Fabrication and perfusion culture of anatomically shaped artificial bone using stereolithography

Because patient bone defects are usually varied and complicated in geometry, it would be preferred to fabricate custom-made artificial bone grafts that are anatomically specific to individual patient defects. Using a rabbit femoral segment as a bone reconstruction model, we successfully produced customized ceramic scaffolds by stereolithography, which not only had an anatomically correct external shape according to computed tomography data but also contained an interconnecting internal network of channels designed for perfusion culture. Rabbit bone marrow stromal cells were isolated and cultured with these scaffolds using a novel oscillatory perfusion system that was stereolithographically fabricated to fit well to the unique scaffold shapes. After five days of three-dimensional culture with oscillatory perfusion, the cells attached and proliferated homogenously in the scaffolds. However, control cells inside the scaffolds cultured under static conditions were dead after prolonged in vitro culture. Cellular DNA content and alkaline phosphatase activities were significantly higher in the perfusion group versus the static group. Therefore, anatomically correct artificial bone can be successfully constructed using stereolithography and oscillatory culture technology, and could be useful for bone engraftment and defect repair.

[1]  Matthias Epple,et al.  Biological and medical significance of calcium phosphates. , 2002, Angewandte Chemie.

[2]  S. Hollister,et al.  Optimal design and fabrication of scaffolds to mimic tissue properties and satisfy biological constraints. , 2002, Biomaterials.

[3]  Krishnendu Roy,et al.  Laser-layered microfabrication of spatially patterned functionalized tissue-engineering scaffolds. , 2005, Journal of biomedical materials research. Part B, Applied biomaterials.

[4]  L. Lanyon,et al.  Mechanical strain and fluid movement both activate extracellular regulated kinase (ERK) in osteoblast-like cells but via different signaling pathways. , 2002, Bone.

[5]  C J Damien,et al.  Bone graft and bone graft substitutes: a review of current technology and applications. , 1991, Journal of applied biomaterials : an official journal of the Society for Biomaterials.

[6]  Swee Hin Teoh,et al.  A biaxial rotating bioreactor for the culture of fetal mesenchymal stem cells for bone tissue engineering. , 2009, Biomaterials.

[7]  Andreas Hein,et al.  Application of stereolithography for scaffold fabrication for tissue engineered heart valves. , 2000 .

[8]  K. Lau,et al.  Fluid flow shear stress stimulates human osteoblast proliferation and differentiation through multiple interacting and competing signal transduction pathways. , 2003, Bone.

[9]  Benjamin Wu,et al.  Customized biomimetic scaffolds created by indirect three-dimensional printing for tissue engineering , 2013, Biofabrication.

[10]  K. Furukawa,et al.  Oscillatory perfusion seeding and culturing of osteoblast-like cells on porous beta-tricalcium phosphate scaffolds. , 2008, Journal of Biomedical Materials Research. Part A.

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

[12]  G. Vunjak‐Novakovic,et al.  Optimizing the medium perfusion rate in bone tissue engineering bioreactors , 2011, Biotechnology and bioengineering.

[13]  Pamela Habibovic,et al.  Osteoinductive biomaterials—properties and relevance in bone repair , 2007, Journal of tissue engineering and regenerative medicine.

[14]  Thomas Boland,et al.  Rapid prototyping of tissue-engineering constructs, using photopolymerizable hydrogels and stereolithography. , 2004, Tissue engineering.

[15]  Robert E Guldberg,et al.  Effects of medium perfusion rate on cell-seeded three-dimensional bone constructs in vitro. , 2003, Tissue engineering.

[16]  Antonios G. Mikos,et al.  Fluid flow increases mineralized matrix deposition in 3D perfusion culture of marrow stromal osteoblasts in a dose-dependent manner , 2002, Proceedings of the National Academy of Sciences of the United States of America.

[17]  A. Goldstein,et al.  Fluid flow stimulates expression of osteopontin and bone sialoprotein by bone marrow stromal cells in a temporally dependent manner. , 2005, Bone.

[18]  G. Vunjak‐Novakovic,et al.  Bone grafts engineered from human adipose-derived stem cells in perfusion bioreactor culture. , 2010, Tissue engineering. Part A.

[19]  Gordana Vunjak-Novakovic,et al.  Bone Tissue Engineering Using Human Mesenchymal Stem Cells: Effects of Scaffold Material and Medium Flow , 2004, Annals of Biomedical Engineering.

[20]  D. Kaplan,et al.  Porosity of 3D biomaterial scaffolds and osteogenesis. , 2005, Biomaterials.

[21]  Yng-Jiin Wang,et al.  Osteogenic enrichment of bone-marrow stromal cells with the use of flow chamber and type I collagen-coated surface. , 2003, Journal of biomedical materials research. Part A.

[22]  K. Furukawa,et al.  Oscillatory Perfusion Culture of CaP-Based Tissue Engineering Bone with and without Dexamethasone , 2008, Annals of Biomedical Engineering.

