Switching it Up: The Promise of Stimuli-Responsive Polymer Systems in Biomedical Science.

Responsive polymer systems have the ability to change properties or behavior in response to external stimuli. The properties of responsive polymer systems can be fine-tuned by adjusting the stimuli, enabling tailored responses for specific applications. These systems have applications in drug delivery, biosensors, tissue engineering, and more, as their ability to adapt and respond to dynamic environments leads to improved performance. However, challenges such as synthesis complexity, sensitivity limitations, and manufacturing issues need to be addressed for successful implementation. In our review, we provide a comprehensive summary on stimuli-responsive polymer systems, delving into the intricacies of their mechanisms and actions. Future developments should focus on precision medicine, multifunctionality, reversibility, bioinspired designs, and integration with advanced technologies, driving the dynamic growth of sensitive polymer systems in biomedical applications.

Thermoresponsive Smart Copolymer Coatings Based on P(NIPAM-co-HEMA) and P(OEGMA-co-HEMA) Brushes for Regenerative Medicine.

The fabrication of multifunctional, thermoresponsive platforms for regenerative medicine based on polymers that can be easily functionalized is one of the most important challenges in modern biomaterials science. In this study, we utilized atom transfer radical polymerization (ATRP) to produce two series of novel smart copolymer brush coatings. These coatings were based on copolymerizing 2-hydroxyethyl methacrylate (HEMA) with either oligo(ethylene glycol) methyl ether methacrylate (OEGMA) or N-isopropylacrylamide (NIPAM). The chemical compositions of the resulting brush coatings, namely, poly(oligo(ethylene glycol) methyl ether methacrylate-co-2-hydroxyethyl methacrylate) (P(OEGMA-co-HEMA)) and poly(N-isopropylacrylamide-co-2-hydroxyethyl methacrylate) (P(NIPAM-co-HEMA)), were predicted using reactive ratios of the monomers. These predictions were then verified using time-of-flight-secondary ion mass spectrometry (ToF-SIMS) and X-ray photoelectron spectroscopy (XPS). The thermoresponsiveness of the coatings was examined through water contact angle (CA) measurements at different temperatures, revealing a transition driven by lower critical solution temperature (LCST) or upper critical solution temperature (UCST) or a vanishing transition. The type of transition observed depended on the chemical composition of the coatings. Furthermore, it was demonstrated that the transition temperature of the coatings could be easily adjusted by modifying their composition. The topography of the coatings was characterized using atomic force microscopy (AFM). To assess the biocompatibility of the coatings, dermal fibroblast cultures were employed, and the results indicated that none of the coatings exhibited cytotoxicity. However, the shape and arrangement of the cells were significantly influenced by the chemical structure of the coating. Additionally, the viability of the cells was correlated with the wettability and roughness of the coatings, which determined the initial adhesion of the cells. Lastly, the temperature-induced changes in the properties of the fabricated copolymer coatings effectively controlled cell morphology, adhesion, and spontaneous detachment in a noninvasive, enzyme-free manner that was confirmed using optical microscopy.

Temperature- and pH-Responsive Schizophrenic Copolymer Brush Coatings with Enhanced Temperature Response in Pure Water.

