Activation of microglia and resulting neuroinflammation are central processes that significantly contribute to neurodegenerative disease progression. Treatments capable of attenuating neuroinflammation are therefore an urgent medical need. Vitis vinifera L., cultivated since ancient times for its fruits, is known for its antioxidant and anti-inflammatory activities. However, polyphenols, the main bioactive molecules in V. vinifera extracts, exhibit considerable variability due to numerous hard-to-control factors, which complicates the production of standardized extracts with consistent biological activity. To address this issue, plant cell culture biotechnology was used to produce a highly standardized V. vinifera phytocomplex (VP), and its anti-neuroinflammatory profile was investigated in LPS-stimulated microglial cells, an in vitro model of neuroinflammation. VP reduced the LPS-induced pro-inflammatory phenotype, improved cell viability and cell number, attenuated NF-κB activation and ERK1/2 phosphorylation, and increased SIRT1 levels. To overcome VP’s poor water solubility, water-soluble cellulose nanocrystal (CNC)-based formulations were developed and tested. VP-CNC formulations markedly reduced the BV2 pro-inflammatory phenotype and increased cell viability under both basal and LPS-stimulated conditions. The nanoformulations also decreased pERK1/2 levels and increased SIRT1 expression, exhibiting biological activities comparable to VP alone. V. vinifera phytocomplex derived from plant cell cultures represents an innovative and standardized product with promising anti-neuroinflammatory properties.

The therapeutic monoclonal antibody bevacizumab is typically purified using protein A affinity chromatography, a highly effective but costly method. Affinity-based precipitation for antibody purification is a lower-cost approach. In this work, a precipitation protocol was developed for bevacizumab purification using a branched peptide (Ac-PHQGQHIG-Ahx3)2-K-Ahx3-PHQGQHIG-NH2, which contains the epitope PHQGQHIG that is responsible for interacting with bevacizumab. The peptide was synthesised by a microwave-assisted solid-phase peptide method, employing LiCl as an additive to prevent aggregation and ensure high purity and yield. Three molecules of 6-aminohexanoic acid were introduced between each epitope branch as spacer arms to promote the formation of cyclic complexes. Bevacizumab purification from cell-free culture broth was achieved through a fractional precipitation process. First, a negative precipitation step using (NH4)2SO4 1.18 M was performed to remove contaminants. Afterwards, 5 moles of peptide per mol of bevacizumab was added to the supernatant, together with additional (NH4)2SO4, to reach a final concentration of 1.20 M. Under these conditions, bevacizumab was recovered in the precipitate with 98% purity and a yield of 73%. In addition to being recyclable, the peptide’s relatively low production cost could enable the development of a single-use purification process, which would be particularly advantageous for biopharmaceutical manufacturing.

Crystalline nanocellulose (CNC) is a readily available nanostructured form of cellulose, characterized by a high biocompatibility and easily functionalized to produce materials with new properties and applications. In this study, CNC was modified to act as a drug delivery system for the auranofin pharmacophore-specifically, the gold(I)-containing [Et3PAu]+ moiety. For this purpose, commercial CNC was first derivatized with propargyl groups and subsequently with lipoic acid. These two different functionalizations were then used to insert a simple glucose moiety (as a selector for tumor cells) using a CuAAC reaction with the propargyl end. Meanwhile the lipoic acid S-S bond was reduced with dithiothreitol to react with two equivalents of the gold complex Et3PAuCl, an analogue of auranofin. The modification of CNC afforded a nanohybrid characterized by a loading of 113.2 mg of Au and 0,41 mmol of selector per g of CNC and by a mean diameter of 290 nm and a Zeta Potential with a value of -51 +/- 5 mV. This work opens the possibility of future studies on the use of Auranofin analogues supported on drug delivery systems for biomedical applications. future studies on the use of Auranofin analogues supported on drug delivery systems for biomedical applications.

The high prevalence of onychomycosis, a common pathology of nails due to fungal infections, has drawn attention to the development of new therapeutic approaches in which systemic secondary effects and the rise of drug resistance are avoided. Photodynamic therapy (PDT) is an approach that can be useful for treatment of onychomycosis, relying on the activation of a photosensitizer with light of a specific wavelength producing cytotoxic reactive oxygen species in situ. The present work describes the use of bacterial nanocellulose (BNC) patches delivering two different photosensitizers: Rose Bengal (RB) and a BODIPY derivative (B-I2), whose differing hydrophilicity necessitated distinct formulation strategies. The experiments performed using vertical Franz cells allowed the optimization of the systems, finally tested in in vitro experiments with Candida albicans cultures. The results showed the efficacy of the BNC patches in the delivery of RB, showing that the presence of a keratolytic agent as urea is vital for the RB permeation through the nail models, with the permeation of RB being the first step for an efficient light-driven disinfection of C. albicans using green-laser light. Similar results were obtained with B-I2 although the high lipophilicity of the dye limited its cytotoxic evaluation in vitro.

