Abstract
Purpose
This work evaluated gelatin microparticles and biodegradable composite scaffolds for the controlled release of vascular endothelial growth factor (VEGF) in vitro and in vivo.
Methods
Gelatin crosslinking, VEGF dose, and buffer type were investigated for their effects on VEGF release. Release was also evaluated from microparticles confined within porous polymer scaffolds (composites). In vitro and in vivo studies were conducted using radiolabeled VEGF.
Results
The effect of VEGF dose on its fractional release from gelatin microparticles in vitro was minimal, but the addition of collagenase to the buffer resulted in a higher cumulative release of VEGF. Gelatin crosslinking extent was a significant factor on release from both microparticles alone and composite scaffolds in vitro and in vivo. VEGF bioactivity from composite scaffolds in vitro was maintained above 90% of the expected bioactivity over 14 days.
Conclusions
VEGF release kinetics were dependent on the extent of gelatin crosslinking and were characteristic of the specific growth factor due to the effects of growth factor size, charge, and conformation on its complexation with gelatin. These studies demonstrate the utility of gelatin microparticles and their composite scaffolds as delivery vehicles for the controlled release of VEGF for tissue engineering applications.
Similar content being viewed by others
Abbreviations
- bFGF:
-
basic fibroblast growth factor
- BMP-2:
-
bone morphogenetic protein-2
- Coll:
-
collagenase-containing phosphate buffered saline
- GPC:
-
gel permeation chromatography
- HUVECs:
-
human umbilical vein endothelial cells
- IGF-1:
-
insulin-like growth factor-1
- IEP:
-
isoelectric point
- microCT:
-
microcomputed tomography
- PBS:
-
phosphate buffered saline
- PPF:
-
poly(propylene fumarate)
- SEM:
-
scanning electron microscopy
- TGF-β1:
-
transforming growth factor-β1
- VEGF:
-
vascular endothelial growth factor
- VOI:
-
volume of interest
References
L. A. Lakey, R. Akella, and J. P. Ranieri. Angiogenesis: Implications for tissue repair. In J. E. Davies (ed.), Bone Engineering, Em Squared Inc, Toronto, 2000, pp. 137–142.
R. Strocchi, G. Orsini, G. Iezzi, A. Scarano, C. Rubini, G. Pecora, and A. Piattelli. Bone regeneration with calcium sulfate: evidence for increased angiogenesis in rabbits. J. Oral. Implantol. 28:273–278 (2002). doi:10.1563/1548-1336(2002)028<0273:BRWCSE0273:BRWCSE>2.3.CO;2.
J. Glowacki. Angiogenesis in fracture repair. Clin. Orthop. 355S:S82–S89 (1998). doi:10.1097/00003086-199810001-00010.
H. P. Gerber, and N. Ferrara. Angiogenesis and bone growth. Trends Cardiovasc. Med. 10:223–228 (2000). doi:10.1016/S1050-1738(00)00074-8.
H. Winet, J. Y. Bao, and R. Moffat. A control model for tibial cortex neovascularization in the bone chamber. J. Bone. Miner. Res. 5:19–30 (1990).
J. Schmid, B. Wallkamm, C. H. Hammerle, S. Gogolewski, and N. P. Lang. The significance of angiogenesis in guided bone regeneration. A case report of a rabbit experiment. Clin. Oral. Implants. Res. 8:244–248 (1997). doi:10.1034/j.1600-0501.1997.080311.x.
Y. Tabata, M. Miyao, M. Ozeki, and Y. Ikada. Controlled release of vascular endothelial growth factor by use of collagen hydrogels. J. Biomater. Sci. Polym. Ed. 11:915–930 (2000). doi:10.1163/156856200744101.
C. Wong, E. Inman, R. Spaethe, and S. Helgerson. Fibrin-based biomaterials to deliver human growth factors. Thromb. Haemost. 89:573–582 (2003).
S. Soker, M. Machado, and A. Atala. Systems for therapeutic angiogenesis in tissue engineering. World J. Urol. 18:10–18 (2000). doi:10.1007/PL00007070.
M. Sheridan, L. D. Shea, M. C. Peters, and D. J. Mooney. Bioabsorbable polymer scaffolds for tissue engineering capable of sustained growth factory delivery. J. Control. Release. 64:91–102 (2000). doi:10.1016/S0168-3659(99)00138-8.
W. I. Murphy, M. C. Peters, D. H. Kohn, and D. J. Mooney. Sustained release of vascular endothelial growth factor from mineralized poly(lactide-co-glycolide) scaffolds for tissue engineering. Biomaterials. 21:2521–2527 (2000). doi:10.1016/S0142-9612(00)00120-4.
K. Y. Lee, M. C. Peters, and D. J. Mooney. Comparison of vascular endothelial growth factor and basic fibroblast growth factor on angiogenesis in SCID mice. J Control Release. 87:49–56 (2003). doi:10.1016/S0168-3659(02)00349-8.
M. Yamamoto, Y. Ikada, and Y. Tabata. Controlled release of growth factors based on biodegradation of gelatin hydrogel. J. Biomater Sci. Polym. Ed. 12:77–88 (2001). doi:10.1163/156856201744461.
T. A. Holland, Y. Tabata, and A. G. Mikos. Dual growth factor delivery from degradable oligo(poly(ethylene glycol) fumarate) hydrogel scaffolds for cartilage tissue engineering. J. Control. Release. 101:111–125 (2005). doi:10.1016/j.jconrel.2004.07.004.
