Flat Polystyrene Research Articles

Protein Binding Leads to Reduced Stability and Solvated Disorder in the Polystyrene Nanoparticle Corona.

Understanding the conformation of proteins in the nanoparticle corona has important implications in how organisms respond to nanoparticle-based drugs. These proteins coat the nanoparticle surface, and their properties will influence the nanoparticle's interaction with cell targets and the immune system. While some coronas are thought to be disordered, two key unanswered questions are the degree of disorder and solvent accessibility. Here, a model is developed for protein corona disorder in polystyrene nanoparticles of varying size. For two different proteins, it is found that binding affinity decreases as nanoparticle size increases. The stoichiometry of binding, along with changes in the hydrodynamic size, supports a highly solvated, disordered protein corona anchored at a small number of attachment sites. The scaling of the stoichiometry versus nanoparticle size is consistent with disordered polymer dimensions. Moreover, it is found that proteins are destabilized less in the presence of larger nanoparticles, and hydrophobic exposure decreases at lower curvatures. The observations hold for proteins on flat polystyrene surfaces, which have the lowest hydrophobic exposure. The model provides an explanation for previous observations of increased amyloid fibrillation rates in the presence of larger nanoparticles, and it may rationalize how cell receptors can recognize protein disorder in therapeutic nanoparticles.

Reducing Cancer Cell Adhesion using Microtextured Surfaces.

For the past century, trypsin has been the primary method of cell dissociation, largely without any major changes to the process. Enzymatic cell detachment strategies for large-scale cell culturing processes are popular but can be labor-intensive, potentially lead to the accumulation of genetic mutations, and produce large quantities of liquid waste. Therefore, engineering surfaces to lower cell adhesion strength could enable the next generation of cell culture surfaces for delicate primary cells and automated, high-throughput workflows. In this study, a process for creating microtextured polystyrene (PS) surfaces to measure the impact of microposts on the adhesion strength of cells is developed. Cell viability and proliferation assays show comparable results in twocancer cell lines between micropost surfaces and standard cell culture vessels. However, cell image analysis on microposts reveals that cell area decreases by half, and leads to an average twofold increase in cell length per area. Using a microfluidic-based method up to a seven times greater percentage of cells are removed from micropost surfaces than the flat control surfaces. These results show that micropost surfaces enable decreased cell adhesion strength while maintaining similar cell viabilities and proliferation as compared to flat PS surfaces.

Novel insight into charge transfer regulation based on carrier density-dependent Ag/ITO composite films

Cosputtering technology was utilized to prepare a Ag and indium tin oxide (ITO) composite on a flat polystyrene (PS) microsphere array. The carrier density estimated by Hall effect testing of different Sn concentrations in the cosputtered films can be tuned from 1018 to 1020 cm−3. The bandgap calculated based on ultraviolet photoelectron spectroscopy can be adjusted within the range of 3.95–4.02 eV. We explored the possible mechanism of charge transfer (CT) by varying the bandgap and explained the causes of the surface-enhanced Raman scattering (SERS). Surprisingly, a synchronous change in the CT process with the carrier density was discovered. This observation suggests that the CT process can be precisely regulated by changes in the composition of the metal-semiconductor nanostructures. Our study provides a reference for the application of Ag/ITO films as alternative near-infrared plasmonic materials.

Polycaprolactone Electrospun Scaffolds Produce an Enrichment of Lung Cancer Stem Cells in Sensitive and Resistant EGFRm Lung Adenocarcinoma.

