Osteosarcoma growth on trabecular bone mimicking structures manufactured via laser direct write

² Biomaterials and Tissue Engineering Group, Department of Materials Science and Engineering, Kroto Research Institute, University of Sheffield, Sheffield S3 7HQ, United Kingdom

IJB 2016, 2(2), 67–77; https://doi.org/10.18063/IJB.2016.02.005

© Invalid date by the Author(s). This article is an open access article distributed under the terms and conditions of the Creative Commons Attribution 4.0 International License ( https://creativecommons.org/licenses/by/4.0/ )

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Abstract

This paper describes the direct laser write of a photocurable acrylate-based PolyHIPE (High Internal Phase Emulsion) to produce scaffolds with both macro- and microporosity, and the use of these scaffolds in osteosarcoma-based 3D cell culture. The macroporosity was introduced via the application of stereolithography to produce a classical “woodpile” structure with struts having an approximate diameter of 200 μm and pores were typically around 500 μm in diameter. The PolyHIPE retained its microporosity after stereolithographic manufacture, with a range of pore sizes typically between 10 and 60 μm (with most pores between 20 and 30 μm). The resulting scaffolds were suitable substrates for further modification using acrylic acid plasma polymerisation. This scaffold was used as a structural mimic of the trabecular bone and in vitro determination of biocompatibility using cultured bone cells (MG63) demonstrated that cells were able to colonise all materials tested, with evidence that acrylic acid plasma polymerisation improved biocompatibility in the long term. The osteosarcoma cell culture on the 3D printed scaffold exhibits different growth behaviour than observed on tissue culture plastic or a flat disk of the porous material; tumour spheroids are observed on parts of the scaffolds. The growth of these spheroids indicates that the osteosarcoma behave more akin to in vivo in this 3D mimic of trabecular bone. It was concluded that PolyHIPEs represent versatile biomaterial systems with considerable potential for the manufacture of complex devices or scaffolds for regenerative medicine. In particular, the possibility to readily mimic the hierarchical structure of native tissue enables opportunities to build in vitro models closely resembling tumour tissue.

