Insulin-like Growth Factor I—Releasing Alginate-Tricalciumphosphate Composites for Bone Regeneration

PurposeDevelopment and characterization of an in situ–forming, osteoconductive, and growth factor–releasing bone implant.MethodsInjectable in situ–forming scaffolds were prepared from a 2% (m/v) alginate solution, tricalciumphosphate (TCP) granules, and poly(lactide-co-glycolide) microspheres (MS), loaded with the osteoinductive growth factor insulin-like growth factor I (IGF-I). Scaffolds were prepared by mixing the components followed by hydrogel formation through calcium carbonate–induced physical cross-linking of the alginate at slightly acidic pH. Physical-chemical properties and cell biocompatibility using osteoblast-like cells (MG-63 and Saos-2) of these scaffolds were investigated.ResultsThe addition of TCP to the alginate resulted in reduced swelling and gelation time and an increase in stiffness. Osteoblast-like cells (MG-63 and Saos-2) did not show toxic reactions and adhered circumferentially to the TCP granules surface. The addition of the IGF-I MS resulted in an up to sevenfold increased proliferation rate of MG-63 cells as compared to scaffold preparations without IGF-I MS. The alkaline phosphate (ALP) activity—a parameter for osteblastic activity—increased with increasing amounts of TCP in Saos-2 loaded composite scaffolds.ConclusionsA prototype in situ–hardening composite system for conformal filling of bone defects supporting osteoblastic activity for further clinical testing in relevant fracture models was developed and characterized.

[1]  C. Farquharson,et al.  Mitogenic action of insulin-like growth factor-I on human osteosarcoma MG-63 cells and rat osteoblasts maintained in situ: the role of glucose-6-phosphate dehydrogenase. , 1993, Bone and mineral.

[2]  M. Longaker,et al.  Gene expression of insulin-like growth factors I and II in rat membranous osteotomy healing. , 1999, Annals of Plastic Surgery.

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

[4]  Athanassios Sambanis,et al.  The role of the CaCl2-guluronic acid interaction on alginate encapsulated betaTC3 cells. , 2004, Biomaterials.

[5]  L. S. Matthews,et al.  The mechanical properties of human tibial trabecular bone as a function of metaphyseal location. , 1983, Journal of biomechanics.

[6]  G. Wesolowski,et al.  Characterization of a human osteosarcoma cell line (Saos-2) with osteoblastic properties. , 1987, Cancer research.

[7]  D J Mooney,et al.  Regulating Bone Formation via Controlled Scaffold Degradation , 2003, Journal of dental research.

[8]  H. Blau,et al.  Induction of angiogenesis by implantation of encapsulated primary myoblasts expressing vascular endothelial growth factor , 2000, The journal of gene medicine.

[9]  J. Jansen,et al.  The effect of poly-L-lactic acid with parallel surface micro groove on osteoblast-like cells in vitro. , 1999, Biomaterials.

[10]  P. Ma,et al.  Ionically crosslinked alginate hydrogels as scaffolds for tissue engineering: part 1. Structure, gelation rate and mechanical properties. , 2001, Biomaterials.

[11]  M. Gowen,et al.  Are MG-63 and HOS TE85 human osteosarcoma cell lines representative models of the osteoblastic phenotype? , 1994, Bone.

[12]  Antonios G Mikos,et al.  Development of an injectable, in situ crosslinkable, degradable polymeric carrier for osteogenic cell populations. Part 2. Viability of encapsulated marrow stromal osteoblasts cultured on crosslinking poly(propylene fumarate). , 2002, Biomaterials.

[13]  S. Dedhar,et al.  The osteoblast-like differentiated phenotype of a variant of MG-63 osteosarcoma cell line correlated with altered adhesive properties. , 1989, Connective tissue research.

[14]  Allan S Hoffman,et al.  Hydrogels for biomedical applications. , 2002, Advanced drug delivery reviews.

[15]  F. Gu,et al.  Sustained delivery of vascular endothelial growth factor with alginate beads. , 2004, Journal of controlled release : official journal of the Controlled Release Society.