Simple SummaryThe culture of lung cancer stem cells (LCSCs) is not possible using traditional flat polystyrene surfaces. The study of these tumor-initiating cells is fundamental due to their key role in the resistance to anticancer therapies, tumor recurrence, and metastasis. Hence, we evaluated the use of polycaprolactone electrospun (PCL-ES) scaffolds for culturing LCSC population in sensitive and resistant EGFR-mutated lung adenocarcinoma models. Our findings revealed that both cell models seeded on PCL-ES structures showed a higher drug resistance, enhanced levels of several genes and proteins related to epithelial-to-mesenchymal process, stemness, and surface markers, and the activation of the Hedgehog pathway. We also determined that the non-expression of CD133 was associated with a low degree of histological differentiation, disease progression, distant metastasis, and worse overall survival in EGFR-mutated non-small cell lung cancer patients. Therefore, we confirmed PCL-ES scaffolds as a suitable three-dimensional cell culture model for the study of LCSC niche. The establishment of a three-dimensional (3D) cell culture model for lung cancer stem cells (LCSCs) is needed because the study of these stem cells is unable to be done using flat surfaces. The study of LCSCs is fundamental due to their key role in drug resistance, tumor recurrence, and metastasis. Hence, the purpose of this work is the evaluation of polycaprolactone electrospun (PCL-ES) scaffolds for culturing LCSCs in sensitive and resistant EGFR-mutated (EGFRm) lung adenocarcinoma cell models. We performed a thermal, physical, and biological characterization of 10% and 15%-PCL-ES structures. Several genes and proteins associated with LCSC features were analyzed by RT-qPCR and Western blot. Vimentin and CD133 tumor expression were evaluated in samples from 36 patients with EGFRm non-small cell lung cancer through immunohistochemistry. Our findings revealed that PC9 and PC9-GR3 models cultured on PCL-ES scaffolds showed higher resistance to osimertinib, upregulation of ABCB1, Vimentin, Snail, Twist, Sox2, Oct-4, and CD166, downregulation of E-cadherin and CD133, and the activation of Hedgehog pathway. Additionally, we determined that the non-expression of CD133 was significantly associated with a low degree of histological differentiation, disease progression, and distant metastasis. To sum up, we confirmed PCL-ES scaffolds as a suitable 3D cell culture model for the study of the LCSC niche.

Ultraviolet germicidal irradiation is effective against SARS-CoV-2 in contaminated makeup powder and lipstick

Severe Acute Respiratory Syndrome Coronavirus 2 (SARS-CoV-2) is mainly transmitted by airborne droplets generated by infected individuals. Since this and many other pathogens are able to remain viable on inert surfaces for extended periods of time, contaminated surfaces play an important role in SARS-CoV-2 fomite transmission. Cosmetic products are destined to be applied on infection-sensitive sites, such as the lips and eyelids. Therefore, special biosafety precautions should be incorporated into the routine procedures of beauty parlors and shops. Indeed, innovative cosmetics companies are currently searching for disinfection protocols that ensure the customers’ safety in makeup testing. Here, we propose an ultraviolet germicidal irradiation (UVGI) strategy that can be used to reduce the odds of COVID-19 fomite transmission by makeup testers. It is well-known that UVGI effectively inactivates pathogens on flat surfaces and clear fluids. However, ultraviolet-C (UVC) radiation at 254 nm penetrates poorly in turbid and porous materials, such as makeup and lipstick formulations. Thus, we investigated the virucidal effect of UVGI against SARS-CoV-2 deposited on such substrates and compared their performance to that of flat polystyrene surfaces, used as controls. Concentrated infectious SARS-CoV-2 inoculum (106 PFU/mL) deposited on lipstick and makeup powder was completely inactivated (>5log10 reduction) following UVC exposures at 1,260 mJ/cm2, while flat plastic surfaces required 10 times less exposure (126 mJ/cm2) to reach the same microbicidal performance. We conclude that UVGI comprises an effective disinfection strategy to promote biosafety for cosmetics testers. However, appropriate UVC dosimetry must be implemented to overcome inefficiencies caused by the optical properties of turbid materials in lipsticks and makeup powders.

A Reproducible Method for Growing Biofilms on Polystyrene Surfaces: Biomass and Bacterial Viability Evolution of Pseudomonas fluorescens and Staphylococcus epidermidis

Since biofilm development represents a crucial issue within industrial, clinical and domestic sectors, innovative technologies/approaches (e.g., light technology for inactivation, antibiofilm coatings) are required to eradicate them. In this multidisciplinary scenario, protocols for the development of biofilms are necessary, particularly, in laboratories (not specialised in biofilm science) lacking in sophisticated devices for their growth. A protocol was developed for growing Pseudomonas fluorescens (Gram-negative) biofilms on wide, flat, polystyrene surfaces within 24 h. Several factors, such as inoculum level, area size and growth medium concentration, were investigated. Biofilm development was studied via viable cells and biomass quantification. A comparative analysis between kinetics and growth parameters, estimated using the Baranyi and Roberts model, was conducted at different inoculum levels (104 and 107 CFU/mL). The inoculum levels did not influence the final population within the 24-h-grown biofilms, but they influenced the total biomass development, which followed different kinetics. Confocal laser scanning microscopy confirmed that overnight growth allowed for development of a densely packed biofilm with its 3D structure. The developed protocol was validated for Staphylococcus epidermidis (Gram-positive). The present work is the first study to develop an easy-to-use protocol to obtain highly reproducible biofilms, on flat polystyrene surfaces, with no need for sophisticated technologies.