Introduction: Bevacizumab, applied in cancer treatment, binds vascular endothelial growth factor (VEGF) inhibiting blood vessels growth, avoiding tumours expansion [...]

The unique physicochemical properties of gold nanoparticles (GNPs) have made them versatile tools for biomedical applications, such as imaging, therapy, and drug delivery. The surface modification of GNPs with polymers or biomolecules can enhance their colloidal stability and facilitate internalization into cells. However, the efficient and biocompatible delivery to the central nervous system remains a major challenge, as many existing nanocarriers show poor capacity to cross the blood-brain barrier. We developed a method to coat GNPs with linear polyethyleneimine (GNP@PEI) through a chemical reduction bottom-up approach, in which linear PEI hydrochloride acts simultaneously as a reducing and stabilizing agent of colloidal dispersion. This strategy yielded monodisperse spherical GNP@PEI nanoparticles with an average diameter of 50 nm. The physicochemical profile, biocompatibility, and capacity for neural uptake of this potentially brain-targeted nanoplatform were then evaluated. GNP@PEI nanoparticles exhibited high biocompatibility in several primary neural cultures and cell lines, with cellular uptake showing clear cell-type-dependent differences. In vivo studies carried out in a murine model demonstrated that after the intranasal or intraperitoneal administrations of GNP@PEI nanoparticles, detectable levels of gold were found in several organs, including the brain. Collectively, these findings highlight the potential of GNP@PEI as a promising nanoplatform for brain-targeted delivery and for advancing the development of therapeutic strategies for neurological disorders.

We report here the use of Tris-BODIPY-OH as a scaffold for the multivalent display of sugar heads. A chloroacetyl thioether ligation reaction easily yields mannosylated BODIPYs, named Man9-BODIPY and (Man-TEG)9-BODIPY, which display nine mannose residues. Regardless of the linker length, both glycoBODIPYs provide an arrangement of mannose heads that allows for proper recognition by the carbohydrate binding domain of concanavalin A (ConA). Moreover, the interactions of Man9-BODIPY with relevant human lectins, i.e. dendritic cell-specific intercellular adhesion molecule-3-grabbing non-integrin (DC-SIGN) and langerin, were further investigated. The approach proposed is versatile and paves the way for the development of multivalent and fluorescent glyco-BODIPY probes useful to interrogate carbohydrate-lectin interactions in different biological contexts.

In mammals, every cell is covered by a sugar coat called the "glycocalyx", a meshwork created by sugar modifications of cell surface proteins and lipids. Essentially all cell membrane proteins and lipids contain oligosaccharide clusters known as "glycan motifs" that confer distinct functional properties on these respective glycoproteins or glycolipids. These motifs are generated by glycosyltransferases that assemble the component monosaccharides in a stereospecific and regiospecific fashion. Glycocalyx motifs bearing l-fucose in α(1→3) linkage to N-acetyl-glucosamine are found on a highly restricted subset of membrane glycoproteins and glycolipids, and changes in α(1→3)-fucosylation levels impact a wide range of physiologic and pathologic processes. Within this biological framework, we herein review the pivotal role of α(1→3)-fucosylation in cell biology and then comprehensively review the evolving chemical strategies to custom-modify α(1→3)-fucosylation of the glycocalyx to achieve highly specific control of human cell surface fucosylated glycan motifs. These efforts serve as a prime example of how the fine control of cell surface fucosylation can enable the generation of glycan-based precision therapeutics, driving forward the field of "translational glycobiology".

The conjugation of fluorescent probes to tumor-targeting molecules represents a promising strategy for the development of precision cancer bioimaging and treatment. Among the different tumor-targeting strategies, the use of d-glucose residues, which exploit the high energy demand of cancer cells, can enable recognition by a broad spectrum of tumors, thus overcoming limitations related to cancer heterogeneity. In this study, we combined the distinctive optical properties of BODIPY-based probes with the known tumor-targeting abilities of d-glucose. We report on the characterization of a glucosylated BODIPY, named Glc-BODIPY, and its ability to target different cancer cell types in both in vitro and in vivo models.

