Silk fibroin-bioactive glass based advanced biomaterials: towards patient-specific bone grafts

A major challenge in bone tissue engineering is to develop patient-specific, defect-site specific grafts capable of triggering specific cell signaling pathways. We could programmably fabricate the 3D printed bone constructs via direct ink writing of silk-gelatin-bioactive glass (SF-G-BG) hybrids using two different compositions of melt-derived bioactive glasses (with and without strontium) and compared against commercial 45S5 Bioglass®. Physico-chemical characterization revealed that released ions from bioactive glasses inhibited the conformational change of Bombyx mori silk fibroin protein (from random coil to β-sheet conformation), affecting printability of the SF-G-BG ink. In-depth molecular investigations showed that strontium containing SF-G-BG constructs demonstrated superior osteogenic differentiation of mesenchymal stem cells (TVA-BMSCs) over 21 days towards osteoblastic (marked by upregulated expression of runt related transcription factor, alkaline phosphatase, osteopontin, osteonectin, integrin bone sialoprotein, osteocalcin) and osteocytic (marked by podoplanin, dentin matrix acidic phosphoprotein, sclerostin) phenotype compared to other BG compositions and silk-gelatin alone. Moreover, ionic release from bioactive glasses in the silk-gelatin ink triggered the activation of signaling pathways (BMP-2, BMP-4 and IHH), which are critical in regulating bone formation in vivo. Overall, the presence of strontium containing bioactive glass in silk-gelatin matrices provided appropriate cues in regulating the development of custom-made 3D in vitro human bone constructs.

[1]  P. Ao,et al.  Deregulation of Bone Forming Cells in Bone Diseases and Anabolic Effects of Strontium-Containing Agents and Biomaterials , 2014, BioMed research international.

[2]  Ivan Martin,et al.  The role of 3D structure and protein conformation on the innate and adaptive immune responses to silk-based biomaterials. , 2013, Biomaterials.

[3]  R. P. Thompson,et al.  Orthosilicic acid stimulates collagen type 1 synthesis and osteoblastic differentiation in human osteoblast-like cells in vitro. , 2003, Bone.

[4]  S. Chawla,et al.  Elucidating role of silk-gelatin bioink to recapitulate articular cartilage differentiation in 3D bioprinted constructs , 2017 .

[5]  Sanskrita Das,et al.  Enhanced redifferentiation of chondrocytes on microperiodic silk/gelatin scaffolds: toward tailor-made tissue engineering. , 2013, Biomacromolecules.

[6]  Swati Midha,et al.  Regulation of Chondrogenesis and Hypertrophy in Silk Fibroin-Gelatin-Based 3D Bioprinted Constructs. , 2016, ACS biomaterials science & engineering.

[7]  Z. Shao,et al.  Effect of metallic ions on silk formation in the Mulberry silkworm, Bombyx mori. , 2005, The journal of physical chemistry. B.

[8]  D. Kaplan,et al.  Silk/chitosan biohybrid hydrogels and scaffolds via green technology , 2014 .

[9]  Zhanghua Li,et al.  Potential mechanisms underlying the Runx2 induced osteogenesis of bone marrow mesenchymal stem cells. , 2015, American journal of translational research.

[10]  R. Spontak,et al.  Swelling behavior and morphological evolution of mixed gelatin/silk fibroin hydrogels. , 2005, Biomacromolecules.

[11]  G. Hunter,et al.  Modulation of crystal formation by bone phosphoproteins: structural specificity of the osteopontin-mediated inhibition of hydroxyapatite formation. , 1994, The Biochemical journal.

[12]  K. Hankenson,et al.  Integration of BMP, Wnt, and notch signaling pathways in osteoblast differentiation , 2011, Journal of cellular biochemistry.

[13]  David L. Kaplan,et al.  Controlling silk fibroin particle features for drug delivery. , 2010, Biomaterials.

[14]  K. Pramanik,et al.  Development of novel silk fibroin/polyvinyl alcohol/sol–gel bioactive glass composite matrix by modified layer by layer electrospinning method for bone tissue construct generation , 2017, Biofabrication.

[15]  A. Terry,et al.  pH induced changes in the rheology of silk fibroin solution from the middle division of Bombyx mori silkworm. , 2004, Biomacromolecules.

