Preclinical study of SZ2080 material 3D microstructured scaffolds for cartilage tissue engineering made by femtosecond direct laser writing lithography

Over the last decade DLW employing ultrafast pulsed lasers has become a well-established technique for the creation of custom-made free-form three-dimensional (3D) microscaffolds out of a variety of materials ranging from proteins to biocompatible glasses. Its potential applications for manufacturing a patient's specific scaffold seem unlimited in terms of spatial resolution and geometry complexity. However, despite few exceptions in which live cells or primitive organisms were encapsulated into a polymer matrix, no demonstration of an in vivo study case of scaffolds generated with the use of such a method was performed. Here, we report a preclinical study of 3D artificial microstructured scaffolds out of hybrid organic-inorganic (HOI) material SZ2080 fabricated using the DLW technique. The created 2.1 × 2.1 × 0.21 mm(3) membrane constructs are tested both in vitro by growing isolated allogeneic rabbit chondrocytes (Cho) and in vivo by implanting them into rabbit organisms for one, three and six months. An ex vivo histological examination shows that certain pore geometry and the pre-growing of Cho prior to implantation significantly improves the performance of the created 3D scaffolds. The achieved biocompatibility is comparable to the commercially available collagen membranes. The successful outcome of this study supports the idea that hexagonal-pore-shaped HOI microstructured scaffolds in combination with Cho seeding may be successfully implemented for cartilage tissue engineering.

[1]  Marcel Zeelenberg,et al.  Data analysis 2 , 2016 .

[2]  R. Wyrwa,et al.  Two‐Photon Polymerization of Biocompatible Photopolymers for Microstructured 3D Biointerfaces , 2011 .

[3]  R. Salter,et al.  The potential for regeneration of articular cartilage in defects created by chondral shaving and subchondral abrasion. An experimental investigation in rabbits. , 1991, The Journal of bone and joint surgery. American volume.

[4]  F. Guilak,et al.  Advanced Material Strategies for Tissue Engineering Scaffolds , 2009, Advanced materials.

[5]  M J Glimcher,et al.  Cell origin and differentiation in the repair of full-thickness defects of articular cartilage. , 1993, The Journal of bone and joint surgery. American volume.

[6]  R Langer,et al.  Neocartilage formation in vitro and in vivo using cells cultured on synthetic biodegradable polymers. , 1993, Journal of biomedical materials research.

[7]  K. Sugioka,et al.  Ultrafast lasers—reliable tools for advanced materials processing , 2014, Light: Science & Applications.

[8]  H. Bursig,et al.  Chondrocyte suspension in fibrin glue , 2010, Cell and Tissue Banking.

[9]  R. Osellame,et al.  Integrated three-dimensional filter separates nanoscale from microscale elements in a microfluidic chip. , 2012, Lab on a chip.

[10]  James J. Yoo,et al.  Hybrid printing of mechanically and biologically improved constructs for cartilage tissue engineering applications , 2012, Biofabrication.

[11]  M. Hincke,et al.  Strategies for articular cartilage lesion repair and functional restoration. , 2010, Tissue engineering. Part B, Reviews.

[12]  Martin Wegener,et al.  Tailored 3D Mechanical Metamaterials Made by Dip‐in Direct‐Laser‐Writing Optical Lithography , 2012, Advanced materials.

[13]  E B Hunziker,et al.  Articular cartilage repair: basic science and clinical progress. A review of the current status and prospects. , 2002, Osteoarthritis and cartilage.

[14]  R. Salter,et al.  The chondrogenic potential of free autogenous periosteal grafts for biological resurfacing of major full-thickness defects in joint surfaces under the influence of continuous passive motion. An experimental investigation in the rabbit. , 1986, The Journal of bone and joint surgery. American volume.

[15]  R. Cancedda,et al.  Three-dimensional cultures of osteogenic and chondrogenic cells: a tissue engineering approach to mimic bone and cartilage in vitro. , 2009, European cells & materials.

[16]  H. J. Mankin,et al.  Instructional Course Lectures, The American Academy of Orthopaedic Surgeons - Articular Cartilage. Part I: Tissue Design and Chondrocyte-Matrix Interactions*† , 1997 .

[17]  T. Klar,et al.  Nano-anchors with single protein capacity produced with STED lithography. , 2013, Nano letters.

[18]  Gabriela A Silva,et al.  Natural-origin polymers as carriers and scaffolds for biomolecules and cell delivery in tissue engineering applications. , 2007, Advanced drug delivery reviews.

[19]  Saulius Juodkazis,et al.  Three-dimensional laser micro-sculpturing of silicone: towards bio-compatible scaffolds. , 2013, Optics express.

[20]  Š. Polák,et al.  Growth factors and chondrogenic differentiation of mesenchymal stem cells. , 2012, Tissue & cell.

