Biological Activity and Chemical Composition of Propolis Extracts with Potential Use in Vulvovaginal Candidiasis Management.

Environmental sustainability is an increasing challenge in the pharmaceutical field, leading to the search for eco-friendly active ingredients. Among natural ingredients, propolis arises as an excellent alternative, being a complex substance with pharmacological properties. This work aims to explore the potential of propolis as a new pharmaceutical ingredient for the replacement of conventional vulvovaginal antifungals. Propolis extracts were obtained by Ultrasound-Assisted Extraction using different solvents (water, water/ethanol (50:50, v/v), and ethanol). Afterwards, the extracts were characterized regarding total phenolic content (TPC), antioxidant/antiradical activities, radical scavenging capacity, antifungal activity against strains of Candida species, and viability effect on two female genital cell lines. The aqueous extract achieved the best TPC result as well as the highest antioxidant/antiradical activities and ability to capture reactive oxygen species. A total of 38 phenolic compounds were identified and quantified by HPLC, among which ferulic acid, phloridzin and myricetin predominated. Regarding the anti-Candida spp. activity, the aqueous and the hydroalcoholic extracts achieved the best outcomes (with MIC values ranging between 128 and 512 µg/mL). The cell viability assays confirmed that the aqueous extract presented mild selectivity, while the hydroalcoholic and alcoholic extracts showed higher toxicities. These results attest that propolis has a deep potential for vulvovaginal candidiasis management, supporting its economic valorization.

Trends in 3D models of inflammatory bowel disease.

Inflammatory bowel disease (IBD) encompasses a set of chronic inflammatory conditions, namely Crohn's disease and ulcerative colitis. Despite all advances in the management of IBD, a definitive cure is not available, largely due to a lack of a holistic understanding of its etiology and pathophysiology. Several in vitro, in vivo, and ex vivo models have been developed over the past few decades in order to abbreviate remaining gaps. The establishment of reliable and predictable in vitro intestinal inflammation models may indeed provide valuable tools to expedite and validate the development of therapies for IBD. Three-dimensional (3D) models provide a more accurate representation of the different layers of the intestine, contributing to a stronger impact on drug screening and research on intestinal inflammation, and bridging the gap between in vitro and in vivo research. This work provides a critical overview on the state-of-the-art on existing 3D models of intestinal inflammation and discusses the remaining challenges, providing insights on possible pathways towards achieving IBD mimetic models. We also address some of the main challenges faced by implementing cell culture models in IBD research while bearing in mind clinical translational aspects.

Chitosan-based sponges containing clotrimazole for the topical management of vulvovaginal candidiasis.

Vulvovaginal candidiasis (VVC) persists as a worrying women's healthcare issue, often relying on suboptimal therapeutics. Novel intravaginal dosage forms focusing on improving patient acceptability and featuring improved biopharmaceutical properties could be interesting alternatives to available antifungal products. Different formulations of sponges based on chitosan (Ch), with or without crosslinking and co-formulated with poly(N-vinylcaprolactam) (PNVCL), were produced for the topical administration of clotrimazole (CTZ) and further tested for physicochemical properties, drug release, cytotoxicity and antifungal activity. Results showed that high amounts of CTZ (roughly 30-50 %) could be incorporated into sponges obtained by using a simple freeze-drying methodology. Cross-linking of Ch with ammonia affected the morphology and mechanical features of sponges and shifted the release profile from sustained (around 20 % and 60 % drug released after 4 h and 24 h, respectively) to fast-releasing (over 90 % at 4 h). The combination of PNVCL with non-crosslinked Ch also allowed tuning drug release, namely by increasing the initial amount of CTZ released in simulated vaginal fluid (roughly 40 % after 4 h), as compared to sponges featuring only non-crosslinked Ch. All formulations displayed low toxicity to cell lines derived from the female genital tract, with viability values kept above 70 % after 24 h incubation with sponge extracts. These also allowed maintaining the rapid onset of the antifungal effects of CTZ at minimum inhibitory concentrations ranging from 0.5 to 16 µg/mL for a panel of six different Candida spp. strains. Overall, proposed sponge formulations appear to be promising alternatives for the safe and effective management of VVC.