[16]  Julian R. Jones.  Reprint of: Review of bioactive glass: From Hench to hybrids. , 2015, Acta biomaterialia.

[17]  Shuping Peng,et al.  Current Progress in Bioactive Ceramic Scaffolds for Bone Repair and Regeneration , 2014, International journal of molecular sciences.

[18]  B. Mandal,et al.  Mimicking Hierarchical Complexity of the Osteochondral Interface Using Electrospun Silk-Bioactive Glass Composites. , 2017, ACS applied materials & interfaces.

[19]  Sourabh Ghosh,et al.  Strategies for replicating anatomical cartilaginous tissue gradient in engineered intervertebral disc. , 2014, ACS applied materials & interfaces.

[20]  S. Chawla,et al.  Differential Regulation of Hedgehog and Parathyroid Signaling in Mulberry and Nonmulberry Silk Fibroin Textile Braids. , 2018, ACS biomaterials science & engineering.

[21]  Thomas Scheibel,et al.  Polymeric materials based on silk proteins , 2008 .

[22]  C. Dayanand,et al.  Structural investigations of phosphate glasses: a detailed infrared study of the x(PbO)-(1−x) P2O5 vitreous system , 1996, Journal of Materials Science.

[23]  Julian R. Jones,et al.  Bioactive Glasses: Frontiers and Challenges , 2015, Front. Bioeng. Biotechnol..

[24]  G. Ciardelli,et al.  Physico-chemical and biological studies on three-dimensional porous silk/spray-dried mesoporous bioactive glass scaffolds , 2016 .

[25]  C. Bain,et al.  External reflection FTIR spectroscopy of the cationic surfactant hexadecyltrimethylammonium bromide (CTAB) on an overflowing cylinder. , 2004, Langmuir : the ACS journal of surfaces and colloids.

[26]  L. Radev,et al.  Silk Fibroin/Calcium Phosphate Silicate Composites: In vitro Bioactivity , 2013 .

[27]  Eileen Gentleman,et al.  Bioactive Glass Scaffolds for Bone Regeneration , 2007 .

[28]  K. Hing,et al.  Strontium substituted bioactive glasses for tissue engineered scaffolds: the importance of octacalcium phosphate , 2015, Journal of Materials Science: Materials in Medicine.

[29]  S. Rath,et al.  Characterization of BMP signaling dependent osteogenesis using a BMP depletable avianized bone marrow stromal cell line (TVA-BMSC). , 2016, Bone.

[30]  Larry L Hench,et al.  Bioactive glasses for in situ tissue regeneration , 2004, Journal of biomaterials science. Polymer edition.

[31]  Julian R. Jones,et al.  Bioglass and Bioactive Glasses and Their Impact on Healthcare , 2016 .

[32]  Julian R. Jones,et al.  Bioactive Glass Foam Scaffolds are Remodelled by Osteoclasts and Support the Formation of Mineralized Matrix and Vascular Networks In Vitro , 2013, Advanced Healthcare Materials.

[33]  Masaki Noda,et al.  Ihh/Gli2 signaling promotes osteoblast differentiation by regulating Runx2 expression and function. , 2007, Molecular biology of the cell.

[34]  Julian R. Jones,et al.  Differentiation of fetal osteoblasts and formation of mineralized bone nodules by 45S5 Bioglass conditioned medium in the absence of osteogenic supplements. , 2009, Biomaterials.

[35]  Keita Ito,et al.  Silk fibroin as biomaterial for bone tissue engineering. , 2016, Acta biomaterialia.

[36]  G. Hunter,et al.  Modulation of crystal formation by bone phosphoproteins: role of glutamic acid-rich sequences in the nucleation of hydroxyapatite by bone sialoprotein. , 1994, The Biochemical journal.

[37]  R. Viana,et al.  Infrared Spectroscopy of Anionic, Cationic, and Zwitterionic Surfactants , 2012 .

[38]  H. Oonishi,et al.  Particulate Bioglass Compared With Hydroxyapatite as a Bone Graft Substitute , 1997, Clinical orthopaedics and related research.

[39]  David L. Kaplan,et al.  Direct‐Write Assembly of Microperiodic Silk Fibroin Scaffolds for Tissue Engineering Applications , 2008 .

