Gene Delivery in Tissue Engineering: A Photopolymer Platform to Coencapsulate Cells and Plasmid DNA
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Purpose. Toward the ultimate goal of developing an engineered tissue capable of mimicking complex natural healing processes, we have designed a photopolymer platform that enables simultaneous encapsulation of cells and plasmid DNA in degradable hydrogels. Photopolymerization enables spatial and temporal control of gel formation under physiological conditions, but the presence of photoinitiator radicals poses challenges for DNA photoencapsulation.
Methods. The effects of photoinitiating conditions (ultraviolet light and photoinitiator radicals) on plasmid DNA were studied. Protection methods were identified. Plasmid DNA was photoencapsulated in photocrosslinked hydrogels, and the quantity and quality of the released DNA were assessed. Plasmid DNA was simultaneously entrapped (coencapsulated) with cells in hydrogels to assess in situ transfection.
Results. Experiments showed that in the absence of other species, plasmid DNA was sensitive to photoinitiator radicals, but the addition of transfection agents and/or antioxidants greatly reduced DNA damage by radicals. Encapsulated plasmid DNA was released from degradable, photocrosslinked hydrogels in active forms (supercoiled and relaxed plasmids) with an overall ∼60% recovery. Released DNA was capable of transfecting both plated and encapsulated cells. Encapsulated cells expressed the encoded gene of the coencapsulated plasmid as the polymer degraded.
Conclusions. This photopolymerization platform allows for the creation of engineered tissues with enhanced control of cell behavior through the spatially and temporally controlled release of plasmid DNA.
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- 1.J. E. Babensee, L. V. McIntire, and A. G. Mikos. Growth factor delivery for tissue engineering. Pharm. Res. 17:497-504 (2000).Google Scholar
- 2.T. P. Richardson, W. L. Murphy, and D. J. Mooney. Polymeric delivery of proteins and plasmid DNA for tissue engineering and gene therapy. Crit. Rev. Eukaryotic Gene Expression 11:47-58 (2001).Google Scholar
- 3.S. J. Lee. Cytokine delivery and tissue engineering. Yonsei Med. J. 41:704-719 (2000).Google Scholar
- 4.C. Andree, M. Kullmer, A. Wenger, D. J. Schaefer, U. Kneser, and G. B. Stark. Gene technology and tissue engineering. Min. Invasive Ther. All. Technol. 11:93-99 (2002).Google Scholar
- 5.L. D. Shea, E. Smiley, J. Bonadio, and D. J. Mooney. DNA delivery from polymer matrices for tissue engineering. Nature Biotechnol. 17:551-554 (1999).Google Scholar
- 6.J. Bonadio. Tissue engineering via local gene delivery: Update and future prospects for enhancing the technology. Adv. Drug Deliv. Rev. 44:185-194 (2000).Google Scholar
- 7.H. Kamiya, H. Tsuchiya, J. Yamazaki, and H. Harashima. Intracellular trafficking and transgene expression of viral and non-viral gene vectors. Adv. Drug Deliv. Rev. 52:153-164 (2001).Google Scholar
- 8.F. Liu and L. Huang. Development of non-viral vectors for systemic gene delivery. J. Control. Rel. 78:259-266 (2002).Google Scholar
- 9.G. Zuber, E. Dauty, M. Nothisen, P. Belguise, and J. P. Behr. Towards synthetic viruses. Adv. Drug Deliv. Rev. 52:245-253 (2001).Google Scholar
- 10.N. Aggarwal, H. HogenEsch, P. X. Guo, A. North, M. Suckow, and S. K. Mittal. Biodegradable alginate microspheres as a delivery system for naked DNA. Can. J. Vet. Res. 63:148-152 (1999).Google Scholar
- 11.C. Aral, S. Ozbas-Turan, L. Kabasakal, M. Keyer-Uysal, and J. Akbuga. Studies of effective factors of plasmid DNA-loaded chitosan microspheres I. Plasmid size, chitosan concentration and plasmid addition techniques. Stp. Pharma Sci. 10: 83-88 (2000).Google Scholar
- 12.S. T. Li. Biologic biomaterials: tissue-derived biomaterials (collagen). In J. D. Brozino (ed), The Biomedical Engineering Handbook, CRC Press, Boca Raton, 1995, pp. 627-647.Google Scholar