Novel brush coatings were fabricated with glass surface-grafted chains copolymerized using surface-initiated atom transfer radical polymerization (SI-ATRP) from 2-(2-methoxyethoxy)ethyl methacrylate (OEGMA188) and acrylamide (AAm), taken in different proportions. P(OEGMA188-co-AAm) brushes with AAm mole fraction >44% (determined with XPS and TOF-SIMS spectroscopy) and nearly constant with the depth copolymer composition (TOF-SIMS profiling) exhibit unusual temperature-induced transformations: The contact angle of water droplets on P(OEGMA188-co-AAm) coatings increases by ∼45° with temperature, compared to 17-18° for POEGMA188 and PAAm. The thickness of coatings immersed in water and the morphology of coatings imaged in air show a temperature response for POEGMA188 (using reflectance spectroscopy and AFM, respectively), but this response is weak for P(OEGMA188-co-AAm) and absent for PAAm. This suggests mechanisms more complex than a simple transition between hydrated loose coils and hydrophobic collapsed chains. For POEGMA188, the hydrogen bonds between the ether oxygens of poly(ethylene glycol) and water hydrogens are formed below the transition temperature Tc and disrupted above Tc when polymer-polymer interactions are favored. Different hydrogen bond structures of PAAm include free amide groups, cis-trans-multimers, and trans-multimers of amide groups. Here, hydrogen bonds between free amide groups and water dominate at T < Tc but structures favored at T > Tc, such as cis-trans-multimers and trans-multimers of amide groups, can still be hydrated. The enhanced temperature-dependent response of wettability for P(OEGMA188-co-AAm) with a high mole fraction of AAm suggests the formation at Tc of more hydrophobic structures, realized by hydrogen bonding between the ether oxygens of OEGMA188 and the amide fragments of AAm, where water molecules are caged. Furthermore, P(OEGMA188-co-AAm) coatings immersed in pH buffer solutions exhibit a 'schizophrenic' behavior in wettability, with transitions that mimic LCST and UCST for pH = 3, LCST for pH = 5 and 7, and any transition blocked for pH = 9.

Passive antifouling and active self-disinfecting antiviral surfaces.

Viruses pose a serious threat to human health and society in general, as virus infections are one of the main causes of morbidity and mortality. Till May 2022, over 513 million people around the world have been confirmed to be infected and more than 6.2 million have died due to SARS-CoV-2. Although the COVID-19 pandemic will be defeated in the near future, we are likely to face new viral threats in the coming years. One of the important instruments to protect from viruses are antiviral surfaces, which are essentially capable of limiting their spread. The formulation of the concept of antiviral surfaces is relatively new. In general, five types of mechanism directed against virus spread can be proposed for antiviral surfaces; involving: direct and indirect actions, receptor inactivation, photothermal effect, and antifouling behavior. All antiviral surfaces can be classified into two main types - passive and active. Passive antiviral surfaces are based on superhydrophobic coatings that are able to repel virus contaminated droplets. In turn, viruses can become biologically inert (e.g., blocked or destroyed) upon contact with active antiviral surfaces, as they contain antiviral agents: metal atoms, synthetic or natural polymers, and small molecules. The functionality of antiviral surfaces can be significantly improved with additional properties, such as temperature- or pH-responsivity, multifunctionality, non-specific action on different virus types, long-term application, high antiviral efficiency and self-cleaning.

Fabrication and Impact of Fouling-Reducing Temperature-Responsive POEGMA Coatings with Embedded CaCO3 Nanoparticles on Different Cell Lines.

In the present work, we have successfully prepared and characterized novel nanocomposite material exhibiting temperature-dependent surface wettability changes, based on grafted brush coatings of non-fouling poly(di(ethylene glycol)methyl ether methacrylate) (POEGMA) with the embedded CaCO3 nanoparticles. Grafted polymer brushes attached to the glass surface were prepared in a three-step process using atom transfer radical polymerization (ATRP). Subsequently, uniform CaCO3 nanoparticles (NPs) embedded in POEGMA-grafted brush coatings were synthesized using biomineralized precipitation from solutions of CaCl2 and Na2CO3. An impact of the low concentration of the embedded CaCO3 NPs on cell adhesion and growth depends strongly on the type of studied cell line: keratinocytes (HaCaT), melanoma (WM35) and osteoblastic (MC3T3-e1). Based on the temperature-responsive properties of grafted brush coatings and CaCO3 NPs acting as biologically active substrate, we hope that our research will lead to a new platform for tissue engineering with modified growth of the cells due to the release of biologically active substances from CaCO3 NPs and the ability to detach the cells in a controlled manner using temperature-induced changes of the brush.

Dewetting of Polymer Films Controlled by Protein Adsorption.

