An additive manufacturing-based PCL–alginate–chondrocyte bioprinted scaffold for cartilage tissue engineering
Joydip Kundu
Department of Mechanical Engineering, Pohang University of Science and Technology (POSTECH), Kyungbuk, South Korea
These authors contributed equally to this study.
Search for more papers by this authorJin-Hyung Shim
Department of Mechanical Engineering, Pohang University of Science and Technology (POSTECH), Kyungbuk, South Korea
These authors contributed equally to this study.
Search for more papers by this authorJinah Jang
Division of Integrative Biosciences and Biotechnology, Pohang University of Science and Technology (POSTECH), Kyungbuk, South Korea
Search for more papers by this authorSung-Won Kim
Department of Otolaryngology-Head and Neck Surgery, The Catholic University of Korea, College of Medicine, Seoul, Korea
Search for more papers by this authorCorresponding Author
Dong-Woo Cho
Department of Mechanical Engineering, Pohang University of Science and Technology (POSTECH), Kyungbuk, South Korea
Division of Integrative Biosciences and Biotechnology, Pohang University of Science and Technology (POSTECH), Kyungbuk, South Korea
Correspondence to: Dong-Woo Cho, Division of Biosciences and Biotechnology and Department of Mechanical Engineering, Pohang University of Science and Technology (POSTECH), San 31, Hyoja-dong, Nam-gu, Pohang, Kyungbuk, 790-784, South Korea. E-mail: [email protected]Search for more papers by this authorJoydip Kundu
Department of Mechanical Engineering, Pohang University of Science and Technology (POSTECH), Kyungbuk, South Korea
These authors contributed equally to this study.
Search for more papers by this authorJin-Hyung Shim
Department of Mechanical Engineering, Pohang University of Science and Technology (POSTECH), Kyungbuk, South Korea
These authors contributed equally to this study.
Search for more papers by this authorJinah Jang
Division of Integrative Biosciences and Biotechnology, Pohang University of Science and Technology (POSTECH), Kyungbuk, South Korea
Search for more papers by this authorSung-Won Kim
Department of Otolaryngology-Head and Neck Surgery, The Catholic University of Korea, College of Medicine, Seoul, Korea
Search for more papers by this authorCorresponding Author
Dong-Woo Cho
Department of Mechanical Engineering, Pohang University of Science and Technology (POSTECH), Kyungbuk, South Korea
Division of Integrative Biosciences and Biotechnology, Pohang University of Science and Technology (POSTECH), Kyungbuk, South Korea
Correspondence to: Dong-Woo Cho, Division of Biosciences and Biotechnology and Department of Mechanical Engineering, Pohang University of Science and Technology (POSTECH), San 31, Hyoja-dong, Nam-gu, Pohang, Kyungbuk, 790-784, South Korea. E-mail: [email protected]Search for more papers by this authorAbstract
Regenerative medicine is targeted to improve, restore or replace damaged tissues or organs using a combination of cells, materials and growth factors. Both tissue engineering and developmental biology currently deal with the process of tissue self-assembly and extracellular matrix (ECM) deposition. In this investigation, additive manufacturing (AM) with a multihead deposition system (MHDS) was used to fabricate three-dimensional (3D) cell-printed scaffolds using layer-by-layer (LBL) deposition of polycaprolactone (PCL) and chondrocyte cell-encapsulated alginate hydrogel. Appropriate cell dispensing conditions and optimum alginate concentrations for maintaining cell viability were determined. In vitro cell-based biochemical assays were performed to determine glycosaminoglycans (GAGs), DNA and total collagen contents from different PCL–alginate gel constructs. PCL–alginate gels containing transforming growth factor-β (TGFβ) showed higher ECM formation. The 3D cell-printed scaffolds of PCL–alginate gel were implanted in the dorsal subcutaneous spaces of female nude mice. Histochemical [Alcian blue and haematoxylin and eosin (H&E) staining] and immunohistochemical (type II collagen) analyses of the retrieved implants after 4 weeks revealed enhanced cartilage tissue and type II collagen fibril formation in the PCL–alginate gel (+TGFβ) hybrid scaffold. In conclusion, we present an innovative cell-printed scaffold for cartilage regeneration fabricated by an advanced bioprinting technology. Copyright © 2013 John Wiley & Sons, Ltd.
References
- Almqvist KF, Dhollander AA, Verdonk PC, et al. 2009; Treatment of cartilage defects in the knee using alginate beads containing human mature allogeneic chondrocytes. Am J Sports Med 37: 1920–1929.
