Biomimetic formation of apatite on the surface of porous gelatin/bioactive glass nanocomposite scaffolds

Abstract There have been several attempts to combine bioactive glasses (BaGs) with biodegradable polymers to create a scaffold material with excellent biocompatibility, bioactivity, biodegradability and toughness. In the present study, the nanocomposite scaffolds with compositions based on gelatin (Gel) and BaG nanoparticles in the ternary SiO 2 –CaO–P 2 O 5 system were prepared. In vitro evaluations of the nanocomposite scaffolds were performed, and for investigating their bioactive capacity these scaffolds were soaked in a simulated body fluid (SBF) at different time intervals. The scaffolds showed significant enhancement in bioactivity within few days of immersion in SBF solution. The apatite formation at the surface of the nanocomposite samples confirmed by Fourier transform infrared spectroscopy (FTIR), scanning electron microscopy (SEM), energy dispersive X-ray spectroscopy (EDX) and X-ray powder diffraction (XRD) analyses. In vitro experiments with osteoblast cells indicated an appropriate penetration of the cells into the scaffold's pores, and also the continuous increase in cell aggregation on the bioactive scaffolds with increase in the incubation time demonstrated the ability of the scaffolds to support cell growth. The SEM observations revealed that the prepared scaffolds were porous with three dimensional (3D) and interconnected microstructure, pore size was 200–500 μm and the porosity was 72–86%. The nanocomposite scaffold made from Gel and BaG nanoparticles could be considered as a highly bioactive and potential bone tissue engineering implant.

[1]  Peter X Ma,et al.  Biomimetic nanofibrous gelatin/apatite composite scaffolds for bone tissue engineering. , 2009, Biomaterials.

[2]  H. Oonishi,et al.  Quantitative comparison of bone growth behavior in granules of Bioglass, A-W glass-ceramic, and hydroxyapatite. , 2000, Journal of biomedical materials research.

[3]  M. Mozafari,et al.  Investigation of the physico-chemical reactivity of a mesoporous bioactive SiO2–CaO–P2O5 glass in simulated body fluid , 2010 .

[4]  J. Lima,et al.  Surface analysis of titanium dental implants with different topographies , 2000 .

[5]  M. Vallet‐Regí,et al.  Influence of composition and surface characteristics on the in vitro bioactivity of SiO(2)-CaO-P(2)O(5)-MgO sol-gel glasses. , 1999, Journal of biomedical materials research.

[6]  N. Chayen,et al.  Experiment and theory for heterogeneous nucleation of protein crystals in a porous medium. , 2006, Proceedings of the National Academy of Sciences of the United States of America.

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

[8]  T. Kokubo,et al.  ROUND-ROBIN TEST OF SBF FOR IN VITRO MEASUREMENT OF APATITE-FORMING ABILITY OF SYNTHETIC MATERIALS , 2004 .

[9]  H. Boedtker,et al.  A Study of Gelatin Molecules, Aggregates and Gels , 1954 .

[10]  F. Moztarzadeh,et al.  Preparation, characterization and mechanical properties of controlled porous gelatin/hydroxyapatite nanocomposite through layer solvent casting combined with freeze-drying and lamination techniques , 2010 .

[11]  B. Peng,et al.  The effect of alginate addition on the structure and morphology of hydroxyapatite/gelatin nanocomposites , 2006 .

[12]  Julian R. Jones,et al.  Extracellular matrix formation and mineralization on a phosphate-free porous bioactive glass scaffold using primary human osteoblast (HOB) cells. , 2007, Biomaterials.

[13]  W. Neuman,et al.  THE CHEMICAL DYNAMICS OF BONE MINERAL , 1959 .

[14]  M. Vallet‐Regí,et al.  Controlled Crystallization of Calcium Phosphate Apatites , 2000 .

[15]  Ayako Oyane,et al.  Preparation and assessment of revised simulated body fluids. , 2003, Journal of biomedical materials research. Part A.

[16]  K. P. Rao,et al.  Synthesis of nanocrystalline fluorinated hydroxyapatite by microwave processing and its in vitro dissolution study , 2006 .

