Hydrogels with dual sensitivity to temperature and pH in physiologically relevant ranges as supports for versatile controlled cell detachment.

Thermosensitive hydrogels based on the N-vinyl caprolactam (VCL), capable of allowing for cell adhesion and proliferation, as well as non-aggressive detachment by controlled temperature drop, were functionalized with 23 % or lower molar percentages of the cationizable hydrophobic unit 2-(diisopropylamino) ethyl methacrylate (DPAEMA), to obtain networks with dual sensitivity to temperature and pH. The swelling analysis of the systems has shown a transition pK (pKb) close to physiological values, dependent on the temperature of the medium (pKb of 6.6 and 6.9 when the temperature of the medium is above and below the transition temperature VPTT, respectively) and little dependence on the degree of functionalization of DPAEMA. In addition, at temperatures below the transition temperature (VPTT), the systems have shown large swelling variations as a function of the pH (i.e. below and above the pKb), exhibiting greater absorption capacity at pHs below pKb, where the DPAEMA units are cationized. Cytocompatibility and transplant capacity have been evaluated using the C166-GFP endothelial cell line. None of the thermosensitive hydrogels with variable DPAEMA content showed a delay with respect to the control without DPAEMA neither in terms of adhesion nor in proliferation. However, by increasing the percentage of DPAEMA functionalization -and decreasing thermosensitivity-, a correlative decrease in mitochondrial activity was obtained in the transplant, with significant differences for the hydrogels with DPAEMA molar percentage of 3 % or higher. Taking advantage of the proximity of the pKb to the physiological value, we have evaluated the cellular response and the capacity for transplantation after lowering the pH to 6.5, below pKb. A direct relationship of the DPAEMA functionalization degree on the detachment efficiency was observed, since the hydrogels with the highest molar load of DPAEMA showed higher mitochondrial metabolic activity after cell detachment.

Hierarchical Hybrid Coatings with Drug-Eluting Capacity for Mg Alloy Biomaterials.

A hierarchical hybrid coating (HHC) comprising a ceramic oxide layer and two biodegradable polymeric (polycaprolactone, PCL) layers has been developed on Mg3Zn0.4Ca cast alloy in order to provide a controlled degradation rate and functionality by creating a favorable porous surface topography for cell adhesion. The inner, ceramic layer formed by plasma electrolytic oxidation (PEO) has been enriched in bioactive elements (Ca, P, Si). The intermediate PCL layer sealed the defect in the PEO layer and the outer microporous PCL layer loaded with the appropriate active molecule, thus providing drug-eluting capacity. Morphological, chemical, and biological characterizations of the manufactured coatings loaded with ciprofloxacin (CIP) and paracetamol (PAR) have been carried out. In vitro assays with cell lines relevant for cardiovascular implants and bone prosthesis (endothelial cells and premyoblasts) showed that the drug-loaded coating allows for cell proliferation and viability. The study of CIP and PAR cytotoxicity and release rate indicated that the porous PCL layer does not release concentrations detrimental to the cells. However, complete system assays revealed that corrosion behavior and increase of the pH negatively affects cell viability. H2 evolution during corrosion of Mg alloy substrate generates blisters in PCL layer that accelerate the corrosion locally in crevice microenvironment. A detailed mechanism of the system degradation is disclosed. The accelerated degradation of the developed system may present interest for its further adaptation to new cancer therapy strategies.

Vat Photopolymerization 3D Printing of Hydrogels with Re-Adjustable Swelling.

