Growing Gold Nanostars on 3D Hydrogel Surfaces.

Nanocomposites comprising hydrogels and plasmonic nanoparticles are attractive materials for tissue engineering, bioimaging, and biosensing. These materials are usually fabricated by adding colloidal nanoparticles to the uncured polymer mixture and thus require time-consuming presynthesis, purification, and ligand-exchange steps. Herein, we introduce approaches for rapid synthesis of gold nanostars (AuNSt) in situ on hydrogel substrates, including those with complex three-dimensional (3D) features. These methods enable selective AuNSt growth at the surface of the substrate, and the growth conditions can be tuned to tailor the nanoparticle size and density (coverage). We additionally demonstrate proof-of-concept applications of these nanocomposites for SERS sensing and imaging. High surface coverage with AuNSt enabled 1-2 orders of magnitude higher SERS signals compared to plasmonic hydrogels loaded with premade colloids. Importantly, AuNSt can be prepared without the addition of any potentially cytotoxic surfactants, thereby ensuring a high biocompatibility. Overall, in situ growth becomes a versatile and straightforward approach for the fabrication of plasmonic biomaterials.

3D Bioprinted Tumor-Stroma Models of Triple-Negative Breast Cancer Stem Cells for Preclinical Targeted Therapy Evaluation.

Breast cancer stem cells (CSCs) play a pivotal role in therapy resistance and tumor relapse, emphasizing the need for reliable in vitro models that recapitulate the complexity of the CSC tumor microenvironment to accelerate drug discovery. We present a bioprinted breast CSC tumor-stroma model incorporating triple-negative breast CSCs (TNB-CSCs) and stromal cells (human breast fibroblasts), within a breast-derived decellularized extracellular matrix bioink. Comparison of molecular signatures in this model with different clinical subtypes of bioprinted tumor-stroma models unveils a unique molecular profile for artificial CSC tumor models. We additionally demonstrate that the model can recapitulate the invasive potential of TNB-CSC. Surface-enhanced Raman scattering imaging allowed us to monitor the invasive potential of tumor cells in deep z-axis planes, thereby overcoming the depth-imaging limitations of confocal fluorescence microscopy. As a proof-of-concept application, we conducted high-throughput drug testing analysis to assess the efficacy of CSC-targeted therapy in combination with conventional chemotherapeutic compounds. The results highlight the usefulness of tumor-stroma models as a promising drug-screening platform, providing insights into therapeutic efficacy against CSC populations resistant to conventional therapies.

SERS in 3D cell models: a powerful tool in cancer research.

Unraveling the cellular and molecular mechanisms underlying tumoral processes is fundamental for the diagnosis and treatment of cancer. In this regard, three-dimensional (3D) cancer cell models more realistically mimic tumors compared to conventional 2D cell cultures and are more attractive for performing such studies. Nonetheless, the analysis of such architectures is challenging because most available techniques are destructive, resulting in the loss of biochemical information. On the contrary, surface-enhanced Raman spectroscopy (SERS) is a non-invasive analytical tool that can record the structural fingerprint of molecules present in complex biological environments. The implementation of SERS in 3D cancer models can be leveraged to track therapeutics, the production of cancer-related metabolites, different signaling and communication pathways, and to image the different cellular components and structural features. In this review, we highlight recent progress in the use of SERS for the evaluation of cancer diagnosis and therapy in 3D tumoral models. We outline strategies for the delivery and design of SERS tags and shed light on the possibilities this technique offers for studying different cellular processes, through either biosensing or bioimaging modalities. Finally, we address current challenges and future directions, such as overcoming the limitations of SERS and the need for the development of user-friendly and robust data analysis methods. Continued development of SERS 3D bioimaging and biosensing systems, techniques, and analytical strategies, can provide significant contributions for early disease detection, novel cancer therapies, and the realization of patient-tailored medicine.

A Scaffold-Assisted 3D Cancer Cell Model for Surface-Enhanced Raman Scattering-Based Real-Time Sensing and Imaging.

