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Abstract Layered double hydroxides (LDHs) have been widely studied for biomedical applications due to their excellent properties, such as good biocompatibility, degradability, interlayer ion exchangeability, high loading capacity, pH‐responsive release, and large specific surface area. Furthermore, the flexibility in the structural composition and ease of surface modification of LDHs makes it possible to develop specifically functionalized LDHs to meet the needs of different applications. In this review, the recent advances of LDHs for biomedical applications, which include LDH‐based drug delivery systems, LDHs for cancer diagnosis and therapy, tissue engineering, coatings, functional membranes, and biosensors, are comprehensively discussed. From these various biomedical research fields, it can be seen that there is great potential and possibility for the use of LDHs in biomedical applications. However, at the same time, it must be recognized that the actual clinical translation of LDHs is still very limited. Therefore, the current limitations of related research on LDHs are discussed by combining limited examples of actual clinical translation with requirements for clinical translation of biomaterials. Finally, an outlook on future research related to LDHs is provided.

Implant associated infections are a serious threat to the well-being of patients, which can be mitigated by taking effective disinfection/sterilization (D/S) methods into account. Nanogels (nGel) are stimuli sensitive polymeric hydrogel particles, which have provided numerous innovative applications in the biomedical field to enhance antifouling, antibacterial properties, or drug delivery, or they can be employed as imaging modalities or can be applied as a coating on biomaterials (implants). Prior to translating their application towards clinical use, nGel-based coated implant materials must undergo an intermediary, pre-requisite process of cleaning, disinfection, and sterilization, in sequence. The interplay among the three crucial pillars- the implant material, the nGel coating (with specific function), and the applied D/S processes influence the fate (success or failure) of medical implant in the host body. In this study, we investigated a previously developed NIPAM-co-APMA core shell nGel coating on various clinically-relevant polymeric and inorganic implant materials and tested them on a diverse range of D/S techniques to assess the retention of the coating quality and antifouling function. The stability and integrity of the nGel coating was analyzed by performing Atomic Force Microscopy and the retention of the antifouling function of the nGel-coating after sterilization was studied by Colony forming units against S. aureus RN4220. Among all the materials that were coated, polymeric materials- polypropylene and polyetheretherketone exhibited exceptional coating stability, post-sterilization while also demonstrating a considerable reduction in bacterial attachment with respect to their ‘uncoated, sterilized’ and ‘coated, non-sterilized’ controls. Although often overlooked, sterilization is an indispensable part of clinical translation, therefore research in this domain is of utmost importance when considering clinical translatability.

Abstract Idiopathic pulmonary fibrosis is a progressive lung disease that causes scarring and loss of lung function. Macrophages play a key role in fibrosis, but their responses to altered morphological and mechanical properties of the extracellular matrix in fibrosis is relatively unexplored. Our previous work showed functional changes in murine fetal liver-derived alveolar macrophages on fibrous or globular collagen morphologies. In this study, we applied differential proteomics to further investigate molecular mechanisms underlying the observed functional changes. Macrophages cultured on uncoated, fibrous, or globular collagen-coated plastic were analyzed by liquid chromatography-mass spectrometry. The presence of collagen affected expression of 77 proteins, while 142 were differentially expressed between macrophages grown on fibrous or globular collagen. Biological process and pathway enrichment analysis revealed that culturing on any type of collagen induced higher expression of enzymes involved in glycolysis. However, this did not lead to a higher rate of glycolysis, probably because of a concomitant decrease in activity of these enzymes. Our data suggest that macrophages sense collagen morphologies and can respond with changes in expression and activity of metabolism-related proteins. These findings suggest intimate interactions between macrophages and their surroundings that may be important in repair or fibrosis of lung tissue.

The superior physicochemical properties of layered double hydroxides (LDHs) sets them apart from traditional polylactic acid (PLA) fillers. However, achieving efficient dispersion of LDHs within the PLA matrix poses a challenge in the fabrication of PLA/LDHs nanocomposites. In order to enhance the dispersion of LDHs in the PLA matrix and improve their compatibility with PLA, in this study sodium stearate (SS) and sodium dodecyl sulfate (SDS) were used as surface modifiers and intercalation modifiers for LDHs, respectively. Notably, the strategy of intercalation-surface combined modification for LDHs nanofillers in PLA was explored for the first time. Furthermore, the resulting PLA/LDHs nanocomposites were prepared via a sonication-assisted solution compounding method. The results indicate that the addition of LDHs modified through different methods and concentrations affects not only the thermal properties and crystallinity of the PLA/LDHs composites, but also improves the tensile strength and elongation at break of PLA. Notably, the PLA_LDH_SDS_SS_9% composite exhibited an elongation at break that was 230% higher than neat PLA. This study fills a research gap in the field of PLA/LDHs nanocomposites and establishes a significant foundation for the future preparation and application of these.