[21]  A Ranella,et al.  Direct laser writing of 3D scaffolds for neural tissue engineering applications , 2011, Biofabrication.

[22]  Mangirdas Malinauskas,et al.  Fabrication of micro-tube arrays in photopolymer SZ2080 by using three different methods of a direct laser polymerization technique , 2012 .

[23]  Farshid Guilak,et al.  Tissue engineering for articular cartilage repair--the state of the art. , 2013, European cells & materials.

[24]  Roberto Osellame,et al.  Optimization of Femtosecond Laser Polymerized Structural Niches to Control Mesenchymal Stromal Cell Fate in Culture , 2014, Micromachines.

[25]  G. Collins,et al.  Challenges In Cartilage Tissue Engineering , 2013 .

[26]  Mangirdas Malinauskas,et al.  Micro-structured polymer scaffolds fabricated by direct laser writing for tissue engineering. , 2012, Journal of biomedical optics.

[27]  Milica Radisic,et al.  Biomaterial based cardiac tissue engineering and its applications , 2015, Biomedical materials.

[28]  Vilmos Vécsei,et al.  The influence of scaffold architecture on chondrocyte distribution and behavior in matrix-associated chondrocyte transplantation grafts. , 2011, Biomaterials.

[29]  K. Bohndorf,et al.  Injuries at the articulating surfaces of bone (chondral, osteochondral, subchondral fractures and osteochondrosis dissecans) , 1996, European journal of radiology.

[30]  B. Chichkov,et al.  Fabrication of microscale medical devices by two-photon polymerization with multiple foci via a spatial light modulator , 2011, Biomedical optics express.

[31]  Satoshi Kawata,et al.  Finer features for functional microdevices , 2001, Nature.

[32]  J. Verhaar,et al.  Multiplication of human chondrocytes with low seeding densities accelerates cell yield without losing redifferentiation capacity. , 2004, Tissue engineering.

[33]  Aleksandr Ovsianikov,et al.  Two‐photon polymerization technique for microfabrication of CAD‐designed 3D scaffolds from commercially available photosensitive materials , 2007, Journal of tissue engineering and regenerative medicine.

[34]  Maria Farsari,et al.  Direct laser writing , 2015 .

[35]  G. Lyons,et al.  Image-inspired 3D multiphoton excited fabrication of extracellular matrix structures by modulated raster scanning. , 2013, Optics express.

[36]  Saulius Juodkazis,et al.  3D Microporous Scaffolds Manufactured via Combination of Fused Filament Fabrication and Direct Laser Writing Ablation , 2014, Micromachines.

[37]  Eric Mazur,et al.  3D Cell‐Migration Studies using Two‐Photon Engineered Polymer Scaffolds , 2008 .

[38]  B. Feeley,et al.  Management of articular cartilage defects of the knee. , 2010, The Journal of bone and joint surgery. American volume.

[39]  L. Koch,et al.  Laser printing of cells into 3D scaffolds , 2010, Biofabrication.

[40]  S. Nicoll,et al.  The Effect of Serial Monolayer Passaging on the Collagen Expression Profile of Outer and Inner Anulus Fibrosus Cells , 2006, Spine.

[41]  A. Abramovici,et al.  Use of cultured embryonal chick epiphyseal chondrocytes as grafts for defects in chick articular cartilage. , 1987, Clinical orthopaedics and related research.

[42]  Jari Hyttinen,et al.  Direct laser writing and geometrical analysis of scaffolds with designed pore architecture for three-dimensional cell culturing , 2012 .

[43]  M. Kino‐oka,et al.  Comprehension of terminal differentiation and dedifferentiation of chondrocytes during passage cultures. , 2011, Journal of bioscience and bioengineering.

[44]  MironovVladimir,et al.  Direct laser writing , 2015 .

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

[46]  Dimitris Karalekas,et al.  Design, Fabrication and Computational Characterization of a 3D Micro-Valve Built by Multi-Photon Polymerization , 2014, Micromachines.

[47]  C. Fotakis,et al.  Ultra-low shrinkage hybrid photosensitive material for two-photon polymerization microfabrication. , 2008, ACS nano.

[48]  Kozo Nakamura,et al.  Screening of chondrogenic factors with a real-time fluorescence-monitoring cell line ATDC5-C2ER: identification of sorting nexin 19 as a novel factor. , 2009, Arthritis and rheumatism.

[49]  Sabrina Schlie-Wolter,et al.  Hyaluronic acid based materials for scaffolding via two-photon polymerization. , 2014, Biomacromolecules.

[50]  T. Mickevičius,et al.  Concomitant autologous chondrocyte implantation with osteochondral grafting for treatment of a massive osteochondral defect in the bilateral knees of a child. , 2011, Medicina.

[51]  Mangirdas Malinauskas,et al.  Nanophotonic lithography: a versatile tool for manufacturing functional three-dimensional micro-/nano-objects , 2012 .