Vat photopolymerization typically prints highly crosslinked networks. Printing hydrogels, which are also networks but with a high swelling capacity in water and therefore with low crosslinking density, is a challenge for this technique. However, it may be of interest in medicine and in other areas, since it would allow for the preparation of this type of 3D-shaped material. In this work, an approach for printing hydrogels via vat photopolymerization that uses a mixture of stable and hydrolysable crosslinkers has been evaluated so that an initial highly crosslinked network can be printed, although after hydrolysis it becomes a network with low crosslinking. This approach has been studied with PEO/PEG-related formulations, that is, with a PEG-dimethacrylate as a stable crosslinker, a PEO-related derivative carrying ß-aminoesters as a degradable crosslinker, and PEG-methyl ether acrylate and hydroxyethyl acrylate as monofunctional monomers. A wide family of formulations has been studied, maintaining the weight percentage of the crosslinkers at 15%. Resins have been studied in terms of viscosity, and the printing process has been evaluated through the generation of Jacobs working curves. It has been shown that this approach allows for the printing of pieces of different shapes and sizes via vat photopolymerization, and that these pieces can re-ajust their water content in a tailored fashion through treatments in different media (PBS or pH 10 buffer).

Clear polyurethane coatings with excellent virucidal properties: Preparation, characterization and rapid inactivation of human coronaviruses 229E and SARS-CoV-2.

Commercial polyurethane (PU) coating formulations have been modified with 1-(hydroxymethyl)-5,5-dimethylhydantoin (HMD) both in bulk (0.5 and 1% w/w) and onto the coatings surface as an N-halamine precursor, to obtain clear coatings with high virucidal activity. Upon immersion in diluted chlorine bleaching, the hydantoin structure on the grafted PU membranes was transformed into N-halamine groups, with a high surface chlorine concentration (40-43µg/cm2). Fourier transform infrared spectroscopy (FTIR) spectroscopy, thermogravimetric analysis (TGA), energy-dispersive X-ray (EDX), X-ray photoelectron spectroscopy (XPS) and iodometric titration were used to characterize the coatings and quantify the chlorine contents of the PU membranes after chlorination. Biological evaluation of their activity against Staphylococcus aureus (Gram-positive bacteria) and human coronaviruses HCoV-229E and SARS-CoV-2 was performed, and high inactivation of these pathogens was observed after short contact times. The inactivation of HCoV-229E was higher than 98% for all modified samples after just 30 minutes, whereas it was necessary 12 hours of contact time for complete inactivation of SARS-CoV-2. The coatings were fully rechargeable by immersion in diluted chlorine bleach (2% v/v) for at least 5 chlorination-dechlorination cycles. Moreover, the performance of the antivirus efficiency of the coatings is considered as long-lasting, because experiments of reinfection of the coatings with HCoV-229E coronavirus did not show any loss of the virucidal activity after three consecutive infection cycles without reactivation of the N-halamine groups.

Evaluation of Poly(N-Ethyl Pyrrolidine Methacrylamide) (EPA) and Derivatives as Polymeric Vehicles for miRNA Delivery to Neural Cells.

MicroRNAs (miRNAs) are endogenous, short RNA oligonucleotides that regulate the expression of hundreds of proteins to control cells' function in physiological and pathological conditions. miRNA therapeutics are highly specific, reducing the toxicity associated with off-target effects, and require low doses to achieve therapeutic effects. Despite their potential, applying miRNA-based therapies is limited by difficulties in delivery due to their poor stability, fast clearance, poor efficiency, and off-target effects. To overcome these challenges, polymeric vehicles have attracted a lot of attention due to their ease of production with low costs, large payload, safety profiles, and minimal induction of the immune response. Poly(N-ethyl pyrrolidine methacrylamide) (EPA) copolymers have shown optimal DNA transfection efficiencies in fibroblasts. The present study aims to evaluate the potential of EPA polymers as miRNA carriers for neural cell lines and primary neuron cultures when they are copolymerized with different compounds. To achieve this aim, we synthesized and characterized different copolymers and evaluated their miRNA condensation ability, size, charge, cytotoxicity, cell binding and internalization ability, and endosomal escape capacity. Finally, we evaluated their miRNA transfection capability and efficacy in Neuro-2a cells and rat primary hippocampal neurons. The results indicate that EPA and its copolymers, incorporating ß-cyclodextrins with or without polyethylene glycol acrylate derivatives, can be promising vehicles for miRNA administration to neural cells when all experiments on Neuro-2a cells and primary hippocampal neurons are considered together.