[23]  M. Kassem,et al.  Flow perfusion culture of human mesenchymal stem cells on silicate-substituted tricalcium phosphate scaffolds. , 2008, Biomaterials.

[24]  P. Fratzl,et al.  Three-dimensional growth behavior of osteoblasts on biomimetic hydroxylapatite scaffolds. , 2007, Journal of biomedical materials research. Part A.

[25]  Moustapha Kassem,et al.  Effect of dynamic 3-D culture on proliferation, distribution, and osteogenic differentiation of human mesenchymal stem cells. , 2008, Journal of biomedical materials research. Part A.

[26]  R. Detsch,et al.  Static and dynamic cultivation of bone marrow stromal cells on biphasic calcium phosphate scaffolds derived from an indirect rapid prototyping technique , 2010, Journal of materials science. Materials in medicine.

[27]  Wei Sun,et al.  Computer‐aided tissue engineering: overview, scope and challenges , 2004, Biotechnology and applied biochemistry.

[28]  K. Dai,et al.  Effects of flow shear stress and mass transport on the construction of a large-scale tissue-engineered bone in a perfusion bioreactor. , 2009, Tissue engineering. Part A.

[29]  Antonios G. Mikos,et al.  Flow Perfusion Culture of Marrow Stromal Cells Seeded on Porous Biphasic Calcium Phosphate Ceramics , 2005, Annals of Biomedical Engineering.

[30]  S. Hollister Porous scaffold design for tissue engineering , 2005, Nature materials.

[31]  A. Mikos,et al.  Effect of convection on osteoblastic cell growth and function in biodegradable polymer foam scaffolds. , 2001, Biomaterials.

[32]  K. Leong,et al.  The design of scaffolds for use in tissue engineering. Part II. Rapid prototyping techniques. , 2002, Tissue engineering.

[33]  Antonios G. Mikos,et al.  Mineralized matrix deposition by marrow stromal osteoblasts in 3D perfusion culture increases with increasing fluid shear forces , 2003, Proceedings of the National Academy of Sciences of the United States of America.

[34]  E. Brunner,et al.  Growth behavior, matrix production, and gene expression of human osteoblasts in defined cylindrical titanium channels. , 2004, Journal of biomedical materials research. Part A.

[35]  K. Hing Bone repair in the twenty–first century: biology, chemistry or engineering? , 2004, Philosophical Transactions of the Royal Society of London. Series A: Mathematical, Physical and Engineering Sciences.

[36]  Takashi Ushida,et al.  3D culture of osteoblast‐like cells by unidirectional or oscillatory flow for bone tissue engineering , 2009, Biotechnology and bioengineering.

[37]  A. Goldstein,et al.  Hydrodynamic shear stimulates osteocalcin expression but not proliferation of bone marrow stromal cells. , 2004, Tissue engineering.

[38]  Dong-Woo Cho,et al.  A new method of fabricating robust freeform 3D ceramic scaffolds for bone tissue regeneration , 2013, Biotechnology and bioengineering.

[39]  E. Kastenbauer,et al.  Clinical aspects and strategy for biomaterial engineering of an auricle based on three-dimensional stereolithography , 2003, European Archives of Oto-Rhino-Laryngology.

[40]  A J Verbout,et al.  Design and fabrication of standardized hydroxyapatite scaffolds with a defined macro-architecture by rapid prototyping for bone-tissue-engineering research. , 2004, Journal of biomedical materials research. Part A.

[41]  Antonios G Mikos,et al.  Influence of the in vitro culture period on the in vivo performance of cell/titanium bone tissue-engineered constructs using a rat cranial critical size defect model. , 2003, Journal of biomedical materials research. Part A.

[42]  Antonios G Mikos,et al.  Flow perfusion culture induces the osteoblastic differentiation of marrow stroma cell-scaffold constructs in the absence of dexamethasone. , 2005, Journal of biomedical materials research. Part A.

[43]  S. E. Feinberg,et al.  Hydroxyapatite implants with designed internal architecture , 2001, Journal of materials science. Materials in medicine.

[44]  M. Kassem,et al.  Flow perfusion culture of human mesenchymal stem cells on coralline hydroxyapatite scaffolds with various pore sizes. , 2011, Journal of biomedical materials research. Part A.

[45]  J A Frangos,et al.  Steady and Transient Fluid Shear Stress Stimulate NO Release in Osteoblasts Through Distinct Biochemical Pathways , 1999, Journal of bone and mineral research : the official journal of the American Society for Bone and Mineral Research.

[46]  Antonios G Mikos,et al.  Design of a flow perfusion bioreactor system for bone tissue-engineering applications. , 2003, Tissue engineering.

[47]  Tzu-Wei Wang,et al.  Regulation of adult human mesenchymal stem cells into osteogenic and chondrogenic lineages by different bioreactor systems. , 2009, Journal of biomedical materials research. Part A.

[48]  Eko Supriyanto,et al.  Tangible nanocomposites with diverse properties for heart valve application , 2015, Science and technology of advanced materials.

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