Three-dimensional bioprinting of multilayered constructs containing human mesenchymal stromal cells for osteochondral tissue regeneration in the rabbit knee joint

The use of cell-rich hydrogels for three-dimensional (3D) cell culture has shown great potential for a variety of biomedical applications. However, the fabrication of appropriate constructs has been challenging. In this study, we describe a 3D printing process for the preparation of a multilayered 3D construct containing human mesenchymal stromal cells with a hydrogel comprised of atelocollagen and supramolecular hyaluronic acid (HA). This construct showed outstanding regenerative ability for the reconstruction of an osteochondral tissue in the knee joints of rabbits. We found that the use of a mechanically stable, host-guest chemistry-based hydrogel was essential and allowed two different types of extracellular matrix (ECM) hydrogels to be easily printed and stacked into one multilayered construct without requiring the use of potentially harmful chemical reagents or physical stimuli for post-crosslinking. To the best of our knowledge, this is the first study to validate the potential of a 3D printed multilayered construct consisting of two different ECM materials (atelocollagen and HA) for heterogeneous tissue regeneration using an in vivo animal model. We believe that this 3D printing-based platform technology can be effectively exploited for regeneration of various heterogeneous tissues as well as osteochondral tissue.

[1]  Shih-Jung Liu,et al.  Enhancement of diabetic wound repair using biodegradable nanofibrous metformin-eluting membranes: in vitro and in vivo. , 2014, ACS applied materials & interfaces.

[2]  Anders Lindahl,et al.  Coculture of Human Embryonic Stem Cells and Human Articular Chondrocytes Results in Significantly Altered Phenotype and Improved Chondrogenic Differentiation , 2009, Stem cells.

[3]  Jae Hyun Kim,et al.  The use of biodegradable PLGA nanoparticles to mediate SOX9 gene delivery in human mesenchymal stem cells (hMSCs) and induce chondrogenesis. , 2011, Biomaterials.

[4]  Diego Mantovani,et al.  Tailoring Mechanical Properties of Collagen-Based Scaffolds for Vascular Tissue Engineering: The Effects of pH, Temperature and Ionic Strength on Gelation , 2010 .

[5]  Rui L Reis,et al.  Vascularization in bone tissue engineering: physiology, current strategies, major hurdles and future challenges. , 2010, Macromolecular bioscience.

[6]  Dong-Woo Cho,et al.  Three-dimensional printing of rhBMP-2-loaded scaffolds with long-term delivery for enhanced bone regeneration in a rabbit diaphyseal defect. , 2014, Tissue engineering. Part A.

[7]  E. Place,et al.  Complexity in biomaterials for tissue engineering. , 2009, Nature materials.

[8]  Mark A Randolph,et al.  Survival of porcine mesenchymal stem cells over the alginate recovered cellular method. , 2011, Journal of biomedical materials research. Part A.

[9]  J. Fallas,et al.  Multi-hierarchical self-assembly of a collagen mimetic peptide from triple helix to nanofibre and hydrogel. , 2011, Nature chemistry.

[10]  D. Cho,et al.  Human turbinate mesenchymal stromal cell sheets with bellows graft for rapid tracheal epithelial regeneration. , 2015, Acta biomaterialia.

[11]  Jaesung Park,et al.  Development of a hybrid scaffold with synthetic biomaterials and hydrogel using solid freeform fabrication technology , 2011, Biofabrication.

[12]  J. Southgate,et al.  Plasticity of in vitro-generated urothelial cells for functional tissue formation. , 2014, Tissue engineering. Part A.

[13]  Dong-An Wang,et al.  Therapeutic cell delivery and fate control in hydrogels and hydrogel hybrids. , 2010, Advanced drug delivery reviews.

[14]  H. Yoshikawa,et al.  Direct Induction of Chondrogenic Cells from Human Dermal Fibroblast Culture by Defined Factors , 2013, PloS one.

[15]  Glenn D Prestwich,et al.  Hyaluronic acid-based hydrogels functionalized with heparin that support controlled release of bioactive BMP-2. , 2012, Biomaterials.

