Development of a 3D cell printed structure as an alternative to autologs cartilage for auricular reconstruction.

Surgical technique using autologs cartilage is considered as the best treatment for cartilage tissue reconstruction, although the burdens of donor site morbidity and surgical complications still remain. The purpose of this study is to apply three-dimensional (3D) cell printing to fabricate a tissue-engineered graft, and evaluate its effects on cartilage reconstruction. A multihead tissue/organ building system is used to print cell-printed scaffold (CPS), then assessed the effect of the CPS on cartilage regeneration in a rabbit ear. The cell viability and functionality of chondrocytes were significantly higher in CPS than in cell-seeded scaffold (CSS) and cell-seeded hybrid scaffold (CSHS) in vitro. CPS was then implanted into a rabbit ear that had an 8 mm-diameter cartilage defect; at 3 months after implantation the CPS had fostered complete cartilage regeneration whereas CSS and autologs cartilage (AC) fostered only incomplete healing. This result demonstrates that cell printing technology can provide an appropriate environment in which encapsulated chondrocytes can survive and differentiate into cartilage tissue in vivo. Moreover, the effects of CPS on cartilage regeneration were even better than those of AC. Therefore, we confirmed the feasibility of CPS as an alternative to AC for auricular reconstruction. © 2016 Wiley Periodicals, Inc. J Biomed Mater Res Part B: Appl Biomater, 105B: 1016-1028, 2017.

[1]  R. Siegert.  Combined reconstruction of congenital auricular atresia and severe microtia , 2003, The Laryngoscope.

[2]  J H Brekke,et al.  Regeneration of articular cartilage--evaluation of osteochondral defect repair in the rabbit using multiphasic implants. , 2005, Osteoarthritis and cartilage.

[3]  Dong-Woo Cho,et al.  Hybrid scaffold composed of hydrogel/3D-framework and its application as a dopamine delivery system. , 2014, Journal of controlled release : official journal of the Controlled Release Society.

[4]  R. Kandel,et al.  Integration of Tissue-engineered Cartilage With Host Cartilage: An In Vitro Model , 2011, Clinical orthopaedics and related research.

[5]  R. Reis,et al.  Cryopreservation of cell/scaffold tissue-engineered constructs. , 2012, Tissue engineering. Part C, Methods.

[6]  O. Muratoglu,et al.  Porous poly(vinyl alcohol)-alginate gel hybrid construct for neocartilage formation using human nasoseptal cells. , 2010, The Journal of surgical research.

[7]  Charles A Vacanti,et al.  Tissue Engineering of Autologous Cartilage for Craniofacial Reconstruction by Injection Molding , 2003, Plastic and reconstructive surgery.

[8]  M. Jolicoeur,et al.  Low calcium levels in serum-free media maintain chondrocyte phenotype in monolayer culture and reduce chondrocyte aggregation in suspension culture. , 2005, Osteoarthritis and cartilage.

[9]  S. Waldman,et al.  Growth Factor Stimulation Improves the Structure and Properties of Scaffold-Free Engineered Auricular Cartilage Constructs , 2014, PloS one.

[10]  Joachim Aigner,et al.  Alginate as a chondrocyte-delivery substance in combination with a non-woven scaffold for cartilage tissue engineering. , 2002, Biomaterials.

[11]  Erik K. Bassett,et al.  Engineering ear constructs with a composite scaffold to maintain dimensions. , 2011, Tissue engineering. Part A.

[12]  R. Spencer,et al.  Hyaline cartilage engineered by chondrocytes in pellet culture: histological, immunohistochemical and ultrastructural analysis in comparison with cartilage explants , 2004, Journal of anatomy.

[13]  Dong-Woo Cho,et al.  Three-dimensional printing of antibiotics-loaded poly-ε-caprolactone/poly(lactic-co-glycolic acid) scaffolds for treatment of chronic osteomyelitis , 2015, Tissue Engineering and Regenerative Medicine.

[14]  D. Cho,et al.  Bioprinting of a mechanically enhanced three-dimensional dual cell-laden construct for osteochondral tissue engineering using a multi-head tissue/organ building system , 2012 .