[40]  Gavin Jell,et al.  The effects of strontium-substituted bioactive glasses on osteoblasts and osteoclasts in vitro. , 2010, Biomaterials.

[41]  Yinghong Zhou,et al.  A comparative study of mesoporous glass/silk and non-mesoporous glass/silk scaffolds: physiochemistry and in vivo osteogenesis. , 2011, Acta biomaterialia.

[42]  L L Hench,et al.  Gene-expression profiling of human osteoblasts following treatment with the ionic products of Bioglass 45S5 dissolution. , 2001, Journal of biomedical materials research.

[43]  Hala Zreiqat,et al.  Design and Fabrication of 3D printed Scaffolds with a Mechanical Strength Comparable to Cortical Bone to Repair Large Bone Defects , 2016, Scientific Reports.

[44]  D. Kaplan,et al.  Apatite-coated silk fibroin scaffolds to healing mandibular border defects in canines. , 2009, Bone.

[45]  Joseph Christakiran Moses,et al.  Multifunctional Cell Instructive Silk‐Bioactive Glass Composite Reinforced Scaffolds Toward Osteoinductive, Proangiogenic, and Resorbable Bone Grafts , 2018, Advanced healthcare materials.

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

[47]  D. Kaplan,et al.  Silk coating on a bioactive ceramic scaffold for bone regeneration: effective enhancement of mechanical and in vitro osteogenic properties towards load‐bearing applications , 2017, Journal of tissue engineering and regenerative medicine.

[48]  Å. Rosengren,et al.  Protein adsorption onto two bioactive glass-ceramics. , 2003, Biomaterials.

[49]  Molly M. Stevens,et al.  Sparse feature selection methods identify unexpected global cellular response to strontium-containing materials , 2015, Proceedings of the National Academy of Sciences.

[50]  Chengtie Wu,et al.  Osteoimmunomodulation for the development of advanced bone biomaterials , 2016 .

[51]  M. R. Kessler,et al.  Study of Physically Transient Insulating Materials as a Potential Platform for Transient Electronics and Bioelectronics , 2014 .

[52]  J. Polak,et al.  Ionic products of bioactive glass dissolution increase proliferation of human osteoblasts and induce insulin-like growth factor II mRNA expression and protein synthesis. , 2000, Biochemical and biophysical research communications.

[53]  J. Papkoff,et al.  Activated β-catenin induces osteoblast differentiation of C3H10T1/2 cells and participates in BMP2 mediated signal transduction , 2003 .

[54]  S. Midha,et al.  Effect of visco‐elastic silk–chitosan microcomposite scaffolds on matrix deposition and biomechanical functionality for cartilage tissue engineering , 2017, Journal of tissue engineering and regenerative medicine.

[55]  Xiaoyan Wang,et al.  Function and mechanism of mesoporous bioactive glass adsorbed epidermal growth factor for accelerating bone tissue regeneration , 2017, Biomedical materials.

[56]  Sharda Nara,et al.  Precise patterning of biopolymers and cells by direct write technique , 2014 .

[57]  S. Mukherjee,et al.  Nonmulberry Silk Fibroin Scaffold Shows Superior Osteoconductivity Than Mulberry Silk Fibroin in Calvarial Bone Regeneration , 2015, Advanced healthcare materials.

[58]  R. Hill,et al.  Influence of strontium and the importance of glass chemistry and structure when designing bioactive glasses for bone regeneration. , 2010, Acta biomaterialia.

[59]  S. Midha,et al.  Elucidation of differential mineralisation on native and regenerated silk matrices. , 2016, Materials science & engineering. C, Materials for biological applications.

[60]  G. Vunjak‐Novakovic,et al.  Silk based biomaterials to heal critical sized femur defects. , 2006, Bone.

[61]  D. Brauer.  Bioactive glasses—structure and properties. , 2015, Angewandte Chemie.

[62]  C. Rey,et al.  The carbonate environment in bone mineral: A resolution-enhanced fourier transform infrared spectroscopy study , 1989, Calcified Tissue International.

[63]  Hassane Oudadesse,et al.  Antioxidative/oxidative effects of strontium-doped bioactive glass as bone graft. In vivo assays in ovariectomised rats , 2012 .