[16]  George M Whitesides,et al.  Polymer-based mesh as supports for multi-layered 3D cell culture and assays. , 2014, Biomaterials.

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

[18]  J. De las Rivas,et al.  Transcriptomic portrait of human Mesenchymal Stromal/Stem cells isolated from bone marrow and placenta , 2014, BMC Genomics.

[19]  Y. Zhou,et al.  Bio-inspired hard-to-soft interface for implant integration to bone. , 2015, Nanomedicine : nanotechnology, biology, and medicine.

[20]  Jung-Bo Huh,et al.  Comparative Efficacies of a 3D-Printed PCL/PLGA/β-TCP Membrane and a Titanium Membrane for Guided Bone Regeneration in Beagle Dogs , 2015 .

[21]  Shoji Takeuchi,et al.  Molding Cell Beads for Rapid Construction of Macroscopic 3D Tissue Architecture , 2011, Advanced materials.

[22]  Sindy K. Y. Tang,et al.  Paper-supported 3D cell culture for tissue-based bioassays , 2009, Proceedings of the National Academy of Sciences.

[23]  Y. Ko,et al.  Reduction-sensitive, robust vesicles with a non-covalently modifiable surface as a multifunctional drug-delivery platform. , 2010, Small.

[24]  J. Lewis,et al.  3D Microperiodic Hydrogel Scaffolds for Robust Neuronal Cultures , 2011, Advanced functional materials.

[25]  A. Saito,et al.  Master Regulator for Chondrogenesis, Sox9, Regulates Transcriptional Activation of the Endoplasmic Reticulum Stress Transducer BBF2H7/CREB3L2 in Chondrocytes* , 2014, The Journal of Biological Chemistry.

[26]  D. Cho,et al.  Development of hybrid scaffolds using ceramic and hydrogel for articular cartilage tissue regeneration. , 2015, Journal of biomedical materials research. Part A.

[27]  Shanshan Xu,et al.  Preparation and optimization of silver nanoparticles embedded electrospun membrane for implant associated infections prevention. , 2013, ACS applied materials & interfaces.

[28]  J. S. Park,et al.  Multilineage differentiation of human-derived dermal fibroblasts transfected with genes coated on PLGA nanoparticles plus growth factors. , 2013, Biomaterials.

[29]  Dong-Woo Cho,et al.  Ornamenting 3D printed scaffolds with cell-laid extracellular matrix for bone tissue regeneration. , 2015, Biomaterials.

[30]  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 .

[31]  A. Lendlein,et al.  Controlling Major Cellular Processes of Human Mesenchymal Stem Cells using Microwell Structures , 2014, Advanced healthcare materials.

[32]  Elizabeth Blaber,et al.  Stem cell health and tissue regeneration in microgravity. , 2014, Stem cells and development.

[33]  Dietmar Werner Hutmacher,et al.  The evaluation of a biphasic osteochondral implant coupled with an electrospun membrane in a large animal model. , 2009, Tissue engineering. Part A.

[34]  Yan Li,et al.  Asymmetric biodegradable microdevices for cell-borne drug delivery. , 2015, ACS applied materials & interfaces.

[35]  G. Prestwich,et al.  Photocrosslinkable hyaluronan-gelatin hydrogels for two-step bioprinting. , 2010, Tissue engineering. Part A.

[36]  A. Baria,et al.  Aggrecan is required for growth plate cytoarchitecture and differentiation. , 2014, Developmental biology.

[37]  Jae Wook Lee,et al.  Cucurbituril homologues and derivatives: new opportunities in supramolecular chemistry. , 2003, Accounts of chemical research.

[38]  Xin Zhang,et al.  Dextran-coated fluorapatite crystals doped with Yb3+/Ho3+ for labeling and tracking chondrogenic differentiation of bone marrow mesenchymal stem cells in vitro and in vivo. , 2015, Biomaterials.

