Bioactive nanoparticles stimulate bone tissue formation in bioprinted three-dimensional scaffold and human mesenchymal stem cells.

Bioprinting based on thermal inkjet printing is a promising but unexplored approach in bone tissue engineering. Appropriate cell types and suitable biomaterial scaffolds are two critical factors to generate successful bioprinted tissue. This study was undertaken in order to evaluate bioactive ceramic nanoparticles in stimulating osteogenesis of printed bone marrow-derived human mesenchymal stem cells (hMSCs) in poly(ethylene glycol)dimethacrylate (PEGDMA) scaffold. hMSCs suspended in PEGDMA were co-printed with nanoparticles of bioactive glass (BG) and hydroxyapatite (HA) under simultaneous polymerization so the printed substrates were delivered with highly accurate placement in three-dimensional (3D) locations. hMSCs interacted with HA showed the highest cell viability (86.62 ± 6.02%) and increased compressive modulus (358.91 ± 48.05 kPa) after 21 days in culture among all groups. Biochemical analysis showed the most collagen production and highest alkaline phosphatase activity in PEG-HA group, which is consistent with gene expression determined by quantitative PCR. Masson's trichrome staining also showed the most collagen deposition in PEG-HA scaffold. Therefore, HA is more effective comparing to BG for hMSCs osteogenesis in bioprinted bone constructs. Combining with our previous experience in vasculature, cartilage, and muscle bioprinting, this technology demonstrates the capacity for both soft and hard tissue engineering with biomimetic structures.

[1]  Guifang Gao,et al.  Human cartilage tissue fabrication using three-dimensional inkjet printing technology. , 2014, Journal of visualized experiments : JoVE.

[2]  C. Devlin,et al.  Dexamethasone induction of osteoblast mRNAs in rat marrow stromal cell cultures , 1991, Journal of cellular physiology.

[3]  Xiaofeng Cui,et al.  Application of inkjet printing to tissue engineering , 2006, Biotechnology journal.

[4]  Xiaofeng Cui,et al.  Thermal inkjet printing in tissue engineering and regenerative medicine. , 2012, Recent patents on drug delivery & formulation.

[5]  Xiaofeng Cui,et al.  Collagen matrix alignment using inkjet printer technology , 2008 .

[6]  T. Boland,et al.  Simultaneous Deposition of Human Microvascular Endothelial Cells and Biomaterials for Human Microvasculature Fabrication Using Inkjet Printing , 2008, NIP & Digital Fabrication Conference.

[7]  J. Elisseeff,et al.  Photoencapsulation of chondrocytes in poly(ethylene oxide)-based semi-interpenetrating networks. , 2000, Journal of biomedical materials research.

[8]  Guifang Gao,et al.  Accelerated myotube formation using bioprinting technology for biosensor applications , 2012, Biotechnology Letters.

[9]  Arndt F Schilling,et al.  High amplitude direct compressive strain enhances mechanical properties of scaffold-free tissue-engineered cartilage. , 2011, Tissue engineering. Part A.

[10]  S. Chizhik,et al.  Atomic force microscopy probing of cell elasticity. , 2007, Micron.

[11]  Leo Q Wan,et al.  A hydrogel-mineral composite scaffold for osteochondral interface tissue engineering. , 2012, Tissue engineering. Part A.

[12]  Uwe Gbureck,et al.  3D powder printed calcium phosphate implants for reconstruction of cranial and maxillofacial defects. , 2010, Journal of cranio-maxillo-facial surgery : official publication of the European Association for Cranio-Maxillo-Facial Surgery.

[13]  A Boyde,et al.  Autologous bone marrow stromal cells loaded onto porous hydroxyapatite ceramic accelerate bone repair in critical-size defects of sheep long bones. , 2000, Journal of biomedical materials research.

[14]  Stephen B Doty,et al.  Development of controlled matrix heterogeneity on a triphasic scaffold for orthopedic interface tissue engineering. , 2006, Tissue engineering.

[15]  D. D’Lima,et al.  Direct human cartilage repair using three-dimensional bioprinting technology. , 2012, Tissue engineering. Part A.

[16]  Hermann Seitz,et al.  A review on 3D micro-additive manufacturing technologies , 2012, The International Journal of Advanced Manufacturing Technology.

