Cell-Laden Gelatin Methacryloyl Bioink for the Fabrication of Z-Stacked Hydrogel Scaffolds for Tissue Engineering
Abstract
:1. Introduction
2. Materials and Methods
2.1. Materials
2.2. Cell Culture
2.3. Gelatin Methacrylate Synthesis
2.4. Preparation and Fabrication of Bioink
2.5. 1H nuclear Magnetic Resonance (1H NMR) Spectroscopy
2.6. Photorheological Analysis
2.7. Printing Accuracy
2.8. Scanning Electron Microscopy (SEM)
2.9. Mechanical Testing
2.10. Cell Adhesion
2.11. Live/Dead Fluorescence Assay
2.12. Cell Proliferation Assay
2.13. Statistical Analysis
3. Results and Discussion
3.1. 1H Nuclear Magnetic Resonance Spectroscopy
3.2. Z-Stacking Strategy for Complex GelMA Scaffolds
3.3. Printing Accuracy Analysis
3.4. Scanning Electron Microscopy (SEM)
3.5. Mechanical Properties
3.6. Cell Adhesion in Z-Stacked Scaffolds
3.7. Cell Viability
3.8. Cell Proliferation Analysis
3.9. Ear-Shaped Z-Stacked Scaffold Printing and Cell Culture
4. Conclusions
Supplementary Materials
Author Contributions
Funding
Conflicts of Interest
References
- Lee, K.Y.; Mooney, D.J. Hydrogels for Tissue Engineering. Chem. Rev. 2001, 101, 1869–1880. [Google Scholar] [CrossRef] [PubMed]
- Hubbell, J.A. Biomaterials in Tissue Engineering. Nat. Biotechnol. 1995, 13, 565–576. [Google Scholar] [CrossRef] [PubMed]
- O’Brien, F.J. Biomaterials & scaffolds for tissue engineering. Mater. Today 2011, 14, 88–95. [Google Scholar] [CrossRef]
- Khademhosseini, A.; Vacanti, J.P.; Langer, R. Progress in Tissue Engineering. Sci. Am. 2009, 300, 64–71. [Google Scholar] [CrossRef]
- Guvendiren, M.; Lu, H.D.; Burdick, J.A. Shear-thinning hydrogels for biomedical applications. Soft Matter 2011, 8, 260–272. [Google Scholar] [CrossRef]
- Stanton, M.M.; Samitier, J.; Sánchez, S. Bioprinting of 3D hydrogels. Lab Chip 2015, 15, 3111–3115. [Google Scholar] [CrossRef] [Green Version]
- Gungor-Ozkerim, P.S.; Inci, I.; Zhang, Y.S.; Khademhosseini, A.; Dokmeci, M.R. Bioinks for 3D bioprinting: An overview. Biomater. Sci. 2018, 6, 915–946. [Google Scholar] [CrossRef] [Green Version]
- Gopinathan, J.; Noh, I. Recent trends in bioinks for 3D printing. Biomater. Res. 2018, 22, 1–15. [Google Scholar] [CrossRef] [Green Version]
- Liu, W.; Zhang, Y.S.; Heinrich, M.A.; De Ferrari, F.; Jang, H.L.; Bakht, S.M.; Alvarez, M.M.; Yang, J.; Li, Y.-C.; Santiago, G.T.-D.; et al. Rapid Continuous Multimaterial Extrusion Bioprinting. Adv. Mater. 2017, 29, 1604630. [Google Scholar] [CrossRef]
- Ozbolat, I.T.; Hospodiuk, M. Current advances and future perspectives in extrusion-based bioprinting. Biomaterials 2016, 76, 321–343. [Google Scholar] [CrossRef] [Green Version]
- Yue, K.; Santiago, G.T.-D.; Alvarez, M.M.; Tamayol, A.; Annabi, N.; Khademhosseini, A. Synthesis, properties, and biomedical applications of gelatin methacryloyl (GelMA) hydrogels. Biomaterials 2015, 73, 254–271. [Google Scholar] [CrossRef] [PubMed] [Green Version]
- Nie, J.; Gao, Q.; Wang, Y.; Zeng, J.; Zhao, H.; Sun, Y.; Shen, J.; Ramezani, H.; Fu, Z.; Liu, Z.; et al. Vessel-on-a-chip with Hydrogel-based Microfluidics. Small 2018, 14, e1802368. [Google Scholar] [CrossRef] [PubMed]
- Nichol, J.W.; Koshy, S.T.; Bae, H.; Hwang, C.M.; Yamanlar, S.; Khademhosseini, A. Cell-laden microengineered gelatin methacrylate hydrogels. Biomaterials 2010, 31, 5536–5544. [Google Scholar] [CrossRef] [PubMed] [Green Version]
- Han, M.-E.; Kang, B.-J.; Kim, S.-H.; Kim, H.D.; Hwang, N.S. Gelatin-based extracellular matrix cryogels for cartilage tissue engineering. J. Ind. Eng. Chem. 2017, 45, 421–429. [Google Scholar] [CrossRef]
- Tabata, Y. Protein release from gelatin matrices. Adv. Drug Deliv. Rev. 1998, 31, 287–301. [Google Scholar] [CrossRef]