Polycaprolactone with multiscale porosity and patterned surface topography prepared using sacrificial 3D printed moulds: Towards tailor-made scaffolds.

Biocompatible three-dimensional porous scaffolds are widely used in multiple biomedical applications. However, the fabrication of tailor-made 3D structures with controlled and combined multiscale macroscopic-microscopic, surface and inner porosities in a straightforward manner is still a current challenge. Herein, we use multimaterial fused deposition modeling (FDM) to generate poly (vinyl alcohol) (PVA) sacrificial moulds filled with poly (Ɛ-caprolactone) (PCL) to generate well defined PCL 3D objects. Further on, the supercritical CO2 (SCCO2) technique, as well as the breath figures mechanism (BFs), were additionally employed to fabricate specific porous structures at the core and surfaces of the 3D PCL object, respectively. The biocompatibility of the resulting multiporous 3D structures was tested in vitro and in vivo, and the versatility of the approach was assessed by generating a vertebra model fully tunable at multiple pore size levels. In sum, the combinatorial strategy to generate porous scaffolds offers unique possibilities to fabricate intricate structures by combining the advantages of additive manufacturing (AM), which provides flexibility and versatility to generate large sized 3D structures, with advantages of the SCCO2 and BFs techniques, which allow to finely tune the macro and micro porosity at material surface and material core levels.

Fabrication of 3D cylindrical thermosensitive hydrogels as supports for cell culture and detachment of tubular cell sheets.

Pseudo interpenetrating vinyl-caprolactam (VCL) based thermosensitive tubular hydrogels with a volume phase transition temperature, VPTT, around 35 °C, have been prepared by combining two different crosslinkers, a di-methacrylate (C1) and a di-vinyl urea (C2). The molar ratio between the two crosslinkers (for a global crosslinker molar percentage of 1.9) has shown to play a key role on the properties of the hydrogel. Increasing the amount of di-vinyl urea, leads to transparent but rather fragile materials and to a lower extent of thermosensitivity, that is, to a lower variation in the hydrogel swelling upon temperature change. However, tubes prepared with a selected crosslinker molar ratio C1/C2 of 65/35 provided a compromise between transparency, thermosensitivity and maneuverability and were, thus, evaluated as supports for cell culture using premyoblastic cells. These hydrogels, used as supports, allow for surface adhesion and cell proliferation until confluence, and eventually an efficient monolayer detachment (and transplant to a 3D-printed polylactic acid (PLA) support) through a controlled drop in temperature. As a result, this method permits to obtain tubular tissue constructs with potential applications in tissue engineering such as in the elaboration of vascular grafts.

Photocurable Thiol-yne Alginate Hydrogels for Regenerative Medicine Purposes.

Every year millions of people worldwide undergo surgical interventions, with the occurrence of mild or severe post-treatment consequences meaning that rehabilitation plays a key role in modern medicine. Considering the cases of burns and plastic surgery, the pressing need for new materials that can be used for wound patches or body fillers and are able to sustain tissue regeneration and promote cell adhesion and proliferation is clear. The challenges facing next-generation implant materials also include the need for improved structural properties for cellular organization and morphogenic guidance together with optimal mechanical, rheological, and topographical behavior. Herein, we propose for the first time a sodium alginate hydrogel obtained by a thiol-yne reaction, easily synthesized using carbodiimide chemistry in a two-step reaction. The hydrogels were formed in all cases within a few minutes of light irradiation, showing good self-standing properties under solicitation. The mechanical, rheological, topographical, and swelling properties of the gels were also tested and reported. Lastly, no cytotoxicity was detected among the hydrogels. Soluble extracts in culture media allowed cell proliferation, and no differences between samples were detected in terms of metabolic activity and DNA content. These results suggest the potential use of these cytocompatible hydrogels in tissue engineering and regenerative medicine.