The stability of the film poly(n-butyl methacrylate) (PnBMA) with different tacticities, prepared on silicon oxide and exposed to aqueous phosphate-buffered saline with different concentrations of bovine serum albumin (CBSA between 0 and 4.5 mg/mL), was examined at temperatures close to the physiological limit (between 4 and 37 °C) with optical microscopy, contact angle measurements, atomic force microscopy, and time-of-flight secondary ion mass spectrometry. For PBS solutions with CBSA = 0, the stability of atactic PnBMA and dewetting of isotactic PnBMA was observed, caused by the interplay between the stabilizing long-range dispersion forces and the destabilizing short-range polar interactions. Analogous considerations of excess free energy cannot explain the retardation of dewetting observed for isotactic PnBMA in PBS solutions with higher CBSA. Instead, formation of a BSA overlayer, adsorbed preferentially but not exclusively to uncovered SiOx regions, is evidenced and postulated to hinder polymer dewetting. Polymer dewetting and protein patterning are obtained in one step, suggesting a simple approach to fabricate biomaterials with micropatterned proteins.

Grafted polymer brush coatings for growth of cow granulosa cells and oocyte-cumulus cell complexes.

In the present work, three types of grafted brush coatings [P4VP, POEGMA246, and P(4VP-co-POEGMA246)] were successfully fabricated using graft polymerization of monomers "from the surface." The composition, thickness, and morphology of the grafted brush coatings were analyzed by TOF-SIMS, ellipsometry, and AFM, respectively. The chemical nature of the polymer surface plays a crucial role in the growth and development of the cow granulosa cells and, therefore, also oocyte-cumulus complexes. In comparison with other coatings, the P(4VP-co-POEGMA246) copolymer coating enables the formation of dispersed and small but numerous cell conglomerates and high cumulus expansion in oocyte-cumulus complexes with highly homogeneous cumulus layers surrounding the oocytes. Moreover, the cellular oxygen uptake for this coating in the presence of NaF (inhibitor glycolysis) was stimulated. This new (4VP-co-POEGMA246) copolymer nanostructured coating is a promising material for granulosa cell and oocyte-cumulus complex cultivation and possibly will have great potential for applications in veterinary and reproductive medicine.

Non-cytotoxic, temperature-responsive and antibacterial POEGMA based nanocomposite coatings with silver nanoparticles.

Non-cytotoxic, temperature-responsive and antibacterial poly(di(ethylene glycol)methyl ether methacrylate) - POEGMA188 based nanocomposite coatings attached to a glass surface were successfully prepared using ATRP polymerization. The thickness, morphology and wettability of the resulting coatings were analyzed using ellipsometry, AFM and contact angle measurements, respectively. The strong impact of the thicknesses of the POEGMA188 grafted brush coatings and content of AgNPs on the morphology and temperature-induced wettability changes of the nanocomposite was demonstrated. In addition to the strong temperature-dependent antibacterial activity, the proposed nanocomposite coatings have no significant cytotoxic effect towards normal cells. Moreover, the slight anti-cancer effect of AgNPs may be suggested.

"Command" surfaces with thermo-switchable antibacterial activity.

In the presented work "smart" antibacterial surfaces based on silver nanoparticles (AgNPs) embedded in temperature-responsive poly(di(ethylene glycol)methyl ether methacrylate) - (POEGMA188) as well as poly(4-vinylpyridine) - (P4VP) coatings attached to a glass surface were successfully prepared. The composition, thickness, morphology and wettability of the resulting coatings were analyzed using ToF-SIMS, XPS, EDX, ellipsometry, AFM, SEM and CA measurements, respectively. Temperature-switched killing of the bacteria was tested against Escherichia coli ATCC 25922 (representative of Gram-negative bacteria) and Staphylococcus aureus ATCC 25923 (representative of Gram-positive bacteria) at 4 and 37 °C. In general at 4 °C no significant difference was observed between the amounts of bacteria accounted on the grafted brush coatings and within the control sample. In contrast, at 37 °C almost no bacteria were visible for temperature-responsive coating with AgNPs, whereas the growth of bacteria remains not disturbed for "pure" coating, indicating strong temperature-dependent antibacterial properties of AgNPs integrated into brushes.

Temperature-Controlled Orientation of Proteins on Temperature-Responsive Grafted Polymer Brushes: Poly(butyl methacrylate) vs Poly(butyl acrylate): Morphology, Wetting, and Protein Adsorption.