- 13.B. Gander, L. Meinel, E. Walter, and H. P. Merkle. Polymers as a platform for drug delivery: Reviewing our current portfolio on poly(lactide-co-glycolide) (PLGA) microspheres. Chimia 55:212-217 (2001).Google Scholar
- 14.L. Lunsford, U. McKeever, V. Eckstein, and M. L. Hedley. Tissue distribution and persistence in mice of plasmid DNA encapsulated in a PLGA-based microsphere delivery vehicle. J. Drug Targeting 8:39-50 (2000).Google Scholar
- 15.Y. Capan, B. H. Woo, S. Gebrekidan, S. Ahmed, and P. P. DeLuca. Preparation and characterization of poly (D,L-lactide-co-glycolide) microspheres for controlled release of poly(L-lysine) complexed plasmid DNA. Pharm. Res. 16:509-513 (1999).Google Scholar
- 16.D. Luo, K. Woodrow-Mumford, N. Belcheva, and W. M. Saltzman. Controlled DNA delivery systems. Pharm. Res. 16:1300-1308 (1999).Google Scholar
- 17.H. Cohen, R. J. Levy, J. Gao, I. Fishbein, V. Kousaev, S. Sosnowski, S. Slomkowski, and G. Golomb. Sustained delivery and expression of DNA encapsulated in polymeric nanoparticles. Gene Ther. 7:1896-1905 (2000).Google Scholar
- 18.S. Hirosue, B. G. Muller, R. C. Mulligan, and R. Langer. Plasmid DNA encapsulation and release from solvent diffusion nanospheres. J. Control. Rel. 70:231-242 (2001).Google Scholar
- 19.D. L. Elbert, A. B. Pratt, M. P. Lutolf, S. Halstenberg, and J. A. Hubbell. Protein delivery from materials formed by self-selective conjugate addition reactions. J. Control. Rel. 76:11-25 (2001).Google Scholar
- 20.L. C. Lu, X. Zhu, R. G. Valenzuela, B. L. Currier, and M. J. Yaszemski. Biodegradable polymer scaffolds for cartilage tissue engineering. Clin. Orthop. Rel. Res. 391:S251-S270 (2001).Google Scholar
- 21.K. T. Nguyen and J. L. West. Photopolymerizable hydrogels for tissue engineering applications. Biomaterials 23:4307-4314 (2002).Google Scholar
- 22.Y. Nakayama, J. Y. Kim, S. Nishi, H. Ueno, and T. Matsuda. Development of high-performance stent: gelatinous photogel-coated stent that permits drug delivery and gene transfer. J. Biomed. Mater. Res. 57:559-566 (2001).Google Scholar
- 23.S. J. Bryant and K. S. Anseth. Hydrogel properties influence ECM production by chondrocytes photoencapsulated in poly(ethylene glycol) hydrogels. J. Biomed. Mater. Res. 59:63-72 (2002).Google Scholar
- 24.A. S. Sawhney, C. P. Pathak, and J. A. Hubbell. Bioerodible hydrogels based on photopolymerized poly(ethylene glycol)-co-poly(alpha-hydroxy acid) diacrylate macromers. Macromolecules 26:581-587 (1993).Google Scholar
- 25.S. Li, M. A. Rizzo, S. Bhattacharya, and L. Huang. Characterization of cationic lipid-protamine-DNA (LPD) complexes for intravenous gene delivery. Gene Ther. 5:930-937 (1998).Google Scholar
- 26.S. J. Bryant and K. S. Anseth. The effects of scaffold thickness on tissue engineered cartilage in photocrosslinked poly(ethylene oxide) hydrogels. Biomaterials 22:619-626 (2001).Google Scholar
- 27.S. J. Bryant, C. R. Nuttelman, and K. S. Anseth. Cytocompatibility of UV and visible light photoinitiating systems on cultured NIH/3T3 fibroblasts in vitro. J. Biomater. Sci. Polymer Ed. 11:439-457 (2000).Google Scholar
- 28.D. Bergan, T. Galbraith, and D. L. Sloane. Gene transfer in vitro and in vivo by cationic lipids is not significantly affected by levels of supercoiling of a reporter plasmid. Pharm. Res. 17:967-973 (2000).Google Scholar
- 29.R. P. Sinha and D. P. Hader. UV-induced DNA damage and repair: a review. Photochem. Photobiol. Sci. 1:225-236 (2002).Google Scholar
- 30.A. P. Breen and J. A. Murphy. Reactions of oxyl radicals with DNA. Free Radical Biol. Med. 18:1033-1077 (1995).Google Scholar
- 31.G. Odian. Principles of Polymerization, John Wiley & Sons, New York, 1991.Google Scholar
- 32.S. C. De Smedt, J. Demeester, and W. E. Hennink. Cationic polymer based gene delivery systems. Pharm. Res. 17:113-126 (2000).Google Scholar