Encapsulation and release of hydrocortisone from proliposomes govern vaginal delivery.

Topical preparations of hydrocortisone can be used for the anti-inflammatory treatment of the female genital area. Although the drug is a low-strength corticosteroid, systemic absorption and distribution of the drug are the most common safety risks associated with this therapy. In the current investigation, we elucidate the physicochemical properties of lipid-based drug carrier systems that govern the local bioavailability of hydrocortisone for intravaginal administration. For this purpose, we compared various proliposome formulations with a commercial cream. Depending on the availability of physiological acceptors, encapsulation and drug release from the lipid phase were found to be the most important drivers of drug bioavailability. The high permeability of hydrocortisone leads to rapid transport of the drug across the mucosal cell layer as indicated by experiments using HEC-1-A and CaSki cell monolayer models. Under sink conditions, differences in the release from the liposomes as determined in the Dispersion Releaser were almost negligible. However, under non-sink conditions, the drug release plateaued at levels corresponding to the encapsulation efficiency. After redispersion, all liposomal formulations performed better than the commercial drug product indicating that the encapsulation into the lipid phase is the main driver sustaining the release.

Antimicrobial peptide-grafted PLGA-PEG nanoparticles to fight bacterial wound infections.

Wound infection treatment with antimicrobial peptides (AMPs) is still not a reality, due to the loss of activity in vivo. Unlike the conventional strategy of encapsulating AMPs on nanoparticles (NPs) leaving activity dependent on the release profile, this work explores AMP grafting to poly(D,L-lactide-co-glycolide)-polyethylene glycol NPs (PLGA-PEG NPs), whereby AMP exposition, infection targeting and immediate action are promoted. NPs are functionalized with MSI-78(4-20), an equipotent and more selective derivative of MSI-78, grafted through a thiol-maleimide (Mal) Michael addition. NPs with different ratios of PLGA-PEG/PLGA-PEG-Mal are produced and characterized, with 40%PLGA-PEG-Mal presenting the best colloidal properties and higher amounts of AMP grafted as shown by surface charge (+8.6 ± 1.8 mV) and AMP quantification (326 µg mL-1, corresponding to 16.3 µg of AMP per mg of polymer). NPs maintain the activity of the free AMP with a minimal inhibitory concentration (MIC) of 8-16 µg mL-1 against Pseudomonas aeruginosa, and 16-32 µg mL-1 against Staphylococcus aureus. Moreover, AMP grafting accelerates killing kinetics, from 1-2 h to 15 min for P. aeruginosa and from 6-8 h to 0.5-1 h for S. aureus. NP activity in a simulated wound fluid is maintained for S. aureus and decreases slightly for P. aeruginosa. Furthermore, NPs do not demonstrate signs of cytotoxicity at MIC concentrations. Overall, this promising formulation helps unleash the full potential of AMPs for the management of wound infections.

MPTHub: An Open-Source Software for Characterizing the Transport of Particles in Biorelevant Media.

The study of particle transport in different environments plays an essential role in understanding interactions with humans and other living organisms. Importantly, obtained data can be directly used for multiple applications in fields such as fundamental biology, toxicology, or medicine. Particle movement in biorelevant media can be readily monitored using microscopy and converted into time-resolved trajectories using freely available tracking software. However, translation into tangible and meaningful parameters is time consuming and not always intuitive. We developed new software-MPTHub-as an open-access, standalone, user-friendly tool for the rapid and reliable analysis of particle trajectories extracted from video microscopy. The software was programmed using Python and allowed to import and analyze trajectory data, as well as to export relevant data such as individual and ensemble time-averaged mean square displacements and effective diffusivity, and anomalous transport exponent. Data processing was reliable, fast (total processing time of less than 10 s), and required minimal memory resources (up to a maximum of around 150 MB in random access memory). Demonstration of software applicability was conducted by studying the transport of different polystyrene nanoparticles (100-200 nm) in mucus surrogates. Overall, MPTHub represents a freely available software tool that can be used even by inexperienced users for studying the transport of particles in biorelevant media.

