Comparison of Primary Rat Hepatocyte Attachment to Collagen and Plasma‐Polymerised Allylamine on Glass

Summary: Cellular attachment to substrata is a crucial stage in cell culture and tissue engineering. Many cells including hepatocytes attach poorly to synthetic substrata and favour attachment to ECM proteins. Currently, cellular attachment to synthetic materials is often optimised by applying ECM protein coatings such as collagen. In this work, ppAAm was examined as an alternative coating method to collagen type-I gel to promote attachment of primary rat hepatocytes to glass. XPS analysis was used to quantify the composition of the nitrogen containing ppAAm surfaces where the nitrogen is present in the form of amine, imine and possibly amide functionalities. Phase contrast microscopy and total cellular protein measurements indicated that hepatocyte attachment to glass was significantly improved by coating with ppAAm, collagen type-I gel or collagen type-I gel on ppAAm. In addition, cytochrome P450 activity and albumin secretion of hepatocytes cultured on ppAAm were similar to the levels observed in hepatocytes plated on collagen type-I gel or collagen type-I gel on ppAAm. In conclusion, the ppAAm coating was shown to improve the attachment of hepatocytes to glass while maintaining similar functionality to cells cultured on collagen type-I gel. Phase contrast images of primary rat hepatocytes plated on collagen type-I gel (top) and plasma polymerised allylamine (bottom) after 24 h in culture.

[1]  K. Shakesheff,et al.  Using a Core–Sheath Distribution of Surface Chemistry through 3D Tissue Engineering Scaffolds to Control Cell Ingress , 2006 .

[2]  Morgan R. Alexander,et al.  Using Plasma Deposits to Promote Cell Population of the Porous Interior of Three‐Dimensional Poly(D,L‐Lactic Acid) Tissue‐Engineering Scaffolds , 2005 .

[3]  Enrico Drioli,et al.  Biotransformation and liver-specific functions of human hepatocytes in culture on RGD-immobilized plasma-processed membranes. , 2005, Biomaterials.

[4]  A. Shard,et al.  The effect of positive ion energy on plasma polymerization: a comparison between acrylic and propionic acids. , 2005, The journal of physical chemistry. B.

[5]  R. A. Talib,et al.  A NEXAFS examination of unsaturation in plasma polymers of allylamine and propylamine , 2004 .

[6]  M. Davidson,et al.  Isopropyl alcohol plasma modification of polystyrene surfaces to influence cell attachment behaviour , 2004 .

[7]  Emile de Heer,et al.  Hepatocyte survival depends on β1‐integrin‐mediated attachment of hepatocytes to hepatic extracellular matrix , 2004, Liver international : official journal of the International Association for the Study of the Liver.

[8]  L. Gribaldo,et al.  Combination of ion beam stabilisation, plasma etching and plasma deposition for the development of tissue engineering micropatterned supports , 2004, Journal of biomaterials science. Polymer edition.

[9]  T. Groth,et al.  Surface properties of and cell adhesion onto allylamine-plasma-coated polyethylenterephtalat membranes. , 2003, Biomaterials.

[10]  T. Groth,et al.  Enhanced tissue-compatibility of polyethylenterephtalat membranes by plasma aminofunctionalisation , 2003 .

[11]  K. Shakesheff,et al.  Spatial confinement of neurite regrowth from dorsal root ganglia within nonporous microconduits. , 2003, Tissue engineering.

[12]  A. Hollander,et al.  Adsorption of vitronectin, collagen and immunoglobulin-G to plasma polymer surfaces by enzyme linked immunosorbent assay (ELISA) , 2002 .

[13]  Shen‐guo Wang,et al.  Enhanced cell affinity of poly (D,L-lactide) by combining plasma treatment with collagen anchorage. , 2002, Biomaterials.

[14]  Junying Chen,et al.  Plasma-surface modification of biomaterials , 2002 .

[15]  M. Schwartz.  Integrin signaling revisited. , 2001, Trends in cell biology.

[16]  T. Groth,et al.  The role of surface wettability on hepatocyte adhesive interactions and function , 2001, Journal of biomaterials science. Polymer edition.

[17]  R. Timmons,et al.  Pulsed plasma deposition of allylamine on polysiloxane: a stable surface for neuronal cell adhesion , 2000, Journal of Neuroscience Methods.

[18]  A. Yamamoto,et al.  Quantitative evaluation of cell attachment to glass, polystyrene, and fibronectin- or collagen-coated polystyrene by measurement of cell adhesive shear force and cell detachment energy. , 2000, Journal of biomedical materials research.

[19]  R. Eberhart,et al.  Enhancing hepatocyte adhesion by pulsed plasma deposition and polyethylene glycol coupling. , 2000, Tissue engineering.

[20]  G. Farrell,et al.  The effects of cell density, attachment substratum and dexamethasone on spontaneous apoptosis of rat hepatocytes in primary culture , 1999, In Vitro Cellular & Developmental Biology - Animal.

[21]  E. Edelman,et al.  Effects of amide and amine plasma-treated ePTFE vascular grafts on endothelial cell lining in an artificial circulatory system. , 1998, Journal of biomedical materials research.

[22]  P. Tresco,et al.  Relative importance of surface wettability and charged functional groups on NIH 3T3 fibroblast attachment, spreading, and cytoskeletal organization. , 1998, Journal of biomedical materials research.

[23]  A. Guillouzo,et al.  Liver cell models in in vitro toxicology. , 1998, Environmental health perspectives.

[24]  Erkki Ruoslahti,et al.  Cell attachment activity of fibronectin can be duplicated by small synthetic fragments of the molecule , 1984, Nature.

[25]  H. Kleinman,et al.  Role of collagenous matrices in the adhesion and growth of cells , 1981, The Journal of cell biology.

[26]  R. Short,et al.  Attachment of human keratinocytes to plasma co-polymers of acrylic acid/octa-1,7-diene and allyl amine/octa-1,7-diene , 1998 .

[27]  M. Alexander,et al.  The chemistry of deposits formed from acrylic acid plasmas , 1998 .

[28]  R. Short,et al.  Radiofrequency-induced plasma polymerisation of propenoic acid and propanoic acid , 1995 .

[29]  A. Guillouzo,et al.  Long-term culture of functional hepatocytes. , 1990, Toxicology in vitro : an international journal published in association with BIBRA.

[30]  P. Seglen.  Preparation of isolated rat liver cells. , 1976, Methods in cell biology.