Yoshihiro Ito Nano Med Eng Lab RIKEN Advanced Science Institute
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1 Immobilization of Growth Factors on Inorganic Materials Yoshihiro Ito Nano Med Eng Lab RIKEN Advanced Science Institute
2 RIKEN Activities
3 Tissue Engineering Cell Scaffold, Matrix Growth factor Fusion Immobilization of growth factor on scaffold
4 Differentiation roliferation Apoptosis Growth Factors Transformation
5 Cell Biosignal molecule Stimulation Immobilized biosignal molecules Long term Y. Ito et al., Biomaterials, 1991; NAS, 1996; Y. Ito, Biomaterials, 1999Soft Matter, 2008
6 Immobilized Growth Factor Work Micropattern-Immobilization of Insulin photomask 100 μm CHO-Tcell
7 Cell hosphorylation of tyrosine residues of signal transduｃtion proteins Substrate receptor
8 Cell Biosignal molecule Stimulation Immobilized biosignal molecules Long term
9 Immobilized growth factor Growth Rate of Cells Soluble growth factor Amount in culture media
10 First time culture Second time culture 100µm Apoptosis Leukocyte Erythoropoetin 100µm
11 List of growth factors immobilized Growth Factors Materials Insulin(1991-) olystyrene Epidermal growth factor () (1996-) oly(methyl methacrylate) Nerve growth factor (NGF) oly(lactic acid) Bone morphogen protein (BM) Vasular endothelial growth factor (V) Titan Fibroblast growth factor (FGF) Stainless steel Insulin growth factor-1 (IGF-1) Gold Transforming growth factor-β1 (TGF-β1) Hapatocyte growth factor (HGF) Hydroxy apatite Leukemia inhibitory factor (LIF) Glass Stem cell growth factor (SCF) Interleukin-2 Interleukin-1 Tumor necrosis factor α (TNF- α ) Erythropoetin Neurotrophin-3 Transferrin E-cadherin Osteopontin -selectin CXCR3 ligand Notch ligand Sonic hedgehog Ephrin-B2 Osteotropic peptide parathyroid hormone (1-34) Y. Ito, Biological Interactions on Materials Surfaces: Understanding and Controlling rotein, Cell, and Tissue Responses ed. by D.uleo, Springer (2009) Y. Ito, Comprehensive Biomaterials, ed. by D. Grainger et al. Elsevier (2011) B. Joddar, Y. Ito J. Mater. Chem., (2011)
12 Stem cell Matrix Growth Factor Fusion Chemical Gene-engineered
13 ECM-Binding Growth Factor Natural Diffusion Injured Sites Collagen Modified Binding Enhancement of regeneration
14 Application for skin ulcerdb/db mouse Day 21 Full-thickness skin wound 1.5cm x 1.5cm in diabetic mouse (db/db) CBD- Injection of growth factors after dressing
15 . Re-vascularization Day0Day10 Vehicle non-treated Necrosis Thermography Recovery Estimation Injection of Growth Factors V artial necrosis CBD-V
16 I Growth Factor Growth Factor Titanium Stainless steel Hydroxyapatite Inorganic surface O Inorganic O Inorganic Growth Factor Inorganic
17 metal ph 4.5 ph 8.5 dopamine coating metal metal dopamine metal immobilization of 3.40 ± 0.49 nm 4.97 ± 0.62 µm dopamine metal
18 Dopamine-coated metal Titanium clean metal Non 1 mg/ml (ph 4.5) 2 mg/ml 1 mg/ml 2 mg/ml (ph 4.5) (ph 8.5) (ph 8.5) overnight dopamine solution Stainless steel dopamine-coated metal Non 1 mg/ml (ph 4.5) 2 mg/ml 1 mg/ml 2 mg/ml (ph 4.5) (ph 8.5) (ph 8.5)
19 Amino group on metal surface FITC S C N dopami ne N H 2 COO H OH O O ph 9.0 dopamine NH C S NH ph 4.5 ph 8.5 COOH titanium stainless steel OH O O
20 Immobilization of on metal surface NH2 Titanium O O metal NHS WSC immobilized (ng/cm 2 ) C O C O C O NH NH NH dopamine metal Stainless steel immobilized (ng/cm 2 )
21 Cell proliferation on modified metal surface Titanium Cell metal immobilized relative cell growth Soluble VS Stainless steel Soluble Cell metal soluble
22 I Growth Factor Growth Factor Titanium Stainless steel Hydroxyapatite Inorganic surface O Inorganic O Inorganic Growth Factor Inorganic Binding sequence has non-canonical amino acids
23 Salivary Statherin Hydroxyapatite (HAp) Binding site (phosphorylated serine) + HAp-Binding or BM Epidermal Growth Factor () Bone Morphogenic rotein (MB)
24 Recombinant MANSD-SECL- SHDGY-CLHDG-VCMYI-EAIDK-YACNC- VVGYI- GERCQ-YRDLK-WWELR-EFLE-G Salivary Statherin DSSEE-KFLRR-IGRFG-YGYG-YQV-EQLY-QYQ-QYQQ-YTF pp N8 CLSH-DGYCL-HDGVC-MYIEA-LDKYA-CNCVV-GYIGE-RCQYR-DLKWW- ECGGDSSEE p p
25 Binding assay Hydroxyapatite N8 inorganic surface N8 immobilized (ng/cm 2 ) VS Titanium inorganic surface
26 Cell proliferation on modified inorganic surface Hydroxyapatite Cell N8 N8 inorganic N8 immobilized VS Titanium Cell inorganic soluble
27 Bioorthogonal rotein Engineering Growth Factor Statherin-spired peptide
28 Binding assay of engineered hbm4 to HA
29 Differentiation on Surface immobilized with BM Immobilized state Solution state Osteogenic marker expressions in C3H10T1/2 cell
30 Nano Medical Engineering Laboratory at RIKEN Advanced Science Institute
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