Mucus-producing 3D cell culture models.

In vitro cell-based models have been used for a long time since they are normally easily obtained and have an advantageous cost-benefit. Besides, they can serve a variety of ends, from studying drug absorption and metabolism to disease modeling. However, some in vitro models are too simplistic, not accurately representing the living tissues. It has been shown, mainly in the last years, that fully mimicking a tissue composition and architecture can be paramount for cellular behavior and, consequently, for the outcomes of the studies using such models. Because of this, 3D in vitro cell models have been gaining much attention, since they are able to better replicate the in vivo environment. In this review we focus on 3D models that contain mucus-producing cells, as mucus can play a pivotal role in drug absorption. Being frequently overlooked, this viscous fluid can have an impact on drug delivery. Thus, the aim of this review is to understand to which extent can mucus affect mucosal drug delivery and to provide a state-of-the-art report on the existing 3D cell-based mucus models.

Effect of antihelminths with contrasting efficacy against gastrointestinal nematodes on the live-weight gain of young Nellore cattle.

We estimated the losses caused by gastrointestinal nematode (GIN) infections in young Nellore cattle born in three consecutive calving seasons (Cycles I, II and III). Three groups of animals: Group 1 - free from GIN using 2.5 mg/kg of albendazole sulfoxide; Group II - free from Oesophagosomum radiatum using 0.2 mg/kg of ivermectin but infected with ivermectin-resistant Haemonchus placei and Cooperia spp.; and Group III - non-treated experimental control animals, infected with all GIN, were observed over a period of 12 months. Male and female calves were evaluated starting before weaning when the animals were an average of approximately four months of age. In Cycle I, only females were evaluated. All the animals continued to graze on pastures of Urochloa spp. (= Brachiaria). All the groups showed median faecal egg counts of fewer than 250 eggs per gram (EPG), and no clinical signs of parasitic gastroenteritis were observed. The blood variables were within the normal ranges, and no calf presented anaemia. In most of the samplings, the median EPGs were significantly lower (P < 0.05) in the group treated with albendazole. Throughout the experiment, the most prevalent parasite observed in the control group was Cooperia spp., followed in decreasing order by Haemonchus spp., O. radiatum and Trichostrongylus spp. All the groups of calves exhibited weight gain throughout the trials with control group displaying the lowest body weight gain. Body weight variables between the albendazole- and ivermectin-treated groups were not significantly different (P > 0.05). After weaning, females animals in albendazole and ivermectin treated groups exhibited higher body weights, 20.4% and 22.7%, respectively, than those of the control group. Likewise, males treated with albendazole and ivermectin exhibited 27.6% and 25.8%, respectively, more body weight gain than animals under control group. Because the main difference between the ivermectin and the control groups was the O. radiatum parasitism, most of the losses in the control group were possibly due to this nematode species. Nevertheless, the other nematodes species that occurred in relatively high intensities in control group could also have an additive effect in such losses.

Lipid Nanocarriers for Anti-HIV Therapeutics: A Focus on Physicochemical Properties and Biotechnological Advances.