[52]  M. Pitman,et al.  Chondrocyte transplantation using a collagen bilayer matrix for cartilage repair. , 1997, The Journal of bone and joint surgery. British volume.

[53]  Boris N. Chichkov,et al.  High-aspect 3D two-photon polymerization structuring with widened objective working range (WOW-2PP) , 2013, Light: Science & Applications.

[54]  M. Wegener,et al.  An elasto-mechanical unfeelability cloak made of pentamode metamaterials , 2014, Nature Communications.

[55]  Maria Farsari,et al.  The effect of porosity on cell ingrowth into accurately defined, laser-made, polylactide-based 3D scaffolds , 2015 .

[56]  E. Quenneville,et al.  Defects in articular cartilage metabolism and early arthritis in fibroblast growth factor receptor 3 deficient mice. , 2006, Human molecular genetics.

[57]  Aleksandr Ovsianikov,et al.  Laser photofabrication of cell-containing hydrogel constructs. , 2014, Langmuir : the ACS journal of surfaces and colloids.

[58]  Boris N. Chichkov,et al.  Materials processing: Two-photon fabrication , 2009 .

[59]  F. Mallein-Gerin,et al.  Enhanced hyaline cartilage matrix synthesis in collagen sponge scaffolds by using siRNA to stabilize chondrocytes phenotype cultured with bone morphogenetic protein-2 under hypoxia. , 2013, Tissue engineering. Part C, Methods.

[60]  Barbara Mazzolai,et al.  The Osteoprint: a bioinspired two-photon polymerized 3-D structure for the enhancement of bone-like cell differentiation. , 2014, Acta biomaterialia.

[61]  A. Giakoumaki,et al.  Tailor-made three-dimensional hybrid scaffolds for cell cultures , 2011, Biomedical materials.

[62]  A. Piskarskas,et al.  Ultrafast laser nanostructuring of photopolymers: a decade of advances , 2013 .

[63]  D. Eyre.  Articular cartilage and changes in Arthritis: Collagen of articular cartilage , 2001, Arthritis research.

[64]  C. Charitidis,et al.  Pre-osteoblastic cell response on three-dimensional, organic-inorganic hybrid material scaffolds for bone tissue engineering. , 2013, Journal of biomedical materials research. Part A.

[65]  A J Palfrey,et al.  Chondrocyte distribution in the articular cartilage of human femoral condyles. , 1988, Journal of anatomy.

[66]  R. Giardino,et al.  Transplantation of chondrocytes seeded on a hyaluronan derivative (hyaff-11) into cartilage defects in rabbits. , 2001, Biomaterials.

[67]  Jason B. Shear,et al.  High‐Resolution Patterning of Hydrogels in Three Dimensions using Direct‐Write Photofabrication for Cell Guidance , 2009 .

[68]  R. Schwartz,et al.  Repair of articular cartilage defects with collagen-chondrocyte allografts. , 1995, Tissue engineering.

[69]  P. Behrens,et al.  We do not have evidence based methods for the treatment of cartilage defects in the knee , 2011, Knee Surgery, Sports Traumatology, Arthroscopy.

[70]  E. Thonar,et al.  Adult human chondrocytes cultured in alginate form a matrix similar to native human articular cartilage. , 1996, The American journal of physiology.

[71]  C. V. van Blitterswijk,et al.  The role of three-dimensional polymeric scaffold configuration on the uniformity of connective tissue formation by adipose stromal cells. , 2010, Biomaterials.

[72]  Boris N. Chichkov,et al.  Three-Dimensional Cell Growth on Structures Fabricated from ORMOCER® by Two-Photon Polymerization Technique , 2007, Journal of biomaterials applications.

[73]  H J Mankin,et al.  Articular cartilage: tissue design and chondrocyte-matrix interactions. , 1998, Instructional course lectures.

[74]  Aleksandr Ovsianikov,et al.  Two-photon polymerization of microneedles for transdermal drug delivery , 2010, Expert opinion on drug delivery.

[75]  Mangirdas Malinauskas,et al.  In vitro and in vivo biocompatibility study on laser 3D microstructurable polymers , 2012 .

[76]  Benjamin Richter,et al.  Micro-engineered 3D scaffolds for cell culture studies. , 2012, Macromolecular bioscience.

[77]  M. Truppe,et al.  Changes in the ratio of type-I and type-II collagen expression during monolayer culture of human chondrocytes. , 2004, The Journal of bone and joint surgery. British volume.

[78]  Michael Sittinger,et al.  Treatment of posttraumatic and focal osteoarthritic cartilage defects of the knee with autologous polymer-based three-dimensional chondrocyte grafts: 2-year clinical results , 2007, Arthritis research & therapy.

[79]  Aleksandr Ovsianikov,et al.  Photo-sensitive hydrogels for three-dimensional laser microfabrication in the presence of whole organisms , 2012, Journal of biomedical optics.