Despite recent advances in the development of scaffold-based three-dimensional (3D) cell models, challenges persist in imaging and monitoring cell behavior within these complex structures due to their heterogeneous cell distribution and geometries. Incorporating sensors into 3D scaffolds provides a potential solution for real-time, in situ sensing and imaging of biological processes such as cell growth and disease development. We introduce a 3D printed hydrogel-based scaffold capable of supporting both surface-enhanced Raman scattering (SERS) biosensing and imaging of 3D breast cancer cell models. The scaffold incorporates plasmonic nanoparticles and SERS tags, for sensing and imaging, respectively. We demonstrate the scaffold's adaptability and modularity in supporting breast cancer spheroids, thereby enabling spatial and temporal monitoring of tumor evolution.

Nanocomposite hydrogel microneedles: a theranostic toolbox for personalized medicine.

Due to the severity and high prevalence of cancer, as well as its complex pathological condition, new strategies for cancer treatment and diagnostics are required. As such, it is important to design a toolbox that integrates multiple functions on a single smart platform. Theranostic hydrogels offer an innovative and personalized method to tackle cancer while also considering patient comfort, thereby facilitating future implementation and translation to the clinic. In terms of theranostic systems used in cancer therapy, nanoparticles are widely used as diagnostic and therapeutic tools. Nanoparticles can achieve systemic circulation, evade host defenses, and deliver drugs and signaling agents at the targeted site, to diagnose and treat the disease at a cellular and molecular level. In this context, hydrogel microneedles have a high potential for multifunctional operation in medical devices, while avoiding the complications associated with the systemic delivery of therapeutics. Compared with oral administration and subcutaneous injection, microneedles offer advantages such as better patient compliance, faster onset of action, and improved permeability and efficacy. In addition, they comprise highly biocompatible polymers with excellent degradability and tunable properties. Nanoparticles and microneedles thus offer the possibility to expand the theranostic potential through combined synergistic use of their respective features. We review herein recent advances concerning processing methods and material requirements within the realm of hydrogel microneedles as theranostic platforms, various approaches toward cancer therapy, and the incorporation of nanoparticles for added functionality.

3D bioprinted breast tumor-stroma models for pre-clinical drug testing.

The use of three-dimensional (3D) bioprinting has been proposed for the reproducible production of 3D disease models that can be used for high-throughput drug testing and personalized medicine. However, most such models insufficiently reproduce the features and environment of real tumors. We report the development of bioprinted in vitro 3D tumor models for breast cancer, which physically and biochemically mimic important aspects of the native tumor microenvironment, designed to study therapeutic efficacy. By combining a mix of breast decellularized extracellular matrix and methacrylated hyaluronic acid with tumor-derived cells and non-cancerous stromal cells of biological relevance to breast cancer, we show that biological signaling pathways involved in tumor progression can be replicated in a carefully designed tumor-stroma environment. Finally, we demonstrate proof-of-concept application of these models as a reproducible platform for investigating therapeutic responses to commonly used chemotherapeutic agents.

Remodeling arteries: studying the mechanical properties of 3D-bioprinted hybrid photoresponsive materials.

3D-printed cell models are currently in the spotlight of medical research. Whilst significant advances have been made, there are still aspects that require attention to achieve more realistic models which faithfully represent the in vivo environment. In this work we describe the production of an artery model with cyclic expansive properties, capable of mimicking the different physical forces and stress factors that cells experience in physiological conditions. The artery wall components are reproduced using 3D printing of thermoresponsive polymers with inorganic nanoparticles (NPs) representing the outer tunica adventitia, smooth muscle cells embedded in extracellular matrix representing the tunica media, and finally a monolayer of endothelial cells as the tunica intima. Cyclic expansion can be induced thanks to the inclusion of photo-responsive plasmonic NPs embedded within the thermoresponsive ink composition, resulting in changes in the thermoresponsive polymer hydration state and hence volume, in a stimulated on-off manner. By changing the thermoresponsive polymer composition, the transition temperature and pulsatility can be efficiently tuned. We show the direct effect of cyclic expansion and contraction on the overlying cell layers by analyzing transcriptional changes in mechanoresponsive mesenchymal genes associated with such microenvironmental physical cues. The technique described herein involving stimuli-responsive 3D printed tissue constructs, also described as four- dimensional (4D) printing, offers a novel approach for the production of dynamic biomodels.

Trends in Tissue Bioprinting, Cell-Laden Bioink Formulation, and Cell Tracking.