Since HIV was first identified, and in a relatively short period of time, AIDS has become one of the most devastating infectious diseases of the 21st century. Classical antiretroviral therapies were a major step forward in disease treatment options, significantly improving the survival rates of HIV-infected individuals. Even though these therapies have greatly improved HIV clinical outcomes, antiretrovirals (ARV) feature biopharmaceutic and pharmacokinetic problems such as poor aqueous solubility, short half-life, and poor penetration into HIV reservoir sites, which contribute to the suboptimal efficacy of these regimens. To overcome some of these issues, novel nanotechnology-based strategies for ARV delivery towards HIV viral reservoirs have been proposed. The current review is focused on the benefits of using lipid-based nanocarriers for tuning the physicochemical properties of ARV to overcome biological barriers upon administration. Furthermore, a correlation between these properties and the potential therapeutic outcomes has been established. Biotechnological advancements using lipid nanocarriers for RNA interference (RNAi) delivery for the treatment of HIV infections were also discussed.

Facts and Figures on Materials Science and Nanotechnology Progress and Investment.

As the twenty-first century unfolds, nanotechnology is no longer just a buzzword in the field of materials science, but rather a tangible reality. This is evident from the surging number of commercial nanoproducts and their corresponding revenue generated in different industry sectors. However, it is important to recognize that sustainable growth of nanotechnology is heavily dependent on government funding and relevant national incentive programs. Consequently, proper analyses on publicly available nanotechnology data sets comprising information on the past two decades can be illuminating, facilitate development, and amend previous strategies as we move forward. Along these lines, classical statistics and machine learning (ML) allow processing large data sets to scrutinize patterns in materials science and nanotechnology research. Herein, we provide an analysis on nanotechnology progress and investment from an unbiased, computational vantage point and using orthogonal approaches. Our data reveal both well-established and surprising correlations in the nanotechnology field and its actors, including the interplay between the number of research institutes-industry, publications-patents, collaborative research, and top contributors to nanoproducts. Overall, data suggest that, supported by incentive programs set out by stakeholders (researchers, funding agencies, policy makers, and industry), nanotechnology could experience an exponential growth and become a centerpiece for economical welfare. Indeed, the recent success of COVID-19 vaccines is also likely to boost public trust in nanotechnology and its global impact over the coming years.

Women-specific routes of administration for drugs: A critical overview.

The woman's body presents a number of unique anatomical features that can constitute valuable routes for the administration of drugs, either for local or systemic action. These are associated with genitalia (vaginal, endocervical, intrauterine, intrafallopian and intraovarian routes), changes occurring during pregnancy (extra-amniotic, intra-amniotic and intraplacental routes) and the female breast (breast intraductal route). While the vaginal administration of drug products is common, other routes have limited clinical application and are fairly unknown even for scientists involved in drug delivery science. Understanding the possibilities and limitations of women-specific routes is of key importance for the development of new preventative, diagnostic and therapeutic strategies that will ultimately contribute to the advancement of women's health. This article provides an overview on women-specific routes for the administration of drugs, focusing on aspects such as biological features pertaining to drug delivery, relevance in current clinical practice, available drug dosage forms/delivery systems and administration techniques, as well as recent trends in the field.

Electrospun fibers for vaginal administration of tenofovir disoproxil fumarate and emtricitabine in the context of topical pre-exposure prophylaxis.

Women are particularly vulnerable to sexual HIV-1 transmission. Oral pre-exposure prophylaxis (PrEP) with tenofovir disoproxil fumarate and emtricitabine (TDF/FTC) is highly effective in avoiding new infections in men, but protection has only been shown to be moderate in women. Such differences have been associated, at least partially, to poor drug penetration of the lower female genital tract and the need for strict adherence to continuous daily oral intake of TDF/FTC. On-demand topical microbicide products could help circumvent these limitations. We developed electrospun fibers based on polycaprolactone (PCL fibers) or liposomes associated to poly(vinyl alcohol) (liposomes-in-PVA fibers) for the vaginal co-delivery of TDF and FTC, and assessed their pharmacokinetics in mice. PCL fibers and liposomes-in-PVA fibers were tested for morphological and physicochemical properties using scanning electron microscopy, differential scanning calorimetry and X-ray diffractometry. Fibers featured organoleptic and mechanical properties compatible with their suitable handling and vaginal administration. Fluorescent quenching of mucin in vitro - used as a proxy for mucoadhesion - was intense for PCL fibers, but mild for liposomes-in-PVA fibers. Both fibers were shown safe in vitro and able to rapidly release drug content (15-30 min) under sink conditions. Liposomes-in-PVA fibers allowed increasing genital drug concentrations after a single intravaginal administration when compared to continuous daily treatment for five days with 25-times higher oral doses. For instance, the levels of tenofovir and FTC in vaginal lavage were around 4- and 29-fold higher, respectively. PCL fibers were also superior to oral treatment, although to a minor extent (approximately 2-fold higher drug concentrations in lavage). Vaginal tissue drug levels were generally low for all treatments, while systemic drug exposure was negligible in the case of fibers. These data suggest that proposed fibers may provide an interesting alternative or an ancillary option to oral PrEP in women.

