Bioglass/ceria nanoparticle hybrids for the treatment of seroma: a comparative long-term study in rats.

Background: Seroma formation is a common postoperative complication. Fibrin-based glues are typically employed in an attempt to seal the cavity. Recently, the first nanoparticle (NP)-based treatment approaches have emerged. Nanoparticle dispersions can be used as tissue glues, capitalizing on a phenomenon known as 'nanobridging'. In this process, macromolecules such as proteins physically adsorb onto the NP surface, leading to macroscopic adhesion. Although significant early seroma reduction has been shown, little is known about long-term efficacy of NPs. The aim of this study was to assess the long-term effects of NPs in reducing seroma formation, and to understand their underlying mechanism. Methods: Seroma was surgically induced bilaterally in 20 Lewis rats. On postoperative day (POD) 7, seromas were aspirated on both sides. In 10 rats, one side was treated with NPs, while the contralateral side received only NP carrier solution. In the other 10 rats, one side was treated with fibrin glue, while the other was left untreated. Seroma fluid, blood and tissue samples were obtained at defined time points. Biochemical, histopathological and immunohistochemical assessments were made. Results: NP-treated sides showed no macroscopically visible seroma formation after application on POD 7, in stark contrast to the fibrin-treated sides, where 60% of the rats had seromas on POD 14, and 50% on POD 21. At the endpoint (POD 42), sides treated with nanoparticles (NPs) exhibited significant macroscopic differences compared to other groups, including the absence of a cavity, and increased fibrous adhesions. Histologically, there were more macrophage groupings and collagen type 1 (COL1) deposits in the superficial capsule on NP-treated sides. Conclusion: NPs not only significantly reduced early manifestations of seroma and demonstrated an anti-inflammatory response, but they also led to increased adhesion formation over the long term, suggesting a decreased risk of seroma recurrence. These findings highlight both the adhesive properties of NPs and their potential for clinical therapy.

Protein Aggregation on Metal Oxides Governs Catalytic Activity and Cellular Uptake.

Engineering of catalytically active inorganic nanomaterials holds promising prospects for biomedicine. Catalytically active metal oxides show applications in enhancing wound healing but have also been employed to induce cell death in photodynamic or radiation therapy. Upon introduction into a biological system, nanomaterials are exposed to complex fluids, causing interaction and adsorption of ions and proteins. While protein corona formation on nanomaterials is acknowledged, its modulation of nanomaterial catalytic efficacy is less understood. In this study, proteomic analyses and nano-analytic methodologies quantify and characterize adsorbed proteins, correlating this protein layer with metal oxide catalytic activity in vitro and in vivo. The protein corona comprises up to 280 different proteins, constituting up to 38% by weight. Enhanced complement factors and other opsonins on nanocatalyst surfaces lead to their uptake into macrophages when applied topically, localizing >99% of the nanomaterials in tissue-resident macrophages. Initially, the formation of the protein corona significantly reduces the nanocatalysts' activity, but this activity can be partially recovered in endosomal conditions due to the proteolytic degradation of the corona. Overall, the research reveals the complex relationship between physisorbed proteins and the catalytic characteristics of specific metal oxide nanoparticles, providing design parameters for optimizing nanocatalysts in complex biological environments.

One-Step Synthesis of Versatile Antimicrobial Nano-Architected Implant Coatings for Hard and Soft Tissue Healing.

Dental implant failure remains a prevalent problem around the globe. The integration of implants at the interface of soft and hard tissues is complex and susceptible to instability and infections. Modifications to the surface of titanium implants have been developed to improve the performance, yet insufficient integration and biofilm formation remain major problems. Introducing nanostructures on the surface to augment the implant-tissue contact holds promise for facilitated implant integration; however, current coating processes are limited in their versatility or costs. We present a highly modular single-step approach to produce multicomponent porous bioactive nanostructured coatings on implants. Inorganic nanoparticle building blocks with complex compositions and architectures are synthesized in situ and deposited on the implants in a single step using scalable liquid-feed flame spray pyrolysis. We present hybrid coatings based on ceria and bioglass, which render the implant surfaces superhydrophilic, promote cell adhesion, and exhibit antimicrobial properties. By modifications to the bioglass/ceria nanohybrid composition and architecture that prevent biomineralization, the coating can instead be tailored toward soft tissue healing. The one-step synthesis of nano-architected tissue-specific coatings has great potential in dental implantology and beyond.

Inorganic nanohybrids combat antibiotic-resistant bacteria hiding within human macrophages.

