A novel 3D printing method for cell alignment and differentiation
The application of bioprinting allows precision deposition of biological materials for bioengineering applications. Here we propose a 2 stage methodology for bioprinting using a back pressure-driven, automated robotic dispensing system. This apparatus can prepare topographic guidance features for cell orientation and then bioprint cells directly onto them. Topographic guidance features generate cues that influence adhered cell morphology and phenotype. The robotic dispensing system was modified to include a sharpened stylus that etched on a polystyrene surface. The same computer-aided design (CAD) software was used for both precision control of etching and bioink deposition. Various etched groove patterns such as linear, concentric circles, and sinusoidal wave patterns were possible. Fibroblasts and mesenchymal stem cells (MSC) were able to sense the grooves, as shown by their elongation and orientation in the direction of the features. The orientated MSCs displayed indications of lineage commitment as detected by fluorescence-activated cell sorting (FACS) analysis. A 2% gelatin bioink was then used to dispense cells onto the etched features using identical, programmed co-ordinates. The bioink allows the cells to contact sense the pattern while containing their deposition within the printed pattern.
1.Irvine SA, Agrawal A, Lee BH, et al. 2015, Printing cell-laden gelatin constructs by free-form fabrication and enzymatic protein crosslinking. Biomed Microdevices. 17(1):16. doi: 10.1007/s10544-014-9915-8
2. Murphy S V and Atala A, 2014, 3D bioprinting of tissues and organs. Nature Biotechnology, vol.32(8):773–785.
http://dx.doi.org/10.1038/nbt.2958
3. Khalil A S, Xie A W and Murphy W L, 2014, Contextclues: the importance of stem cell-material interactions.ACS Chemical Biology, vol.9(1): 45–56.http://dx.doi.org/10.1021/cb400801m.
4. Tay C Y, Irvine S A, Boey F Y C, et al. 2011, Micro-/nano-engineered cellular responses for soft tissue engineering and biomedical applications. Small, vol.7(10):1361–1378.http://dx.doi.org/10.1002/smll.201100046.
5. Subramony SD, Dargis BR, Castillo M, et al. 2013, The guidance of stem cell differentiation by substrate alignment and mechanical stimulation. Biomaterials. 34(8):1942-53 http://dx.doi.org/10.1016/j.biomaterials.2012.11.012
6. Mathieu P S and Loboa E G, 2012, Cytoskeletal and focal adhesion influences on mesenchymal stem cell shape, mechanical properties, and differentiation down osteogenic, adipogenic, and chondrogenic pathways.Tissue Engineering Part B: Reviews, vol.18(6): 436–444.
http://dx.doi.org/10.1089/ten.TEB.2012.0014.
7. Li Y H, Huang G Y, Zhang X H, et al. 2014, Engineering cell alignment in vitro. Biotechnology Advance,vol.32(2): 347–365.
http://dx.doi.org/10.1016/j.biotechadv.2013.11.007.
8. Chang S, Song S, Lee J, et al. 2014, Phenotypic modulation of primary vascular smooth muscle cells by short-term culture on micropatterned substrate. PLoS ONE, vol.9(2): e88089. http://dx.doi.org/10.1371/journal.pone.0088089.
9. Glawe J D, Hill J B, Mills D K, et al. 2005, Influence of channel width on alignment of smooth muscle cells by high-aspect-ratio microfabricated elastomeric cell culture scaffolds. Journal of Biomedical Materials Research Part A, vol.75A(1): 106–114. http://dx.doi.org/10.1002/jbm.a.30403.
10. Hao J, Zhang Y L, Jing D, et al. 2015, Mechanobiology of mesenchymal stem cells: perspective into the mechanically induced MSC fate. Acta Biomaterialia, vol.20(1): 1–9. http://dx.doi.org/10.1016/j.actbio.2015.04.008.
11. Marklein R A and Burdick J A, 2010, Controlling stem cell fate with material design. Advanced Materials, vol.22(2): 175–189.
http://dx.doi.org/10.1002/adma.200901055.
12. Gamboa J R, Mohandes S, Tran P L, et al. 2013, Linear fibroblast alignment on sinusoidal wave micropatterns. Colloids and Surface B: Biointerfaces, vol.104: 318–325. http://dx.doi.org/10.1016/j.colsurfb.2012.11.035.
13. Harrison R G, 1914, The reaction of embryonic cells to solid structures. Journal of Experimental Zoology,vol.17(4): 521–544.
http://dx.doi.org/10.1002/jez.1400170403.
14. Bourget J-M, Guillemette M, Veres T, et al. 2013, Alignment of cells and extracellular matrix within tissue- engineered substitutes, in Advances in Biomaterials Science and Biomedical Applications. InTech, Rijeka. http://dx.doi.org/10.5772/54142.
15. Kolind K, Leong K W, Besenbacher F, et al. 2012, Guidance of stem cell fate on 2D patterned surfaces. Biomaterials, vol.33(28): 6626–6633.
http://dx.doi.org/10.1016/j.biomaterials.2012.05.070.