[39]  W. Grayson,et al.  Oxygen delivery from hyperbarically loaded microtanks extends cell viability in anoxic environments. , 2015, Biomaterials.

[40]  Jie Zheng,et al.  Design of LVFFARK and LVFFARK-functionalized nanoparticles for inhibiting amyloid β-protein fibrillation and cytotoxicity. , 2015, ACS applied materials & interfaces.

[41]  P. Conget,et al.  Intraarticular Administration of Dexamethasone after Mesenchymal Stem Cells Implantation Does Not Improve Significantly the Treatment of Preestablished Full-Thickness Chondral Defect in a Rabbit Model , 2013, Cartilage.

[42]  S. Hsu,et al.  The use of peptide-delivery to protect human adipose-derived adult stem cells from damage caused by the internalization of quantum dots. , 2008, Biomaterials.

[43]  Stephen D. Thorpe,et al.  The Response of Bone Marrow-Derived Mesenchymal Stem Cells to Dynamic Compression Following TGF-β3 Induced Chondrogenic Differentiation , 2010, Annals of Biomedical Engineering.

[44]  P. Bártolo,et al.  Additive manufacturing of tissues and organs , 2012 .

[45]  J. Temenoff,et al.  Characterization of a multilayer heparin coating for biomolecule presentation to human mesenchymal stem cell spheroids. , 2014, Biomaterials science.

[46]  L. Lanier,et al.  Antigen-specific expansion and differentiation of natural killer cells by alloantigen stimulation , 2014, The Journal of experimental medicine.

[47]  T. Young,et al.  Interactive effects of mechanical stretching and extracellular matrix proteins on initiating osteogenic differentiation of human mesenchymal stem cells , 2009, Journal of cellular biochemistry.

[48]  W. Gu,et al.  Mesenchymal stem cells alleviate airway inflammation and emphysema in COPD through down-regulation of cyclooxygenase-2 via p38 and ERK MAPK pathways , 2015, Scientific Reports.

[49]  J. Rhie,et al.  Secretory factors of human chorion-derived stem cells enhance activation of human fibroblasts. , 2015, Cytotherapy.

[50]  K. Kaestner.  An epigenomic road map for endoderm development. , 2015, Cell stem cell.

[51]  Dietmar W Hutmacher,et al.  Biomaterials offer cancer research the third dimension. , 2010, Nature materials.

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

[53]  Dong-Woo Cho,et al.  Solid freeform fabrication technology applied to tissue engineering with various biomaterials , 2012 .

[54]  Wim E Hennink,et al.  25th Anniversary Article: Engineering Hydrogels for Biofabrication , 2013, Advanced materials.

[55]  S. Varghese,et al.  Biomineralized matrices dominate soluble cues to direct osteogenic differentiation of human mesenchymal stem cells through adenosine signaling. , 2015, Biomacromolecules.

[56]  Kimoon Kim,et al.  3D tissue engineered supramolecular hydrogels for controlled chondrogenesis of human mesenchymal stem cells. , 2014, Biomacromolecules.

[57]  Kun Zhang,et al.  Nanosecond Pulsed Electric Fields (nsPEFs) Regulate Phenotypes of Chondrocytes through Wnt/β-catenin Signaling Pathway , 2014, Scientific Reports.

[58]  F. Wang,et al.  The internalized CdSe/ZnS quantum dots impair the chondrogenesis of bone marrow mesenchymal stem cells. , 2006, Journal of biomedical materials research. Part B, Applied biomaterials.

[59]  M. Eiraku,et al.  Generation of a ciliary margin-like stem cell niche from self-organizing human retinal tissue , 2015, Nature Communications.

[60]  Deok-Ho Kim,et al.  Microfluidics-assisted in vitro drug screening and carrier production. , 2013, Advanced drug delivery reviews.

[61]  R. Cancedda,et al.  Recruitment of a host's osteoprogenitor cells using exogenous mesenchymal stem cells seeded on porous ceramic. , 2009, Tissue engineering. Part A.

