Insulin-like growth factor I releasing silk fibroin scaffolds induce chondrogenic differentiation of human mesenchymal stem cells.

Growth factor releasing scaffolds are an emerging alternative to autologous or allogenous implants, providing a biologically active template for tissue (re)-generation. The goal of this study is to evaluate the feasibility of controlled insulin-like growth factor I (IGF-I) releasing silk fibroin (SF) scaffolds in the context of cartilage repair. The impact of manufacturing parameters (pH, methanol treatment and drug load) was correlated with IGF-I release kinetics using ELISA and potency tests. Methanol treatment induced water insolubility of SF scaffolds, allowed the control of bioactive IGF-I delivery and did not affect IGF-I potency. The cumulative drug release correlated linearly with the IGF-I load. To evaluate the chondrogenic potential of the scaffolds, hMSC were seeded on unloaded and IGF-I loaded scaffolds in TGF-beta supplemented medium. Chondrogenic differentiation of hMSC was observed on IGF-I loaded scaffolds, starting after 2 weeks and more strongly after 3 weeks, whereas no chondrogenic responses were observed on unloaded control scaffolds. IGF-I loaded porous SF scaffolds have the potential to provide chondrogenic stimuli to hMSC. Evidence for in vivo cartilage (re)generation must be demonstrated by future, pre-clinical proof of concept studies.

[1]  David L Kaplan,et al.  The inflammatory responses to silk films in vitro and in vivo. , 2005, Biomaterials.

[2]  David L Kaplan,et al.  Silk-based biomaterials. , 2003, Biomaterials.

[3]  C. Polychronakos,et al.  Insulinlike growth factor I: a potent mitogen for human osteogenic sarcoma. , 1990, Journal of the National Cancer Institute.

[4]  K. Stewart,et al.  IGF-I does not affect the proliferation or early osteogenic differentiation of human marrow stromal cells. , 2003, Bone.

[5]  Junzo Tanaka,et al.  Growth factor combination for chondrogenic induction from human mesenchymal stem cell. , 2004, Biochemical and biophysical research communications.

[6]  D. Kaplan,et al.  Silk microspheres for encapsulation and controlled release. , 2007, Journal of controlled release : official journal of the Controlled Release Society.

[7]  Lorenz Meinel,et al.  Insulin-like Growth Factor I—Releasing Alginate-Tricalciumphosphate Composites for Bone Regeneration , 2005, Pharmaceutical Research.

[8]  C. Yu,et al.  Influence of backbone conformation on protein aggregation. , 2002, Journal of the American Chemical Society.

[9]  Yasuhiko Tabata,et al.  Collagenous matrices as release carriers of exogenous growth factors. , 2004, Biomaterials.

[10]  Ralph Müller,et al.  Effect of scaffold design on bone morphology in vitro. , 2006, Tissue engineering.

[11]  P. Conget,et al.  Phenotypical and functional properties of human bone marrow mesenchymal progenitor cells , 1999, Journal of cellular physiology.

[12]  Miss A.O. Penney.  (b) , 1974, The New Yale Book of Quotations.

[13]  M. Tsukada,et al.  Attachment and growth of fibroblast cells on silk fibroin. , 1995, Biochemical and biophysical research communications.

[14]  V. Rosen,et al.  Novel regulators of bone formation: molecular clones and activities. , 1988, Science.

[15]  Jörg A Auer,et al.  Localized insulin-like growth factor I delivery to enhance new bone formation. , 2003, Bone.

[16]  David L Kaplan,et al.  The support of adenosine release from adenosine kinase deficient ES cells by silk substrates. , 2006, Biomaterials.

[17]  J. Jansen,et al.  Degradable hydrogel scaffolds for in vivo delivery of single and dual growth factors in cartilage repair. , 2007, Osteoarthritis and cartilage.

[18]  E. Solheim.  Growth factors in bone , 1998, International Orthopaedics.

[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]  Z. Hochberg,et al.  Insulin-like growth factor I accelerates proliferation and differentiation of cartilage progenitor cells in cultures of neonatal mandibular condyles. , 1993, Acta endocrinologica.

[21]  J. Wozney,et al.  The bone morphogenetic protein family and osteogenesis , 1992, Molecular reproduction and development.

[22]  G. Vunjak‐Novakovic,et al.  Engineering bone-like tissue in vitro using human bone marrow stem cells and silk scaffolds. , 2004, Journal of biomedical materials research. Part A.

[23]  S. Mohan,et al.  Bone growth factors. , 1991, Clinical orthopaedics and related research.

[24]  J. Jansen,et al.  Growth factor-loaded scaffolds for bone engineering. , 2005, Journal of controlled release : official journal of the Controlled Release Society.

[25]  T. Peng,et al.  Charge-selective recognition at fibroin-modified electrodes for analytical application , 2005, Analytical and bioanalytical chemistry.

[26]  Lorenz Meinel,et al.  Localized delivery of growth factors for bone repair. , 2004, European journal of pharmaceutics and biopharmaceutics : official journal of Arbeitsgemeinschaft fur Pharmazeutische Verfahrenstechnik e.V.

[27]  T. Aigner,et al.  Collagens--major component of the physiological cartilage matrix, major target of cartilage degeneration, major tool in cartilage repair. , 2003, Advanced drug delivery reviews.

[28]  A. Göpferich,et al.  Insulin in Tissue Engineering of Cartilage: A Potential Model System for Growth Factor Application , 2001, Journal of drug targeting.

[29]  Gordana Vunjak-Novakovic,et al.  Engineering cartilage‐like tissue using human mesenchymal stem cells and silk protein scaffolds , 2004, Biotechnology and bioengineering.