Bacterial infections are one of the main health concerns humanity faces today and bacterial resistances and protection mechanisms are set to aggravate the issue in the coming years. An increasing number of bacterial strains evades antibiotic treatment by hiding inside cells. Conventional antimicrobial agents are unable to penetrate or be retained in the infected mammalian cells. Recent approaches to overcome these limitations have focused on load-carrier systems, requiring a triggered discharge leading to complex release kinetics. The unison of potent antimicrobial activity with high mammalian cell compatibility is a prerequisite for intracellular activity, which is not well-met by otherwise well-established inorganic systems, such as silver-based nanoparticles. In this work, load and carrier are combined into one functional inorganic nanoparticle system, which unites antimicrobial activity with mammalian cell compatibility. These multicomponent nanohybrids based on cerium oxide are produced in one step, yet unite complex materials. The nanoparticles form suprastructures of similar size and surface charge as bacteria, therefore facilitating the uptake into the same subcellular compartments, where they unleash their antibacterial effect. Such intrinsically antibacterial nanohybrids significantly reduce bacterial survival inside macrophages without harming the latter. Furthermore, blocking of nanoparticle endocytosis and subcellular electron microscopy elucidate the mechanism of action. Taken together, this work presents the first demonstration of antibacterial activity of ceria-based nanoparticles inside of mammalian cells and offers a route to straightforward and robust intracellular antibacterial agents that do not depend on payload delivery or biological constituents.

Uniting Drug and Delivery: Metal Oxide Hybrid Nanotherapeutics for Skin Wound Care.

Wound care and soft tissue repair have been a major human concern for millennia. Despite considerable advancements in standards of living and medical abilities, difficult-to-heal wounds remain a major burden for patients, clinicians and the healthcare system alike. Due to an aging population, the rise in chronic diseases such as vascular disease and diabetes, and the increased incidence of antibiotic resistance, the problem is set to worsen. The global wound care market is constantly evolving and expanding, and has yielded a plethora of potential solutions to treat poorly healing wounds. In ancient times, before such a market existed, metals and their ions were frequently used in wound care. In combination with plant extracts, they were used to accelerate the healing of burns, cuts and combat wounds. With the rise of organic chemistry and small molecule drugs and ointments, researchers lost their interest in inorganic materials. Only recently, the advent of nano-engineering has given us a toolbox to develop inorganic materials on a length-scale that is relevant to wound healing processes. The robustness of synthesis, as well as the stability and versatility of inorganic nanotherapeutics gives them potential advantages over small molecule drugs. Both bottom-up and top-down approaches have yielded functional inorganic nanomaterials, some of which unite the wound healing properties of two or more materials. Furthermore, these nanomaterials do not only serve as the active agent, but also as the delivery vehicle, and sometimes as a scaffold. This review article provides an overview of inorganic hybrid nanotherapeutics with promising properties for the wound care field. These therapeutics include combinations of different metals, metal oxides and metal ions. Their production, mechanism of action and applicability will be discussed in comparison to conventional wound healing products.

Multiscale Analysis of Metal Oxide Nanoparticles in Tissue: Insights into Biodistribution and Biotransformation.

Metal oxide nanoparticles have emerged as exceptionally potent biomedical sensors and actuators due to their unique physicochemical features. Despite fascinating achievements, the current limited understanding of the molecular interplay between nanoparticles and the surrounding tissue remains a major obstacle in the rationalized development of nanomedicines, which is reflected in their poor clinical approval rate. This work reports on the nanoscopic characterization of inorganic nanoparticles in tissue by the example of complex metal oxide nanoparticle hybrids consisting of crystalline cerium oxide and the biodegradable ceramic bioglass. A validated analytical method based on semiquantitative X-ray fluorescence and inductively coupled plasma spectrometry is used to assess nanoparticle biodistribution following intravenous and topical application. Then, a correlative multiscale analytical cascade based on a combination of microscopy and spectroscopy techniques shows that the topically applied hybrid nanoparticles remain at the initial site and are preferentially taken up into macrophages, form apatite on their surface, and lead to increased accumulation of lipids in their surroundings. Taken together, this work displays how modern analytical techniques can be harnessed to gain unprecedented insights into the biodistribution and biotransformation of complex inorganic nanoparticles. Such nanoscopic characterization is imperative for the rationalized engineering of safe and efficacious nanoparticle-based systems.

Tailoring the Colloidal Stability, Magnetic Separability, and Cytocompatibility of High-Capacity Magnetic Anion Exchangers.