Coatings are essential for biomedical applications antifouling and antimicrobial properties, supporting cell adhesion and tissue integration and particularly interesting in this field are nanogel (nGel)‐based coatings. Since biomaterials differ in physiochemical properties, specific nGel‐coating strategies need to be developed for every distinct material, leading to complex coating strategies. Hence, the solution lies in adopting a universal strategy to apply the same nGel coating with the same function on a wide range of implant surfaces. To this end, a universal nGel‐based coating approach provides the same coating using a single method on implant materials including stiff polymer materials, metals, ceramics, glass, and elastomers. The coating formation is achieved by electrostatic interactions between oxygen plasma–activated surfaces and positively charged nGels using a spray‐deposition method. Fluorescent labels are introduced into the nGels as a model for post‐modification capabilities to increase the functionality of the coating. The coating is highly stable under in vitro physiological conditions with the retention of its function on different clinically relevant materials. Meanwhile, the in vivo study indicates that the nGel coating on a polyvinylidene fluoride hernia mesh is stable and biocompatible, therefore, making the coating and the coating strategy, a highly impactful approach for future clinical developments.

Layered double hydroxides (LDHs) have been extensively studied as drug delivery systems due to their favorable characteristics, including biocompatibility, high loading efficiency, and pH-responsive release. However, the current research predominantly focuses on LDHs as carriers for various anionic drugs, while there are only limited reports on LDHs as carriers for hydrophobic drugs. In this study, we successfully achieved the loading of a hydrophobic drug mimic, Nile red (NR), into LDHs using sodium dodecyl sulfate (SDS) as an intermediate storage medium. Furthermore, we optimized the experimental methods and varied the SDS/NR molar ratio to optimize this intercalation system. With an increase in the SDS/NR molar ratio from 2/1 to 32/1, the loading efficiency of LDH-SDS-NR for NR initially increased from 1.32% for LDH-SDS-NR_2/1 to 4.46% for LDH-SDS-NR_8/1. Then, the loading efficiency slightly decreased to 3.64% for LDH-SDS-NR_16.8/1, but then increased again to 6.31% for LDH-SDS-NR_32/1. We believe that the established method and the obtained results in this study broaden the application scope of LDHs as delivery systems for hydrophobic drugs and contribute to the further expansion of the application scope of LDHs.

Abstract The loading capacity and strategy of layered double hydroxides (LDHs) determine their suitability for drug delivery applications. In this study, two sizes of MgAl‐CO3 LDHs are converted into MgAl‐NO3 LDHs and MgAl‐Cl LDHs by an acid‐salt method. The influence of the size and type of interlayer anions of LDHs on their loading capacity are studied and compared. Here, Methylene Orange (MO) served as the anionic structure to determine the loading capacity. Due to the strongest anion‐exchange capacity of NO3− and larger interlayer distance of MgAl‐NO3 LDHs, they have impressive high loading capacities, which make these useful as absorbents or drug carriers. Additionally, the pH‐dependent release behaviors of MO‐loaded MgAl‐NO3 LDHs are investigated, which dictates both the release profile and maximum release. Above all, based on the study, it is found that the release of LDHs with agents loaded under mildly acidic environments is not based on its partial dissolution, which is commonly indicated. This study provides a better understanding of the loading and release process of LDHs, which allows for better incorporation of LDHs in biomedical‐related applications.