Poly( n-butyl methacrylate) (PBMA) or poly( n-butyl acrylate) (PBA)-grafted brush coatings attached to glass were successfully prepared using atom-transfer radical polymerization "from the surface". The thicknesses and composition of the PBMA and PBA coatings were examined using ellipsometry and time-of-flight secondary ion mass spectrometry (ToF-SIMS), respectively. For PBMA, the glass-transition temperature constitutes a range close to the physiological limit, which is in contrast to PBA, where the glass-transition temperature is around -55 °C. Atomic force microscopy studies at different temperatures suggest a strong morphological transformation for PBMA coatings, in contrast to PBA, where such essential changes in the surface morphology are absent. Besides, for PBMA coatings, protein adsorption depicts a strong temperature dependence. The combination of bovine serum albumin and anti-IgG structure analysis with the principal component analysis of ToF-SIMS spectra revealed a different orientation of proteins adsorbed to PBMA coatings at different temperatures. In addition, the biological activity of anti-IgG molecules adsorbed at different temperatures was evaluated through tracing the specific binding with goat IgG.

pH-Controlled fluorescence switching in water-dispersed polymer brushes grafted to modified boron nitride nanotubes for cellular imaging.

pH-Switchable, fluorescent, hybrid, water-dispersible nanomaterials based on boron nitride nanotubes (BNNTs) and grafted copolymer brushes (poly(acrylic acid-co-fluorescein acrylate) - P(AA-co-FA)) were successfully fabricated in a two-step process. The functionalization of BNNTs was confirmed by spectroscopic, gravimetric and imaging techniques. In contrast to "pure" BNNTs, P(AA-co-FA)-functionalized BNNTs demonstrate intense green fluorescence emission at 520 nm. Under neutral or alkaline pH values, P(AA-co-FA)-functionalized BNNTs are highly emissive in contrast to acidic pH conditions where the fluorescent intensity is absent or low. No increase in the absorption was observed when the suspension pH was increased from 7 to 10. The functionalized BNNTs are easily taken up by human normal prostate epithelium (PNT1A) and human prostate cancer cell lines (DU145) and are suitable for further evaluation in cellular imaging applications.

Composition, thickness and properties of grafted copolymer brush coatings determined by ellipsometry: calculation and prediction.

The composition, thickness and properties of poly(4-vinylpyridine-co-oligo(ethylene glycol)ethyl ether methacrylate246) [P(4VP-co-OEGMA246)] copolymer grafted brush coatings attached to glass were studied in the dry and swollen states using ellipsometry. These measured data are in good agreement with predicted (estimated) changes in the amount of water, refractive index and thickness of the grafted copolymer brush coatings on swelling. For POEGMA brushes the thickness of the coatings on swelling at 20 °C can be more than double, in contrast to P4VP where those changes are insignificant. The presence of 4VP units in the structure of the P(4VP-co-OEGMA246) copolymer grafted brushes significantly decreases the hydration degree even for coatings with very low concentrations of 4VP units.

Glass transition in temperature-responsive poly(butyl methacrylate) grafted polymer brushes. Impact of thickness and temperature on wetting, morphology, and cell growth.

Poly(n-butyl methacrylate) (PBMA) grafted polymer brushes attached to glass were fabricated in a three-step process involving surface initiated atom transfer radical polymerization. The surface properties of the coatings after subsequent fabrication steps were confirmed using ToF-SIMS and ellipsometry. Measurements of water contact angle and AFM revealed temperature-induced changes in the hydrophobicity and morphology of the coating. The glass transition temperatures (Tg) of the PBMA coatings with different thicknesses were determined from the AFM measurements. For the PBMA grafted brush coatings with thicknesses less than 62 nm, Tg increases sharply with increasing thickness. The PBMA grafted coatings of thickness equal to 86 nm and 43 nm as well as control glass substrates were used as substrates for culturing a urinary bladder cancer HTB-5 cell line. After 144 h of culturing, a well-developed monocellular layer may be observed on the PBMA coating of thickness equal to 86 nm. In turn, the cells incubated on thinner (43 nm) PBMA coatings as well as on a control glass sample only start to form a confluent layer.