Extracorporeal blood purification has been applied to artificially support kidney or liver function. However, convection and diffusion based blood purification systems have limited removal rates for high molecular weight and hydrophobic molecules. This limitation is due to the finite volume of infusion and limited membrane permeability, respectively. Adsorption provides an attractive alternative for the removal of higher molecular weight compounds. The use of adsorption resins containing ion exchanging groups to capture specific molecules has become well-established. Instead of stationary adsorption resins, however, ion exchanging polymers may be immobilized on magnetic particles and serve as freely diffusing, mobile, high capacity solid phase of ion exchange chromatography. While small beads with high surface area are attractive in terms of mass transfer and binding, unifying high capturing capacity with rapid and quantitative bead recovery remains an issue. Therefore, most of the current magnetic ion exchangers are based on micron-sized beads or require long times to separate. In addition to unfavorable magnetic recovery rates, the usually poor cytocompatibility limits their applicability in biomedicine. Here, we report on the synthesis and performance of polycationic polymer coated magnetic nanoflowers (MNF) for highly efficacious anion capturing. We demonstrate accurate control over the polymer content and composition on the beads and show its direct influence on colloidal stability, capturing capacity and magnetic separability. We present the removal of clinically relevant targets by capturing bilirubin with capacities 2-fold higher than previous work as well as quantitative heparin removal. Additionally, we illustrate how copolymerization of poly(2-dimethylaminoethyl methacrylate) (PDMAEMA) with poly(ethylene glycol) methyl ether methacrylate (PEGMEMA) leads to improved cytocompatibility of the polymer-coated MNF capturing agents while retaining high capturing capacities. Taken together, we present a nanoparticle/polymer material, which upon future in vivo validation, unifies high binding capacities and magnetic separability for rapid toxin capturing and hence fulfills key requirements of clinical utility.

Mussel-Inspired Injectable Hydrogel Adhesive Formed under Mild Conditions Features Near-Native Tissue Properties.

Injectable hydrogel adhesives, especially those that can strongly adhere to tissues and feature near-native tissue mechanical properties, are desirable biomaterials for tissue repair. Compared to nonadhesive injectable hydrogels for minimally invasive delivery of therapeutic agents, they can better retain the delivered agents at targeted tissue locations and provide additional local physical barriers. However, regardless of recent advances, an ideal injectable hydrogel adhesive with both proper adhesion and mechanical matching between hydrogels and tissues is yet to be demonstrated with cytocompatible and efficient in situ curing methods. Inspired by marine mussels, where different mussel foot proteins (Mfps) function cooperatively to achieve excellent wet adhesion, we herein report a dual-mode-mimicking strategy by modifying gelatin (Gel) biopolymers with a single-type thiourea-catechol (TU-Cat) functionality to mimic two types of Mfps and their mode of action. This strategy features a minor, yet impactful modification of biopolymers, which gives access to collective properties of an ideal injectable hydrogel adhesive. Specifically, with TU-Cat functionalization of only ∼0.4-1.2 mol % of total amino acid residues, the Mfp-mimetic gelatin biopolymer (Gel-TU-Cat) can be injected and cured rapidly under mild and cytocompatible conditions, giving rise to tissue adhesive hydrogels with excellent matrix ductility, proper wet adhesion, and native tissue-like stress relaxation behaviors. Such a set of properties originating from our novel dual-mode-mimicking strategy makes the injectable hydrogel adhesive a promising platform for cell delivery and tissue repair.

Ultrabright and Stable Luminescent Labels for Correlative Cathodoluminescence Electron Microscopy Bioimaging.

The mechanistic understanding of structure-function relationships in biological systems heavily relies on imaging. While fluorescence microscopy allows the study of specific proteins following their labeling with fluorophores, electron microscopy enables holistic ultrastructural analysis based on differences in electron density. To identify specific proteins in electron microscopy, immunogold labeling has become the method of choice. However, the distinction of immunogold-based protein labels from naturally occurring electron dense granules and the identification of several different proteins in the same sample remain challenging. Correlative cathodoluminescence electron microscopy (CCLEM) bioimaging has recently been suggested to provide an attractive alternative based on labels emitting characteristic light. While luminescence excitation by an electron beam enables subdiffraction imaging, structural damage to the sample by high-energy electrons has been identified as a potential obstacle. Here, we investigate the feasibility of various commonly used luminescent labels for CCLEM bioimaging. We demonstrate that organic fluorophores and semiconductor quantum dots suffer from a considerable loss of emission intensity, even when using moderate beam voltages (2 kV) and currents (0.4 nA). Rare-earth element-doped nanocrystals, in particular Y2O3:Tb3+ and YVO4:Bi3+,Eu3+ nanoparticles with green and orange-red emission, respectively, feature remarkably high brightness and stability in the CCLEM bioimaging setting. We further illustrate how these nanocrystals can be readily differentiated from morphologically similar naturally occurring dense granules based on optical emission, making them attractive nanoparticle core materials for molecular labeling and (multi)color CCLEM.