Presence of biofilms in drinking water distribution systems (DWDS) can be a nuisance, leading to several operational and maintenance issues (i.e., increased secondary disinfectants demand, pipe damage or increased flow resistance), and so far, no single control practice was found to be sufficiently effective. Here, we propose poly (sulfobetaine methacrylate) (P(SBMA))-based hydrogel coating application as a biofilm control strategy in DWDS. The P(SBMA) coating was synthetized through photoinitiated free radical polymerization on polydimethylsiloxane with different combinations of SBMA as a monomer, and N, N′-methylenebis (acrylamide) (BIS) as a cross-linker. The most stable coating in terms of its mechanical properties was obtained using 20% SBMA with a 20:1 SBMA:BIS ratio. The coating was characterized using Scanning Electron Microscopy, Energy Dispersive X-Ray Spectroscopy, and water contact angle measurements. The anti-adhesive performance of the coating was evaluated in a parallel-plate flow chamber system against adhesion of four bacterial strains representing genera commonly identified in DWDS biofilm communities, Sphingomonas and Pseudomonas. The selected strains exhibited varying adhesion behaviors in terms of attachment density and bacteria distribution on the surface. Despite these differences, after 4 h, presence of the P(SBMA)-based hydrogel coating significantly reduced the number of adhering bacteria by 97%, 94%, 98% and 99%, for Sphingomonas Sph5, Sphingomonas Sph10, Pseudomonas extremorientalis and Pseudomonas aeruginosa, respectively, compared to non-coated surfaces. These findings motivate further research into a potential application of a hydrogel anti-adhesive coating as a localized biofilm control strategy in DWDS, especially on materials known to promote excessive biofilm growth.

The implementation of nanotechnology to develop efficient antimicrobial systems has a significant impact on the prospects of the biomedical field. Nanogels are soft polymeric particles with an internally cross-linked structure, which behave as hydrogels and can be reversibly hydrated/dehydrated (swollen/shrunken) by the dispersing solvent and external stimuli. Their excellent properties, such as biocompatibility, colloidal stability, high water content, desirable mechanical properties, tunable chemical functionalities, and interior gel-like network for the incorporation of biomolecules, make them fascinating in the field of biological/biomedical applications. In this review, various approaches will be discussed and compared to the newly developed nanogel technology in terms of efficiency and applicability for determining their potential role in combating infections in the biomedical area including implant-associated infections.

Introduction: Idiopathic pulmonary fibrosis (IPF) is a fibrotic lung disease characterized by excess deposition and altered structure of extracellular matrix (ECM) in the lungs. The fibrotic ECM is paramount in directing resident cells toward a profibrotic phenotype. Collagens, an important part of the fibrotic ECM, have been shown to be structurally different in IPF. To further understand the disease to develop better treatments, the signals from the ECM that drive fibrosis need to be identified. Adipose tissue-derived stromal cell conditioned medium (ASC-CM) has demonstrated antifibrotic effects in animal studies but has not been tested in human samples yet. In this study, the collagen structural integrity in (fibrotic) lung tissue, its interactions with fibroblasts and effects of ASC-CM treatment hereon were studied.Methods: Native and decellularized lung tissue from patients with IPF and controls were stained for denatured collagen using a collagen hybridizing peptide. Primary lung fibroblasts were seeded into decellularized matrices from IPF and control subjects and cultured for 7 days in the presence or absence of ASC-CM. Reseeded matrices were fixed, stained and analyzed for total tissue deposition and specific protein expression.Results: In both native and decellularized lung tissue, more denatured collagen was observed in IPF tissue compared to control tissue. Upon recellularization with fibroblasts, the presence of denatured collagen was equalized in IPF and control matrices, whereas total ECM was higher in IPF matrices than in the control. Treatment with ASC-CM resulted in less ECM deposition, but did not alter the levels of denatured collagen.Discussion: Our data showed that ASC-CM can inhibit fibrotic ECM-induced profibrotic behavior of fibroblasts. This process was independent of collagen structural integrity. Our findings open up new avenues for ASC-CM to be explored as treatment for IPF.

Hygiene is a basic necessity to prevent infections, and though it is regarded as vital in general, its importance has been stressed again during the pandemic. Microbes may spread through touch and aerosols and thereby find their way from host to host. Cleaning and disinfection of possibly contaminated surfaces prevents microbial spread, thus reducing potential illnesses. One item that is used by several people in a way that promotes close contact by touch and aerosol formation is the microphone. A microphone is a complex piece of equipment with respect to shape and various materials used to fabricate it and, hence, its disinfection is challenging. A new device has been developed to efficiently sterilize microphones by using UV-C and a biological assessment has been done to identify its efficacy and translatability. For this investigation, a contamination procedure was developed by using M13 bacteriophage as a model to illustrate the effectiveness of the disinfection. The susceptibility to UV-C irradiation of M13 in solution was compared to that of the PR8 H1N1 influenza virus, which has a similar UV-C susceptibility as SARS-CoV-2. It was found that 10 min of UV-C treatment reduced the percentage of infectious M13 by 99.3% based on whole microphone inoculation and disinfection. UV-C susceptibility of M13 and influenza in suspension were found to be very similar, indicating that the microphone sterilization method and device function are highly useful and broadly applicable.