The convergence of glycochemistry and glycobiology is enabling the creation of new therapeutic approaches with unprecedented capacity to alter cell and organismic biology using strategies that can uniquely and specifically custom-modify the expression of key cell surface glycan motifs. We define this evolving field of chemical biology as 'glycan-motif editing', and one of the principal targets of this glycoengineering effort is the sialofucosylated terminal lactosaminyl glycan known as sLeX (CD15s). This tetrasaccharide structure plays pivotal roles in both steady-state and malignant hematopoiesis, in regulation of the immune response, and in cancer metastasis. Within this biological framework, we discuss the immense potential of glycan-motif editing in enabling precision therapeutics that will profoundly improve outcomes for patients suffering from a wide variety of disabling and life-threatening conditions, particularly cancer.

We report here on a straightforward methodology to synthesize a new water-soluble fluorescent probe Tris-BODIPY-OH 1 that contains three pH-independent hydrophilic arms. This probe has been prepared by exploiting a synthetic strategy that includes as a key step the combination of a Cu-(I)-catalyzed azide-alkyne cycloaddition (CuAAC) and a Sonogashira cross-coupling in a sequential one-pot approach. Tris-BODIPY-OH 1 provides a significant advancement in the field by expanding the BODIPY toolbox with a biocompatible water-soluble probe, which can be used to specifically label and assess the function of the endoplasmic reticulum.

A hybrid cellulose-based programmable nanoplatform for applications in precision radiation oncology is described. Here, sugar heads work as tumor targeting moieties and steer the precise delivery of radiosensitizers, i.e. gold nanoparticles (AuNPs) into triple negative breast cancer (TNBC) cells. This "Trojan horse" approach promotes a specific and massive accumulation of radiosensitizers in TNBC cells, thus avoiding the fast turnover of small-sized AuNPs and the need for high doses of AuNPs for treatment. Application of X-rays resulted in a significant increase of the therapeutic effect while delivering the same dose, showing the possibility to use roughly half dose of X-rays to obtain the same radiotoxicity effect. These data suggest that this hybrid nanoplatform acts as a promising tool for applications in enhancing cancer radiotherapy effects with lower doses of X-rays. A hybrid cellulose-based programmable nanoplatform for applications in precision radiation oncology is described.

Liquid-phase exfoliation (LPE) in aqueous solutions provides a simple, scalable, and green approach to produce 2D materials. By combining atomistic simulations with exfoliation experiments, the interaction between a surfactant and a 2D layer at the molecular scale can be better understood. In this work, two different dyes, corresponding to rhodamine B base (Rbb) and to a phenylboronic acid BODIPY (PBA-BODIPY) derivative, are employed as dispersants to exfoliate graphene and hexagonal boron nitride (hBN) through sonication-assisted LPE. The exfoliated 2D sheets, mostly as few-layers, exhibit good quality and high loading of dyes. Using molecular dynamics (MD) simulations, the binding free energies are calculated and the arrangement of both dyes on the layers are predicted. It has been found that the dyes show a higher affinity toward hBN than graphene, which is consistent with the higher yields of exfoliated hBN. Furthermore, it is demonstrated that the adsorption behavior of Rbb molecules on graphene and hBN is quite different compared to PBA-BODIPY.

In this work, we have discovered that the Gal-α-(1→3)-Gal-β-(1→3)-GlcNAc trisaccharide, a fragment of the B antigen Type-1, is a new ligand of two C-type lectin receptors (CLRs) i. e. DCAR and Mincle which are key players in different types of autoimmune diseases. Accordingly, we report here on a straightforward methodology to access pure Gal-α-(1→3)-Gal-β-(1→3)-GlcNAc trisaccharide. A spacer with a terminal primary amine group was included at the reducing end of the GlcNAc residue thus ensuring the further functionalization of the trisaccharide Gal-α-(1→3)-Gal-β-(1→3)-GlcNAc.

The reversibility of the covalent interaction between boronic acids and 1,2- or 1,3-diols has put the spotlight on this reaction for its potential in the development of sensors and for the fishing of bioactive glycoconjugates. In this work, we describe the investigation of this reaction for the reversible functionalization of the surface of CdSe/ZnS Quantum Rods (QRs). With this in mind, we have designed a turn-off Förster resonance energy transfer (FRET) system that ensures monitoring the extent of the reaction between the phenyl boronic residue at the meso position of a BODIPY probe and the solvent-exposed 1,2-diols on QRs’ surface. The reversibility of the corresponding boronate ester under oxidant conditions has also been assessed, thus envisioning the potential sensing ability of this system.

