Controlling Primary Hepatocyte Adhesion and Spreading on Protein-Free Polyelectrolyte Multilayer Films

Abstract

The development of new methods for fabricating thin films that provide precise control of the three-dimensional topography and cell adhesion could lead to significant advances in the fields of tissue engineering and biosensors. This Communication describes the successful attachment and spreading of primary hepatocytes on polyelectrolyte multilayer (PEM) films without the use of adhesive proteins such as collagen or fibronectin. We demonstrate that the attachment and spreading of primary hepatocytes can be controlled using this layer-by-layer deposition of ionic polymers. In our study, we used synthetic polymers, namely poly(diallyldimethylammonium chloride) (PDAC) and sulfonated poly(styrene) (SPS) as the polycation and polyanion, respectively, to build the multilayers. Primary hepatocytes attached and spread preferentially on SPS surfaces over PDAC surfaces. SPS patterns were formed on PEM surfaces, either by microcontact printing of SPS onto PDAC surfaces or vice versa, to obtain patterns of primary hepatocytes. PEM is a useful technique for fabricating controlled co-cultures with specified cell-cell and cell-surface interactions on a protein-free environment, thus providing flexibility in designing cell-specific surfaces for tissue engineering applications.

Cell Adhesion on Polyelectrolyte Multilayer Coated Polydimethylsiloxane Surfaces with Varying Topographies

“…We reported that primary hepatocytes attached and spread better on SPS surfaces than on PDAC surfaces. 12 In the previous study, we observed that variation in cell adhesion depended on surface chemistry. In contrast, here we studied the effect of surface topography on cell attachment while maintaining the same surface chemistry.…”

“…[4][5][6][7] This study aimed to gain a better understanding of the cellular response to micro-patterns (i.e., periodic microstructures), which is of significance to the design and application of biomaterials. Many studies have demonstrated that different surface chemistries of materials affect cell attachment, [8][9][10][11][12][13] but the role of surface topographical features on cell growth has been less well studied. Microtextured surfaces affect how the cells attach, spread, and proliferate on these surfaces.…”

Primary Neuron/Astrocyte Co‐Culture on Polyelectrolyte Multilayer Films: A Template for Studying Astrocyte‐Mediated Oxidative Stress in Neurons

“…The choice of SPS and PDAC was based on previous studies, wherein the latter material was shown to be resistant to attachment by primary hepatocytes, [44] smooth muscle cells, [45] and primary neuronal cells, [46] while SPS was cytophilic for all the cell types evaluated. The combination of these two polymers facilitated the patterning of co-cultures of primary neurons and astrocytes, thus allowing direct contact between the cells, more akin to the cellcell contact which exists in the brain than can be achieved with random co-cultures.…”

“…[27,35,36,[41][42][43] We previously reported that primary hepatocytes attached and spread on PEM films. [42,44] Here we report that primary neurons can be cultured on PEM films and further co-cultured with astrocytes in patterned co-cultures and used for studying the effect of saturated FFAs on neuronal cell function.…”

Influences of surface chemistry and swelling of salt-treated polyelectrolyte multilayers on migration of smooth muscle cells

“…Cell adhesion, proliferation and differentiation behaviours on substrates are influenced by surface physico-chemical properties such as topography or roughness, hydrophobicity and hydrophilicity, charge, chemical groups, types of ligands and stiffness [1][2][3][4][5][6]. However, cell migration behaviours on different materials, which are essential for understanding the tissue regeneration process and for developing regenerative materials, are less understood.…”