Abstract
Although 3D bio-printing technology has great potential in creating complex tissues with multiple cell types and matrices, maintaining the viability of thick tissue construct for tissue growth and maturation after the printing is challenging due to lack of vascular perfusion. Perfused capillary network can be a solution for this issue; however, construction of a complete capillary network at single cell level using the existing technology is nearly impossible due to limitations in time and spatial resolution of the dispensing technology. To address the vascularization issue, we developed a 3D printing method to construct larger (lumen size of ~1 mm) fluidic vascular channels and to create adjacent capillary network through a natural maturation process, thus providing a feasible solution to connect the capillary network to the large perfused vascular channels. In our model, microvascular bed was formed in between two large fluidic vessels, and then connected to the vessels by angiogenic sprouting from the large channel edge. Our bio-printing technology has a great potential in engineering vascularized thick tissues and vascular niches, as the vascular channels are simultaneously created while cells and matrices are printed around the channels in desired 3D patterns.
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References
Adams, R. H., and K. Alitalo. Molecular regulation of angiogenesis and lymphangiogenesis. Nat. Rev. Mol. Cell Biol. 8:464–478, 2007.
Boland, T., V. Mironov, A. Gutowska, E. A. Roth, and R. R. Markwald. Cell and organ printing 2: fusion of cell aggregates in three-dimensional gels. Anat. Rec. A 272:497–502, 2003.
Borenstein, J. T., E. L. I. J. Weinberg, B. K. Orrick, C. Sundback, M. R. Kaazempur-mofrad, and J. P. Vacanti. Microfabrication of three-dimensional engineered scaffolds. Tissue Eng. 13:1837–1844, 2007.
Carmeliet, P. Blood vessels and nerves: common signals, pathways and diseases. Nature 4:710–720, 2003.
Carmeliet, P. Angiogenesis in health and disease. Nat. Med. 9:653–660, 2003.
Carmeliet, P., and R. K. Jain. Angiogenesis in cancer and other diseases. Nature 407:249–257, 2000.
Carmeliet, P., and R. K. Jain. Molecular mechanisms and clinical applications of angiogenesis. Nature 473:298–307, 2011.
Chen, X., et al. Prevascularization of a fibrin-based tissue construct accelerates the formation of functional anastomosis with host vasculature. Tissue Eng. Part A 15:1363–1371, 2009.
Chiu, D. T., et al. Patterned deposition of cells and proteins onto surfaces by using three-dimensional microfluidic systems. Proc. Natl. Acad. Sci. U.S.A. 97:2408–2413, 2000.
Chrobak, K. M., D. R. Potter, and J. Tien. Formation of perfused, functional microvascular tubes in vitro. Microvasc. Res. 71:185–196, 2006.
Conway, E. M., D. Collen, and P. Carmeliet. Molecular mechanisms of blood vessel growth. Cardiovasc. Res. 49:507–521, 2001.
Cui, X., and T. Boland. Human microvasculature fabrication using thermal inkjet printing technology. Biomaterials 30:6221–6227, 2009.
Davis, G. E., and K. J. Bayless. An integrin and rho GTPase-dependent pinocytic vacuole mechanism controls capillary lumen formation in collagen and fibrin matrices. Microcirculation 10:27–44, 2003.
Davis, G. E., W. Koh, and A. N. Stratman. Mechanisms controlling human endothelial lumen formation and tube assembly in three-dimensional extracellular matrices. Birth Defects Res. C 81:270–285, 2007.
Davis, G. E., and D. R. Senger. Endothelial extracellular matrix: biosynthesis, remodeling, and functions during vascular morphogenesis and neovessel stabilization. Circ. Res. 97:1093–1107, 2005.
Fidkowski, C., M. R. Kaazempur-Mofrad, J. Borenstein, J. P. Vacanti, R. Langer, and Y. Wang. Endothelialized microvasculature based on a biodegradable elastomer. Tissue Eng. 11:302–309, 2005.
Ghajar, C. M., K. S. Blevins, C. C. W. Hughes, S. C. George, and A. J. Putnam. Mesenchymal stem cells enhance angiogenesis early matrix metalloproteinase upregulation. Tissue Eng. 12:2875–2888, 2006.
