Two-dimensional effects of hydrogel self-organized from IKVAV-containing peptides on growth and differentiation of NSCs

Abstract

The neural stem cells (NSCs) were seeded in the surface layer of hydrogels made of IKVAV-containing peptide amphiphile. Two-dimensional effects of hydrogel on growth and differentiation of NSCs were investigated. Peptide was synthesized in solid way. Cells were harvested from the cerebral cortex of neonatal mice, identified by immunohistochemical methods. Cells were incubated in the surface layer of self-assembled peptide hydrogel and coverslips for seven days respectively, detected immunocytochemically for NF and GFAP. The molecular weight (M _W) of Peptide was 1438 and purity was 95.22%. Cells were identified as Nestin-positive NSCs. TEM showed that hydrogel was composed of interactive nanofibers. NSCs extended processes, and were able to be differentiated into NF-positive neurons with red fluorescence and GFAP-positive astrocytes with green one in the surface of hydrogel. However, NSCs only formed undifferentiated neurospheres in the surface layer of coverslips. Results indicate that the self-assembled hydrogel from peptide amphiphile has good cyto-compatibility to NSCs and induced their differentiation.

Keywords

nerve tissue engineering / hydrogel / self-assembly / neural stem cells

References

[1] Flynn L., Dalton P.D., Shoichet M.S. Fiber Templating of Poly (2-hydroxyethyl methacrylate) for Neural Tissue Engineering[J]. Biomaterials, 2003, 24(23): 4265-4272.

[2] Miller M.J., Patrick C.W. Jr Tissue Engineering[J]. Clin. Plast. Surg., 2003, 30(1): 91-103.

[3] Schmidt C.E., Leach J.B. Neural Tissue Engineering: Strategies for Repair and Regeneration[J]. Annu. Rev. Biomed. Eng., 2003, 5: 293-347.

[4] Takezawa T. A Strategy for the Development of Tissue Engineering Scaffolds that Regulate Cell Behavior[J]. Biomaterials, 2003, 24(13): 2267-2275.

[5] Vats A., Tolley N.S., Polak J.M., . Scaffolds and Biomaterials for Tissue Engineering: a Review of Clinical Applications, Scaffolds and Biomaterials for Tissue Engineering: a Review of Clinical Applications[J]. Clin. Otolaryngol. Allied Sci., 2003, 28(3): 165-172.

[6] Atala A. Tissue Engineering of Artificial Organs[J]. J. Endourol., 2000, 14(1): 49-57.

[7] Ibarra C., Koski J.A., Warren R.F. Tissue Engineering Meniscus: Cells and Matrix[J]. Orthop. Clin. North Am., 2000, 31(3): 411-418.

[8] Lanza R.P., Langer R.S., Vacanti J. Principles of Tissue Engineering[M], 2000 San Diego Academic Press 1-73.

[9] Aarum J., Sandberg K., Haeberlein S.L., . Migration and Differentiation of Neural Precursor Cells can be Directed by Microglia[J]. Proc. Natl. Acad. Sci. USA, 2003, 100(26): 15983-15988.

[10] Lavik E., Teng Y.D., Snyder E., . Seeding Neural Stem Cells on Scaffolds of PGA, PLA, and Their Copolymers[J]. Methods Mol. Biol., 2002, 198: 89-97.

[11] Park K.I., Teng Y.D., Snyder E.Y. The Injured Brain Interacts Reciprocally with Neural Stem Cells Supported by Scaffolds to Reconstitute Lost Tissue[J]. Nat. Biotechnol., 2002, 20(11): 1111-1117.

[12] Liu C.Y., Apuzzo M.L., Tirrell D.A. Engineering of the Extracellular Matrix: Working Toward Neural Stem Cell Programming and Neurorestoration-concept and Progress Report[J]. Neurosurgery, 2003, 52(5): 1154-1165.

[13] Levenberg S., Huang N.F., Lavik E., . Differentiation of Human Embryonic Stem Cells on Three-dimensional Polymer Scaffolds[J]. Proc. Natl. Acad. Sci. USA, 2003, 100(22): 12741-12746.

