Development of a synthetic tissue engineered three‐dimensional printed bioceramic‐based bone graft with homogenously distributed osteoblasts and mineralizing bone matrix in vitro

Over the last decade there have been increasing efforts to develop three‐dimensional (3D) scaffolds for bone tissue engineering from bioactive ceramics with 3D printing emerging as a promising technology. The overall objective of the present study was to generate a tissue engineered synthetic bone graft with homogenously distributed osteoblasts and mineralizing bone matrix in vitro, thereby mimicking the advantageous properties of autogenous bone grafts and facilitating usage for reconstructing segmental discontinuity defects in vivo. To this end, 3D scaffolds were developed from a silica‐containing calcium alkali orthophosphate, using, first, a replica technique – the Schwartzwalder–Somers method – and, second, 3D printing, (i.e. rapid prototyping). The mechanical and physical scaffold properties and their potential to facilitate homogenous colonization by osteogenic cells and extracellular bone matrix formation throughout the porous scaffold architecture were examined. Osteoblastic cells were dynamically cultured for 7 days on both scaffold types with two different concentrations of 1.5 and 3 × 109 cells/l. The amount of cells and bone matrix formed and osteogenic marker expression were evaluated using hard tissue histology, immunohistochemical and histomorphometric analysis. 3D‐printed scaffolds (RPS) exhibited more micropores, greater compressive strength and silica release. RPS seeded with 3 × 109 cells/l displayed greatest cell and extracellular matrix formation, mineralization and osteocalcin expression. In conclusion, RPS displayed superior mechanical and biological properties and facilitated generating a tissue engineered synthetic bone graft in vitro, which mimics the advantageous properties of autogenous bone grafts, by containing homogenously distributed terminally differentiated osteoblasts and mineralizing bone matrix and therefore is suitable for subsequent in vivo implantation for regenerating segmental discontinuity bone defects. Copyright © 2016 John Wiley & Sons, Ltd.

[1]  A. Rack,et al.  Effect of a Rapidly Resorbable Calcium Alkali Phosphate Bone Grafting Material on Osteogenesis after Sinus Floor Augmentation in Humans , 2017 .

[2]  A. Bandyopadhyay,et al.  Understanding of dopant-induced osteogenesis and angiogenesis in calcium phosphate ceramics. , 2013, Trends in biotechnology.

[3]  C. Please,et al.  Pore Geometry Regulates Early Stage Human Bone Marrow Cell Tissue Formation and Organisation , 2013, Annals of Biomedical Engineering.

[4]  Scott J Hollister,et al.  Effect of polycaprolactone scaffold permeability on bone regeneration in vivo. , 2011, Tissue engineering. Part A.

[5]  John P Fisher,et al.  Tubular perfusion system for the long-term dynamic culture of human mesenchymal stem cells. , 2011, Tissue engineering. Part C, Methods.

[6]  A. Rack,et al.  Effect of beta-tricalcium phosphate particles with varying porosity on osteogenesis after sinus floor augmentation in humans. , 2008, Biomaterials.

[7]  P. Ducheyne,et al.  Effect of rapidly resorbable bone substitute materials on the temporal expression of the osteoblastic phenotype in vitro. , 2008, Journal of biomedical materials research. Part A.

[8]  P. Ducheyne,et al.  RGDS peptides immobilized on titanium alloy stimulate bone cell attachment, differentiation and confer resistance to apoptosis. , 2007, Journal of biomedical materials research. Part A.

[9]  M. Sayer,et al.  Silicon substitution in the calcium phosphate bioceramics. , 2007, Biomaterials.

[10]  Jun-Ying Sun,et al.  The effect of the ionic products of Bioglass® dissolution on human osteoblasts growth cycle in vitro , 2007, Journal of tissue engineering and regenerative medicine.

[11]  Katherine D Kavlock,et al.  Synthesis and characterization of segmented poly(esterurethane urea) elastomers for bone tissue engineering. , 2007, Acta biomaterialia.

[12]  DW Hutmacher,et al.  Concepts of scaffold-based tissue engineering—the rationale to use solid free-form fabrication techniques , 2007, Journal of cellular and molecular medicine.

[13]  K. Anselme,et al.  Influence of hydroxyapatite microstructure on human bone cell response. , 2006, Journal of biomedical materials research. Part A.

[14]  H. Zreiqat,et al.  A method for immunohistochemical detection of osteogenic markers in undecalcified bone sections , 2006, Biotechnic & histochemistry : official publication of the Biological Stain Commission.

[15]  G. Reilly,et al.  Solution-mediated effect of bioactive glass in poly (lactic-co-glycolic acid)-bioactive glass composites on osteogenesis of marrow stromal cells. , 2005, Journal of biomedical materials research. Part A.

[16]  P. A. Revell,et al.  Microporosity enhances bioactivity of synthetic bone graft substitutes , 2005, Journal of materials science. Materials in medicine.

