Anatomical meniscus construct with zone specific biochemical composition and structural organization.

A PCL/hydrogel construct that would mimic the structural organization, biochemistry and anatomy of meniscus was engineered. The compressive (380 ± 40 kPa) and tensile modulus (18.2 ± 0.9 MPa) of the PCL scaffolds were increased significantly when constructs were printed with a shifted design and circumferential strands mimicking the collagen organization in native tissue (p < 0.05). Presence of circumferentially aligned PCL strands also led to elongation and alignment of the human fibrochondrocytes. Gene expression of the cells in agarose (Ag), gelatin methacrylate (GelMA), and GelMA-Ag hydrogels was significantly higher than that of cells on the PCL scaffolds after a 21-day culture. GelMA exhibited the highest level of collagen type I (COL1A2) mRNA expression, while GelMA-Ag exhibited the highest level of aggrecan (AGG) expression (p < 0.001, compared to PCL). GelMA and GelMA-Ag exhibited a high level of collagen type II (COL2A1) expression (p < 0.05, compared to PCL). Anatomical scaffolds with circumferential PCL strands were impregnated with cell-loaded GelMA in the periphery and GelMA-Ag in the inner region. GelMA and GelMA-Ag hydrogels enhanced the production of COL 1 and COL 2 proteins after a 6-week culture (p < 0.05). COL 1 expression increased gradually towards the outer periphery, while COL 2 expression decreased. We were thus able to engineer an anatomical meniscus with a cartilage-like inner region and fibrocartilage-like outer region.

[1]  B. Owens,et al.  Current Concepts in Meniscus Tissue Engineering and Repair , 2018, Advanced healthcare materials.

[2]  M. Dunn,et al.  Tissue-Engineered Total Meniscus Replacement With a Fiber-Reinforced Scaffold in a 2-Year Ovine Model , 2018, The American journal of sports medicine.

[3]  A. Adesida,et al.  Biomimetic 3D printed scaffolds for meniscus tissue engineering , 2017 .

[4]  P. Sharpe,et al.  Regulation and role of Sox9 in cartilage formation , 1999, Developmental dynamics : an official publication of the American Association of Anatomists.

[5]  Rohan A. Shirwaiker,et al.  Label free process monitoring of 3D bioprinted engineered constructs via dielectric impedance spectroscopy , 2018, Biofabrication.

[6]  H. Cheung,et al.  Distribution of type I, II, III and V in the pepsin solubilized collagens in bovine menisci. , 1987, Connective tissue research.

[7]  D. Kelly,et al.  A comparison of different bioinks for 3D bioprinting of fibrocartilage and hyaline cartilage , 2016, Biofabrication.

[8]  V. Hasırcı,et al.  Effects of microarchitecture and mechanical properties of 3D microporous PLLA-PLGA scaffolds on fibrochondrocyte and L929 fibroblast behavior , 2018, Biomedical materials.

[9]  S. Bryant,et al.  Heterogeneity is key to hydrogel-based cartilage tissue regeneration. , 2017, Soft matter.

[10]  F. Melchels,et al.  Focal adhesion signaling affects regeneration by human nucleus pulposus cells in collagen- but not carbohydrate-based hydrogels. , 2018, Acta biomaterialia.

[11]  Sanjay Kumar,et al.  Microscale mechanisms of agarose-induced disruption of collagen remodeling. , 2011, Biomaterials.

[12]  F. Mallein-Gerin,et al.  Type X collagen in rabbit and human meniscus. , 1999, Osteoarthritis and cartilage.

[13]  Gokhan Bahcecioglu,et al.  Hydrogels of agarose, and methacrylated gelatin and hyaluronic acid are more supportive for in vitro meniscus regeneration than three dimensional printed polycaprolactone scaffolds. , 2019, International journal of biological macromolecules.

[14]  J. Machan,et al.  Quantitative Magnetic Resonance Imaging Detects Changes in Meniscal Volume In Vivo After Partial Meniscectomy , 2010, The American journal of sports medicine.

[15]  U. Demirci,et al.  Quantification of Type, Timing, and Extent of Cell Body and Nucleus Deformations Caused by the Dimensions and Hydrophilicity of Square Prism Micropillars , 2016, Advanced healthcare materials.

[16]  J. Lotz,et al.  Chondrogenic differentiation of human mesenchymal stem cells in three-dimensional alginate gels. , 2008, Tissue engineering. Part A.

[17]  C. V. van Donkelaar,et al.  Polymers in Cartilage Defect Repair of the Knee: Current Status and Future Prospects , 2016, Polymers.

[18]  Wolf Petersen,et al.  Collagenous fibril texture of the human knee joint menisci , 1998, Anatomy and Embryology.

[19]  A. M. Ahmed,et al.  Tensile stress-strain characteristics of the human meniscal material. , 1995, Journal of biomechanics.

[20]  Zhiguo Yuan,et al.  Fabrication and characterization of electrospun nanofibers composed of decellularized meniscus extracellular matrix and polycaprolactone for meniscus tissue engineering. , 2017, Journal of materials chemistry. B.