[17]  H R Allcock,et al.  A highly porous 3-dimensional polyphosphazene polymer matrix for skeletal tissue regeneration. , 1996, Journal of biomedical materials research.

[18]  J. Feijen,et al.  Glutaraldehyde as a crosslinking agent for collagen-based biomaterials , 1995 .

[19]  D. Greenspan,et al.  Effect of Surface Area to Volume Ratio on In Vitro Surface Reactions of Bioactive Glass Particulates , 1994 .

[20]  J. Bateman,et al.  Matrix deposition by a calcifying human osteogenic sarcoma cell line (SAOS-2). , 1995, Bone.

[21]  I. Kangasniemi,et al.  Bonelike Hydroxyapatite Induction by a Gel‐Derived Titania on a Titanium Substrate , 1994 .

[22]  H. Oudadesse,et al.  Investigation of the surface reactivity of a sol-gel derived glass in the ternary system SiO2-CaO-P2O5 , 2008 .

[23]  M. Neo,et al.  A comparative study between in vivo bone ingrowth and in vitro apatite formation on Na2O-CaO-SiO2 glasses. , 2003, Biomaterials.

[24]  Junjun Tan,et al.  Preparation of Gelatin coated hydroxyapatite nanorods and the stability of its aqueous colloidal , 2008 .

[25]  Y Ikada,et al.  Fabrication of porous gelatin scaffolds for tissue engineering. , 1999, Biomaterials.

[26]  S A Goldstein,et al.  Skeletal repair by in situ formation of the mineral phase of bone. , 1995, Science.

[27]  T Kitsugi,et al.  Solutions able to reproduce in vivo surface-structure changes in bioactive glass-ceramic A-W. , 1990, Journal of biomedical materials research.

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

[29]  Y. Ikada,et al.  Bioadhesion of gelatin films crosslinked with glutaraldehyde. , 1999, Journal of biomedical materials research.

[30]  K. Onuma,et al.  Formation and growth of clusters in conventional and new kinds of simulated body fluids. , 2003, Journal of biomedical materials research. Part A.

[31]  Antonios G Mikos,et al.  Biomimetic materials for tissue engineering. , 2003, Biomaterials.

[32]  Larry L. Hench,et al.  The story of Bioglass® , 2006, Journal of materials science. Materials in medicine.

[33]  W. Douglas,et al.  Conformational change of hydroxyapatite/gelatin nanocomposite by glutaraldehyde. , 2003, Biomaterials.

[34]  M. Mozafari,et al.  Development of 3D Bioactive Nanocomposite Scaffolds Made from Gelatin and Nano Bioactive Glass for Biomedical Applications , 2010 .

[35]  J. J. Mecholsky,et al.  Bioactivity of tape cast and sintered bioactive glass-ceramic in simulated body fluid. , 2002, Biomaterials.

[36]  X. Chatzistavrou,et al.  Investigation of the Hydroxyapatite Growth on Bioactive Glass Surface , 2007 .

[37]  P. Hess,et al.  Infrared spectra of photochemically grown suboxides at the Si/SiO2 interface , 2003 .

[38]  Jian Shen,et al.  Covalent immobilization of O-butyrylchitosan with a photosensitive hetero-bifunctional crosslinking reagent on biopolymer substrate surface and bloodcompatibility characterization , 2003, Journal of biomaterials science. Polymer edition.

[39]  W. Douglas,et al.  Preparation of hydroxyapatite-gelatin nanocomposite. , 2003, Biomaterials.

[40]  M. Vallet‐Regí,et al.  Bioactivity of three CaO-P2O5-SiO2 sol-gel glasses. , 2002, Journal of biomedical materials research.

[41]  Julian R. Jones,et al.  Nodule formation and mineralisation of human primary osteoblasts cultured on a porous bioactive glass scaffold. , 2004, Biomaterials.

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

[43]  P. V. von Hippel,et al.  The structure of collagen and gelatin. , 1961, Advances in protein chemistry.

[44]  J. Faure,et al.  Synthesis and characterisation of sol gel derived bioactive glass for biomedical applications , 2006 .

[45]  Y. Doi,et al.  Sintered carbonate apatites as bioresorbable bone substitutes. , 1998, Journal of biomedical materials research.