C1q recognizes antigen-bound IgG in a curvature-dependent manner

C1q is an important recognition protein in the complement system, which is a major protein cascade in the innate immune system. Upon recognition of a target by C1q, the target is marked for opsonization and destruction. C1q recognizes many pathogenic patterns directly, but an important target is the Fc domain of antibodies binding to their antigen. In this paper, the curvature-dependence of the interaction between IgG and C1q is studied by surface plasmon resonance and quartz crystal microbalance. IgG is organized in similar surface coverage densities on flat polystyrene surfaces and polystyrene nanoparticles of different sizes, and the amount of C1q binding to the IgG is investigated. Nanoparticles in solution were found to aggregate upon incubation with IgG, and therefore a new technique utilizing nanoparticles binding to antifouling polymer brush functionalized surfaces was used to prepare surfaces with nanoparticles for measurements with surface plasmon resonance. Interestingly antigen-bound IgG at the curved surface of nanoparticles showed 5.6 times lower binding of C1q compared to at matched flat surfaces. There was no significant difference between the binding at 100 and 200 nm polystyrene particles. These findings are important for designing drug delivery systems to evade the complement system.

A versatile cancer cell trapping and 1D migration assay in a microfluidic device.

Highly migratory cancer cells often lead to metastasis and recurrence and are responsible for the high mortality rates in many cancers despite aggressive treatment. Recently, the migratory behavior of patient-derived glioblastoma multiforme cells on microtracks has shown potential in predicting the likelihood of recurrence, while at the same time, antimetastasis drugs have been developed which require simple yet relevant high-throughput screening systems. However, robust in vitro platforms which can reliably seed single cells and measure their migration while mimicking the physiological tumor microenvironment have not been demonstrated. In this study, we demonstrate a microfluidic device which hydrodynamically seeds single cancer cells onto stamped or femtosecond laser ablated polystyrene microtracks, promoting 1D migratory behavior due to the cells' tendency to follow topographical cues. Using time-lapse microscopy, we found that single U87 glioblastoma multiforme cells migrated more slowly on laser ablated microtracks compared to stamped microtracks of equal width and spacing (p < 0.05) and exhibited greater directional persistence on both 1D patterns compared to flat polystyrene (p < 0.05). Single-cell morphologies also differed significantly between flat and 1D patterns, with cells on 1D substrates exhibiting higher aspect ratios and less circularity (p < 0.05). This microfluidic platform could lead to automated quantification of single-cell migratory behavior due to the high predictability of hydrodynamic seeding and guided 1D migration, an important step to realizing the potential of microfluidic migration assays for drug screening and individualized medicine.

Three-dimensional nanofibrous polystyrene scaffolds modify macrophage phenotypes and activate macrophage angiogenic potential.

3D scaffolds represent an attractive substrate for studying macrophage activation and modification since they mimic extracellular matrix (ECM). However, macrophage response to such materials, particularly with respect to angiogenic potential is still poorly recognized. Therefore, we investigated the effect of 3D nanofibrous polystyrene scaffolds (NPSs) versus tissue culture polystyrene (TCPS) on THP-1-derived macrophages in various environmental conditions, for example, standard (m0), pro-inflammatory (m1), or anti-inflammatory (m2) with respect to pro-angiogenic potential. There were no differences in the expression of TNF-α and IL-10 mRNAs and respective proteins in cells cultured on NPSs compared with flat polystyrene (TCPS), however, NPSs induced an increased VEGF production by macrophages cultured in m0 and m1 media. Cells cultured in m1, and m2 conditions secreted elevated amounts of TNF-α and IL-10, respectively, irrespective of substrate surface geometry. Each macrophage population contains large, medium, and small cells. Moreover, there were significant differences in the proportion of large to small macrophages depending on the medium composition, that is, in m0, m1, and m2 media these proportions were 1:4, 1:3, and 1:10, respectively. The ultrastructure and the immunoexpression of TNF-α and IL-10 were analyzed under a confocal microscope. The results demonstrated differences in cell ultrastructure and suggested that the larger cells were pro-inflammatory macrophages, while the smaller cells were anti-inflammatory macrophages. In conclusion, NPSs activate macrophage pro-angiogenic potential. In addition, an increase in the proportion of pro-inflammatory macrophages relative to anti-inflammatory ones in a given population favors this potential.