Lanthanide-Doped Hafnia Nanoparticles for Multimodal Theranostics: Tailoring the Physicochemical Properties and Interactions with Biological Entities.

High-Z metal oxide nanoparticles hold promise as imaging probes and radio-enhancers. Hafnium dioxide nanoparticles have recently entered clinical evaluation. Despite promising early clinical findings, the potential of HfO2 as a matrix for multimodal theranostics is yet to be developed. Here, we investigate the physicochemical properties and the potential of HfO2-based nanoparticles for multimodal theranostic imaging. Undoped and lanthanide (Eu3+, Tb3+, and Gd3+)-doped HfO2 nanoparticles were synthesized and functionalized with various moieties including poly(vinylpyrrolidone) (PVP), (3-aminopropyl)triethoxysilane (APTES), and folic acid (FA). We show that different synthesis routes, including direct precipitation, microwave-assisted synthesis, and sol-gel chemistry, allow preparation of hafnium dioxide particles with distinct physicochemical properties. Sol-gel based synthesis allows preparation of uniform nanoparticles with dopant incorporation efficiencies superior to the other two methods. Both luminescence and contrast properties can be tweaked by lanthanide doping. We show that MRI contrast can be unified with radio-enhancement by incorporating lanthanide dopants in the HfO2 matrix. Importantly, ion leaching from the HfO2 host matrix in lysosomal-like conditions was minimal. For Gd:HfO2 nanoparticles, leaching was reduced >10× compared to Gd2O3, and no relevant cytotoxic effects have been observed in monocyte-derived macrophages for nanoparticle concentrations up to 250 µg/mL. Chemical surface modification allows further tailoring of the cyto- and hemocompatibility and enables functionalization with molecular targeting entities, which lead to enhanced cellular uptake. Taken together, the present study illustrates the manifold properties of HfO2-based nanomaterials with prospective clinical utility beyond radio-enhancement.

Engineering the Bioactivity of Flame-Made Ceria and Ceria/Bioglass Hybrid Nanoparticles.

Despite its use as a highly efficient and reusable catalyst in research and industrial settings, cerium oxide nanoparticles or nanoceria have yet to gain a foothold in the biomedical field. A variety of beneficial effects of nanoceria have been demonstrated, including its use as an inorganic nanoenzyme to mimic antioxidant enzymes, to protect mammalian cells, and to suppress microbial growth. While these properties are of high interest for wound-management applications, the literature offers contradicting reports on toxicity and enzymatic activity of nanoceria. These discrepancies can be attributed to differences between synthesis methods and insufficient physicochemical characterization, leading to incomparable studies. The activity of nanoceria is mostly governed by its Ce3+/Ce4+ ratio which needs to be controlled to compare different nanoceria systems. In this work, we demonstrate that liquid-feed flame spray pyrolysis offers excellent control over the oxidation state in a one-step synthesis of nanoceria. This control allows a comprehensive comparison of different types of ceria nanoparticles. We connect physicochemical characteristics to biomedically relevant properties such as superoxide dismutase and catalase mimicry, human monocyte and macrophage protection, and antimicrobial activity. Furthermore, we demonstrate how the synthesis method also allows tailoring the properties of ceria/bioglass hybrid nanoparticles, thus creating nanoparticles with manifold biomedical prospects.

Developing a tissue glue by engineering the adhesive and hemostatic properties of metal oxide nanoparticles.

Despite decades of research, wound complications remain a major cause of postoperative mortality, especially in the face of multiple comorbidities. Addressing the issue of anastomotic leakages and impaired wound healing from a new angle is of great interest with the prospect of having direct impact on patient outcome. Recently, aqueous suspensions of silica and iron oxide nanoparticles have been employed to connect biological tissue by serving as an adhesive layer eventually leading to macroscopic gluing of tissue. In this work, we explore the prospects of this effect by introducing bioactive tissue adhesives composed of nanoparticles produced via scalable and sterile flame spray pyrolysis. We investigate six different metal oxides on cytocompatibility, hemostatic activity and adhesive properties in a small intestine lap joint model. While bioglass nanoparticles show exceptionally strong procoagulant and adhesive properties, the cell membrane integrity is impaired at high particle concentrations. Interestingly, when bioglass is combined with ceria, a material that has well-documented cytoprotective effects, the resulting hybrid particles exhibit the same beneficiary effects as bioglass while featuring superior cytocompatibility. Taken together, we demonstrate highly modular synthesis of nanoparticles expressing adhesive properties in conjunction with tailored bioactivity. Such bioactive nanoparticles as adhesion nuclei in wound healing have a wide range of potential applications in surgical wound care and regenerative medicine.