Production and Characterization of Anti-CCR5 siRNA-Loaded Polycaprolactone Nanoparticles for Topical Pre-exposure Prophylaxis.

Topical posttranscriptional silencing of host factors involved in HIV-1 sexual transmission, such as CCR5, presents the potential to prevent new cases of infection. However, issues concerning proper engineering of safe and effective delivery systems for anti-CCR5 siRNA may impair the ability to yield suitable silencing at the mucosal level. Here we describe the production protocol of anti-CCR5 siRNA-loaded polycaprolactone-based nanoparticles (≈100 nm). Furthermore, we present data regarding the physicochemical and in vitro biological characterization of obtained nanosystems, which support their potential as microbicide candidates for topical pre-exposure prophylaxis of HIV-1 infection.

Clotrimazole-loaded N-(2-hydroxy)-propyl-3-trimethylammonium, O-palmitoyl chitosan nanoparticles for topical treatment of vulvovaginal candidiasis.

Vulvovaginal candidiasis (VVC) represents a considerable health burden for women. Despite the availability of a significant array of antifungal drugs and topical products, the management of the infection is not always effective, and new approaches are needed. Here, we explored cationic N-(2-hydroxy)-propyl-3-trimethylammonium, O-palmitoyl chitosan nanoparticles (NPs) as carriers of clotrimazole (CLT) for the topical treatment of VVC. CLT-NPs with approximately 280 nm in diameter were obtained by self-assembly in water and subsequent stabilization by ionic crosslinking with tripolyphosphate. The nanosystem featured pH-independent sustained drug release up to 24 h, which affected both in vitro anti-Candida activity and cytotoxicity. The CLT-loaded nanostructured platform yielded favorable selectivity index values for a panel of standard strains and clinical isolates of Candida spp. and female genital tract cell lines (HEC-1-A, Ca Ski and HeLa), as compared to the free drug. CLT-NPs also improved in vitro drug permeability across HEC-1-A and Ca Ski cell monolayers, thus suggesting that the nanocarrier may provide higher mucosal tissue levels of the active compound. Overall, data support that CLT-NPs may be a valuable asset for the topical treatment of VVC. STATEMENT OF SIGNIFICANCE: Topical azoles such as clotrimazole (CLT) are first line antifungal drugs for the management of vulvovaginal candidiasis (VVC), but their action may be limited by issues such as toxicity and poor capacity to penetrate the genital mucosa. Herein, we report on the ability of a new cationic N-(2­hydroxy)-propyl-3-trimethylammonium, O-dipalmitoyl chitosan derivative (DPCat35) to yield tripolyphosphate-reinforced micelle-like nanostructures that are suitable carriers for CLT. In particular, these nanosystems were able to improve the in vitro selectivity index of the drug and to provide enhanced epithelial drug permeability when tested in cell monolayer models. These data support that CLT-loaded DPCat35 nanoparticles feature favorable properties for the development of new nanomedicines for the topical management of VVC.