Control of Wettability Using Regularly Ordered Two-Dimensional Polymeric Wavy Substrates

Two-dimensional poly(dimethylsiloxane) (PDMS) films with wavy patterns were studied in order to investigate reversible and irreversible wetting effects. Pre-strained, surface oxidized layers of PDMS were used to form relieved wavy geometries, on which hydrophobic functionalization was carried out in order to produce irreversible wetting effects. Wavy-patterned PDMS films showed time-dependent reversible wetting effects. The degree of surface wettability could be tuned by the choice of wavy groove geometries. And the groove geometries were controlled via O2 plasma treatment and mechanical pre-straining. The pre-strained, buckled PDMS films were applied to the fabrication of hydrophobic polystyrene nano-patterns using colloidal self-assembly, where the colloids were arrayed in two-dimensional way. The wavy polystyrene films were found to be more hydrophobic relative to flat polystyrene films. The grooving methodology used in this study could be applied to enhancing the hydrophobicity of other types of polymeric thin films, eliminating the need for chemical treatment.

Fabrication and assessment of an electrospun polymeric microfiber-based platform under bulk flow conditions with rapid and efficient antigen capture.

This study investigated the fabrication and proof of concept design demonstrating rapid and highly sensitive antigen capture utilizing electrospun polystyrene (PS) microfiber mat substrates paired with vacuum pump pressurization to induce bulk flow. In comparison with conventional flat PS surfaces used for immunoassay purposes, this system optimizes the increased surface area of the electrospun polystyrene (ESPS) fiber mat substrates and the accelerated propagation of the antigen through the detection platform by using a vacuum pump to enable efficient and rapid antigen capture. The novelty of this work was demonstrated through a parametric study detailing how a fiber substrate can capture antigen sensitively and at high speeds. In terms of sensitivity, the current system is comparable to the conventionally used flat PS substrates. Additionally, the amount of antigen captured on a flat PS substrate in 60 minutes was surpassed in under 5 seconds when utilizing the ESPS-vacuum system. Three-dimensional ESPS fiber mats were then noted as a comparison between Damkohler numbers and between flat PS and ESPS-vacuum systems. The bulk flow of the ESPS-vacuum system allows for a Damkohler number of 0.37 indicating a balance between the flow rate and the reaction rate as opposed to a PS flat platform of 5.80 × 104 which illustrates a diffusion rate limited system. Finally, the overall ESPS-vacuum system was tested for its immunoassay capability. A sandwich fluorescence-based immunoassay was performed on both PS flat-diffusion and ESPS-vacuum systems. The ESPS-vacuum system indicated a wider detection range capability from 5 to 1000 ng mL-1 in comparison with the PS flat-diffusion system at 5 to 100 ng mL-1.

Waterborne, all-polymeric, colloidal ‘raspberry’ particles with controllable hydrophobicity and water droplet adhesion properties

We present a superhydrophobic coating made from waterborne, all-polymeric ‘raspberry’ particles, composed of a micrometric spherical core decorated with a corona of nanometric spherical particles. As-cast particles produced hydrophobic coatings that were highly adhesive to almost-spherical droplets of water, resembling the properties of some types of rose petals. The coatings could be made slippery to spherical water droplets, like the lotus leaf, by surface activation with air plasma followed by reaction with an alkyl-trichlorosilane. The silanisation of films of latex particles was investigated on two model surfaces (a flat polystyrene film and a monolayer of polystyrene waterborne microparticles) by X-ray photoelectron spectroscopy and water contact angle measurements, and applied to our recently-developed ‘raspberry’ particles to produce a superhydrophobic coating.

Fabrication of ZnO Nanorod/polystyrene Nanosphere Hybrid Nanostructures by Hydrothermal Method for Energy Generation Applications

We report on the successful fabrication of ZnO nanorod (NR)/polystyrene (PS) nanosphere hybrid nanostructure by combining drop coating and hydrothermal methods. Especially, by adopting an atomic layer deposition method for seed layer formation, very uniform ZnO NR structure is grown on the complicated PS surfaces. By using zinc nitrate hexahydrate and hexamine as sources for Zn and O in hydrothermal process, hexagonal shaped single crystal ZnO NRs are synthesized without dissolution of PS in hydrothermal solution. X-ray diffraction results show that the ZnO NRs are grown along c-axis with single crystalline structure and there is no trace of impurities or unintentionally formed intermetallic compounds. Photoluminescence spectrum measured at room temperature for the ZnO NRs on flat Si and PS show typical two emission bands, which are corresponding to the band-edge and deep level emissions in ZnO crystal. Based on these structural and optical investigations, we confirm that the ZnO NRs can be grown well even on the complicated PS surface morphology to form the chestnut-shaped hybrid nanostructures for the energy generation and storage applications.