Keywords

High Internal Phase Emulsion

PolyHIPEs

scaffold

emulsion templating

photopolymerisation

bone cells

MG63

References

1. Bokhari M, Carnachan R J, Przyborski S A, et al., 2007, Emulsion-templated porous polymers as scaffolds for three dimensional cell culture: effect of synthesis parameters on scaffold formation and homogeneity. Journal of Materials Chemistry, vol.17(38): 4088–4094. http://dx.doi.org/10.1039/B707499A
2. Kimmins S D and Cameron N R, 2011, Functional porous polymers by emulsion templating: recent advances. Advanced Functional Materials, vol.21(2): 211–225. http://dx.doi.org/10.1002/adfm.201001330
3. Barbetta A and Cameron N R, 2004, Morphology and surface area of emulsion-derived (PolyHIPE) solid foams prepared with oil-phase soluble porogenic solvents: span 80 as surfactant. Macromolecules, vol.37(9): 3188–3201. http://dx.doi.org/10.1021/ma0359436
4. Kimmins S D, Wyman P and Cameron N R, 2012, Photopolymerised methacrylate-based emulsion-templated porous polymers. Reactive and Functional Polymers, vol.72(12): 947–954. http://dx.doi.org/10.1016/j.reactfunctpolym.2012.06.015
5. Foudazi R, Gokun P, Feke D L, et al., 2013, Chemorheology of Poly(high internal phase emulsions). Macromolecules, vol.46(13): 5393–5396. http://dx.doi.org/10.1021/ma401157b
6. Gitli T and Silverstein M S, 2008, Bicontinuous hydrogel-hydrophobic polymer systems through emulsion templated simultaneous polymerizations. Soft Matter, vol.4(12): 2475–2485. http://dx.doi.org/10.1039/B809346F
7. Moghbeli M R and Shahabi M, 2011, Morphology and mechanical properties of an elastomeric poly(HIPE) nanocomposite foam prepared via an emulsion template. Iranian Polymer Journal, vol.20(5): 343–355.
8. Cummins D, Wyman P, Duxbury C J, et al., 2007, Synthesis of functional photopolymerized macroporous polyHIPEs by atom transfer radical polymerization surface grafting. Chemistry of Materials, vol.19(22): 5285–5292. http://dx.doi.org/10.1021/cm071511o
9. Hayward A S, Sano N, Przyborski S A, et al. 2013, Acrylic-acid-functionalized PolyHIPE scaffolds for use in 3D cell culture. Macromolecular Rapid Communications, vol.34(23–24): 1844–1849. http://dx.doi.org/10.1002/marc.201300709
10. Pierre S J, Thies J C, Dureault A, et al., 2006, Covalent enzyme immobilization onto photopolymerized highly porous monoliths. Advanced Materials, vol.18(14): 1822–1826. http://dx.doi.org/10.1002/adma.200600293
11. Sušec M, Ligon S C, Stampfl J, et al., 2013, Hierarchically porous materials from layer-by-layer photopolymerization of high internal phase emulsions. Macromolecular Rapid Communications, vol.34(11): 938–943. http://dx.doi.org/10.1002/marc.201300016
12. Caldwell S, Johnson D W, Didsbury M P, et al., 2012, Degradable emulsion-templated scaffolds for tissue engineering from thiol-ene photopolymerisation. Soft Matter, vol. 8(40): 10344–10351. http://dx.doi.org/10.1039/C2SM26250A
13. Johnson D W, Sherborne C, Didsbury M P, et al., 2013, Macrostructuring of emulsion-templated porous polymers by 3D laser patterning. Advanced Materials, vol.25(23): 3178–3181. http://dx.doi.org/10.1002/adma.201300552
14. Lovelady E, Kimmins S D, Wu J, et al., 2011, Preparation of emulsion-templated porous polymers using thiol-ene and thiol-yne chemistry. Polymer Chemistry, vol.2(3): 559–562. http://dx.doi.org/10.1039/C0PY00374C
15. Huš S and Krajnc P, 2014, PolyHIPEs from methyl methacrylate: hierarchically structured microcellular polymers with exceptional mechanical properties. Polymer, vol.55(17): 4420–4424. http://dx.doi.org/10.1016/j.polymer.2014.07.007
16. Causa F, Netti P A and Ambrosio L, 2007, A multi-functional scaffold for tissue regeneration: the need to engineer a tissue analogue. Biomaterials, vol.28(34): 5093–5099. http://dx.doi.org/10.1016/j.biomaterials.2007.07.030
17. Griffith L G and Swartz M A, 2006, Capturing complex 3D tissue physiology in vitro. Nature Reviews Molecular Cell Biology, vol.7: 211–224. http://dx.doi.org/10.1038/nrm1858
18. Dalton P D, Vaquette C, Farrugia B L, et al., 2013, Electrospinning and additive manufacturing: converging technologies. Biomaterials Science, vol.1: 171–185. http://dx.doi.org/10.1039/C2BM00039C
19. Ortega I, Ryan A J, Deshpande P, et al., 2013, Combined microfabrication and electrospinning to produce 3-D architectures for corneal repair. Acta Biomaterialia, vol.9(3): 5511–5520. http://dx.doi.org/10.1016/j.actbio.2012.10.039
20. Lee M, Dunn J C Y and Wu B M, 2005, Scaffold fabrication by indirect three-dimensional printing. Biomaterials, vol.26(20): 4281–4289. http://dx.doi.org/10.1016/j.biomaterials.2004.10.040
21. Mohanty S, Sanger K, Heiskanen A, et al., 2016, Fabrication of scalable tissue engineering scaffolds with dual-pore microarchitecture by combining 3D printing and particle leaching. Materials Science and Engineering: C, vol.61: 180–189. http://dx.doi.org/10.1016/j.msec.2015.12.032
22. Ortega I, Dew L, Kelly A G, et al., 2015, Fabrication of biodegradable synthetic perfusable vascular networks via a combination of electrospinning and robocasting. Biomaterials Science, vol.3(4): 592–596. http://dx.doi.org/10.1039/C4BM00418C
23. Jeffries E M, Nakamura S, Lee K W, et al., 2014, Micropatterning electrospun scaffolds to create intrinsic vascular networks. Macromolecular Bioscience, vol.14(11): 1514–1520. http://dx.doi.org/10.1002/mabi.201400306
24. Owen R, Sherborne C, Paterson T, et al., 2015, Emulsion templated scaffolds with tunable mechanical properties for bone tissue engineering. Journal of Mechanical Behavior of Biomedical Materials, vol.54: 159–172. http://dx.doi.org/10.1016/j.jmbbm.2015.09.019
25. Wang A, Paterson T, Owen R, et al., 2016, Photocurable high internal phase emulsions (HIPEs) containing hydroxyapatite for additive manufacture of tissue engineering scaffolds with multi-scale porosity. Materials Sci-ence and Engineering: C, vol.67: 51–58. http://dx.doi.org/10.1016/j.msec.2016.04.087
26. Lumelsky Y, Lalush-Michael I, Levenberg S, et al., 2009, A degradable, porous, emulsion-templated polyacrylate. Journal of Polymer Science Part A Polymer Chemistry, vol.47(24): 7043–7053. http://dx.doi.org/10.1002/pola.23744
27. Silverstein M S, 2014, Emulsion-templated porous polymers: a retrospective perspective. Polymer, vol.55(1): 304–320. http://dx.doi.org/10.1016/j.polymer.2013.08.068
28. Chen P Y and McKittrick J, 2011, Compressive mechanical properties of demineralized and deproteinized cancellous bone. Journal of Mechanical Behavior of Biomedical Materials, vol.4(7): 961–973. http://dx.doi.org/10.1016/j.jmbbm.2011.02.006
29. Santini M T, Rainaldi G, Romano R, et al., 2004, MG-63 human osteosarcoma cells grown in monolayer and as three-dimensional tumor spheroids present a different metabolic profile: a 1H NMR study. FEBS Letters, vol.557(1–3): 148–154. http://dx.doi.org/10.1016/S0014-5793(03)01466-2
30. Trojani C, Weiss P, Michiels J F, et al., 2005, Three-dimensional culture and differentiation of human osteogenic cells in an injectable hydroxypropylmethylcellulose hydrogel. Biomaterials, vol.26(27): 5509–5517. http://dx.doi.org/10.1016/j.biomaterials.2005.02.001
31. Kale S, Biermann S, Edwards C, et al., 2000, Three-dimensional cellular development is essential for ex vivo formation of human bone. Nature Biotechnology, vol.18: 954–958. http://dx.doi.org/10.1038/79439

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