Wrinkling on Stimuli-Responsive Functional Polymer Surfaces as a Promising Strategy for the Preparation of Effective Antibacterial/Antibiofouling Surfaces.

Biocompatible smart interfaces play a crucial role in biomedical or tissue engineering applications, where their ability to actively change their conformation or physico-chemical properties permits finely tuning their surface attributes. Polyelectrolytes, such as acrylic acid, are a particular type of smart polymers that present pH responsiveness. This work aims to fabricate stable hydrogel films with reversible pH responsiveness that could spontaneously form wrinkled surface patterns. For this purpose, the photosensitive reaction mixtures were deposited via spin-coating over functionalized glasses. Following vacuum, UV, or either plasma treatments, it is possible to spontaneously form wrinkles, which could increase cell adherence. The pH responsiveness of the material was evaluated, observing an abrupt variation in the film thickness as a function of the environmental pH. Moreover, the presence of the carboxylic acid functional groups at the interface was evidenced by analyzing the adsorption/desorption capacity using methylene blue as a cationic dye model. The results demonstrated that increasing the acrylic acid in the microwrinkled hydrogel effectively improved the adsorption and release capacity and the ability of the carboxylic groups to establish ionic interactions with methylene blue. Finally, the role of the acrylic acid groups and the surface topography (smooth or wrinkled) on the final antibacterial properties were investigated, demonstrating their efficacy against both gram-positive and gram-negative bacteria model strains (E. coli and S. Aureus). According to our findings, microwrinkled hydrogels presented excellent antibacterial properties improving the results obtained for planar (smooth) hydrogels.

Anticorrosion and Cytocompatibility Assessment of Graphene-Doped Hybrid Silica and Plasma Electrolytic Oxidation Coatings for Biomedical Applications.

Magnesium AZ31 alloy substrates were coated with different coatings, including sol-gel silica-reinforced with graphene nanoplatelets, sol-gel silica, plasma electrolytic oxidation (PEO), and combinations of them, to improve cytocompatibility and control the corrosion rate. Electrochemical corrosion tests, as well as hydrogen evolution tests, were carried out using Hanks' solution as the electrolyte to assess the anticorrosion behavior of the different coating systems in a simulated body fluid. Preliminary cytocompatibility assessment of the different coating systems was carried out by measuring the metabolic activity, deoxyribonucleic acid quantification, and the cell growth of premyoblastic C2C12-GFP cell cultures on the surface of the different coating systems. Anticorrosion behavior and cytocompatibility were improved with the application of the different coating systems. The use of combined PEO + SG and PEO + SG/GNP coatings significantly decreased the degradation of the specimens. The monolayer sol-gel coatings, with and without GNPs, presented the best cytocompatibility improvement.

Wheat germin-like protein: Studies on chitin/chitosan matrix for tissue engineering applications.

Advances in tissue engineering require the development of new biomaterials with adequate properties of cell attachment and growth. The properties of biomaterials can be improved by incorporation of bioactive molecules to enhance in vitro and/or in vivo functions. In this work, we study the role of a wheat germin-like protease inhibitor (GLPI), free or immobilized in biocompatible matrices to improve cell-attachment ability on different mammalian cell lines. The phylogenetic relationships and functional diversity of the GLPI were analyzed among diverse genera to get insights into sequence motif conservations. The cytocompatibility effect of free GLPI on C2C12 premyoblastic cells and B16 cells as tumoral model has been tested. GLPI promoted proliferation and metabolic activity of both cell types on in vitro models, not showing cytotoxic effects. Furthermore, GLPI was immobilized in chitin microparticles and in chitosan films; we demonstrated an accelerated cell adhesion process in both biomaterials.