T. A. Holland, J. K. Tessmar, Y. Tabata, and A. G. Mikos. Transforming growth factor-beta 1 release from oligo(poly(ethylene glycol) fumarate) hydrogels in conditions that model the cartilage wound healing environment. J. Control. Release. 94:101–114 (2004). doi:10.1016/j.jconrel.2003.09.007.
R. G. Payne, J. S. McGonigle, M. J. Yaszemski, A. W. Yasko, and A. G. Mikos. Development of an injectable, in situ crosslinkable, degradable polymeric carrier for osteogenic cell populations. Part 3. Proliferation and differentiation of encapsulated marrow stromal osteoblasts cultured on crosslinking poly(propylene fumarate). Biomaterials. 23:4381–4387 (2002). doi:10.1016/S0142-9612(02)00186-2.
T. A. Holland, and A. G. Mikos. Advances in drug delivery for articular cartilage. J. Control. Release. 86:1–14 (2003). doi:10.1016/S0168-3659(02)00373-5.
T. A. Holland, Y. Tabata, and A. G. Mikos. In vitro release of transforming growth factor-beta1 from gelatin microparticles encapsulated in biodegradable, injectable oligo(poly(ethylene glycol) fumarate) hydrogels. J. Control. Release. 91:299–313 (2003). doi:10.1016/S0168-3659(03)00258-X.
P. R. Salacinski, C. McLean, J. E. Sykes, V. V. Clement-Jones, and P. J. Lowry. Iodination of proteins, glycoproteins, and peptides using a solid-phase oxidizing agent, 1,3,4,6-tetrachloro-3 alpha,6 alpha-diphenyl glycoluril (Iodogen). Anal Biochem. 117:136–146 (1981). doi:10.1016/0003-2697(81)90703-X.
A. K. Shung, E. Behravesh, S. Jo, and A. G. Mikos. Crosslinking characteristics of and cell adhesion to an injectable poly(propylene fumarate-co-ethylene glycol) hydrogel using a water-soluble crosslinking system. Tissue Eng. 9:243–254 (2003). doi:10.1089/107632703764664710.
B. D. Porter, J. B. Oldham, S. L. He, M. E. Zobitz, R. G. Payne, K. N. An, B. L. Currier, A. G. Mikos, and M. J. Yaszemski. Mechanical properties of a biodegradable bone regeneration scaffold. J. Biomech. Eng. 122:286–288 (2000). doi:10.1115/1.429659.
E. L. Hedberg, H. C. Kroese-Deutman, C. K. Shih, R. S. Crowther, D. H. Carney, A. G. Mikos, and J. A. Jansen. Effect of varied release kinetics of the osteogenic thrombin peptide TP508 from biodegradable, polymeric scaffolds on bone formation in vivo. J. Biomed. Mater. Res. A. 72:343–353 (2005). doi:10.1002/jbm.a.30265.
Z. S. Patel. Controlled delivery of angiogenic and osteogenic growth factors for bone regeneration. Rice University, Houston, 2008.
M. C. Peters, B. C. Isenberg, J. A. Rowley, and D. J. Mooney. Release from alginate enhances the biological actiivty of vascular endothelial growth factor. J. Biomater. Sci. Polym. Ed. 9:1267–1278 (1998).
Y. Ikada, and Y. Tabata. Protein release from gelatin matrices. Adv. Drug. Deliv. Rev. 31:287–301 (1998). doi:10.1016/S0169-409X(97)00125-7.
Y. A. Muller, B. Li, H. W. Christinger, J. A. Wells, B. C. Cunningham, and A. M. de Vos. Vascular endothelial growth factor: crystal structure and functional map** of the kinase domain receptor binding site. Proc. Natl. Acad. Sci. U. S. A. 94:7192–7197 (1997). doi:10.1073/pnas.94.14.7192.
W. C. Parks. Matrix metalloproteinases in repair. Wound Repair Regen. 7:423–432 (1999). doi:10.1046/j.1524-475X.1999.00423.x.
Y. Tabata, and Y. Ikada. Vascularization effect of basic fibroblast growth factor released from gelatin hydrogels with different biodegradabilities. Biomaterials. 20:2169–2175 (1999). doi:10.1016/S0142-9612(99)00121-0.
T. P. Richardson, M. C. Peters, A. B. Ennett, and D. J. Mooney. Polymeric system for dual growth factor delivery. Nat. Biotechnol. 19:1029–1034 (2001). doi:10.1038/nbt1101-1029.
A. Lode, A. Reinstorf, A. Bernhardt, C. Wolf-Brandstetter, U. Konig, and M. Gelinsky. Heparin modification of calcium phosphate bone cements for VEGF functionalization. J. Biomed. Mater. Res. A. 27:27 (2007).
J. E. Park, G. A. Keller, and N. Ferrara. The vascular endothelial growth factor (VEGF) isoforms: differential deposition into the subepithelial extracellular matrix and bioactivity of extracellular matrix-bound VEGF. Mol. Biol. Cell. 4:1317–1326 (1993).
Acknowledgments
The authors would like to acknowledge support of this work by a grant from the National Institutes of Health (R01-DE15164) (AGM) and by a National Science Foundation Graduate Research Fellowship (ZSP).
Author information
Authors and Affiliations
Corresponding author
Rights and permissions
About this article
Cite this article
Patel, Z.S., Ueda, H., Yamamoto, M. et al. In Vitro and In Vivo Release of Vascular Endothelial Growth Factor from Gelatin Microparticles and Biodegradable Composite Scaffolds. Pharm Res 25, 2370–2378 (2008). https://doi.org/10.1007/s11095-008-9685-1
Received:
Accepted:
Published:
Issue Date:
DOI: https://doi.org/10.1007/s11095-008-9685-1