[14] Recknor J.B., Recknor J.C., Sakaguchi D.S., . Oriented Astroglial Cell Growth on Micropatterned Polystyrene Substrates[J]. Biomaterials, 2004, 25(14): 2753-2767.

[15] Bottai D., Fiocco R., Hydrogelain F., . Neural Stem Cells in the Adult Nervous System[J]. J. Hematother. Stem. Cell Res., 2003, 12(6): 655-670.

[16] Sakamoto N., Iwahana M., Tanaka N.G., . Inhibition of Angiogenesis and Tumor Growth by A Synthetic Laminin Peptide, GDPGYIGSR-NH2[J]. Cancer Res., 1991, 51: 903-906.

[17] Grant D.S., Tashirom K.I., Segui-Real B., . Two Different Laminin Domains Mediate the Differentiation of Human Endothelial Cells into Capillary-like Structures in Vitro[J]. Cell, 1989, 58: 933-943.

[18] Ponce M.L., Nomizu M., Delgado M.C., . Identification of Endothelial Cell Binding Sites on the Laminin g1 Chain[J]. Circ. Res., 1999, 84: 688-694.

[19] Malinda K.M., Nomizu M., Chung M., . Identification of laminin a1 and b1 Chain Peptides Active for Endothelial Cell Adhesion, Tube Formation and Aortic Sprouting[J]. FASEB J, 1999, 13: 53-62.

[20] Kubota Y., Kleinman H.K., Martin G.R., . Role of Laminin and Basement Membrane in the Morphological Differentiation of Human Endothelial Cells into Capillary-like Structures[J]. J. Cell Biol., 1988, 107: 1589-1598.

[21] Graf J., Ogle R.C., Robey F.A., . A Pentapeptide from the Laminin b1 Chain Mediates Cell Adhesion and Binds the 67000 Laminin Receptor[J]. Biochemistry, 1987, 26: 6896-6900.

[22] Davis G.E., Black S.M., Bayless K.J. Capillary Morphogenesis during Human Endothelial Cell Invation of Three-dimensional Collagen Matrices[J]. In Vitro Cell Dev. Biol. Animal., 2000, 36: 513-519.

[23] Davis G.E., Bayless K.J., Mavila A. Molecular Basis of Endothelial Cell Morphogenesis in Three-dimensional Extracellular Matrices[J]. Anat. Rec., 2002, 268: 252-275.

[24] Bell S.E., Mavila A., Salazar R., . Differential Gene Expression during Capillary Morphogenesis in 3D Collagen Matrices: Regulated Expression of Genes Involved in Basement Membrane Matrix Assembly, Cell Cycle Progression, Cellular Differentiation and G-protein Signalling[J]. J. Cell Science, 2001, 114: 2755-2773.

[25] Kreis T., Vale R. Guidebook of the Extracellular, Anchor and Adhesion Proteins[M], 1999 Second ed. UK A Sabrook & Tooze publication at Oxford University Press

[26] Tashiro K., Sephel G.C., Weeks B., . A Synthetic Peptide Containing the IKVAV Sequence from the A Chain of Laminin Mediates Cell Attachment, Migration, and Neurite Outgrowth[J]. J. Biol. Chem., 1989, 264(27): 16174-16182.

[27] Yamada Y., Kleinman H.K. Functional Domains of Cell Adhesion Molecules[J]. Curr. Opin. Cell Biol., 1992, 4(5): 819-823.

[28] Hartgerink J.D., Beniash E., Stupp S.I. Self-assembly and Mineralization of Peptide-amphiphile Nanofibers[J]. Science, 2001, 294: 1684-1688.

[29] Silva GA, Czeisler C, Niece KL, et al. Selective Differentiation of Neuronal Progenitor Cells by Highepitope Density Nanofibers[J]. Science Express, 2004

[30] Holmes T.C., de Lacalle S., Su X., . Extensive Neurite Outgrowth and Active Synapse Formation on Self-assembling Peptide Scaffolds[J]. Proc. Natl. Acad. Sci. USA, 2000, 97(12): 6728-6733.