[15]  R. Bank,et al.  Synthesis of collagen by bovine chondrocytes cultured in alginate; posttranslational modifications and cell-matrix interaction. , 1997, Experimental cell research.

[16]  Dong-Woo Cho,et al.  A comparative study on collagen type I and hyaluronic acid dependent cell behavior for osteochondral tissue bioprinting , 2014, Biofabrication.

[17]  D. Cho,et al.  Biomimetic 3D tissue printing for soft tissue regeneration. , 2015, Biomaterials.

[18]  C. V. van Blitterswijk,et al.  Design of porous scaffolds for cartilage tissue engineering using a three-dimensional fiber-deposition technique. , 2004, Biomaterials.

[19]  B Brent,et al.  Technical advances in ear reconstruction with autogenous rib cartilage grafts: personal experience with 1200 cases. , 1999, Plastic and reconstructive surgery.

[20]  Woo Seob Kim,et al.  Endoscopic treatment of vesicoureteral reflux with a chondrocyte-alginate suspension. , 1994, The Journal of urology.

[21]  Charles A. Vacanti,et al.  Injectable cartilage. Discussion , 1995 .

[22]  B. Oakes,et al.  Effects of growth factors on cell proliferation and matrix synthesis of low‐density, primary bovine chondrocytes cultured in collagen I gels , 2002, Journal of orthopaedic research : official publication of the Orthopaedic Research Society.

[23]  J. Verhaar,et al.  Tissue-engineered cartilage using serially passaged articular chondrocytes. Chondrocytes in alginate, combined in vivo with a synthetic (E210) or biologic biodegradable carrier (DBM). , 2000, Biomaterials.

[24]  K. Leong,et al.  Scaffolding in tissue engineering: general approaches and tissue-specific considerations , 2008, European Spine Journal.

[25]  Peter Müller,et al.  Relationship between cell shape and type of collagen synthesised as chondrocytes lose their cartilage phenotype in culture , 1977, Nature.

[26]  Rajan S Uppal,et al.  Donor-Site Morbidity after Autologous Costal Cartilage Harvest in Ear Reconstruction and Approaches to Reducing Donor-Site Contour Deformity , 2008, Plastic and reconstructive surgery.

[27]  D J Mooney,et al.  Alginate hydrogels as synthetic extracellular matrix materials. , 1999, Biomaterials.

[28]  W. Davis,et al.  Absorption of autogenous cartilage grafts in man. , 1956, British journal of plastic surgery.

[29]  A. Atala,et al.  Engineered cartilage covered ear implants for auricular cartilage reconstruction. , 2011, Biomacromolecules.

[30]  D. Cho,et al.  3D printing of composite tissue with complex shape applied to ear regeneration , 2014, Biofabrication.

[31]  K. Arden,et al.  Expression of the human chondrocyte phenotype in vitro , 1989, In Vitro Cellular & Developmental Biology.

[32]  Michael C. McAlpine,et al.  3D Printed Bionic Ears , 2013, Nano letters.

[33]  P. Gillet,et al.  The effect of alginate, hyaluronate and hyaluronate derivatives biomaterials on synthesis of non-articular chondrocyte extracellular matrix , 2005, Journal of materials science. Materials in medicine.

[34]  C. Maas,et al.  Dorsal onlay cartilage autografts: comparing resorption in a rabbit model. , 2003, Archives of facial plastic surgery.

[35]  M Kon,et al.  Auricular reconstruction using biofabrication-based tissue engineering strategies , 2015, Biofabrication.

[36]  J. McPherson,et al.  Synergistic action of transforming growth factor-beta and insulin-like growth factor-I induces expression of type II collagen and aggrecan genes in adult human articular chondrocytes. , 1997, Experimental cell research.

[37]  J. Bonaventure,et al.  Reexpression of cartilage-specific genes by dedifferentiated human articular chondrocytes cultured in alginate beads. , 1994, Experimental cell research.

[38]  G. Vasios,et al.  Enhanced proliferation and differentiation of human articular chondrocytes when seeded at low cell densities in alginate In Vitro , 2000, Journal of orthopaedic research : official publication of the Orthopaedic Research Society.

[39]  P. Benya,et al.  Dedifferentiated chondrocytes reexpress the differentiated collagen phenotype when cultured in agarose gels , 1982, Cell.