- Baker SC, Rohman G, Southgate J, et al. 2009; The relationship between the mechanical properties and cell behavior on PLGA and PCL scaffolds for bladder tissue engineering. Biomaterials 30: 1321–1328.
- Barron JA, Ringeisen BR, Kim H, et al. 2004; Application of laser printing to mammalian cells. Thin Solid Films 453: 383–387.
- Boland T, Xu T, Damon B, et al. 2006; Application of inkjet printing to tissue engineering. Biotechnol J 1: 910–917.
- Elisseeff JH, Lee A, Kleinman HK, et al. 2002; Biological response of chondrocytes to hydrogels. Ann NY Acad Sci 961: 118–122.
- Fan H, Hu Y, Qin L, et al. 2006; Porous gelatin–chondroitin–hyaluronate tri-copolymer scaffolds containing microspheres loaded with TGF. J Biomed Mater Res A 77: 785–794.
- Farndale R, Buttle D, Barrett A. 1986; Improved quantitation and discrimination of sulphated glycosaminoglycans by the use of dimethylmethylene blue. Biochim Biophys Acta 883: 173–177.
- Fedorovich NE, Alblas J, Hennink WE, et al. 2011; Organ printing: the future of bone regeneration. Trend Biotechnol 29: 601–606.
- Guillermot F, Souquet A, Catros S, et al. 2010; High-throughput laser printing of cells and biomaterials for tissue engineering. Acta Biomater 6: 2494–2500.
- Haraguchi Y, Shimizu T, Yamato M, et al. 2011; Regenerative therapies using cell sheet-based tissue engineering for cardiac disease. Cardiol Res Pract 2011: 845170.
- Hauselmann HJ, Fernandes RJ, Mok SS, et al. 1994; Phenotypic stability of bovine articular chondrocytes after long-term culture in alginate beads. J Cell Sci 107: 17–27.
- Hoffman AS. 2001; Hydrogels for biomedical applications. Ann NY Acad Sci 944: 62–73.
- Hollister SJ. 2005; Porous scaffold design for tissue engineering. Nat Mater 4: 518–524.
- Hutmacher DW, Sittinger M, Risnud MV. 2004; Scaffold based tissue engineering: rationale for computer aided design and solid free form fabrication systems. Trends Biotechnol 22: 354–362.
- Hutmacher DW. 2001; Scaffold design and fabrication technologies for engineering tissues – state of the art and future perspectives. J Biomater Sci Polym Ed 12: 107–124.
- Jeon O, Bouhadir KH, Mansour JM, et al. 2009; Photocrosslinked alginate hydrogels with tunable biodegradation rates and mechanical properties. Biomaterials 30: 2724–2734.
- Jung JW, Kang HW, Kang TY, et al. 2012; Projection image-generation algorithm for fabrication of a complex structure using projection-based microstereolithography. Int J Precis Eng Manuf 13: 445–449.
- Kang S-W, Lee S-J, Kim J-S, et al. 2011; Effect of a scaffold fabricated thermally from acetylated PLGA on the formation of engineered cartilage. Macromol Biosci 11: 267–274.
- Khalil S, Sun W. 2009; Bioprinting endothelial cells with alginate for 3D tissue constructs. J Biomech Eng 131: 111002–111009.
- Kim TK, Sharma B, Williams CG, et al. 2003; Experimental model for cartilage tissue engineering to regenerate the zonal organization of articular cartilage. Osteoarthr Cartilage 11, 653–664.
- Kim Y, Sah RL, Doong JY, et al. 1988; Fluorometric assay of DNA in cartilage explants using Hoechst 33258. Anal Biochem 174: 168.
- Klein TJ, Malda J, Sah RL, et al. 2009; Tissue engineering of articular cartilage with biomimetic zones. Tissue Eng 15: 143–157.
- Langer R, Vacanti JP. 1993; Tissue engineering. Science 260: 920–926.
- Lee JS, Kim B, Kim MS, et al. 2006; The effect of 3D construction culture of human chondrocytes using alginate sponge. Key Eng Mat 326: 883–888.
- Lee J-W, Ahn G, Kim J-Y, et al. 2010a; Evaluating cell proliferation based on internal pore size and 3D scaffold architecture fabricated using solid freeform fabrication technology. J Mater Sci Mater M 21: 3195–3205.