[17]  Dietmar W Hutmacher,et al.  Assessment of bone ingrowth into porous biomaterials using MICRO-CT. , 2007, Biomaterials.

[18]  B. Riggs,et al.  Isolation and characterization of osteoblast precursor cells from human bone marrow , 1996, Journal of bone and mineral research : the official journal of the American Society for Bone and Mineral Research.

[19]  E B Hunziker,et al.  Functional barrier principle for growth-factor-based articular cartilage repair. , 2003, Osteoarthritis and cartilage.

[20]  T. Boland,et al.  Human microvasculature fabrication using thermal inkjet printing technology. , 2009, Biomaterials.

[21]  T. Boland,et al.  Cell damage evaluation of thermal inkjet printed Chinese hamster ovary cells , 2010, Biotechnology and bioengineering.

[22]  S. Bruder,et al.  Mesenchymal stem cells: building blocks for molecular medicine in the 21st century. , 2001, Trends in molecular medicine.

[23]  Xiaofeng Cui,et al.  Synergistic action of fibroblast growth factor‐2 and transforming growth factor‐beta1 enhances bioprinted human neocartilage formation , 2012, Biotechnology and bioengineering.

[24]  Sheng Lin-Gibson,et al.  Synthesis and characterization of PEG dimethacrylates and their hydrogels. , 2004, Biomacromolecules.

[25]  S. Bryant,et al.  Hydrogel properties influence ECM production by chondrocytes photoencapsulated in poly(ethylene glycol) hydrogels. , 2002, Journal of biomedical materials research.

[26]  Gerard A. Ateshian,et al.  Bioactive Stratified Polymer Ceramic-Hydrogel Scaffold for Integrative Osteochondral Repair , 2010, Annals of Biomedical Engineering.

[27]  D. D’Lima,et al.  92 DIRECT HUMAN CARTILAGE REPAIR USING THERMAL INKJET PRINTING TECHNOLOGY , 2011 .

[28]  D. D’Lima,et al.  Structured three‐dimensional co‐culture of mesenchymal stem cells with meniscus cells promotes meniscal phenotype without hypertrophy , 2012, Biotechnology and bioengineering.

[29]  Benjamin M. Wu,et al.  Human mesenchymal stem cell proliferation and osteogenic differentiation in fibrin gels in vitro. , 2006, Tissue engineering.

[30]  J. Triffitt.  Osteogenic stem cells and orthopedic engineering: summary and update. , 2002, Journal of biomedical materials research.

[31]  A. Boccaccini,et al.  Biodegradable and bioactive porous polymer/inorganic composite scaffolds for bone tissue engineering. , 2006, Biomaterials.

[32]  R. Oreffo,et al.  Future potentials for using osteogenic stem cells and biomaterials in orthopedics. , 1999, Bone.

[33]  John P Fisher,et al.  Characterization of cyclic acetal hydroxyapatite nanocomposites for craniofacial tissue engineering. , 2010, Journal of biomedical materials research. Part A.

[34]  Aldo R Boccaccini,et al.  Bone tissue engineering therapeutics: controlled drug delivery in three-dimensional scaffolds , 2010, Journal of The Royal Society Interface.

[35]  S. Bruder,et al.  Growth kinetics, self‐renewal, and the osteogenic potential of purified human mesenchymal stem cells during extensive subcultivation and following cryopreservation , 1997, Journal of cellular biochemistry.

[36]  T. Komori.  Regulation of osteoblast differentiation by Runx2. , 2010, Advances in experimental medicine and biology.

[37]  Stephen B Doty,et al.  In vivo evaluation of a multiphased scaffold designed for orthopaedic interface tissue engineering and soft tissue-to-bone integration. , 2008, Journal of biomedical materials research. Part A.

[38]  Xiaofeng Cui,et al.  Image based printing of structured biomaterials for realizing complex 3D cardiovascular constructs , 2006, NIP & Digital Fabrication Conference.

[39]  Dietmar W. Hutmacher,et al.  A Tissue Engineering Solution for Segmental Defect Regeneration in Load-Bearing Long Bones , 2012, Science Translational Medicine.

[40]  H. Seitz,et al.  Three-dimensional printing of porous ceramic scaffolds for bone tissue engineering. , 2005, Journal of biomedical materials research. Part B, Applied biomaterials.