[21]  G. Viano,et al.  Building the basis for patient-specific meniscal scaffolds: From human knee MRI to fabrication of 3D printed scaffolds , 2016 .

[22]  T. H. Haut Donahue,et al.  Dynamic compression of human and ovine meniscal tissue compared with a potential thermoplastic elastomer hydrogel replacement. , 2017, Journal of biomedical materials research. Part A.

[23]  D. Elliott,et al.  Microstructural heterogeneity directs micromechanics and mechanobiology in native and engineered fibrocartilage. , 2016, Nature materials.

[24]  Jos Malda,et al.  Reinforcement of hydrogels using three-dimensionally printed microfibres , 2015, Nature Communications.

[25]  A. Pennings,et al.  Porous implants for knee joint meniscus reconstruction: a preliminary study on the role of pore sizes in ingrowth and differentiation of fibrocartilage. , 1993, Clinical materials.

[26]  Marcia Simon,et al.  Hydrogels for Regenerative Medicine , 2016 .

[27]  Shaochen Chen,et al.  Digital micromirror device projection printing system for meniscus tissue engineering. , 2013, Acta biomaterialia.

[28]  S. Fan,et al.  The amelioration of cartilage degeneration by photo-crosslinked GelHA hydrogel and crizotinib encapsulated chitosan microspheres , 2017, Oncotarget.

[29]  Jeremy J. Mao,et al.  Protein-releasing polymeric scaffolds induce fibrochondrocytic differentiation of endogenous cells for knee meniscus regeneration in sheep , 2014, Science Translational Medicine.

[30]  P. Scott,et al.  Isolation and characterization of small proteoglycans from different zones of the porcine knee meniscus. , 1997, Biochimica et biophysica acta.

[31]  Heike Walles,et al.  Tissue Mimicry in Morphology and Composition Promotes Hierarchical Matrix Remodeling of Invading Stem Cells in Osteochondral and Meniscus Scaffolds , 2018, Advanced materials.

[32]  V. Hasırcı,et al.  Mimicking corneal stroma using keratocyte‐loaded photopolymerizable methacrylated gelatin hydrogels , 2018, Journal of tissue engineering and regenerative medicine.

[33]  Gokhan Bahcecioglu,et al.  A 3D printed PCL/hydrogel construct with zone-specific biochemical composition mimicking that of the meniscus , 2019, Biofabrication.

[34]  Cynthia A. Reinhart-King,et al.  3D Bioprinting of Spatially Heterogeneous Collagen Constructs for Cartilage Tissue Engineering. , 2016, ACS biomaterials science & engineering.

[35]  M. Heberer,et al.  Specific growth factors during the expansion and redifferentiation of adult human articular chondrocytes enhance chondrogenesis and cartilaginous tissue formation in vitro , 2001, Journal of cellular biochemistry.

[36]  D. Kaplan,et al.  Multilayered silk scaffolds for meniscus tissue engineering. , 2011, Biomaterials.

[37]  Brendon M. Baker,et al.  Dynamic tensile loading improves the functional properties of mesenchymal stem cell-laden nanofiber-based fibrocartilage. , 2011, Tissue engineering. Part A.

[38]  A. Khademhosseini,et al.  Hydrogels in Regenerative Medicine , 2009, Advanced materials.

[39]  Yunxiao Liu,et al.  A biomimetic hydrogel based on methacrylated dextran-graft-lysine and gelatin for 3D smooth muscle cell culture. , 2010, Biomaterials.

[40]  V. Hasırcı,et al.  A multilayer tissue engineered meniscus substitute , 2014, Journal of Materials Science: Materials in Medicine.

[41]  Jianxun Ding,et al.  Role of scaffold mean pore size in meniscus regeneration. , 2016, Acta biomaterialia.

[42]  M. Hull,et al.  Compressive moduli of the human medial meniscus in the axial and radial directions at equilibrium and at a physiological strain rate , 2008, Journal of orthopaedic research : official publication of the Orthopaedic Research Society.

[43]  I. Cengiz,et al.  Building the Basis for Patient-Specific Meniscal Scaffolds , 2017 .

[44]  P. Roughley,et al.  Limitations of using aggrecan and type X collagen as markers of chondrogenesis in mesenchymal stem cell differentiation , 2006, Journal of orthopaedic research : official publication of the Orthopaedic Research Society.

[45]  Jian-Xun Ding,et al.  3D-Printed Poly(ε-caprolactone) Scaffold Augmented With Mesenchymal Stem Cells for Total Meniscal Substitution: A 12- and 24-Week Animal Study in a Rabbit Model , 2017, The American journal of sports medicine.

[46]  V. Hasırcı,et al.  Construction and in vitro testing of a multilayered, tissue-engineered meniscus , 2014 .

[47]  K. Athanasiou,et al.  Regional variation in the mechanical role of knee meniscus glycosaminoglycans. , 2011, Journal of applied physiology.