[30]  K. Mark,et al.  Insulin enhances the growth of cartilage in organ and tissue cultures of mouse neonatal mandibular condyle , 1993, Calcified Tissue International.

[31]  G. Vunjak‐Novakovic,et al.  Silk fibroin as an organic polymer for controlled drug delivery. , 2006, Journal of controlled release : official journal of the Controlled Release Society.

[32]  V. Rosen,et al.  Novel regulators of bone formation: molecular clones and activities. , 1988 .

[33]  Hamidreza Ghandehari,et al.  In vitro and in vivo evaluation of recombinant silk-elastinlike hydrogels for cancer gene therapy. , 2004, Journal of controlled release : official journal of the Controlled Release Society.

[34]  David L. Kaplan,et al.  Mechanical and thermal properties of dragline silk from the spider Nephila clavipes , 1994 .

[35]  M. Mastrogiacomo,et al.  Effect of different growth factors on the chondrogenic potential of human bone marrow stromal cells. , 2001, Osteoarthritis and cartilage.

[36]  David L Kaplan,et al.  Porous silk fibroin 3-D scaffolds for delivery of bone morphogenetic protein-2 in vitro and in vivo. , 2006, Journal of biomedical materials research. Part A.

[37]  M. Urist,et al.  Bone: Formation by Autoinduction , 1965, Science.

[38]  C. Rosen,et al.  Insulin‐like growth factor binding proteins in femoral and vertebral bone marrow stromal cells: Expression and regulation by thyroid hormone and dexamethasone , 2001, Journal of cellular biochemistry.

[39]  A. Schulz,et al.  Insulin-like growth factor I (IGF-I) stimulates proliferation but also increases caspase-3 activity, Annexin-V binding, and DNA-fragmentation in human MG63 osteosarcoma cells: co-activation of pro- and anti-apoptotic pathways by IGF-I. , 2003, Hormone and metabolic research = Hormon- und Stoffwechselforschung = Hormones et metabolisme.

[40]  J. Wallace,et al.  Mutations in the B-domain of insulin-like growth factor-I influence the oxidative folding to yield products with modified biological properties. , 1995, The Biochemical journal.

[41]  L. Meinel,et al.  Stabilizing insulin-like growth factor-I in poly(D,L-lactide-co-glycolide) microspheres. , 2001, Journal of controlled release : official journal of the Controlled Release Society.

[42]  M B McCarthy,et al.  Functionalized silk-based biomaterials for bone formation. , 2001, Journal of biomedical materials research.

[43]  D. Leroith,et al.  Clinical relevance of systemic and local IGF-I. , 2005, Endocrine development.

[44]  D. Zurakowski,et al.  Overexpression of human insulin-like growth factor-I promotes new tissue formation in an ex vivo model of articular chondrocyte transplantation , 2001, Gene Therapy.

[45]  Robert Langer,et al.  Controlled‐release of IGF‐I and TGF‐β1 in a photopolymerizing hydrogel for cartilage tissue engineering , 2001, Journal of orthopaedic research : official publication of the Orthopaedic Research Society.

[46]  Hyun D Kim,et al.  Retention and activity of BMP-2 in hyaluronic acid-based scaffolds in vitro. , 2002, Journal of biomedical materials research.

[47]  K. Shakesheff,et al.  Growth factor release from tissue engineering scaffolds , 2001, The Journal of pharmacy and pharmacology.

[48]  W. Lopaczynski,et al.  Insulin-like growth factor-I (IGF-I) dependent phosphorylation of the IGF-I receptor in MG-63 cells , 1993, Regulatory Peptides.

[49]  D. Streiner.  Maintaining Standards: Differences between the Standard Deviation and Standard Error, and When to Use Each , 1996, Canadian journal of psychiatry. Revue canadienne de psychiatrie.

[50]  Hamidreza Ghandehari,et al.  Controlled Release of Plasmid DNA from a Genetically Engineered Silk-Elastinlike Hydrogel , 2002, Pharmaceutical Research.

[51]  G. S. Nadiger,et al.  Crystallinity in silk fibers: Partial acid hydrolysis and related studies , 1980 .

[52]  H. Moses,et al.  Effect of IGF‐I in the Chondrogenesis of Bone Marrow Mesenchymal Stem Cells in the Presence or Absence of TGF‐β Signaling , 2005, Journal of bone and mineral research : the official journal of the American Society for Bone and Mineral Research.

[53]  G. Kopen,et al.  Characterization of mesenchymal stem cells isolated from murine bone marrow by negative selection , 2003, Journal of cellular biochemistry.

[54]  S. -. Lee,et al.  Cytokine delivery and tissue engineering. , 2000, Yonsei medical journal.

[55]  T. Mittlmeier,et al.  IGF-I and TGF-beta1 application by a poly-(D,L-lactide)-coated cage promotes intervertebral bone matrix formation in the sheep cervical spine. , 2002, Spine.

[56]  F. Rose,et al.  Delivery systems for bone growth factors — the new players in skeletal regeneration , 2004, The Journal of pharmacy and pharmacology.

[57]  S. Mohan,et al.  Insulin-like growth factors I and II are present in the skeletal tissues of ten vertebrates. , 1990, Metabolism: clinical and experimental.

[58]  David L Kaplan,et al.  Macrophage responses to silk. , 2003, Biomaterials.

[59]  R W Farndale,et al.  A direct spectrophotometric microassay for sulfated glycosaminoglycans in cartilage cultures. , 1982, Connective tissue research.