Use of three-dimensional bioprinting for the in vitro engineering of tissues has boomed during the past five years. An increasing number of commercial bioinks are available, with suitable mechanical and rheological characteristics and excellent biocompatibility. However, cell-laden bioinks based on a single polymer do not properly mimic the complex extracellular environment needed to tune cell behavior, as required for tissue and organ formation. Processes such as cell aggregation, migration, and tissue patterning should be dynamically monitored, and progress is being made in these areas, most prominently derived from nanoscience. We review recent developments in tissue bioprinting, cellularized bioink formulation, and cell tracking, from both chemistry and cell biology perspectives. We conclude that an interdisciplinary approach including expertise in polymer science, nanoscience, and cell biology/tissue engineering is required to drive further advancements in this field toward clinical application.

Macroporous Silica Foams Fabricated via Soft Colloid Templating.

Macroporous materials with controlled pore sizes are of high scientific and technological interest, due to their low specific weight, as well as unique acoustic, thermal, or optical properties. Solid foams made of titania, silica, or silicon, as representative materials, have been previously obtained with several hundred nanometer pore sizes, by using sacrificial templates such as spherical emulsion droplets or colloidal particles. Macroporous structures in particular are excellent candidates as photonic materials with applications in structural coloration and photonic bandgap devices. However, whereas using spherical building blocks as templates may provide tight control over pore shape and size, it results in materials with an often unfavorable local topology. Templating dry-foam or compressed-emulsion structures appear as attractive alternatives, but have not been demonstrated so far for submicron pore sizes. Herein, the use of soft, flexible microgel colloids decorated with silica nanoparticles as templates of macroporous foams is reported. These purposely synthesized core-shell colloids are assembled at ultra-high effective volume fractions by centrifuging and thermal swelling, thereby resulting in uniform disordered materials with facetted pores, mimicking dry foams. After removal of the polymer component via calcination, lightweight pure silica structures are obtained with a well-defined cellular or network topology.

Nanocomposite Scaffolds for Monitoring of Drug Diffusion in Three-Dimensional Cell Environments by Surface-Enhanced Raman Spectroscopy.

Monitoring dynamic processes in complex cellular environments requires the integration of uniformly distributed detectors within such three-dimensional (3D) networks, to an extent that the sensor could provide real-time information on nearby perturbations in a non-invasive manner. In this context, the development of 3D-printed structures that can function as both sensors and cell culture platforms emerges as a promising strategy, not only for mimicking a specific cell niche but also toward identifying its characteristic physicochemical conditions, such as concentration gradients. We present herein a 3D cancer model that incorporates a hydrogel-based scaffold containing gold nanorods. In addition to sustaining cell growth, the printed nanocomposite inks display the ability to uncover drug diffusion profiles by surface-enhanced Raman scattering, with high spatiotemporal resolution. We additionally demonstrate that the acquired information could pave the way to designing novel strategies for drug discovery in cancer therapy, through correlation of drug diffusion with cell death.

Colloidal systems toward 3D cell culture scaffolds.

Three-dimensional porous scaffolds are essential for the development of tissue engineering and regeneration, as biomimetic supports to recreate the microenvironment present in natural tissues. To successfully achieve the growth and development of a specific kind of tissue, porous matrices should be able to influence cell behavior by promoting close cell-cell and cell-matrix interactions. To achieve this goal, the scaffold must fulfil a set of conditions, including ordered interconnected porosity to promote cell diffusion and vascularization, mechanical strength to support the tissue during continuous ingrowth, and biocompatibility to avoid toxicity. Among various building approaches to the construction of porous matrices, selected strategies afford hierarchical scaffolds with such defined properties. The control over porosity, microstructure or morphology, is crucial to the fabrication of high-end, reproducible scaffolds for the target application. In this review, we provide an insight into recent advances toward the colloidal fabrication of hierarchical scaffolds. After identifying the main requirements for scaffolds in biomedical applications, conceptual building processes are introduced. Examples of tissue regeneration applications are provided for different scaffold types, highlighting their versatility and biocompatibility. We finally provide a prospect about the current state of the art and limitations of porous scaffolds, along with challenges that are to be addressed, so these materials consolidate in the fields of tissue engineering and drug delivery.