Glioblastoma multiforme (GBM) is the most aggressive brain tumor in adults. In addition to genetic causes, the tumor microenvironment (TME), including stiffening of the extracellular matrix (ECM), is a main driver of GBM progression. Mechano-transduction and the unfolded protein response (UPR) are essential for tumor-cell adaptation to harsh TME conditions. Here, we studied the effect of a variable stiff ECM on the morphology and malignant properties of GBM stem cells (GSCs) and, moreover, examined the possible involvement of the UPR sensor PERK herein. For this, stiffness-tunable human blood plasma (HBP)/alginate hydrogels were generated to mimic ECM stiffening. GSCs showed stiffness-dependent adaptation characterized by elongated morphology, increased proliferation, and motility which was accompanied by F-Actin cytoskeletal remodeling. Interestingly, in PERK-deficient GSCs, stiffness adaptation was severely impaired, which was evidenced by low F-Actin levels, the absence of F-Actin remodeling, and decreased cell proliferation and migration. This impairment could be linked with Filamin-A (FLN-A) expression, a known interactor of PERK, which was strongly reduced in PERK-deficient GSCs. In conclusion, we identified a novel PERK/FLNA/F-Actin mechano-adaptive mechanism and found a new function for PERK in the cellular adaptation to ECM stiffening.

Gene delivery holds great promise for bioengineering, biomedical applications, biosensors, diagnoses, and gene therapy. In particular, the influence of topography on gene delivery is considered to be an attractive approach due to low toxicity and localized delivery properties. Even though many gene vectors and transfection systems have been developed to enhance transfection potential and combining it with other forms of stimulations could even further enhance it. Topography is an interesting surface property that has been shown to stimulate differentiation, migration, cell morphology, and cell mechanics. Therefore, it is envisioned that topography might also be able to stimulate transfection. In this study, we tested the hypothesis “topography is able to regulate transfection efficiency”, for which we used nano- and microwave-like topographical substrates with wavelengths ranging from 500 nm to 25 µm and assessed the transfectability of human bone marrow-derived mesenchymal stem cells (hBM-MSCs) and myoblasts. For transfection, Lipofectamine 2000 and a gene encoding plasmid for red-fluorescent protein (m-Cherry) were used and topography-induced cell morphology and transfection efficiency was analyzed. As a result, topography directs cell spreading, elongation, and proliferation as well as the transfection efficiency, which were investigated but were found not to be correlated and dependent on the cell type. A 55% percent improvement of transfection efficiency was identified for hBM-MSCs grown on 2 µm wrinkles (24.3%) as compared to hBM-MSCs cultured on flat controls (15.7%). For myoblast cells, the highest gene-expression efficiency (46.1%) was observed on the 10 µm topography, which enhanced the transfection efficiency by 64% as compared to the flat control (28.1%). From a qualitative assessment, it was observed that the uptake capacity of cationic complexes of TAMRA-labeled oligodeoxynucleotides (ODNs) was not topography-dependent but that the intracellular release was faster, as indicated by the positively stained nuclei on 2 μm for hBM-MSCs and 10 μm for myoblasts. The presented results indicate that topography enhances the gene-delivery capacity and that the responses are dependent on cell type. This study demonstrates the important role of topography on cell stimulation for gene delivery as well as understanding the uptake capacity of lipoplexes and may be useful for developing advanced nonviral gene delivery strategies.