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

[17]  M. Jasionowski,et al.  Injectable gels for tissue engineering , 2001, The Anatomical record.

[18]  Jason A Burdick,et al.  Delivery of osteoinductive growth factors from degradable PEG hydrogels influences osteoblast differentiation and mineralization. , 2002, Journal of controlled release : official journal of the Controlled Release Society.

[19]  D. Bidwell,et al.  Formation , 2006, Revue Francophone d'Orthoptie.

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

[21]  Patrick Soon-Shiong,et al.  Induction of Cytokine Production from Human Monocytes Stimulated with Alginate , 1991, Journal of immunotherapy : official journal of the Society for Biological Therapy.

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

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

[24]  Thomas A Einhorn,et al.  Differential Temporal Expression of Members of the Transforming Growth Factor β Superfamily During Murine Fracture Healing , 2002, Journal of bone and mineral research : the official journal of the American Society for Bone and Mineral Research.

[25]  David Byrom.  Biomaterials: Novel Materials from Biological Sources , 1991 .

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

[27]  H. Sussman,et al.  Characterization of a human osteoblastic osteosarcoma cell line (SAOS‐2) with high bone alkaline phosphatase activity , 1987, Journal of bone and mineral research : the official journal of the American Society for Bone and Mineral Research.

[28]  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.

[29]  D. Mooney,et al.  Designing alginate hydrogels to maintain viability of immobilized cells. , 2003, Biomaterials.

[30]  J. V. Van Wyk,et al.  Purification of somatomedin-C from human plasma: chemical and biological properties, partial sequence analysis, and relationship to other somatomedins. , 1980, Biochemistry.

[31]  D J Mooney,et al.  Alginate hydrogels as synthetic extracellular matrix materials. , 1999, Biomaterials.

[32]  Wayne R. Gombotz,et al.  Protein release from alginate matrices. , 1998, Advanced drug delivery reviews.

[33]  H. Senoo,et al.  Effects of ascorbic acid and ascorbic acid 2‐phosphate, a long‐acting vitamin C derivative, on the proliferation and differentiation of human osteoblast‐like cells , 2004, Cell biology international.

[34]  H K Koerten,et al.  Degradation of calcium phosphate ceramics. , 1999, Journal of biomedical materials research.

[35]  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.

[36]  J. Drysdale,et al.  Isoelectric points of proteins: a table. , 1978, Analytical biochemistry.

[37]  H. Blau,et al.  Angiogenesis monitored by perfusion with a space-filling microbead suspension. , 2000, Molecular therapy : the journal of the American Society of Gene Therapy.

[38]  F. Lin,et al.  The effects of calcium phosphate particles on the growth of osteoblasts. , 1997, Journal of biomedical materials research.

[39]  Y. M. Elçin,et al.  Extensive in vivo angiogenesis following controlled release of human vascular endothelial cell growth factor: implications for tissue engineering and wound healing. , 2001, Artificial organs.

[40]  B. Spargo,et al.  Recruitment of tissue resident cells to hydrogel composites: in vivo response to implant materials. , 1994, Biomaterials.

[41]  P. Dynes,et al.  Relationship between viscoelastic properties and gelation in thermosetting systems , 1982 .

[42]  E. Schoenle,et al.  Insulin-like growth factors I and II: some biological actions and receptor binding characteristics of two purified constituents of nonsuppressible insulin-like activity of human serum. , 1978, European journal of biochemistry.

[43]  J. Pearlman,et al.  Local perivascular delivery of basic fibroblast growth factor in patients undergoing coronary bypass surgery: results of a phase I randomized, double-blind, placebo-controlled trial. , 1999, Circulation.

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

[45]  D J Mooney,et al.  Release from alginate enhances the biological activity of vascular endothelial growth factor. , 1998, Journal of biomaterials science. Polymer edition.

[46]  Antonios G Mikos,et al.  Development of an injectable, in situ crosslinkable, degradable polymeric carrier for osteogenic cell populations. Part 1. Encapsulation of marrow stromal osteoblasts in surface crosslinked gelatin microparticles. , 2002, Biomaterials.