.org/10.1007/s11517-010-0632-z

[19]   Kumar, G., Tison, C.K., Chatterjee, K., et al. (2011) The Determination of Stem Cell Fate by 3D Scaffold Structures through the Control of Cell Shape. Biomaterials, 32, 9188-9196. http://dx.doi.org/10.1016/j.biomaterials.2011.08.054

[20]   Sahoo, S., Ang, L.T., Goh, J.C. and Toh, S.L. (2010) Growth Factor Delivery through Electrospun Nanofibers in Scaffolds for Tissue Engineering Applications. Journal of Biomedical Materials Research Part B: Applied Biomaterials, 93, 1539-1550.

[21]   Zhang, X., Baughman, C.B. and Kaplan, D.L. (2008) In Vitro Evaluation of Electrospun Silk Fibroin Scaffolds for Vascular Cell Growth. Biomaterials, 29, 2217-2227. http://dx.doi.org/10.1016/j.biomaterials.2008.01.022

[22]   Limbourg, A., Korff, T., Napp, L.C., Schaper, W., Drexler, H. and Limbourg, F.P. (2009) Evaluation of Postnatal Arteriogenesis and Angiogenesis in a Mouse Model of Hind-Limb Ischemia. Nature Protocols, 4, 1737-1748.

[23]   Cao, L. and Mooney, D.J. (2007) Spatiotemporal Control over Growth Factor Signaling for Therapeutic Neovascularization. Advanced Drug Delivery Reviews, 59, 1340-1350. http://dx.doi.org/10.1016/j.addr.2007.08.012

[24]   Jakobsson, A. and Nilsson, G.E. (1993) Prediction of Sampling Depth and Photon Pathlength in Laser Doppler Flowmetry. Medical & Biological Engineering & Computing, 31, 301-307. http://dx.doi.org/10.1007/BF02458050

[25]   Chalothorn, D., Clayton, J.A., Zhang, H., Pomp, D. and Faber, J.E. (2007) Collateral Density, Remodeling, and VEGF —An Expression Differ Widely between Mouse Strains. Physiological Genomics, 30, 179-191.

[26]   Li, Y., Song, Y., Zhao, L., Gaidosh, G., Laties, A.M. and Wen, R. (2008) Direct Labeling and Visualization of Blood Vessels with Lipophilic Carbocyanine Dye DiI. Nature Protocols, 3, 1703-1708.

[27]   Hasan, M.R., Herz, J., Hermann, D.M. and Doeppner, T.R. (2012) Visualization of Macroscopic Cerebral Vessel Anatomy—A New and Reliable Technique in Mice. Journal of Neuroscience Methods, 204, 249-253.

[28]   Schmidt, C., Bezuidenhout, D., Beck, M., Van der Merwe, E., Zilla, P. and Davies, N. (2009) Rapid Three-Dimensional Quantification of VEGF-Induced Scaffold Neovascularisation by Microcomputed Tomography. Biomaterials, 30, 5959-5968. http://dx.doi.org/10.1016/j.biomaterials.2009.07.044