[64]  D. Kaplan,et al.  Mandibular repair in rats with premineralized silk scaffolds and BMP-2-modified bMSCs. , 2009, Biomaterials.

[65]  M. Lindner,et al.  The effect of crystallization of bioactive bioglass 45S5 on apatite formation and degradation. , 2013, Dental materials : official publication of the Academy of Dental Materials.

[66]  Jun Magoshi,et al.  Rheology and dynamic light scattering of silk fibroin solution extracted from the middle division of Bombyx mori silkworm. , 2002, Biomacromolecules.

[67]  S. Midha,et al.  Osteogenic signaling on silk-based matrices. , 2016, Biomaterials.

[68]  B. Mandal,et al.  Biomimetic, Osteoconductive Non-mulberry Silk Fiber Reinforced Tricomposite Scaffolds for Bone Tissue Engineering. , 2016, ACS applied materials & interfaces.

[69]  Julian R Jones,et al.  Review of bioactive glass: from Hench to hybrids. , 2013, Acta biomaterialia.

[70]  Yufeng Zhang,et al.  The Osteogenic Potential of Mesoporous Bioglasses/Silk and Non-Mesoporous Bioglasses/Silk Scaffolds in Ovariectomized Rats: In vitro and In vivo Evaluation , 2013, PloS one.

[71]  L Geris,et al.  Current views on calcium phosphate osteogenicity and the translation into effective bone regeneration strategies. , 2012, Acta biomaterialia.

[72]  E. Vuorio,et al.  Silica-based bioactive glasses modulate expression of bone morphogenetic protein-2 mRNA in Saos-2 osteoblasts in vitro. , 2001, Biomaterials.

[73]  David L Kaplan,et al.  Direct‐Write Assembly of 3D Silk/Hydroxyapatite Scaffolds for Bone Co‐Cultures , 2012, Advanced healthcare materials.

[74]  Kalim Deshmukh,et al.  Characterization of polyvinyl alcohol/gelatin blend hydrogel films for biomedical applications , 2008 .

[75]  H. Cheung,et al.  Recent Advances in Hydroxyapatite Scaffolds Containing Mesenchymal Stem Cells , 2015, Stem cells international.

[76]  T. Yoneda,et al.  Core-binding factor alpha 1 (Cbfa1) induces osteoblastic differentiation of C2C12 cells without interactions with Smad1 and Smad5. , 2002, Bone.

[77]  S. Midha,et al.  Nonmulberry Silk Braids Direct Terminal Osteocytic Differentiation through Activation of Wnt-Signaling. , 2017, ACS biomaterials science & engineering.

[78]  N. Artigas,et al.  Extracellular calcium promotes bone formation from bone marrow mesenchymal stem cells by amplifying the effects of BMP-2 on SMAD signalling , 2017, PloS one.

[79]  L. Ye,et al.  The Hedgehog signalling pathway in bone formation , 2015, International Journal of Oral Science.

[80]  S. Hesaraki,et al.  The effect of Sr concentration on bioactivity and biocompatibility of sol-gel derived glasses based on CaO-SrO-SiO2-P2O5 quaternary system , 2010 .

[81]  Sourabh Ghosh,et al.  Modulation of Self-Assembly Process of Fibroin: An Insight for Regulating the Conformation of Silk Biomaterials. , 2015, Biomacromolecules.

[82]  L. Ghasemi‐Mobarakeh,et al.  Structural properties of scaffolds: Crucial parameters towards stem cells differentiation. , 2015, World journal of stem cells.

[83]  Julian R. Jones,et al.  Compressive Strength of Bioactive Sol–Gel Glass Foam Scaffolds , 2016 .

[84]  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.

[85]  Julian R. Jones,et al.  Preconditioned 70S30C bioactive glass foams promote osteogenesis in vivo. , 2013, Acta biomaterialia.

[86]  N. Naaman,et al.  Bioavailability of Strontium Ions from Bioactive Glasses In Vivo: A Micro-PIXE Study of Trace Elements at the Bone Interface , 2013 .

[87]  F. Vollrath,et al.  Transition to a beta-sheet-rich structure in spidroin in vitro: the effects of pH and cations. , 2004, Biochemistry.