Synthesis and behavior of click cross-linked alginate hydrogels: Effect of cross-linker length and functionality.

Various bismaleimides and trismaleimides of varying molar masses, chemical architectures and functionalities were explored as cross-linkers for furan-modified alginate chains via Diels-Alder click reactions. An environmentally friendly approach is described for the preparation of hydrogels based on naturally occurring biomacromolecules, without catalysts. The behavior of the resulting polysaccharides-based hydrogels was analyzed in terms of swelling, rheological properties and drug-release efficiency, in connection with potential biomedical applications. The use of the different cross-linkers allows tuning the mechanical properties as well as the pulsatile swelling behavior of the hydrogels. When using trifunctional cross-linkers stiffer hydrogels were formed with high storage modulus whereas the chain length and the composition of the cross-linker clearly influence the swelling of the hydrogel network. In connection with drug delivery applications, release of vanillin as a traceable aromatic biobased model drug was also monitored as a function of hydrogel composition. To the best of our knowledge, for the first-time furan-modified alginates were reacted and studied with polyethylene glycol-based bis or trismaleimides with different molar masses and architectures, resulting in advanced hydrogels with different behavior.

Carbonaceous Filler Type and Content Dependence of the Physical-Chemical and Electromechanical Properties of Thermoplastic Elastomer Polymer Composites.

Graphene, carbon nanotubes (CNT), and carbon nanofibers (CNF) are the most studied nanocarbonaceous fillers for polymer-based composite fabrication due to their excellent overall properties. The combination of thermoplastic elastomers with excellent mechanical properties (e.g., styrene-b-(ethylene-co-butylene)-b-styrene (SEBS)) and conductive nanofillers such as those mentioned previously opens the way to the preparation of multifunctional materials for large-strain (up to 10% or even above) sensor applications. This work reports on the influence of different nanofillers (CNT, CNF, and graphene) on the properties of a SEBS matrix. It is shown that the overall properties of the composites depend on filler type and content, with special influence on the electrical properties. CNT/SEBS composites presented a percolation threshold near 1 wt.% filler content, whereas CNF and graphene-based composites showed a percolation threshold above 5 wt.%. Maximum strain remained similar for most filler types and contents, except for the largest filler contents (1 wt.% or more) in graphene (G)/SEBS composites, showing a reduction from 600% for SEBS to 150% for 5G/SEBS. Electromechanical properties of CNT/SEBS composite for strains up to 10% showed a gauge factor (GF) varying from 2 to 2.5 for different applied strains. The electrical conductivity of the G and CNF composites at up to 5 wt.% filler content was not suitable for the development of piezoresistive sensing materials. We performed thermal ageing at 120 °C for 1, 24, and 72 h for SEBS and its composites with 5 wt.% nanofiller content in order to evaluate the stability of the material properties for high-temperature applications. The mechanical, thermal, and chemical properties of SEBS and the composites were identical to those of pristine composites, but the electrical conductivity decreased by near one order of magnitude and the GF decreased to values between 0.5 and 1 in aged CNT/SEBS composites. Thus, the materials can still be used as large-deformation sensors, but the reduction of both electrical and electromechanical response has to be considered.

Ionic-Liquid-Based Printable Materials for Thermochromic and Thermoresistive Applications.

Smart materials exhibiting thermochromic and themoresistive properties based on the electroactive polymer poly(vinylidene fluoride) (PVDF) and the ionic liquid (IL) bis(1-butyl-3-methylimidazolium) tetrachloronickelate ([Bmim]2[NiCl4]) have been developed with different contents of [Bmim]2[NiCl4] (10, 20, and 40 wt %) within the polymer matrix. The morphology of the composites is studied, and the thermochromic and thermoresistive properties are evaluated. Independently of the IL content, the PVDF/[Bmim]2[NiCl4] composites present a porous morphology and thermochromic response, revealed by the color change of the composites from transparent to dark blue, attributed to the tetrahedral complex NiCl42- formed after a dehydration process. Further, the electrical conductivity increases with increasing IL content and decreases with increasing temperature. It is also shown that the incorporation of the IL into the PVDF matrix leads to an increase in the electroactive ß phase and a decrease in the degree of crystallinity and thermal stability with increasing [Bmim]2[NiCl4] content. The printability and applicability of the developed materials as sensors are also demonstrated.