Efficient and Low Cytotoxicity Gene Carriers Based on Amine-Functionalized Polyvinylpyrrolidone.

Non-viral vectors are a safety tool for gene therapy to deliver therapeutic genes. Among the different non-viral vectors, polyvinylpyrrolidone (PVP), a well-known hydrosoluble, neutral, and non-toxic polymer, satisfies the requirements and becomes a suitable candidate for gene delivery. In this study, we describe the preparation of polyvinylpyrrolidones decorated with pyrrolidine, piperidine, and piperazine groups, and evaluate them in vitro as non-viral gene carriers. The properties of these new systems are compared with those of hyperbranched polyethyleneimine (PEI) used as a positive control. Their ability to complex DNA at different N/P molar ratios, from 1:1 up to 10:1, was studied through agarose gel electrophoresis and dynamic light scattering. The resulting complexes (polyplexes) were characterized and evaluated in vitro with murine fibroblast (Swiss 3T3) as non-viral gene carriers, using luciferase as the reporter gene and a calcein cytocompatibility assay. All the copolymers condensed DNA to a particle average size between 100-400 nm when used at N/P ratios of 4:1 or higher. The copolymers with piperidine groups showed higher transfection efficiency than the pyrrolidine and piperazine modified copolymers, and even higher than the positive control of PEI at N/P ratios of 4:1 or higher. All the synthesized polyplexes from an aminated PVP displayed a general tendency of high cytocompatibility (75-95%) in comparison with the positive control PEI (55%).

Biocompatible fluorinated wrinkled hydrogel films with antimicrobial activity.

Surface-modified hydrogel films were designed to control the bacterial colonization on their surface and to promote cell proliferation through the gradual insertion of highly hydrophobic functional monomers. These hydrogel films were deposited via spin-coating technique, using muscovite mica as a substrate. These samples were then exposed to different external stimuli to produce wrinkled patterns. The relationship between the monomers which compose the hydrogel, was varied to alter the hydrophobic/hydrophilic balance of the final composite. Contact angle and confocal Raman spectroscopy measurements were carried out to characterize the surface and the bulk of the hydrogel film. Cell proliferation and antimicrobial tests were performed using premyoblastic murine cells (C2C12-GFP) and RAW 264.7 (ATCC® TIB-71) macrophagic cell lines, and also for bacteria strains, Staphylococcus aureus and Escherichia coli. The results indicate that the inclusion of the TFPMA produces an increase in cell proliferation, together with a decrease in living bacterial colonies after 48 h, both for Gram-positive or Gram-negative species.

Hierarchical Functionalized Polymeric-Ceramic Coatings on Mg-Ca Alloys for Biodegradable Implant Applications.

Magnesium-based implants present several advantages for clinical applications, in particular due to their biocompatibility and degradability. However, degradation products can affect negatively the cell activity. In this work, a combined coating strategy to control the implant degradation and cell regulation processes is evaluated, including plasma electrolytic oxidation (PEO) that produces a 13 µm-thick Ca, P, and Si containing ceramic coating with surface porosity, and breath figures (BF) approach that produces a porous polymeric poly(ε-caprolactone) surface. The degradation of PCL-PEO-coated Mg hierarchical scaffold can be tailored to promote cell adhesion and proliferation into the porous structure. As a result, cell culture can colonize the inner PEO-ceramic coating structure where higher amount of bioelements are present. The Mg/PEO/PCL/BF scaffolds exhibit equally good or better premyoblast cell adhesion and proliferation compared with Ti CP control. The biological behavior of this new hierarchical functionalized scaffold can improve the implantation success in bone and cardiovascular clinical applications.

Microwrinkled pH-sensitive hydrogel films and their role on the cell adhesion/proliferation.