Controlled Three-Dimensional Hierarchical Structuring by Memory-Based, Sequential Wrinkling.

This paper describes how a memory-based, sequential wrinkling process can transform flat polystyrene sheets into multiscale, three-dimensional hierarchical textures. Multiple cycles of plasma-mediated skin growth followed by directional strain relief of the substrate resulted in hierarchical architectures with characteristic generational (G) features. Independent control over wrinkle wavelength and wrinkle orientation for each G was achieved by tuning plasma treatment time and strain-relief direction for each cycle. Lotus-type superhydrophobicity was demonstrated on three-dimensional G1-G2-G3 hierarchical wrinkles as well as tunable superhydrophilicity on these same substrates after oxygen plasma. This materials system provides a general approach for nanomanufacturing based on bottom-up sequential wrinkling that will benefit a diverse range of applications and especially those that require large area (>cm(2)), multiscale, three-dimensional patterns.

Stability of Polymer Ultrathin Films (&lt;7 nm) Made by a Top-Down Approach

In polymer physics, the dewetting of spin-coated polystyrene ultrathin films on silicon remains mysterious. By adopting a simple top-down method based on good solvent rinsing, we are able to prepare flat polystyrene films with a controlled thickness ranging from 1.3 to 7.0 nm. Their stability was scrutinized after a classical annealing procedure above the glass transition temperature. Films were found to be stable on oxide-free silicon irrespective of film thickness, while they were unstable (<2.9 nm) and metastable (>2.9 nm) on 2 nm oxide-covered silicon substrates. The Lifshitz-van der Waals intermolecular theory that predicts the domains of stability as a function of the film thickness and of the substrate nature is now fully reconciled with our experimental observations. We surmise that this reconciliation is due to the good solvent rinsing procedure that removes the residual stress and/or the density variation of the polystyrene films inhibiting thermodynamically the dewetting on oxide-free silicon.

The effect of nanoscale surface curvature on the oligomerization of surface-bound proteins.

The influence of surface topography on protein conformation and association is used routinely in biological cells to orchestrate and coordinate biomolecular events. In the laboratory, controlling the surface curvature at the nanoscale offers new possibilities for manipulating protein-protein interactions and protein function at surfaces. We have studied the effect of surface curvature on the association of two proteins, α-lactalbumin (α-LA) and β-lactoglobulin (β-LG), which perform their function at the oil-water interface in milk emulsions. To control the surface curvature at the nanoscale, we have used a combination of polystyrene (PS) nanoparticles (NPs) and ultrathin PS films to fabricate chemically pure, hydrophobic surfaces that are highly curved and are stable in aqueous buffer. We have used single-molecule force spectroscopy to measure the contour lengths Lc for α-LA and β-LG adsorbed on highly curved PS surfaces (NP diameters of 27 and 50 nm, capped with a 10 nm thick PS film), and we have compared these values in situ with those measured for the same proteins adsorbed onto flat PS surfaces in the same samples. The Lc distributions for β-LG adsorbed onto a flat PS surface contain monomer and dimer peaks at 60 and 120 nm, respectively, while α-LA contains a large monomer peak near 50 nm and a dimer peak at 100 nm, with a tail extending out to 200 nm, corresponding to higher order oligomers, e.g. trimers and tetramers. When β-LG or α-LA is adsorbed onto the most highly curved surfaces, both monomer peaks are shifted to much smaller values of Lc. Furthermore, for β-LG, the dimer peak is strongly suppressed on the highly curved surface, whereas for α-LA the trimer and tetramer tail is suppressed with no significant change in the dimer peak. For both proteins, the number of higher order oligomers is significantly reduced as the curvature of the underlying surface is increased. These results suggest that the surface curvature provides a new method of manipulating protein-protein interactions and controlling the association of adsorbed proteins, with applications to the development of novel biosensors.