16. Teh T K H, Toh S-L and Goh J C H, 2011, Aligned hy-brid silk scaffold for enhanced differentiation of mesenchymal stem cells into ligament fibroblasts. Tissue Engineering Part C: Methods, vol.17(6): 687–703.http://dx.doi.org/10.1089/ten.tec.2010.0513.
17. Jiang X, Cao H Q, Shi L Y, et al. 2012, Nanofiber topography and sustained biochemical signaling enhance human mesenchymal stem cell neural commitment. Acta Biomaterialia, vol.8(3): 1290–1302. http://dx.doi.org/10.1016/j.actbio.2011.11.019.
18. Li H Q, Wen F, Wong Y S, et al. 2012. Direct laser machining-induced topographic pattern promotes upregulation of myogenic markers in human mesenchymal stem cells. Acta Biomater, vol.8(2): 531–539. http://dx.doi.org/10.1016/j.actbio.2011.09.029.
19. Peterbauer T, Yakunin S, Siegel J, et al. 2011. Dynamics of spreading and alignment of cells cultured in vitro on a grooved polymer surface. Journal of Nanomaterials, vol.2011, 413079. http://dx.doi.org/10.1155/2011/413079.
20. Curtis A and Wilkinson C, 1997. Topographical control of cells. Biomaterials, vol.18(24): 1573–1583. http://dx.doi.org/10.1016/S0142-9612(97)00144-0.
21. Clark P, Connolly P, Curtis A S G, et al. 1990, Topographical control of cell behaviour: II. multiple grooved substrata. Development, vol.108(4): 635–644.
22. Fröhlich E, Bonstingl G, Höflerb A, et al. 2013, Comparison of two in vitro systems to assess cellular effects of nanoparticles-containing aerosols. Toxicology in Vitro, vol.27(1): 409–417. http://dx.doi.org/10.1016/j.tiv.2012.08.008.
23. Safaeijavan R, Soleimani M, Divsalar A, et al. 2014,Comparison of random and aligned PCL nanofibrous electrospun scaffolds on cardiomyocyte differentiation of human adipose-derived stem cells. Iranian Journal of Basic Medical Sciences, vol.17(11): 903–911.
24. Ma Z, Liu Q Y, Yang H X, et al. 2013, Laser patterning for the study of MSC cardiogenic differentiation at the single-cell level. Light: Science & Applications, vol.2:e68. http://dx.doi.org/10.1038/lsa.2013.24.
25. Cao Y, Poon Y F, Feng J, et al. 2010, Regulating orientation and phenotype of primary vascular smooth muscle cells by biodegradable films patterned with arrays ofmicrochannels and discontinuous microwalls. Biomaterials, vol.31(24): 6228–6238.
http://dx.doi.org/10.1016/j.biomaterials.2010.04.059
26. Anene-Nzelu C G, Peh K Y, Fraiszudeen A, et al. 2013,Scalable alignment of three-dimensional cellular constructs in a microfluidic chip. Lab on a Chip, vol.13(20):4124–4133.http://dx.doi.org/10.1039/c3lc50730k.
27. Zhou X T, Hu J, Li J J, et al. 2012, Patterning of two-level topographic cues for observation of competitive guidance of cell alignment. ACS Applied Materials & Interfaces, vol.4(8):3888–3892. http://dx.doi.org/10.1021/am301237j.
28. Hosseini V, Ahadian S, Ostrovidov S, et al. 2012, Engineered contractile skeletal muscle tissue on a microgrooved methacrylated gelatin substrate. Tissue Engineering Part A. vol.18(23–24): 2453–65. http://dx.doi.org/10.1089/ten.TEA.2012.0181.
29. Lu J, Rao M P, MacDonald N C, et al. 2008, Improved endothelial cell adhesion and proliferation on patterned titanium surfaces with rationally designed, micrometer to nanometer features. Acta Biomater, vol.4(1): 192–201. http://dx.doi.org/10.1016/j.actbio.2007.07.008.
30. Ma Z, Pirlo R K, Yun J X, et al. 2010, Laser guidance-based cell micropatterning, in Cell and Organ Printing. Springer Netherlands, Dordrecht, 137–159. http://dx.doi.org/10.1007/978-90-481-9145-1_8.
31. Smerup M, Nielsen E, Agger P, et al. 2009, The three-dimensional arrangement of the myocytes aggregated together within the mammalian ventricular myocardium, Anatomical Record, vol.292(1): 1–11. http://dx.doi.org/10.1002/ar.20798.
32. Pfister G, Stroh C M, Perschinka H, et al. 2005, Detection of HSP60 on the membrane surface of stressed human endothelial cells by atomic force and confocal microscopy, Journal of Cell Science, vol.118(8): 1587–1594. http://dx.doi.org/10.1242/jcs.02292.
33. DeSilva M N, 2010, Patterning cells on complex curved surface by precision spraying of polymers, in Cell and Organ Printing. Springer Netherlands, Dordrecht, 187–202. http://dx.doi.org/10.1007/978-90-481-9145-1_11.