In this work, hydrogels based on HEMA and DMAEMA (pH-sensitive monomer) were used to form biocompatible films which present microwrinkled patterns in their surface, with the focus of exploring the role of chemical composition on cell adhesion and proliferation. Three different pH (5.4, 7.4, and 8.3) were employed to prepare these hydrogels. The pre-polymerized hydrogel mixtures were deposited via spin coating, then exposed to vacuum for deswelling the films and finally, to UV-light to spontaneously generate the wrinkled pattern. By following this procedure, is possible to form a thin rigid layer on the top of the soft and incompletely polymerized hydrogel film which generates, in turn, a wrinkled pattern due to strain mismatch in the interface. FE-SEM and AFM micrographs allowed us to characterize the wrinkled pattern dimensions. The results evidenced that chemical composition is directly related to the surface pattern morphologies obtained, not so in the case of pH variation, which does not generate relevant changes in the pattern morphology. Interestingly, these pH variations resulted in significant alterations on the interface-cell interactions. More precisely, a premyoblastic cell monolayer was cultured over the wrinkled pattern, showing an optimal cell proliferation at neutral pH. Also, the variation of DMAEMA amount on the monomer feed composition employed for the preparation of the wrinkle surfaces revealed that a certain amount is required to favor cell attachment and growth.

Design and fabrication of biocompatible wrinkled hydrogel films with selective antibiofouling properties.

In this article, we explored the selective antibiofouling capacity acquired by functional wrinkled hydrogel films via a fine tuning of their chemical structure through the gradual insertion of hydrophobic radical groups in their network. The hydrogel consists of three main components: hydroxyethyl methacrylate (HEMA, amphiphilic monomer), trifluoroethyl methacrylate (TFMA, hydrophobic monomer), and poly(ethylene glycol) diacrylate (PEGDA, hydrophilic crosslinking agent). Interestingly, the manipulation of the chemical composition affects both, surface morphology and physicochemical characteristics of the patterns, inducing transitions between different surface microstructures, i.e. from wrinkles to creases, to folds, and to crumples. Contact angle measurements show that the insertion of TFMA produces a slight decrease in surface wettability, remaining however highly hydrophilic. By using confocal Raman spectroscopy, important information about wrinkle formation mechanism could be obtained. The procedure presented in this article involves two consecutive thermal and photopolymerization steps, generating a "pseudo" two-layer system, which contracts at different extents when is exposed to external stimuli, leading to the formation of wrinkled surfaces. Finally, bacterial and cellular adhesion/proliferation studies were carried out, evidencing that the amount of TFMA included clearly reduce the bacterial adhesion while mammalian cells are able to still proliferate.

Wrinkled Hydrogel Surfaces with Modulated Surface Chemistry and Topography: Evaluation As Supports for Cell Growth and Transplant.

We report a straightforward procedure to simultaneously functionalize hydrophobic PC supports with vinylpyrrolidone (VP)-based hydrogels with both variable ionic load as well as surface topography, forming wrinkles. The strategy involves three consecutive steps: first, a contact of the polymeric support (PC) with a photopolymerizable solution comprising vinylic monomers is established. Second, UV-light exposure curing of the solution and finally, the third step involes the swelling of the hydrogel network that finally provokes its surface detachment. Interestingly, a wrinkled hybrid PC/hydrogel interface remains after this detachment. Several experimental parameters permitted us to finely control the wrinkle characteristics such as amplitude and period. The experimental parameters that can be varied, herein we will focus on the variation of the elapsed time (i.e., time of contact between the support and the photosensitive monomer mixture, or the solvent (type and amount) included in the monomer mixture. Equally, the nature of the additional ionic methacrylate monomers (M) employed plays a key role on the final topography. According to confocal raman microscopy results, we evidenced that a monomer diffusion into the PC substrate before the UV irradiation step modifies the interfacial (hydrogel/substrate) chemical composition and leads upon UV irradiation to the formation of a thin hydrogel surface layer. The surface chemical composition and structural characteristics were demonstrated to significantly change the surface interaction with different cell lines, affecting cell adhesion, proliferation, or transplantation.