Back Cover, Volume 131, Issue 14

Journal of Applied Polymer ScienceVolume 131, Issue 14 Cover ImageFree Access Back Cover, Volume 131, Issue 14 First published: 16 April 2014 https://doi.org/10.1002/app.40550AboutPDF ToolsRequest permissionExport citationAdd to favoritesTrack citation ShareShare Give accessShare full text accessShare full-text accessPlease review our Terms and Conditions of Use and check box below to share full-text version of article.I have read and accept the Wiley Online Library Terms and Conditions of UseShareable LinkUse the link below to share a full-text version of this article with your friends and colleagues. Learn more.Copy URL Share a linkShare onFacebookTwitterLinked InRedditWechat Abstract To transform standard flat polystyrene surfaces into textured microenvironments for three-dimensional (3D) cell cultures, Michael E. DeRosa, Yulong Hong, Ronald A. Faris, and Hongwei Rao describe a solvent treatment process. Cell culture experiments typically use thermoplastic plates that have 2D well bottoms. A 3D cell culture plate made of polystyrene in a conventional, inexpensive format could be an attractive technology since cultures could then be conducted on the same material as cell biologists have been using for decades. The study demonstrates that microtextured polystyrene surfaces for 3D cell cultures can easily be made with off-the-shelf microplates by a simple solvent treatment process. DOI: 10.1002/app.40181 Volume131, Issue14Special Issue: Bioactive Surface FunctionalizationJuly 15, 2014 RelatedInformation

Investigation on kinetic processes of fabrication of honeycomb-patterned polystyrene film and adsorption of surfactant by quartz crystal microbalance in impedance analysis method

In this work, the kinetic processes of fabrication of honeycomb-patterned polystyrene (PS) porous film by a breath figures method and the adsorption of sodium dodecyl benzene sulfonate (SDBS) onto the PS films were monitored by a quartz crystal microbalance technique. Three distinct viscoelastic behaviors of PS film were observed. The viscosity of PS film was increased slightly in the initial liquid film stage, reached a maximum in the middle of the viscous film stage, then decreased significantly in the end of rigid film stage. Compared with a flat PS film, the porous PS film had a shorter liquid film stage but a longer viscous film stage. The adsorption processes of SDBS onto the PS films were approximated favorably by the pseudo-second-order kinetic model. The adsorption rate constants onto flat film are 9.8–27 times of those onto porous film. The adsorption isotherms were well fitted by the Langmuir isotherm model. The adsorption equilibrium constant and capacity are 1.29×104L/mol and 29.7mg/g onto flat PS film, 6.85×103L/mol and 375.8mg/g onto porous PS film, respectively.

Microtextured polystyrene surfaces for three‐dimensional cell culture made by a simple solvent treatment method

ABSTRACTThis article describes a solvent treatment process for transforming standard flat polystyrene surfaces into textured microenvironments for three‐dimensional (3D) cell culture. The process can be used with off‐the‐shelf microplates having 6, 12, 96, or even 384‐well formats and completed in a few steps. The surface microtexture resembles tightly packed, non‐interconnected micropores having a median pore size of either 115 or 19 μm depending on the strength of the solvent mixture. Hansen solubility parameter analysis was used to map out the polymer–solvent interaction conditions necessary for creating the surface texture. Primary human hepatocytes cultured on two‐dimensional control surfaces attached with a flat monolayer morphology while those cultured on surfaces having 115 μm‐sized pores exhibited a round 3D morphology. The cells also showed more in‐vivo like behavior on the porous surface by exhibiting significantly higher basal enzymatic activity and expressing higher levels of several drug metabolism‐related genes. © 2013 Wiley Periodicals, Inc. J. Appl. Polym. Sci. 2014, 131, 40181.

Fabrication of polystyrene microscale porous substrate and its effects on HL-7702 cells behaviors

A novel polystyrene (PS) substrate with microscale porous structure was facilely fabricated by crystalline-controlled casting method using mixed solvent [N,N-dimethylformamide and ethyl alcohol (v/v)] based on the nonsolvent induced phase separation process. The substrate surfaces exhibited a bi-continuous microscale porous morphology with high porosity, large pore size and pore-pore connection structure. Moreover, behaviors of the normal human liver cell line (HL-7702) seeded on this substrate surface were carefully investigated. The results indicated that the cell adhesion, spread and cell-cell connection on the surface with subcellular pore size (∼ 10 μm) were similar to the cells proliferated on the flat PS surface. However, the number of HL-7702 cells proliferated on the PS microscale porous surface was higher than cells on the conventional PS flat surface, suggesting that the pore-pore structure was conducive to HL-7702 cell proliferation. Furthermore, hematoxylin and eosin staining and micronucleus test were performed. The results showed that fewer damages for nuclear and cytoplasm and less cell genotoxicity were caused by the microscale porous structure within the scope of pore size (∼ 10 μm) than that of the flat surface.