Temperature-Controlled Three-Stage Switching of Wetting, Morphology, and Protein Adsorption.

The novel polymeric coatings of oligoperoxide-graft-poly(4-vinylpyridine-co-oligo(ethylene glycol)ethyl ether methacrylate246) [oligoperoxide-graft-P(4VP-co-OEGMA246)] attached to glass were successfully fabricated. The composition, thickness, morphology, and wettability of resulting coatings were analyzed using X-ray photoelectron spectroscopy, ellipsometry, atomic force microscopy, and contact angle measurements, respectively. In addition, adsorption of the bovine serum albumin was examined with fluorescence microscopy. The thermal response of wettability and morphology of the coatings followed by that of protein adsorption revealed two distinct transitions at 10 and 23 °C. For the first time, three stage switching was observed not only for surface wetting but also for morphology and protein adsorption. Moreover, the influence of the pH on thermo-sensitivity of modified surfaces was shown.

Cholesterol-Based Grafted Polymer Brushes as Alignment Coating with Temperature-Tuned Anchoring for Nematic Liquid Crystals.

Novel alignment coating with temperature-tuned anchoring for nematic liquid crystals (NLCs) was successfully fabricated in three step process, involving polymerization of poly(cholesteryl methacrylate) (PChMa) from oligoproxide grafted to the glass surface premodified with 3-aminopropyltriethoxysilane. Molecular composition, thickness, wettability of the PChMa coating and its alignment action for a NLC were examined with time of flight-secondary ion mass spectrometry, ellipsometry, contact angle measurements, polarization optical microscopy and commercially produced PolScope technique allowing for mapping of the optic axis and optical retardance within the microscope field view. We find that the PChMa coating provides a specific monotonous increase (decrease) in the tilt angle of the NLC director with respect to the substrates normal upon heating (cooling) referred to as anchoring tuning.

Water-dispersed thermo-responsive boron nitride nanotubes: synthesis and properties.

In this study, water-dispersed thermo-responsive boron nitride nanotubes (BNNTs) were prepared in a simple two-step process, where on the first step oligoperoxide was grafted via the interaction of amino groups (defects) of BNNTs with pyromellitic chloroanhydride fragments in oligoperoxide molecules. The second step involves N-isopropylacrylamide (NIPAM) graft polymerization 'from the surface' of oligoperoxide-functionalized BNNTs resulting in poly(N-isopropylacrylamide) (PNIPAM) coating. The pristine and functionalized BNNTs were characterized by thermogravimetric analysis, Fourier transform infrared spectroscopy, ultraviolet-visible spectrophotometry, dynamic light scattering, scanning electron microscopy and atomic force microscopy. PNIPAM-functionalized BNNTs exhibit excellent dispersibility in water and possess thermo-responsive properties. The water-dispersion of thermo-responsive PNIPAM-functionalized BNNTs can help their potential use in biomedical applications as 'smart' surfaces, nanotransducers and nanocarriers.

Synthesis and Postpolymerization Modification of Thermoresponsive Coatings Based on Pentaerythritol Monomethacrylate: Surface Analysis, Wettability, and Protein Adsorption.

Properties of novel temperature-responsive hydroxyl-containing poly(pentaerythritol monomethacrylate) (PPM) coatings, polymerized from oligoperoxide grafted to glass surface premodified with (3-aminopropyl)triethoxysilane, are presented. Molecular composition, chemical state, thickness, and wettability are examined with time of flight-secondary ion mass spectrometry (ToF-SIMS), X-ray photoelectron spectroscopy (XPS), ellipsometry, and contact angle measurements, respectively. Temperature-induced changes in hydrophobicity of grafted PPM brushes are revealed by water contact angle and ellipsometric measurements. Partial postpolymerization modification of hydroxyl groups (maximum a few percent), performed with acetyl chloride or pyromellitic acid chloride, is demonstrated to preserve thermal response of coatings. Adsorption of bovine serum albumin to PPM brushes, observed with fluorescence microscopy, is higher than on glass in contrast to similar hydroxyl-containing layers reported as nonfouling. Enhanced and temperature-controlled protein adsorption is obtained after postpolymerization modification with pyromellitic acid chloride.