Combining Breath Figures and Supercritical Fluids To Obtain Porous Polymer Scaffolds.

Supercritical fluids technology is a clean methodology to foam polymeric materials. However, this technique provides only the formation of inner porosity, whereas the so-called skin layer is commonly observed at the polymer surface. This article describes a new method for the preparation of outer and inner porous poly(ε-caprolactone) (PCL) scaffolds by combination of supercritical CO2 (SCCO2) foaming and the breath figures technique. In the first step, experiments with a SCCO2 reactor were performed at 35-45 °C, 100-250 bar, and 1-20 min depressurization time. The effect of these parameters in the formation of inner porosity was investigated for an adequate optimization. In a late stage, to provide also surface porosity to the polymeric samples and remove the skin layer, the breath figures technique was employed. The evaluation of porosity was determined by scanning electronic microscopy, mercury porosimetry, and micro X-ray computerized tomography scanning processing the images obtained with the ImageJ software. The results of this study using these two complementary techniques showed the existence of interconnectivity between inner and outer porosity of the samples. Furthermore, thermal transitions and crystallinity of the PCL samples have been analyzed by differential scanning calorimetry. Finally, a preliminary biological evaluation of the resulting scaffolds with mouse endothelial cells (C166-GFP) was performed to assess their biocompatibility and cellular viability.

Fabrication of biocompatible and efficient antimicrobial porous polymer surfaces by the Breath Figures approach.

We designed and fabricated highly efficient and selective antibacterial substrates, i.e. surface non-cytotoxic against mammalian cells but exhibiting strong antibacterial activity. For that purpose, microporous substrates (pore sizes in the range of 3-5 µm) were fabricated using the Breath Figures approach (BFs). These substrates have additionally a defined chemical composition in the pore cavity (herein either a poly(acrylic acid) or the antimicrobial peptide Nisin) while the composition of the rest of the surface is identical to the polymer matrix. As a result, considering the differences in size of bacteria (1-4 µm) in comparison to mammalian cells (above 10 µm) the bacteria were able to enter in contact with the inner part of the pores where the antimicrobial functionality has been placed. On the opposite, mammalian cells remain in contact with the top surface thus preventing cytotoxic effects and enhancing the biocompatibility of the substrates. The resulting antimicrobial surfaces were exposed to Staphylococcus aureus as a model bacteria and murine endothelial C166-GFP cells. Superior antibacterial performance while maintaining an excellent biocompatibility was obtained by those surfaces prepared using PAA while no evidence of significant antibacterial activity was observed at those surfaces prepared using Nisin.

Highly Efficient Antibacterial Surfaces Based on Bacterial/Cell Size Selective Microporous Supports.

We report on the fabrication of efficient antibacterial substrates selective for bacteria, i.e., noncytotoxic against mammalian cells. The strategy proposed is based on the different size of bacteria (1-4 µm) in comparison with mammalian cells (above 20 µm) that permit the bacteria to enter in contact with the inner part of micrometer-sized pores where the antimicrobial functionality are placed. On the contrary, mammalian cells, larger in terms of size, remain at the top surface, thus reducing adverse cytotoxic effects and improving the biocompatibility of the substrates. For this purpose, we fabricated well-ordered functional microporous substrates (3-5 µm) using the breath figures approach that enabled the selective functionalization of the pore cavity, whereas the rest of the surface remained unaffected. Microporous surfaces were prepared from polymer blends comprising a homopolymer (i.e., polystyrene) and a block copolymer (either polystyrene-b-poly(dimethylaminoethyl methacrylate) (PDMAEMA) or a quaternized polystyrene-b-poly(dimethylaminoethyl methacrylate)). As a result, porous surfaces with a narrow size distribution and a clear enrichment of the PDMAEMA or the quaternized PDMAEMA block inside the pores were obtained that, in the case of the quaternized PDMAEMA, provided an excellent antimicrobial activity to the films.