The biology and physics underlying glioblastoma is not yet completely understood, resulting in the limited efficacy of current clinical therapy. Recent studies have indicated the importance of mechanical stress on the development and malignancy of cancer. Various types of mechanical stress activate adaptive tumor cell responses that include alterations in the extracellular matrix (ECM) which have an impact on tumor malignancy. In this review, we describe and discuss the current knowledge of the effects of ECM alterations and mechanical stress on GBM aggressiveness. Gradual changes in the brain ECM have been connected to the biological and physical alterations of GBM cells. For example, increased expression of several ECM components such as glycosaminoglycans (GAGs), hyaluronic acid (HA), proteoglycans and fibrous proteins result in stiffening of the brain ECM, which alters inter- and intracellular signaling activity. Several mechanosensing signaling pathways have been identified that orchestrate adaptive responses, such as Hippo/YAP, CD44, and actin skeleton signaling, which remodel the cytoskeleton and affect cellular properties such as cell–cell/ECM interactions, growth, and migration/invasion of GBM cells. In vitro, hydrogels are used as a model to mimic the stiffening of the brain ECM and reconstruct its mechanics, which we also discuss. Overall, we provide an overview of the tumor microenvironmental landscape of GBM with a focus on ECM stiffening and its associated adaptive cellular signaling pathways and their possible therapeutic exploitation.

Understanding how endothelial cell phenotype is affected by topography could improve the design of new tools for tissue engineering as many tissue engineering approaches make use of topography-mediated cell stimulation. Therefore, we cultured human pulmonary microvascular endothelial cells (ECs) on a directional topographical gradient to screen the EC vascular-like network formation and alignment response to nano to microsized topographies. The cell response was evaluated by microscopy. We found that ECs formed unstable vascular-like networks that aggregated in the smaller topographies and flat parts whereas ECs themselves aligned on the larger topographies. Subsequently, we designed a mixed topography where we could explore the network formation and proliferative properties of these ECs by live imaging for three days. Vascular-like network formation continued to be unstable on the topography and were only produced on the flat areas and a fibronectin coating did not improve the network stability. However, an instructive adipose tissue-derived stromal cell (ASC) coating provided the correct environment to sustain the vascular-like networks, which were still affected by the topography underneath. It was concluded that large microsized topographies inhibit vascular endothelial network formation but not proliferation and flat and nano/microsized topographies allow formation of early networks that can be stabilized by using an ASC instructive layer.

Gene therapy is a promising treatment for hereditary diseases, as well as acquired genetic diseases, including cancer. Facing the complicated physiological and pathological environment in vivo, developing efficient non-viral gene vectors is needed for their clinical application. Here, poly(N-isopropylacrylamide) (p(NIPAM)) nanogels are presented with either protonatable tertiary amine groups or permanently charged quaternized ammonium groups to achieve DNA complexation ability. In addition, a quaternary ammonium-functionalized nanogel was further provided with an aliphatic moiety using 1-bromododecane to add a membrane-interacting structure to ultimately facilitate intracellular release of the genetic material. The ability of the tertiary amine-, quaternized ammonium-, and aliphatic quaternized ammonium-functionalized p(NIPAM) nanogels (i.e., NGs, NGs-MI, and NGs-BDD, respectively) to mediate gene transfection was evaluated by fluorescence microscopy and flow cytometry. It is observed that NGs-BDD/pDNA complexes exhibit efficient gene loading, gene protection ability, and intracellular uptake similar to that of NGs-MI/pDNA complexes. However, only the NGs-BDD/pDNA complexes show a notable gene transfer efficiency, which can be ascribed to their ability to mediate DNA escape from endosomes. We conclude that NGs-BDD displays a cationic lipid-like behavior that facilitates endosomal escape by perturbing the endosomal/lysosomal membrane. These findings demonstrate that the presence of aliphatic chains within the nanogel is instrumental in accomplishing gene delivery, which provides a rationale for the further development of nanogel-based gene delivery systems.

Fluorescent nanodiamonds are a useful for biosensing of intracellular signaling networks or environmental changes (such as temperature, pH or free radical generation). HeLa cells are interesting to study with these nanodiamonds since they are a model cell system that is widely used to study cancer-related diseases. However, they only internalize low numbers of nanodiamond particles very slowly via the endocytosis pathway. In this work, we show that pH-sensitive, dextran-coated fluorescent nanodiamonds can be used to visualise this pathway. Additionally, this coating improved diamond uptake in HeLa cells by 5.3 times (*** p < 0.0001) and decreased the required time for uptake to only 30 min. We demonstrated further that nanodiamonds enter HeLa cells via endolysosomes and are eventually expelled by cells.

