Evaluating apatite formation and osteogenic activity of electrospun composites for bone tissue engineering

Significant interest has been in examining calcium phosphate ceramics, specifically β‐tricalcium phosphate (β‐TCP) (Ca3(PO4)2) and synthetic hydroxyapatite (HA) (Ca10(PO4)6(OH)2), in composites and more recently, in fibrous composites formed using the electrospinning technique for bone tissue engineering applications. Calcium phosphate ceramics are sought because they can be bone bioactive, which means an apatite forms on their surface that facilitates bonding to bone tissue, and are osteoconductive. However, studies examining the bioactivity of electrospun composites containing calcium phosphates and their corresponding osteogenic activity have been limited. In this study, electrospun composites consisting of (20/80) HA/TCP nanoceramics and poly (ϵ‐caprolactone) (PCL) were fabricated. Solvent and solvent combinations were evaluated to form scaffolds with a maximum concentration and dispersion of ceramic and pore sizes large enough for cell infiltration and tissue growth. PCL was dissolved in either methylene chloride (Composite‐MC) or a combination of methylene chloride (80%) and dimethylformamide (20%; Composite‐MC + DMF). Composites were evaluated in vitro for degradation, apatite formation, and osteogenic differentiation of human mesenchymal stem cells (MSCs) with an emphasis on temporal gene expression of osteogenic markers and the pluripotent gene Sox‐2. Apatite formation and the osteogenic differentiation was the greatest for Composite‐MC as determined by gene expression, protein production and biochemical markers, even without the presence of osteoinductive factors in the media, in comparison to Composite‐MC + DMF and unfilled PCL mats. Sox‐2 levels also reduced over time. The results of this study demonstrate that the solvent or solvent combination used in preparing the electrospun composite mats plays a critical role in determining their bioactivity which may, in turn, affect cell behavior. Biotechnol. Biotechnol. Bioeng. 2014;111: 1000–1017. © 2013 Wiley Periodicals, Inc.

[1]  Larry L. Hench,et al.  An Introduction to Bioceramics , 2013 .

[2]  Yingjun Wang,et al.  Poly(lactide-co-glycolide)/hydroxyapatite nanofibrous scaffolds fabricated by electrospinning for bone tissue engineering , 2011, Journal of materials science. Materials in medicine.

[3]  M. Gümüşderelioğlu,et al.  Synthesis, characterization and osteoblastic activity of polycaprolactone nanofibers coated with biomimetic calcium phosphate. , 2009, Acta biomaterialia.

[4]  A. Day,et al.  Prevalence, complications, and hospital charges associated with use of bone-morphogenetic proteins in spinal fusion procedures. , 2009, JAMA.

[5]  M. Bohner,et al.  Can bioactivity be tested in vitro with SBF solution? , 2009, Biomaterials.

[6]  M. J. Arellano-Jimenez,et al.  Synthesis and hydrolysis of octacalcium phosphate and its characterization by electron microscopy and X-ray diffraction , 2009 .

[7]  A. Caplan,et al.  Dexamethasone inhibition of confluence‐induced apoptosis in human mesenchymal stem cells , 2009, Journal of orthopaedic research : official publication of the Orthopaedic Research Society.

[8]  R. Hu,et al.  Effects of structure and composition of the CaP composite coatings on apatite formation and bioactivity in simulated body fluid , 2009 .

[9]  Antonios G Mikos,et al.  The influence of an in vitro generated bone-like extracellular matrix on osteoblastic gene expression of marrow stromal cells. , 2008, Biomaterials.

[10]  Fang Yang,et al.  Biomimetic calcium phosphate coating on electrospun poly(ɛ-caprolactone) scaffolds for bone tissue engineering , 2008 .

[11]  W. Stark,et al.  Cotton wool-like nanocomposite biomaterials prepared by electrospinning: in vitro bioactivity and osteogenic differentiation of human mesenchymal stem cells. , 2008, Journal of biomedical materials research. Part B, Applied biomaterials.

[12]  Hong Bu,et al.  A long-term evaluation of osteoinductive HA/β-TCP ceramics in vivo: 4.5 years study in pigs , 2007, Journal of materials science. Materials in medicine.

[13]  Thomas A. Einhorn,et al.  Current and future clinical applications of bone morphogenetic proteins in orthopaedic trauma surgery , 2007, International Orthopaedics.

[14]  I Harvey,et al.  Clinical effectiveness and cost-effectiveness of bone morphogenetic proteins in the non-healing of fractures and spinal fusion: a systematic review. , 2007, Health technology assessment.

[15]  T. Buckland,et al.  Comparative performance of three ceramic bone graft substitutes. , 2007, The spine journal : official journal of the North American Spine Society.

[16]  R. Tuan,et al.  Comparison of Proliferative and Multilineage Differentiation Potential of Human Mesenchymal Stem Cells Derived from Umbilical Cord and Bone Marrow , 2007, Stem cells.

[17]  Z. Trajanoski,et al.  Gene expression profiling of human mesenchymal stem cells derived from bone marrow during expansion and osteoblast differentiation , 2007, BMC Genomics.

[18]  H. Kim,et al.  Electrospinning biomedical nanocomposite fibers of hydroxyapatite/poly(lactic acid) for bone regeneration. , 2006, Journal of biomedical materials research. Part A.

[19]  S. Bhaduri,et al.  In vitro testing of calcium phosphate (HA, TCP, and biphasic HA-TCP) whiskers. , 2006, Journal of biomedical materials research. Part A.

[20]  K. Solntsev,et al.  Bioceramic granules with controlled resorbability for bone tissue therapy , 2006 .