- Lee J-W, Kang K-S, Lee SH, et al. 2011; Bone regeneration using a microstereolithography produced customized poly(propylene fumarate) diethyl fumarate photopolymer 3D scaffold incorporating BMP-2 loaded PLGA microspheres. Biomaterials 32: 744–752.
- Lee J-W, Kim J-Y, Cho D-W. 2010b; Solid free-form fabrication technology and its application to bone tissue engineering. Inter J Stem Cells 3: 85–93.
- Ma PX, Elisseeff JH. 2006; Scaffolding in Tissue Engineering. CRC Press: Boca Raton, FL, USA.
- Mironov V, Boland T, Trusk T, et al. 2003; Organ printing: computer-aided jet based 3D tissue engineering. Trends Biotechnol 21: 157–161.
- Mironov V, Prestwich G, Forgacs G. 2007; Bioprinting living structures. J Mater Chem 17: 2054–2060.
- Mironov V, Visconti RP, Kasyanov V, et al. 2009; Organ printing: tissue spheroids as building blocks. Biomaterials 30: 2164–2174.
- Mueller-Rath R, Gavénis K, Gravius S, et al. 2007; In vivo cultivation of human articular chondrocytes in a nude mouse-based contained defect organ culture model. Biomed Mater Eng 17: 357–366.
- Nair K, Gandhi M, Khalil S, et al. 2009; Characterization of cell viability during bioprinting processes. Biotechnol J 4: 1168–1177.
- Nakamura M, Nishiyama Y, Henmi C, et al. 2006; Inkjet bioprinting as an effective tool for tissue fabrication. Proc Digital Fabrication 1: 89–92.
- Peltola SM, Melchels FPW, Grijpma DW, et al. 2008; A review on the rapid prototyping techniques for tissue engineering scaffolds. Ann Med 40: 268–280.
- Peppas N, Hilt JZ, Khademhosseini A, et al. 2006; Hydrogels in biology and medicine. Adv Mater 18: 1–17.
- Pfister A, Landers R, Laib A, et al. 2004; Biofunctional rapid prototyping for tissue-engineering applications: 3D bioplotting versus 3D printing. J Polym Sci Polym Chem 42: 624–638.
- Rosen DW. 2007; Computer-aided design for additive manufacturing of cellular structures. Comput Aided Des Appl 4: 585–594.
10.1080/16864360.2007.10738493 Google Scholar
- Rouwkema J, Rivron NC, Blitterswijk CA. 2008; Vascularization in tissue engineering. Trends Biotechnol 26: 434–441.
- Sanders JE, Goldstein BS. 2001; Collagen fibril diameters increase and fibril densities decrease in skin subjected to repetitive compressive and shear stresses. J Biomech 34: 1581–1587.
- Schuurman W, Khristov V, Pot MW, et al. 2011; Bioprinting of hybrid tissue constructs with tailorable mechanical properties. Biofabrication 3: 021001–021008.
- Shim J-H, Lee J-S, Kim JY, et al. 2012; Bioprinting of a mechanically enhanced three-dimensional dual cell-laden construct for osteochondral tissue engineering using a multi-head tissue/organ building system. J Micromech Microeng 22: 085014–085024.
- Shim JH, Kim JY, Park M, et al. 2011; Development of a hybrid scaffold with synthetic biomaterials and hydrogel using solid freeform fabrication technology. Biofabrication 3: 034102–034111.
- Shirazi R, Shirazi-Adl A, Hurtig M. 2008; Role of cartilage collagen fibrils networks in knee joint biomechanics under compression. J Biomech 41: 3340–3348.
- Silva MMCG, Cyster LA, Barry JJ, et al. 2006; The effect of anisotropic architecture on cell and tissue infiltration into tissue engineered scaffolds. Biomaterials 27: 5909–5917.
- Simpson RL, Wiria FE, Amis AA, et al. 2008; Development of a 95/5 poly(l-lactide-co-glycolide)/hydroxyapatite and β-tricalciumphosphate scaffold as bone replacement material via selective laser sintering. J Biomed Mater Res B App Biomat 84: 17–25.
- Wiria FE, Yong JMS, Lim PN, et al. 2010; Printing of titanium implant prototype. Mater Design 31: S101–S105.
- Xu W, Ma J, Jabbari E. 2010; Material properties and osteogenic differentiation of marrow stromal cells on fiber-reinforced laminated hydrogel nanocomposites. Acta Biomater 6: 1992–2002.