Grinnell, F. Fibroblast–collagen–matrix contraction: growth-factor signalling and mechanical loading. Trends Cell Biol. 10:362–365, 2000.
Grinnell, F. Fibroblast biology in three-dimensional collagen matrices. Trends Cell Biol. 13:264–269, 2003.
Hsu, Y.-H., M. L. Moya, P. Abiri, C. C. W. Hughes, S. C. George, and A. P. Lee. Full range physiological mass transport control in 3D tissue cultures. Lab Chip 13:81–89, 2013.
Iruela-Arispe, M. L., and G. E. Davis. Cellular and molecular mechanisms of vascular lumen formation. Dev. Cell 16:222–231, 2009.
Kachgal, S., and A. J. Putnam. Mesenchymal stem cells from adipose and bone marrow promote angiogenesis via distinct cytokine and protease expression mechanisms. Angiogenesis 14:47–59, 2011.
Kamei, M., W. B. Saunders, K. J. Bayless, L. Dye, G. E. Davis, and B. M. Weinstein. Endothelial tubes assemble from intracellular vacuoles in vivo. Nature 442:453–456, 2006.
Khademhosseini, A., R. Langer, J. Borenstein, and J. P. Vacanti. Microscale technologies for tissue engineering and biology. Proc. Natl. Acad. Sci. USA 103:2480–2487, 2006.
Koh, W., A. N. Stratman, A. Sacharidou, and G. E. Davis. In vitro three dimensional collagen matrix models of endothelial lumen formation during vasculogenesis and angiogenesis. Methods Enzymol. 443:83–101, 2008.
Langer, R. S., and J. P. Vacanti. Tissue engineering: the challenges ahead. Sci. Am. 280:86–89, 1999.
Lee, V., and G. Dai. Micro and nanotechnology in vascular regeneration. In: Tissue and Organ Regeneration—Advances in Micro- and Nanotechnology, edited by G. L. Zhang, T. Webster, and A. Khademhosseini. Singapore: Pan Stanford Publishing, 2014.
Lee, W., et al. Multi-layered culture of human skin fibroblasts and keratinocytes through three-dimensional freeform fabrication. Biomaterials 30:1587–1595, 2009.
Lee, W., et al. On-demand three-dimensional freeform fabrication of multi-layered hydrogel scaffold with fluidic channels. Biotechnol. Bioeng. 105:1178–1186, 2010.
Lee, V. K., et al. Design and fabrication of human skin by 3D bioprinting. Tissue Eng. Part C 20:473–484, 2014.
Leong, M. F., et al. Patterned prevascularised tissue constructs by assembly of polyelectrolyte hydrogel fibres. Nat. Commun. 4:2353, 2013.
Li, Y.-S. J., J. H. Haga, and S. Chien. Molecular basis of the effects of shear stress on vascular endothelial cells. J. Biomech. 38:1949–1971, 2005.
Liu Tsang, V., et al. Fabrication of 3D hepatic tissues by additive photopatterning of cellular hydrogels. FASEB J. 21:790–801, 2007.
Miller, J. S., et al. Rapid casting of patterned vascular networks for perfusable engineered three-dimensional tissues. Nat. Mater. 11:768–774, 2012.
Mironov, V., R. P. Visconti, V. Kasyanov, G. Forgacs, C. J. Drake, and R. R. Markwald. Organ printing: tissue spheroids as building blocks. Biomaterials 30:2164–2174, 2009.
Moya, M. L., Y. Hsu, A. P. Lee, C. C. W. Hughes, and S. C. George. In vitro perfused human capillary networks. Tissue Eng. Part C 19:730–737, 2013.
Nahmias, Y., R. E. Schwartz, C. M. Verfaillie, and D. J. Odde. Laser-guided direct writing for three-dimensional tissue engineering. Biotechnol. Bioeng. 92:129–136, 2005.
Nakatsu, M. N., and C. C. W. Hughes. An optimized three-dimensional in vitro model for the analysis of angiogenesis. Methods Enzymol. 443:65–82, 2008.
Nakatsu, M. N., et al. Angiogenic sprouting and capillary lumen formation modeled by human umbilical vein endothelial cells (HUVEC) in fibrin gels: the role of fibroblasts and Angiopoietin-1. Microvasc. Res. 66:102–112, 2003.