Temperature-responsive peptide-mimetic coating based on poly(N-methacryloyl-l-leucine): properties, protein adsorption and cell growth.

Poly(N-methacryloyl-l-leucine) (PNML) coatings were successfully fabricated via polymerization from peroxide initiator grafted to premodified glass substrate. Chemical composition and thickness of PNML coatings were determined using time of flight-secondary ion mass spectrometry (TOF- SIMS) and ellipsometry, respectively. PNML coatings exhibit thermal response of the wettability, between 4 and 28°C, which indicates a transition between hydrated loose coils and hydrophobic collapsed chains. Morphology of the PNML coating was observed with the AFM, transforming with increasing temperature from initially relatively smooth surface to rough and more structured surface. Protein adsorption observed by fluorescence microscopy for model proteins (bovine serum albumin and lentil lectin labeled with fluorescein isothiocyanate) at transition from 5 to 25°C, showed high affinity of PNML coating to proteins at all investigated temperatures and pH. Thus, PNML coating have significant potential for medical and biotechnological applications as protein capture agents or functional replacements of antibodies ("plastic antibodies"). The high proliferation growth of the human embryonic kidney cell (HEK 293) onto PNML coating was demonstrated, indicating its excellent cytocompatibility.

Temperature and pH dual-responsive POEGMA-based coatings for protein adsorption.

Poly(oligo(ethylene glycol)ethyl ether methacrylate (POEGMA246) coatings were successfully fabricated using novel approach via polymerization from oligoperoxide grafted to premodified glass substrate. Wettability, content and composition of coatings fabricated with different polymerization times were determined using contact angle measurements, ellipsometry and Time of Flight-Secondary Ion Mass Spectrometry (TOF-SIMS). Thermo- and pH-responsive properties of POEGMA246 coatings were found to depend significantly on concentration of the grafted POEGMA246. Coatings fabricated with polymerization time 30 h exhibit not only temperature- but also pH-dependence of wettability. Thermal response of wettability, measured between 20 and 32°C, was prominent at pH 9 and 7 and diminished or was absent at pH 5 and 3, indicating a transition between hydrated loose coils and hydrophobic collapsed chains, blocked at low pH. Protein adsorption, observed by fluorescence microscopy and analyzed semi-quantitatively using integral geometry approach, decreased dramatically for model protein (lentil lectin labeled with fluorescein isothiocyanate) at transition from pH 5 to pH 9, showing only very weak thermal-dependence. Strong protein adsorption response to pH and very weak one to temperature was confirmed by TOF-SIMS and Principal Component Analysis.

Temperature and pH dual-responsive coatings of oligoperoxide-graft-poly(N-isopropylacrylamide): wettability, morphology, and protein adsorption.

Poly(N-isopropylacrylamide) (PNIPAM) coatings attached to glass with novel approach involving polymerization from oligoperoxide grafted to surface with (3-aminopropyl)triethoxysilane exhibit not only temperature- but also pH-dependence of wettability and protein adsorption. Wettability and composition of coatings, fabricated with different polymerization times, were determined using contact angle measurements and Time Of Flight-Secondary Ion Mass Spectrometry (TOF-SIMS), respectively. Thermal response of wettability, measured between 20 and 40°C, was prominent at pH 9 and 7 and diminished or absent at pH 5 and 3. This indicates a transition between hydrated loose coils and hydrophobic collapsed chains that is blocked at low pH. Higher surface roughness and dramatically increased adsorption of model protein (lentil lectin labeled with fluorescein isothiocyanate) were observed with AFM and fluorescence microscopy to occur in hydrophobic phases (at pH 3, for pH varied at constant temperature of 22°C and at ∼33°C, for temperature varied at constant pH 9). Protein adsorption response to pH was confirmed by TOF-SIMS and Principal Component Analysis.