C3A Cell Behaviors on Micropatterned Chitosan— Collagen

Abstract

The influence of the properties and surface micropatterning of chitosan—collagen—gelatin (CCG) blended membranes on C3A cell's activities has been investigated. It is aimed to guide the cell growth and improve the growth rate in vitro for the application in tissue engineering. Masters with micropatterns are prepared on stainless steel plates by photolithography. The CCG membranes with surface micropatterns are then fabricated by soft lithography and dry—wet phase inversion techniques. The morphology and metabolic activity of cultured C3A cells on the membranes are recorded. When the C3A cells are seeded on the membranes with micropattern spacing of 200 μm width and 80 μm depth, they adhere and aggregate in the groove of the membranes in a few minutes. The aggregated cells migrate up to the surface of the ridge later. This phenomenon, however, is not found on membranes with a micropattern spacing of 500 μm width. In addition, it is demonstrated that the cells on the CCG membranes with micropatterns have higher metabolism and growth rates than those on the flat CCG membranes and on T-flask discs. Micropatterning on the membrane surface can affect the distribution of cells and the communication among cells, and results in a difference in cell adhesion, morphology, mobility, and growth activity.

Keywords

biomaterials cell—material interactions chitosan— collagen—gelatin and micropatterning.

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References

Lee, L.J. (2003). BioMEMs and Micro-/Nano-processing of Polymers — An Overview, J. Chin. Inst. Chem. Engrs., 34(1): 25—46.

Nikolovski, J. and Mooney, D.J. (2000). Smooth Muscle Cell Adhesion to Tissue Engineering Scaffolds, Biomaterials , 21(20): 2025—2032.

Griffith, L.G. and Noughton, G. (2002). Tissue Engineering-Current Challenges and Expanding Opportunities, Science, 295(5557): 1009—1014.

Manwaring, M.E. , Walsh, J.F. and Tresco, P.A. (2004). Contact Guidance Induced Organization of Extracellular Matrix, Biomaterials, 25(17): 3631— 3638.

Yamato, M. , Konno, C. , Utsumi, M. , Kikuchi, A. and Okano, T. (2002). Thermally Responsive Polymer-Grafted Surfaces Facilitate Patterned Cell Seeding and Co-Culture, Biomaterial , 23(2): 561—567.

Massia, S.P. and Hubbell, J.A. (1992). Immobilized Amines and Basic Amino Acids as Mimetic Heparin-Binding Domains for Cell Surface Proteoglycan-Mediated Adhesion, J. Biological Chem., 267(14): 10133—10141.

Grinnell, F. , Milam, M. and Spree, P.A. (1973). Attachment of Normal and Transformed Hamster Kidney Cells to Substrata Varying in Chemical Composition , Biochem. Med., 7(1): 87—90.

Anselme, K. (2000). Osteoblast Adhesion on Biomaterials, Biomaterials, 21(7): 667—681.

Krasteva, N. , Harms, U. , Albrecht, W. , Seifert, B. , Hopp, M. , Altankov, G. and Groth, T. (2002). Membranes for Biohybrid Liver Support Systems-Investigations on Hepatocyte Attachment, Morphology and Growth, Biomaterials , 23(12): 2467—2478.

10.

Griffith, L.G. (2000). Polymeric Biomaterials , Acta Mater., 48(1): 263—277.

11.

Mao, J.S. , Zhao, L.G. , Yin, Y.J. and Yao, K.D. (2003). Structure and Properties of Bilayer Chitosan-Gelatin Scaffolds, Biomaterial, 24(6): 1067—1074.

12.

Zhao, F. , Yin, Y. , Lu, W.W. , Leong, J.C. , Zhang, W. , Zhang, J. , Zhang, M. and Yao, K. (2002). Preparation and Histological Evaluation of Biomimetic Three-Dimensional Hydroxyapatite/Chitosan-Gelatin Network Composite Scaffolds , Biomaterials, 23(15): 3227—3234.

13.

Yin, Y.J. , Yao, K.D. , Cheng, G.X. and Ma, J.B. (1999). Properties of Polyelectrolyte Complex Films of Chitosan and Gelatin, Polym. Int., 48(16): 420—432.

14.