Nguyen, D.-H. T., et al. Biomimetic model to reconstitute angiogenic sprouting morphogenesis in vitro. Proc. Natl. Acad. Sci. USA 110:6712–6717, 2013.
Ozturk, M. S., V. K. Lee, L. Zhao, G. Dai, and X. Intes. Mesoscopic fluorescence molecular tomography of reporter genes in bioprinted thick tissue. J. Biomed. Opt. 18:100501, 2013.
Potente, M., H. Gerhardt, and P. Carmeliet. Basic and therapeutic aspects of angiogenesis. Cell 146:873–887, 2011.
Price, G. M., and J. Tien. Chapter 17: methods for forming human microvascular tubes in vitro and measuring their macromolecular permeability. In: Biological Microarrays: Methods and Protocols, Methods in Molecular Biology, edited by A. Khademhosseini, K.-Y. Suh, and M. Zourob. Totowa, NJ: Humana Press, 2011, pp. 281–293.
Raghavan, S., C. M. Nelson, J. D. Baranski, E. Lim, and C. S. Chen. Geometrically controlled endothelial tubulogenesis in micropatterned gels. Tissue Eng. 16:2255–2263, 2010.
Roth, E. A., T. Xu, M. Das, C. Gregory, J. J. Hickman, and T. Boland. Inkjet printing for high-throughput cell patterning. Biomaterials 25:3707–3715, 2004.
Rouwkema, J., N. C. Rivron, and C. A. van Blitterswijk. Vascularization in tissue engineering. Trends Biotechnol. 26:434–441, 2008.
Saunders, W. B., et al. Coregulation of vascular tube stabilization by endothelial cell TIMP-2 and pericyte TIMP-3. J. Cell Biol. 175:179–191, 2006.
Sekine, H., et al. In vitro fabrication of functional three-dimensional tissues with perfusable blood vessels. Nat. Commun. 4:1399, 2013.
Shin, Y., et al. In vitro 3D collective sprouting angiogenesis under orchestrated ANG-1 and VEGF gradients. Lab Chip 11:2175–2181, 2011.
Stratman, A. N., K. M. Malotte, R. D. Mahan, M. J. Davis, and G. E. Davis. Pericyte recruitment during vasculogenic tube assembly stimulates endothelial basement membrane matrix formation. Blood 114:5091–5101, 2009.
Wong, K. H. K., J. M. Chan, R. D. Kamm, and J. Tien. Microfluidic models of vascular functions. Annu. Rev. Biomed. Eng. 14:205–230, 2012.
Xu, T., J. Jin, C. Gregory, J. J. J. J. Hickman, and T. Boland. Inkjet printing of viable mammalian cells. Biomaterials 26:93–99, 2005.
Yancopoulos, G. D., S. Davis, N. W. Gale, J. S. Rudge, S. J. Wiegand, and J. Holash. Vascular-specific growth factors and blood vessel formation. Nature 14:407, 2000.
Zhao, L., V. K. Lee, S–. S. Yoo, G. Dai, and X. Intes. The integration of 3-D cell printing and mesoscopic fluorescence molecular tomography of vascular constructs within thick hydrogel scaffolds. Biomaterials 33:5325–5332, 2012.
Zheng, Y., et al. In vitro microvessels for the study of angiogenesis and thrombosis. Proc. Natl. Acad. Sci. USA 109:9342–9347, 2012.
Acknowledgments
This work was supported by NIHR01HL118245, NSF CBET-1263455, CBET-1350240 and New York Capital Region Research Alliance grant.
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Vivian K. Lee, Alison M. Lanzi, Haygan, Ngo, Seung-SchikYoo, Peter A. Vincent, Guohao Dai declare that they have no conflicts of interest.
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No human studies were carried out by the authors for this article. No animal studies were carried out by the authors for this article.
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Associate Editor Anubhav Tripathi oversaw the review of this article.
This article has been designated as a 2013 BMES Outstanding Contribution.
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Lee, V.K., Lanzi, A.M., Ngo, H. et al. Generation of Multi-scale Vascular Network System Within 3D Hydrogel Using 3D Bio-printing Technology. Cel. Mol. Bioeng. 7, 460–472 (2014). https://doi.org/10.1007/s12195-014-0340-0
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DOI: https://doi.org/10.1007/s12195-014-0340-0