Influence of Plasticizers on the pH-Dependent Drug Release and Cellular Interactions of Hydroxypropyl Methylcellulose/Zein Vaginal Anti-HIV Films Containing Tenofovir.

Vaginal films featuring the pH-dependent release of tenofovir (TFV) were developed for the prevention of sexual transmission of human immunodeficiency syndrome (HIV). Films based on hydroxypropyl methylcellulose and zein were prepared incorporating different plasticizers [oleic acid, lactic acid, glycerol, and polyethylene glycol 400 (PEG)] and evaluated for in vitro drug release in an acidic simulated vaginal fluid (pH 4.2) and a slightly alkaline mixture of simulated seminal and vaginal fluids (pH 7.5). Results revealed that optimal biphasic TFV release was possible with proper combination of plasticizers (PEG and oleic acid, 1:7 w/w) and by adjusting the plasticizer/matrix-forming material ratio. The films had similar or higher levels of TFV associated with genital epithelial cells (Ca Ski or HEC-1-A cells) but lower drug permeability compared to the free drug. These data confirm that films have the potential to achieve suitable mucosal levels of TFV with low systemic exposure. The films developed could protect women from HIV sexual transmission.

Development of pH-sensitive vaginal films based on methacrylate copolymers for topical HIV-1 pre-exposure prophylaxis.

Interest is growing in "smart" vaginal microbicides as a strategy to protect women from sexual transmission of human immunodeficiency virus (HIV). The concept is based on the development of products featuring low drug release in acidic media such as vaginal fluid but switch to a fast release profile when the medium becomes neutral or slightly alkaline. This mimics the surge in pH occurring in the vagina after sexual intercourse due to the seminal fluid. Semen is the main vehicle for HIV-1, and increasing antiretroviral drug levels in the vagina upon ejaculation may contribute to enhanced protection against viral sexual transmission. This work explores the use of different pharmaceutical-grade methacrylic acid-based polymers (EudragitⓇ RL, RS, L and S) for developing vaginal films allowing the pH-dependant release of the antiretroviral drug tenofovir (TFV). EudragitⓇ L 100 and EudragitⓇ S 100, containing triethyl citrate as plasticiser, proved to be suitable for manufacturing films with optimal dual in vitro drug-release behaviour. TFV-release can be sustained for several days after film administration and all the drug is released in a few hours in conditions simulating ejaculation. The films' mechanical properties were also deemed suitable for comfortable vaginal administration. Two optimized films were further assessed using HEC-1-A and Ca Ski cell monolayer models and were found to possess favourable drug permeability profiles and drug levels associated to cell monolayer as compared to free TFV. Overall, pH-dependant films containing tenofovir may constitute promising candidates for "smart" vaginal microbicides to protect women from sexual HIV transmission.

Design, fabrication and characterisation of drug-loaded vaginal films: State-of-the-art.

Films have undoubtedly seen the most significant advances in their development in recent years of all the pharmaceutical forms for the vaginal administration of drugs. Films combine the advantages of gels and solid pharmaceutical forms, and their great versatility is largely determined by the numerous polymers that can be used for their fabrication. They may be based on many natural polymers, and cellulose derivatives, polyvinyl alcohol or acrylic derivatives - among others - are also frequently used. The combination of different polymers and the inclusion of plasticizing agents makes them extremely versatility for responding to a range of therapeutic needs. The techniques used to produce films have also undergone substantial development. Although the solvent casting technique is by far the most widely used in fabrication, alternative options have also emerged such as electrospinning, moulding extrusion and 3D printing. Various research groups have proposed a proliferation of assays to characterise vaginal films in recent years, which highlight the importance of the preliminary evaluation and determination of the films' uniformity, in addition to tests to determine their permeability and hydrophilic/hydrophobic coefficient and their mechanical properties, the application of imaging techniques and thermal analysis, and especially the evaluation of the mucoadhesive properties and control over the drug release. This article offers a critical overview of the advances in the development and fabrication of films intended for vaginal drug delivery, and summarises current clinical applications for vaginal films.