[17]  P. Ducheyne,et al.  Apoptosis and Survival of Osteoblast-like Cells Are Regulated by Surface Attachment* , 2005, Journal of Biological Chemistry.

[18]  Sangeeta N Bhatia,et al.  Three-dimensional tissue fabrication. , 2004, Advanced drug delivery reviews.

[19]  H. Eufinger,et al.  Growth and transplantation of a custom vascularised bone graft in a man , 2004, The Lancet.

[20]  L. Silvio,et al.  Porosity variation in hydroxyapatite and osteoblast morphology: a scanning electron microscopy study , 2004, Journal of microscopy.

[21]  J. Skepper,et al.  Effect of sintered silicate-substituted hydroxyapatite on remodelling processes at the bone-implant interface. , 2004, Biomaterials.

[22]  C. R. Howlett,et al.  Effect of rapidly resorbable calcium phosphates and a calcium phosphate bone cement on the expression of bone-related genes and proteins in vitro. , 2004, Journal of biomedical materials research. Part A.

[23]  R. Langer,et al.  Designing materials for biology and medicine , 2004, Nature.

[24]  F. Peters,et al.  Functional Materials for Bone Regeneration from Beta‐Tricalcium Phosphate , 2004 .

[25]  J. Chevalier,et al.  Effect of micro- and macroporosity of bone substitutes on their mechanical properties and cellular response , 2003, Journal of materials science. Materials in medicine.

[26]  C. R. Howlett,et al.  The Effect of Magnesium Ions on Bone Bonding to Hydroxyapatite Coating on Titanium Alloy Implants , 2003 .

[27]  G. Berger,et al.  Determination of the Internal Surface of Spongiosa-Like Ceramic Scaffolds using Light Microscopy and X-Ray Refraction Technique , 2002 .

[28]  R J Composto,et al.  RGD Peptides Immobilized on a Mechanically Deformable Surface Promote Osteoblast Differentiation , 2002, Journal of bone and mineral research : the official journal of the American Society for Bone and Mineral Research.

[29]  H. Takita,et al.  Geometry of Carriers Controlling Phenotypic Expression in BMP-Induced Osteogenesis and Chondrogenesis , 2001, The Journal of bone and joint surgery. American volume.

[30]  D. Hutmacher,et al.  Scaffolds in tissue engineering bone and cartilage. , 2000, Biomaterials.

[31]  P Ducheyne,et al.  Bioactive ceramics: the effect of surface reactivity on bone formation and bone cell function. , 1999, Biomaterials.

[32]  P. Ducheyne,et al.  Effect of serum proteins on osteoblast adhesion to surface‐modified bioactive glass and hydroxyapatite , 1999, Journal of orthopaedic research : official publication of the Orthopaedic Research Society.

[33]  D Buser,et al.  Localized ridge augmentation with autografts and barrier membranes. , 1999, Periodontology 2000.

[34]  G. Vunjak‐Novakovic,et al.  Culture of organized cell communities. , 1998, Advanced drug delivery reviews.

[35]  G. Boering,et al.  Morbidity from iliac crest bone harvesting. , 1996, Journal of oral and maxillofacial surgery : official journal of the American Association of Oral and Maxillofacial Surgeons.

[36]  Robert Langer,et al.  Biodegradable Polymer Scaffolds for Tissue Engineering , 1994, Bio/Technology.

[37]  S. Furner,et al.  Musculoskeletal Conditions in the United States , 1992 .

[38]  S F Hulbert,et al.  Potential of ceramic materials as permanently implantable skeletal prostheses. , 1970, Journal of biomedical materials research.

[39]  R. Gildenhaar,et al.  Effect of a rapidly resorbable calcium alkali phosphate bone grafting material on bone formation and osteogenic marker expression after sinus floor augmentation in humans , 2016 .

[40]  L. Kuhn,et al.  Design and characterization of calcium phosphate ceramic scaffolds for bone tissue engineering. , 2016, Dental materials : official publication of the Academy of Dental Materials.

[41]  P. Ducheyne,et al.  Dental Graft Materials , 2011 .

[42]  P. Ducheyne,et al.  Cellular response to bioactive ceramics , 2008 .

[43]  Anna Tampieri,et al.  Biomimetic Mg-substituted hydroxyapatite: from synthesis to in vivo behaviour , 2008, Journal of materials science. Materials in medicine.

[44]  D. Deligianni,et al.  Effect of surface roughness of hydroxyapatite on human bone marrow cell adhesion, proliferation, differentiation and detachment strength. , 2001, Biomaterials.

[45]  W C Hayes,et al.  Evolution of bone transplantation: molecular, cellular and tissue strategies to engineer human bone. , 1996, Biomaterials.

[46]  R. Gildenhaar,et al.  Rapid resorbable, glassy crystalline materials on the basis of calcium alkali orthophosphates. , 1995, Biomaterials.

[47]  M. Schneider,et al.  Investigations of phase relations in the system CaONa2OK2OP2O5 , 1994 .