The strength of the extracellular matrix (ECM) is that it is hierarchical in terms of matrix built-up, matrix density and fiber structure, which allows for hormones, cytokines, and other small biomolecules to be stored within its network. The ECM-like hydrogels that are currently used do not possess this ability, and long-term storage, along with the need for free diffusion of small molecules, are generally incompatible requirements. Nanogels are able to fulfill the additional requirements upon successful integration. Herein, a stable hierarchical nanogel–gelatin methacryloyl (GelMA) composite hydrogel system is provided by covalently embedding nanogels inside the micropore network of GelMA hydrogel to allow a controlled local functionality that is not found in a homogenous GelMA hydrogel. Nanogels have emerged as a powerful tool in nanomedicine and are highly versatile, due to their simplicity of chemical control and biological compatibility. In this study, an N-isopropylacrylamide-based nanogel with primary amine groups on the surface was modified with methacryloyl groups to obtain a photo-cross-linking ability similar to GelMA. The nanogel-GelMA composite hydrogel was formed by mixing the GelMA and the photo-initiator within the nanogel solution through UV irradiation. The morphology of the composite hydrogel was observed by scanning electron microscopy, which clearly showed the nanogel wrapped within the GelMA network and covering the surface of the pore wall. A release experiment was conducted to prove covalent bonding and the stability of the nanogel inside the GelMA hydrogel. In addition, 3D printability studies showed that the nanogel-GelMA composite ink is printable. Therefore, the suggested stable hierarchical nanogel-GelMA composite hydrogel system has great potential to achieve the in situ delivery and controllable release of bioactive molecules in 3D cell culture systems.

Summary: The relevance of topographic cues for commitment of induced pluripotent stem cells (iPSCs) is largely unknown. In this study, we demonstrate that groove-ridge structures with a periodicity in the submicrometer range induce elongation of iPSC colonies, guide the orientation of apical actin fibers, and direct the polarity of cell division. Elongation of iPSC colonies impacts also on their intrinsic molecular patterning, which seems to be orchestrated from the rim of the colonies. BMP4-induced differentiation is enhanced in elongated colonies, and the submicron grooves impact on the spatial modulation of YAP activity upon induction with this morphogen. Interestingly, TAZ, a YAP paralog, shows distinct cytoskeletal localization in iPSCs. These findings demonstrate that topography can guide orientation and organization of iPSC colonies, which may affect the interaction between mechanosensors and mechanotransducers in iPSCs. : Topography can impact on growth and differentiation of iPSCs. Abagnale et al. used laser beam interference to generate submicron grooves that guide the shape of iPSC colonies, thereby modulating the spatial distribution of pluripotency markers, induction with BMP4, in vitro differentiation, and subcellular localization of the mechanotransducers YAP and TAZ. Keywords: iPSC, topography, biomaterials, microtopography, laser beam interference, pattern, mechanobiology, cytoskeleton, YAP, TAZ

Introduction. Progenitor cells cultured on biomaterials with optimal physical-topographical properties respond with alignment and differentiation. Stromal cells from connective tissue can adversely differentiate to profibrotic myofibroblasts or favorably to smooth muscle cells (SMC). We hypothesized that myogenic differentiation of adipose tissue-derived stromal cells (ASC) depends on gradient directional topographic features. Methods. Polydimethylsiloxane (PDMS) samples with nanometer and micrometer directional topography gradients (wavelength w=464-10, 990 nm; amplitude a=49-3, 425 nm) were fabricated. ASC were cultured on patterned PDMS and stimulated with TGF-β1 to induce myogenic differentiation. Cellular alignment and adhesion were assessed by immunofluorescence microscopy after 24 h. After seven days, myogenic differentiation was examined by immunofluorescence microscopy, gene expression, and immunoblotting. Results. Cell alignment occurred on topographies larger than w=1758 nm/a=630 nm. The number and total area of focal adhesions per cell were reduced on topographies from w=562 nm/a=96 nm to w=3919 nm/a=1430 nm. Focal adhesion alignment was increased on topographies larger than w=731 nm/a=146 nm. Less myogenic differentiation of ASC occurred on topographies smaller than w=784 nm/a=209 nm. Conclusion. ASC adherence, alignment, and differentiation are directed by topographical cues. Our evidence highlights a minimal topographic environment required to facilitate the development of aligned and differentiated cell layers from ASC. These data suggest that nanotopography may be a novel tool for inhibiting fibrosis.