Starch/graphene hydrogels via click chemistry with relevant electrical and antibacterial properties.

Starch-based hydrogels were performed by Diels-Alder cross-linking reactions between furan-modified starch and a water soluble bismaleimide, with improving conducting properties by using graphene layers as active nanofillers. The characterization results demonstrated that the Diels-Alder reaction and the corresponding conditions for the hydrogel formation were appropriate. The effect of increasing the furan/maleimide ratio on the architecture of the hydrogels and on the morphological, rheological and swelling properties were thoroughly evaluated. Effective network structure was obtained by increasing the cross-linker content leading to decreasing pore size and increasing storage modulus value of the final material. It was shown that the swelling behavior of hydrogels was mainly governed by the hydrophilic character of bismaleimide. Graphene nanosheets were added for the synthesis of nanocomposite hydrogel and it was characterized in terms of rheological properties, electrical conductivity and antimicrobial activity. The nanocomposite hydrogel presented enhanced mechanical performance, antimicrobial activity and increased conductivity values, up to a decade, indicating that conductive and active hydrogels could be satisfactory obtained, for a large range of potential applications such as biomed.

Synthesis and evaluation of functional alginate hydrogels based on click chemistry for drug delivery applications.

Environment-sensitive alginate-based hydrogels for drug delivery applications are receiving increasing attention. However, most work in this field involves traditional cross-linking strategies which led to hydrogels with poor long-term stability. Herein, a series of chemically cross-linked alginate hydrogels was synthesized via click chemistry using Diels-Alder reaction by reacting furan-modified alginate and bifunctional cross-linkers. Alginate was successfully functionalized with furfurylamine. Then, 3D architectures were synthesized with water-soluble bismaleimides. Different substitution degrees were achieved in order to study the effect of alginate modification and the cross-linking extent over the behaviour of the hydrogels. The ensuing hydrogels were analysed in terms of microstructure, swelling, structure modification and rheological behaviour. The materials response to external stimuli such as pH was also investigated, revealing a pulsatile behaviour in a large pH range (1-13) and a clear pH-dependent swelling. Finally, vanillin release studies were conducted to demonstrate the potential of these biobased materials for drug delivery applications.

Cellulose nanocrystals reinforced environmentally-friendly waterborne polyurethane nanocomposites.

Focusing on eco-friendly materials, cellulose nanocrystals (CNC) have gained attention as nanoreinforcement due to their exceptional properties conferred by the elevated length/diameter aspect ratio and high specific mechanical properties. Furthermore, their water dispersibility makes them suitable nanoreinforcements for their incorporation in waterborne polyurethanes (WBPU). The possibility of tailoring the properties by varying the composition and nature of the reagents, opens the opportunity for a wide range of applications. Therefore, in this work a WBPU was synthesized for the preparation of nanocomposite films with different CNC content and the properties of the films were analyzed. The effective incorporation of CNC resulted in an increase in moduli and stress at yield besides in an increased thermomechanical stability, reaching the percolation threshold at a 3wt% CNC as determined theoretically. Nevertheless, above the percolation threshold, the presence of agglomerates reduced slightly these values. The prepared nanocomposites showed increased hydrophilicity after CNC addition.

Biocompatible hydrogel nanocomposite with covalently embedded silver nanoparticles.

Bionanocomposite materials, combining the properties of biopolymers and nanostructured materials, are attracting interest of the wider scientific community due to their potential application in design of implants, drug delivery systems, and tissue design platforms. Herein, we report on the use of maleimide-coated silver nanoparticles (Ag NPs) as cocross-linkers for the preparation of a bionanocomposite gelatin based hydrogel. Diels-Alder cycloaddition of benzotriazole maleimide (BTM) functionalized Ag NPs and furan containing gelatin in combination with additional amide coupling resulted in stable and biocompatible hybrid nanocomposite. The storage moduli values for the hydrogel are nearly three times higher than that of control hydrogel without NPs indicating a stabilizing role of the covalently bound NPs. Finally, the swelling and drug release properties of the materials as well as the biocompatibility and toxicity tests indicate the biomedical potential of this type of material.