Mao, J.S. , Cui, Y.L. , Wang, X.H. , Sun, Y. , Yin, Y.J. and Zhao, H.M. (2004). A Preliminary Study on Chitosan and Gelatin Polyelectrolyte Complex Cytocompatibility by Cell Cycle and Apoptosis Analysis, Biomaterials, 25(18): 3973—3981.

15.

Huang, Y. , Onyeri, S. , Siewe, M. , Moshfeghian, A. and Madihally, S.V. (2005). In vitro Characterization of Chitosan-Gelatin Scaffolds for Tissue Engineering, Biomaterials, 26(36): 7616—7626.

16.

Chen, C.S. , Mrksich, M. , Huang, S. , Whitesides, G.M. and Ingber, D.E. (1997). Geometric Control of Cell Life and Death, Science, 276(5317): 1425—1428.

17.

Zhang, S. , Yan, L. , Altman, M. , Lassle, M. , Nugent, H. , Frankel, F. , Lauffenburger, D.A. , Whitesides, G.M. and Rich, A. (1999). Biological Surface Engineering: A Simple System for Cell Pattern Formation, Biomaterials, 20(13): 1213—1220.

18.

Kane, R.S. , Takayama, S. , Ostuni, E. , Ingber, D.E. and Whitesides, G.M. (1999). Patterning Proteins and Cells Using Soft Lithography, Biomaterials, 20(23—24): 2363—2376.

19.

Wang, L. , Sun, J. , Li, L. , Mears, D. , Horvat, M. and Sheil, A.R. (1998). Comparison of Porcine Hepatocytes with Human Hepatoma (C3A) Cells for Use in a Bioartifical liver support system, Cell Transplant., 7(5): 459—468.

20.

Dalby, M.J. , Childs, S. , Riehle , M.O. , Johnstone, H.J.H. , Affrossman , S. and Curtis, A.S.G. (2003). Fibroblast Reaction to Island Topography: Changes in Cytoskeleton and Morphology with Time, Biomaterials, 24(16): 927—935.

21.

Rajaraman, R. , Rounds, D.E. , Yen , S.P.S. and Rembaum , A. (1974). A Scanning Electron Microscopy Study of Cell Adhesion and Spreading in vitro , Experimental Cell Research, 88(2): 327—339.

22.

Hughes, R.D. and Williams, R. (1996). Assessment of Bio-Artificial Liver Support in Acute Liver Failure, Int. J. Artif. Organs , 19(1): 3—6.

23.

Chen, J.P. , Yu, S.C. , Hsu, B.R.S. , Fu, S.H. and Lin, H.S. (2003). Loofa Sponge as a Scaffold for the Culture of Human Hepatocyte Cell Line, Biotechnol. Prog., 19(2): 522—527.

24.

Kang, I.K. , Kim, G.J. , Kwon, O.H. and Ito, Y. (2004). Co-culture of Hepatocytes and Fibroblasts by Micropatterned Immobilization of β-galactose Derivatives, Biomaterials , 25(18): 4225—4232.

25.

Fukuda, J. , Khademhosseini, A. , Yeh, J. , Eng, G. , Cheng, J. , Farokhzad, O.C. and Langer, R. (2006). Micropatterned Cell Co-Cultures Using Layer-by-layer Deposition of Extracellular Matrix Components, Biomaterials , 27(8): 1479—1486.

26.

Mosmann, T. (1983). Rapid Colorimetric Assay for Cellular Growth and Survival: Application to Proliferation and Cytotoxicity Assays, J. Immuno. Methods, 65(1—2): 55—63.

27.

Lobner, D. (2000). Comparison of the LDH and MTT Assays for Quantifying Cell Death: Validity for Neuronal Apoptosis?, J. Neurosci. Meth., 96(2): 147—152.

28.

Ren, J.G. , Zheng, R.L. , Shi, Y.M. , Gong, B. and Li, J.F. (1998). Apoptosis, Redifferentiation and Arresting Proliferation Simultaneously Triggered by Oxidative Stress in Human Hepatoma Cells, Cell Biology Int., 22(1): 41—49.

C3A Cell Behaviors on Micropatterned Chitosan— Collagen—Gelatin Membranes

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