Molecular and cellular cues governing nanomaterial-mucosae interactions: from nanomedicine to nanotoxicology.

Mucosal tissues constitute the largest interface between the body and the surrounding environment and they regulate the access of molecules, supramolecular structures, particulate matter, and pathogens into it. All mucosae are characterized by an outer mucus layer that protects the underlying cells from physicochemical, biological and mechanical insults, a mono-layered or stratified epithelium that forms tight junctions and controls the selective transport of solutes across it and associated lymphoid tissues that play a sentinel role. Mucus is a gel-like material comprised mainly of the glycoprotein mucin and water and it displays both hydrophilic and hydrophobic domains, a net negative charge, and high porosity and pore interconnectivity, providing an efficient barrier for the absorption of therapeutic agents. To prolong the residence time, absorption and bioavailability of a broad spectrum of active compounds upon mucosal administration, mucus-penetrating and mucoadhesive particles have been designed by tuning the chemical composition, the size, the density, and the surface properties. The benefits of utilizing nanomaterials that interact intimately with mucosae by different mechanisms in the nanomedicine field have been extensively reported. To ensure the safety of these nanosystems, their compatibility is evaluated in vitro and in vivo in preclinical and clinical trials. Conversely, there is a growing concern about the toxicity of nanomaterials dispersed in air and water effluents that unintentionally come into contact with the airways and the gastrointestinal tract. Thus, deep understanding of the key nanomaterial properties that govern the interplay with mucus and tissues is crucial for the rational design of more efficient drug delivery nanosystems (nanomedicine) and to anticipate the fate and side-effects of nanoparticulate matter upon acute or chronic exposure (nanotoxicology). This review initially overviews the complex structural features of mucosal tissues, including the structure of mucus, the epithelial barrier, the mucosal-associated lymphatic tissues and microbiota. Then, the most relevant investigations attempting to identify and validate the key particle features that govern nanomaterial-mucosa interactions and that are relevant in both nanomedicine and nanotoxicology are discussed in a holistic manner. Finally, the most popular experimental techniques and the incipient use of mathematical and computational models to characterize these interactions are described.

Zein nanoparticles as low-cost, safe, and effective carriers to improve the oral bioavailability of resveratrol.

The clinical translation of the multiple pharmacological effects of resveratrol (RSV) found in preclinical studies has been impaired by its poor bioavailability, due to poor solubility and rapid metabolism and elimination. The inclusion of this molecule in medicines or functional food products will be ineffective unless suitable systems are developed. Zein protein may constitute an inexpensive, safe, and effective choice to produce nanoparticles (NPs) to incorporate hydrophobic molecules and overcome the bioavailability issues of RSV. In this work, we loaded RSV into zein NPs by using a nanoprecipitation method. Unloaded and RSV-loaded NPs presented average diameter values in the range of 120-180 nm, narrow size distribution (polydispersity index < 0.150), and zeta potential of around + 20 mV. The association efficiency of the drug was equal to or greater than 77% for different initial drug loads. Scanning electron microscopy imaging revealed that zein NPs were round-shaped and presented a smooth surface. Aqueous suspensions of zein NPs were stable for at least 1 month when stored at 4 °C. The freeze-drying of zein NPs using sucrose as cryoprotectant allowed an easy re-suspension of NPs in water without significantly changing the initial colloidal properties. RSV-loaded NPs presented low cytotoxicity to the human colorectal Caco-2 and HT29-MTX cell lines. Finally, permeability studies of RSV across Caco-2 and Caco-2/HT29-MTX evidenced some ability of zein NPs to protect RSV from metabolism events. However, further investigation is needed in order to confirm the possible role of zein NPs in the metabolic stability of RSV. Overall, zein NPs may present the potential to circumvent bioavailability issues of RSV. Graphical abstract.