[21]  D. Zhao,et al.  The in-vitro bioactivity of mesoporous bioactive glasses. , 2006, Biomaterials.

[22]  Tadashi Kokubo,et al.  How useful is SBF in predicting in vivo bone bioactivity? , 2006, Biomaterials.

[23]  Byung-Soo Kim,et al.  Poly(lactide-co-glycolide)/hydroxyapatite composite scaffolds for bone tissue engineering. , 2006, Biomaterials.

[24]  Helen H. Lu,et al.  Compositional effects on the formation of a calcium phosphate layer and the response of osteoblast-like cells on polymer-bioactive glass composites. , 2005, Biomaterials.

[25]  Yang Leng,et al.  A comparative study of calcium phosphate formation on bioceramics in vitro and in vivo. , 2005, Biomaterials.

[26]  G. Daculsi,et al.  A comparative study of biphasic calcium phosphate ceramics for human mesenchymal stem-cell-induced bone formation. , 2005, Biomaterials.

[27]  C. Choong,et al.  Simple surface modification of poly(ε-caprolactone) for apatite deposition from simulated body fluid , 2005 .

[28]  C. Shaffrey,et al.  Iliac Crest Bone Graft Donor Site Pain After Anterior Lumbar Interbody Fusion: A Prospective Patient Satisfaction Outcome Assessment , 2005, Journal of spinal disorders & techniques.

[29]  Rui L Reis,et al.  Bone tissue engineering: state of the art and future trends. , 2004, Macromolecular bioscience.

[30]  O. Kwon,et al.  Nanofabrication of microbial polyester by electrospinning promotes cell attachment , 2004 .

[31]  K. Kraus,et al.  Allogeneic mesenchymal stem cells regenerate bone in a critical-sized canine segmental defect. , 2003, The Journal of bone and joint surgery. American volume.

[32]  Roland Baron,et al.  BMP‐2 Controls Alkaline Phosphatase Expression and Osteoblast Mineralization by a Wnt Autocrine Loop , 2003, Journal of bone and mineral research : the official journal of the American Society for Bone and Mineral Research.

[33]  Y. Leng,et al.  TEM study of calcium phosphate precipitation on HA/TCP ceramics. , 2003, Biomaterials.

[34]  Zuzana Dobbie,et al.  Processing of gene expression data generated by quantitative real-time RT-PCR. , 2002, BioTechniques.

[35]  G. Sukhikh,et al.  Mesenchymal Stem Cells , 2002, Bulletin of Experimental Biology and Medicine.

[36]  David G Simpson,et al.  Electrospinning of collagen nanofibers. , 2002, Biomacromolecules.

[37]  G. H. Nancollas,et al.  Constant composition dissolution of mixed phases. II. Selective dissolution of calcium phosphates. , 2001, Journal of colloid and interface science.

[38]  A. Meunier,et al.  Tissue-engineered bone regeneration , 2000, Nature Biotechnology.

[39]  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.

[40]  L L Hench,et al.  Bioactive materials: the potential for tissue regeneration. , 1998, Journal of biomedical materials research.

[41]  K. Nakanishi,et al.  Apatite formation on silica gel in simulated body fluid: effects of structural modification with solvent-exchange , 1998, Journal of materials science. Materials in medicine.

[42]  S. Bruder,et al.  Osteogenic differentiation of purified, culture‐expanded human mesenchymal stem cells in vitro , 1997, Journal of cellular biochemistry.

[43]  V. Goldberg,et al.  Factors Affecting Bone Graft Incorporation , 1996, Clinical orthopaedics and related research.

[44]  R. V. Griend.  The effect of internal fixation on the healing of large allografts. , 1994 .

[45]  H. Aro,et al.  Clinical use of bone allografts. , 1993, Annals of medicine.

[46]  J. Aubin,et al.  Inorganic phosphate added exogenously or released from beta-glycerophosphate initiates mineralization of osteoid nodules in vitro. , 1992, Bone and mineral.

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

[48]  Arnold I. Caplan,et al.  Mesenchymal Stem Cells , 1991, Journal of orthopaedic research : official publication of the Orthopaedic Research Society.

[49]  Larry L. Hench,et al.  Bioceramics: From Concept to Clinic , 1991 .

[50]  T Kitsugi,et al.  Ca,P-rich layer formed on high-strength bioactive glass-ceramic A-W. , 1990, Journal of biomedical materials research.

[51]  J. A. Pask,et al.  Wetting and reactions in the lead borosilicate glass-precious metal systems , 1983 .

[52]  A. Patlolla,et al.  Solvent-dependent properties of electrospun fibrous composites for bone tissue regeneration. , 2010, Acta biomaterialia.

[53]  K. Karbalaei,et al.  ELECTROSPUN POLY (EPSILON-CAPROLACTONE) NANOFIBER MAT AS EXTRACELLULAR MATRIX , 2008 .

[54]  S. Teoh,et al.  The degradation profile of novel, bioresorbable PCL-TCP scaffolds: an in vitro and in vivo study. , 2008, Journal of biomedical materials research. Part A.

[55]  L. Ghasemi‐Mobarakeh,et al.  Electrospun Poly (ε-Caprolactone) Nanofiber Mat as Extracellular Matrix , 2008 .

[56]  C. Choong,et al.  Simple surface modification of poly(epsilon-caprolactone) for apatite deposition from simulated body fluid. , 2005, Biomaterials.

[57]  R. V. Vander Griend.  The effect of internal fixation on the healing of large allografts. , 1994, The Journal of bone and joint surgery. American volume.

[58]  A I Caplan,et al.  Characterization of cells with osteogenic potential from human marrow. , 1992, Bone.