- Gornall, J.L.; Terentjev, E.M. Helix–coil transition of gelatin: Helical morphology and stability. Soft Matter 2008, 4, 544–549. [Google Scholar] [CrossRef]
- Xu, W.; Molino, B.Z.; Cheng, F.; Molino, P.; Yue, Z.; Su, D.; Wang, X.; Willför, S.; Xu, C.; Wallace, G.G. On Low-Concentration Inks Formulated by Nanocellulose Assisted with Gelatin Methacrylate (GelMA) for 3D Printing toward Wound Healing Application. ACS Appl. Mater. Interfaces 2019, 11, 8838–8848. [Google Scholar] [CrossRef] [Green Version]
- Sun, Y.; Yu, K.; Nie, J.; Sun, M.; Fu, J.; Wang, H.; He, Y. Modeling the printability of photocuring and strength adjustable hydrogel bioink during projection based 3D bioprinting. Biofabrication 2020. [Google Scholar] [CrossRef]
- Bulcke, A.I.V.D.; Bogdanov, B.; De Rooze, N.; Schacht, E.H.; Cornelissen, M.; Berghmans, H. Structural and Rheological Properties of Methacrylamide Modified Gelatin Hydrogels. Biomacromolecules 2000, 1, 31–38. [Google Scholar] [CrossRef]
- Melchels, F.P.; Feijen, J.; Grijpma, D.W. A poly(d,l-lactide) resin for the preparation of tissue engineering scaffolds by stereolithography. Biomaterials 2009, 30, 3801–3809. [Google Scholar] [CrossRef] [Green Version]
- Kolb, C.; Lindemann, N.; Wolter, H.; Sextl, G. 3D-printing of highly translucent ORMOCER ®-based resin using light absorber for high dimensional accuracy. J. Appl. Polym. Sci. 2020, 49691. [Google Scholar] [CrossRef]
- Ge, L.; Dong, L.; Wang, D.; Ge, Q.; Gu, G. A digital light processing 3D printer for fast and high-precision fabrication of soft pneumatic actuators. Sens. Actuators A Phys. 2018, 273, 285–292. [Google Scholar] [CrossRef]
- Ding, R.; Du, Y.; Goncalves, R.B.; Francis, L.F.; Reineke, T.M. Sustainable near UV-curable acrylates based on natural phenolics for stereolithography 3D printing. Polym. Chem. 2019, 10, 1067–1077. [Google Scholar] [CrossRef]
- Burdick, J.A.; Vunjak-Novakovic, G. Engineered Microenvironments for Controlled Stem Cell Differentiation. Tissue Eng. Part A 2009, 15, 205–219. [Google Scholar] [CrossRef] [PubMed]
- Engler, A.J.; Sen, S.; Sweeney, H.L.; Discher, D.E. Matrix Elasticity Directs Stem Cell Lineage Specification. Cell 2006, 126, 677–689. [Google Scholar] [CrossRef] [Green Version]
- Loh, Q.L.; Choong, C. Three-Dimensional Scaffolds for Tissue Engineering Applications: Role of Porosity and Pore Size. Tissue Eng. Part B Rev. 2013, 19, 485–502. [Google Scholar] [CrossRef] [Green Version]
- Liu, J.; Li, L.; Suo, H.; Yan, M.; Yin, J.; Fu, J. 3D printing of biomimetic multi-layered GelMA/nHA scaffold for osteochondral defect repair. Mater. Des. 2019, 171, 107708. [Google Scholar] [CrossRef]
- Muhammad, R.; Sarah, W.C.; Patricia, A.C.; Thomas, L.W.; Evelyn, K.F.Y. Effect of sterilization treatment on mechanical properties, biodegradation, bioactivity and printability of GelMA hydrogels. Biomed. Mater. 2020, 15, 065017. [Google Scholar] [CrossRef]
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Seo, J.W.; Moon, J.H.; Jang, G.; Jung, W.K.; Park, Y.H.; Park, K.T.; Shin, S.R.; Hwang, Y.-S.; Bae, H. Cell-Laden Gelatin Methacryloyl Bioink for the Fabrication of Z-Stacked Hydrogel Scaffolds for Tissue Engineering. Polymers 2020, 12, 3027. https://doi.org/10.3390/polym12123027
Seo JW, Moon JH, Jang G, Jung WK, Park YH, Park KT, Shin SR, Hwang Y-S, Bae H. Cell-Laden Gelatin Methacryloyl Bioink for the Fabrication of Z-Stacked Hydrogel Scaffolds for Tissue Engineering. Polymers. 2020; 12(12):3027. https://doi.org/10.3390/polym12123027
Chicago/Turabian StyleSeo, Jeong Wook, Joon Ho Moon, Goo Jang, Woo Kyung Jung, Yong Ho Park, Kun Taek Park, Su Ryon Shin, Yu-Shik Hwang, and Hojae Bae. 2020. "Cell-Laden Gelatin Methacryloyl Bioink for the Fabrication of Z-Stacked Hydrogel Scaffolds for Tissue Engineering" Polymers 12, no. 12: 3027. https://doi.org/10.3390/polym12123027