[40]  M. Craigmyle.  Studies of cartilage autografts and homografts in the rabbit. , 1955, British journal of plastic surgery.

[41]  R. Eavey,et al.  Engineering Autogenous Cartilage in the Shape of a Helix Using an Injectable Hydrogel Scaffold , 2000, The Laryngoscope.

[42]  Junzo Tanaka,et al.  Rapid and Large-Scale Formation of Chondrocyte Aggregates by Rotational Culture , 2003, Cell transplantation.

[43]  E. Ippolito,et al.  Histochemical properties of cartilage proteoglycans. , 1983, The journal of histochemistry and cytochemistry : official journal of the Histochemistry Society.

[44]  Y. Jang,et al.  Outcomes following rhinoplasty using autologous costal cartilage. , 2012, Archives of facial plastic surgery.

[45]  Joseph P Vacanti,et al.  The tissue-engineered auricle: past, present, and future. , 2012, Tissue engineering. Part B, Reviews.

[46]  Dong-Woo Cho,et al.  An additive manufacturing‐based PCL–alginate–chondrocyte bioprinted scaffold for cartilage tissue engineering , 2015, Journal of tissue engineering and regenerative medicine.

[47]  James M. Anderson,et al.  Foreign body reaction to biomaterials. , 2008, Seminars in immunology.

[48]  Y. Qinghua,et al.  Technical innovations in ear reconstruction using a skin expander with autogenous cartilage grafts. , 2008, Journal of plastic, reconstructive & aesthetic surgery : JPRAS.

[49]  D. Cho,et al.  3D-Printed Drug/Cell Carrier Enabling Effective Release of Cyclosporin a for Xenogeneic Cell-Based Therapy , 2015, Cell transplantation.

[50]  Guangdong Zhou,et al.  Engineering ear-shaped cartilage using electrospun fibrous membranes of gelatin/polycaprolactone. , 2013, Biomaterials.

[51]  D. Hungerford,et al.  Human chondrocytes proliferate and produce matrix components in microcarrier suspension culture. , 1996, Biomaterials.

[52]  P. Gatenholm,et al.  3D Bioprinting Human Chondrocytes with Nanocellulose-Alginate Bioink for Cartilage Tissue Engineering Applications. , 2015, Biomacromolecules.

[53]  Andrés J. García,et al.  Chondrocyte phenotypes on different extracellular matrix monolayers. , 2004, Biomaterials.

[54]  W. Liu,et al.  Regeneration of human-ear-shaped cartilage by co-culturing human microtia chondrocytes with BMSCs. , 2014, Biomaterials.

[55]  Dong-Woo Cho,et al.  Bioprintable, cell-laden silk fibroin-gelatin hydrogel supporting multilineage differentiation of stem cells for fabrication of three-dimensional tissue constructs. , 2015, Acta biomaterialia.

[56]  Hwa-Chang Liu,et al.  Gelatin-chondroitin-hyaluronan tri-copolymer scaffold for cartilage tissue engineering. , 2003, Biomaterials.

[57]  J Malda,et al.  Bioprinting of hybrid tissue constructs with tailorable mechanical properties , 2011, Biofabrication.

[58]  B Brent,et al.  Auricular Repair with Autogenous Rib Cartilage Grafts: Two Decades of Experience with 600 Cases , 1992, Plastic and reconstructive surgery.

[59]  Jason A. Spector,et al.  High-Fidelity Tissue Engineering of Patient-Specific Auricles for Reconstruction of Pediatric Microtia and Other Auricular Deformities , 2013, PloS one.

[60]  M. Neumeister,et al.  Vascularized Tissue-Engineered Ears , 2006, Plastic and reconstructive surgery.

[61]  Anthony Atala,et al.  3D bioprinting of tissues and organs , 2014, Nature Biotechnology.

[62]  C. Archer,et al.  Cartilage integration: evaluation of the reasons for failure of integration during cartilage repair. A review. , 2008, European cells & materials.

[63]  R. Eavey,et al.  Tissue Engineering of a Human Sized And Shaped Auricle Using a Mold , 2004, The Laryngoscope.