[62]  Tianjiao Ji,et al.  "Triple-punch" strategy for triple negative breast cancer therapy with minimized drug dosage and improved antitumor efficacy. , 2015, ACS nano.

[63]  Young Ho Ko,et al.  Functionalized cucurbiturils and their applications. , 2007, Chemical Society reviews.

[64]  Hansoo Park,et al.  Co-delivery of Cbfa-1-targeting siRNA and SOX9 protein using PLGA nanoparticles to induce chondrogenesis of human mesenchymal stem cells. , 2014, Biomaterials.

[65]  A. Hoffman,et al.  In situ supramolecular assembly and modular modification of hyaluronic acid hydrogels for 3D cellular engineering. , 2012, ACS nano.

[66]  W. Khan,et al.  Clinical applications of mesenchymal stem cells in the treatment of fracture non-union and bone defects. , 2012, Current stem cell research & therapy.

[67]  Krishnendu Roy,et al.  Engineering articular cartilage with spatially-varying matrix composition and mechanical properties from a single stem cell population using a multi-layered hydrogel. , 2011, Biomaterials.

[68]  Smadar Cohen,et al.  The effect of immobilized RGD peptide in macroporous alginate scaffolds on TGFbeta1-induced chondrogenesis of human mesenchymal stem cells. , 2010, Biomaterials.

[69]  J. Mao,et al.  Stem cell‐based composite tissue constructs for regenerative medicine , 2005, Biotechnology and bioengineering.

[70]  A. Khanna,et al.  Effect of HSA coated iron oxide labeling on human umbilical cord derived mesenchymal stem cells , 2015, Nanotechnology.

[71]  J. S. Park,et al.  SOX9 gene plus heparinized TGF-β 3 coated dexamethasone loaded PLGA microspheres for inducement of chondrogenesis of hMSCs. , 2012, Biomaterials.

[72]  C. Ha,et al.  Mesenchymal stem/progenitor cells developed in cultures from UC blood. , 2004, Cytotherapy.

[73]  C. Maes.  Role and Regulation of Vascularization Processes in Endochondral Bones , 2013, Calcified Tissue International.

[74]  Phuc Van Pham,et al.  Production of good manufacturing practice-grade human umbilical cord blood-derived mesenchymal stem cells for therapeutic use. , 2015, Methods in molecular biology.

[75]  H. Varma,et al.  Short-term studies using ceramic scaffolds in lapine model for osteochondral defect amelioration , 2012, Biomedical materials.

[76]  P. Dubruel,et al.  The 3D printing of gelatin methacrylamide cell-laden tissue-engineered constructs with high cell viability. , 2014, Biomaterials.

[77]  Julian R. Jones,et al.  Differentiation of fetal osteoblasts and formation of mineralized bone nodules by 45S5 Bioglass conditioned medium in the absence of osteogenic supplements. , 2009, Biomaterials.

[78]  Dong-Woo Cho,et al.  Human Inferior Turbinate , 2012, Otolaryngology--head and neck surgery : official journal of American Academy of Otolaryngology-Head and Neck Surgery.

[79]  David J Mooney,et al.  Alginate hydrogels as biomaterials. , 2006, Macromolecular bioscience.

[80]  Jianwu Dai,et al.  Enhanced proliferation and osteogenic differentiation of mesenchymal stem cells on graphene oxide-incorporated electrospun poly(lactic-co-glycolic acid) nanofibrous mats. , 2015, ACS applied materials & interfaces.

[81]  Kimoon Kim,et al.  Cucurbituril-based nanoparticles: a new efficient vehicle for targeted intracellular delivery of hydrophobic drugs. , 2009, Chemical communications.

[82]  Jerry C. Hu,et al.  Unlike Bone, Cartilage Regeneration Remains Elusive , 2012, Science.

[83]  D. Hutmacher,et al.  The return of a forgotten polymer : Polycaprolactone in the 21st century , 2009 .

[84]  M. Teixeira,et al.  Adult stem cells in neural repair: Current options, limitations and perspectives. , 2015, World journal of stem cells.