Understanding the response of endothelial cells to aligned myotubes is important to create an appropriate environment for tissue-engineered vascularized skeletal muscle. Part of the native tissue environment is the extracellular matrix (ECM). The ECM is a supportive scaffold for cells and allows cellular processes such as proliferation, differentiation, and migration. Interstitial matrix and basal membrane both comprise proteinaceous and polysaccharide components for strength, architecture, and volume retention. Virtually all cells are anchored to their basal lamina. One of the physical factors that affects cell behavior is topography, which plays an important role on cell alignment. We tested the hypothesis that topography-driven aligned human myotubes promote and support vascular network formation as a prelude to in vitro engineered vascularized skeletal muscle. Therefore, we used a PDMS-based topography substrate to investigate the influence of pre-aligned myotubes on the network formation of microvascular endothelial cells. The aligned myotubes produced a network of collagen fibers and laminin. This network supported early stages of endothelial network formation.

Here we describe the combined process of surface modification with electrochemical atom transfer radical polymerization (e-ATRP) initiated from the surface of a modified gold-electrode in a pure aqueous solution without any additional supporting electrolyte. This approach allows for a very controlled growth of the polymer chains leading towards a steady increase in film thickness. Electrochemical quartz crystal microbalance displayed a highly regular increase in surface confined mass only after the addition of the pre-copper catalyst which is reduced in situ and transformed into the catalyst. Even after isolation and washing of the modified electrode surface, reinitiation was achieved with retention of the controlled electrochemical ATRP reaction. This reinitiation after isolation proves the livingness of the polymerization. This approach has interesting potential for smart thin film materials and offers also the possibility of post-modification via additional electrochemical induced reactions.

Protein aggregation and protein self-assembly is an important occurrence in natural systems, and is in some form or other dictated by biopolymers. Very obvious influences of biopolymers on protein assemblies are, e.g., virus particles. Viruses are a multi-protein assembly of which the morphology is dictated by poly-nucleotides namely RNA or DNA. This “biopolymer” directs the proteins and imposes limitations on the structure like the length or diameter of the particle. Not only do these bionanoparticles use polymer-directed self-assembly, also processes like amyloid formation are in a way a result of directed protein assembly by partial unfolded/misfolded biopolymers namely, polypeptides. The combination of proteins and synthetic polymers, inspired by the natural processes, are therefore regarded as a highly promising area of research. Directed protein assembly is versatile with respect to the possible interactions which brings together the protein and polymer, e.g., electrostatic, v.d. Waals forces or covalent conjugation, and possible combinations are numerous due to the large amounts of different polymers and proteins available. The protein-polymer interacting behavior and overall morphology is envisioned to aid in clarifying protein-protein interactions and are thought to entail some interesting new functions and properties which will ultimately lead to novel bio-hybrid materials.

Polydimethylsiloxane (PDMS) is a silicone elastomer-based material that is used in various applications, including coatings, tubing, microfluidics, and medical implants. PDMS has been modified with hydrogel coatings to prevent fouling, which can be done through UV-mediated free radical polymerization using benzophenone. However, to the best of our knowledge, the properties of hydrogel coatings and their influence on the bulk properties of PDMS under various preparation conditions, such as the type and concentration of monomers, and UV treatment time, have never been investigated. Acrylate-based monomers were used to perform free radical polymerization on PDMS surfaces under various reaction conditions. This approach provides insights into the relationship between the hydrogel coating and bulk properties of PDMS. Altering the UV polymerization time and the monomer concentration resulted in different morphologies with different roughness and thickness of the hydrogel coating, as well as differences in the bulk material stiffness. The surface morphology of the coated PDMS was characterized by AFM. The cross section and thickness of the coatings were examined using scanning electron microscopy coupled with energy-dispersive X-ray spectroscopy. The dependence of coating development on the monomer type and concentration used was evaluated by surface hydrophilicity, as measured by water contact angle. Elongation-until-break analysis revealed that specific reaction conditions affected the bulk properties and made the coated PDMS brittle. Therefore, boundary conditions have been identified to enable high quality hydrogel coating formation without affecting the bulk properties of the material.