Biomedical applications of molecules that are able to modulate β-adrenergic signaling have become increasingly attractive over the last decade, revealing that β-adrenergic receptors (β-ARs) are key targets for a plethora of therapeutic interventions, including cancer. Despite successes in β-AR drug discovery, identification of β-AR ligands that are useful as selective chemical tools in pharmacological studies of the three β-AR subtypes, or lead compounds for drug development is still a highly challenging task. This is mainly due to the intrinsic plasticity of β-ARs as G protein-coupled receptors in conjunction with the requirement for functional receptor subtype selectivity, tissue specificity and minimal off-target effects. With the aim to provide insight into structure-activity relationships for the three β-AR subtypes, we have synthesized and obtained the pharmacological profile of a series of structurally diverse compounds (named MC) that were designed based on the aryloxy-propanolamine scaffold of SR59230A. Comparative analysis of their predicted binding mode within the active and inactive states of the receptors in combination with their pharmacological profile revealed key structural elements that control their activity as agonists or antagonists, in addition to clues about substituents that mediate selectivity for one receptor subtype over the others. We anticipate that these results will facilitate selective β-AR drug development efforts.

Cellulose nanocrystal and gold nanoparticles are assembled, in a unique way, to yield a novel modular glyconanomaterial whose surface is then easily engineered with one or two different headgroups, by exploiting a robust click chemistry route. We demonstrate the potential of this approach by conjugating monosaccharide headgroups to the glyconanomaterial and show that the sugars retain their binding capability to C-type lectin receptors, as also directly visualized by cryo-TEM.

Covalent functionalization of graphene oxide (GO) with boron dipyrromethenes (BODIPYs) was achieved through a facile synthesis, affording two different GO-BODIPY conjugates where the main difference lies in the nature of the spacer and the type of bonds between the two components. The use of a long but flexible spacer afforded strong electronic GO-BODIPY interactions in the ground state. This drastically altered the light absorption of the BODIPY structure and impeded its selective excitation. In contrast, the utilisation of a short, but rigid spacer based on boronic esters resulted in a perpendicular geometry of the phenyl boronic acid BODIPY (PBA-BODIPY) with respect to the GO plane, which enables only minor electronic GO-BODIPY interactions in the ground state. In this case, selective excitation of PBA-BODIPY was easily achieved, allowing to investigate the excited state interactions. A quantitative ultrafast energy transfer from PBA-BODIPY to GO was observed. Furthermore, due to the reversible dynamic nature of the covalent GO-PBA-BODIPY linkage, some PBA-BODIPY is free in solution and, hence, not quenched from GO. This resulted in a weak, but detectable fluorescence from the PBA-BODIPY that will allow to exploit GO-PBA-BODIPY for slow release and imaging purposes.

The increasing resistance of bacteria to conventional antibiotics represents a severe global emergency for human health. The broad-spectrum antibacterial activity of silver has been known for a long time, and silver at the nanoscale shows enhanced antibacterial activity. This has prompted research into the development of silver-based nanomaterials for applications in clinical settings. In this work, the synthesis of three different silver nanoparticles (AgNPs) hybrids using both organic and inorganic supports with intrinsic antibacterial properties is described. The tuning of the AgNPs' shape and size according to the type of bioactive support was also investigated. Specifically, the commercially available sulfated cellulose nanocrystal (CNC), the salicylic acid functionalized reduced graphene oxide (rGO-SA), and the commercially available titanium dioxide (TiO2) were chosen as organic (CNC, rGO-SA) and inorganic (TiO2) supports. Then, the antimicrobial activity of the AgNP composites was assessed on clinically relevant multi-drug-resistant bacteria and the fungus Candida albicans. The results show how the formation of Ag nanoparticles on the selected supports provides the resulting composite materials with an effective antibacterial activity.

Carbohydrates are ubiquitous molecules expressed on the surface of nearly all living cells, and their interaction with carbohydrate-binding proteins is critical to many immunobiological processes. Carbohydrates are utilized as antigens in many licensed vaccines against bacterial pathogens. More recently, they have also been considered as adjuvants. Interestingly, unlike other types of vaccines, adjuvants have improved immune response to carbohydrate-based vaccine in humans only in a few cases. Furthermore, despite the discovery of many new adjuvants in the last years, aluminum salts, when needed, remain the only authorized adjuvant for carbohydrate-based vaccines. In this review, we highlight historical and recent advances on the use of glycans either as vaccine antigens or adjuvants, and we review the use of currently available adjuvants to improve the efficacy of carbohydrate-based vaccines. A better understanding of the mechanism of carbohydrate interaction with innate and adaptive immune cells will benefit the design of a new generation of glycan-based vaccines and of immunomodulators to fight both longstanding and emerging diseases.