[31] Huang Y.Y., Wu X.N., Du X.J. Preparation of a Polyclonal Antibody Against N-terminal of 1A6/DRIM with MAPs Method[J]. Beijing Da Xue Xue Bao, 2004, 36(4): 390-393.

[32] Ren F.R., Lv Q.S., Zhuang H., . Significance of the Signal-to-cutoff Ratios of Anti-hepatitis C Virus Enzyme Immunoassays in Screening of Chinese Blood Donors[J]. Transfusion, 2005, 45(11): 1816-1822.

[33] Lin Y.C., Wu M.Y., Li D.R., . Prognostic and Clinicopathological Features of E-cadherin, Alpha-catenin, Beta-catenin, Gamma-catenin and Cyclin D1 Expression in Human Esophageal Squamous Cell Carcinoma[J]. World J. Gastroenterol, 2004, 10(22): 3235-3239.

[34] Kumar M., Liu Z.R., Thapa L., . Antiangiogenic Effect of Somatostatin Receptor Subtype 2 on Pancreatic Cancer Cell Line: Inhibition of Vascular Endothelial Growth Factor and Matrix Metalloproteinase-2 Expression in Vitro[J]. World J. Gastroenterol, 2004, 10(3): 393-399.

[35] Zhang L., Yuan S.Z. Expression of c-erbB-2 Oncogene Protein, Epidermal Growth Factor Receptor, and TGF-beta1 in Human Pancreatic Ductal Adenocarcinoma[J]. Hepatobiliary Pancreat Dis. Int., 2002, 1(4): 620-623.

[36] Leers M.P., Hoop J.G., Nap M. Her2/neu Analysis in Formalin-fixed, Paraffin-embedded Breast Carcinomas: Comparison of Immunohistochemistry and Multiparameter DNA Flow Cytometry[J]. Anticancer Res., 2003, 23(2A): 999-1006.

[37] Hasui K., Takatsuka T., Sakamoto R., . Improvement of Supersensitive Immunohistochemistry with an Autostainer: a Simplified Catalysed Signal Amplification System[J]. Histochem. J., 2002, 34(5): 215-222.

[38] Sabattini E., Bisgaard K., Ascani S., . The EnVision++ system: A New Immunohistochemical Method for Diagnostics and Research. Critical Comparison with the APAAP, ChemMate, CSA, LABC, and SABC Techniques[J]. J. Clin. Pathol., 1998, 51(7): 506-511.

[39] Sgubin D., Aztiria E., Perin A., . Activation of Endogenous Neural Stem Cells in the Adult Human Brain Following Subarachnoid Hemorrhage[J]. J. Neurosci. Res., 2007, 85(8): 1647-1655.

[40] Takezawa T. A Strategy for the Development of Tissue Engineering Scaffolds that Regulate Cell Behavior[J]. Biomaterials, 2003, 24(13): 2267-2275.

[41] Taha M.O., Mueller S.F., Fraga M.M., . Morphologic Analysis of the Fibular Nerve Repaired with Fibrin Adhesive[J]. Transplant Proc., 2004, 36(2): 401-403.

[42] Matthews B.D., Overby D.R., Alenghat F.J., . Mechanical Properties of Individual Focal Adhesions Probed with A Magnetic Microneedle[J]. Biochem. Biophys. Res. Commun., 2004, 313(3): 758-764.

[43] Polte T.R., Eichler G.S., Wang N., . Extracellular Matrix Controls Myosin Light Chain Phosphorylation and Cell Contractility Through Modulation of Cell Shape and Cytoskeletal Prestress[J]. Am. J. Physiol. Cell Physiol., 2004, 286(3): C518-C528.

[44] Xu Y., Kimura K., Matsumoto N., . Isolation of Neural Stem Cells from the Forebrain of Deceased Early Postnatal and Adult Rats with Protracted Post-mortem Intervals[J]. J. Neurosci. Res., 2003, 74(4): 533-540.

[45] Weber L.M., Hayda K.N., Haskins K., . The Effects of Cell-matrix Interactions on Encapsulated Beta-cell Function within Hydrogels Functionalized with Matrix-derived Adhesive Peptides[J]. Biomaterials, 2007, 28(19): 3004-3011.