Fever-Range Hyperthermia Promotes Macrophage Polarization towards Regulatory Phenotype M2b.

Fever-range hyperthermia (FRH) is utilized in chronic disease treatment and serves as a model for fever's thermal component investigation. Macrophages, highly susceptible to heat, play a pivotal role in various functions determined by their polarization state. However, it is not well recognized whether this process can be modulated by FRH. To address this, we used two different macrophage cell lines that were treated with FRH. Next, to define macrophage phenotype, we examined their functional surface markers CD80 and CD163, intracellular markers such as inducible nitric oxide synthase (iNOS), arginase-1 (Arg-1), and the expression of interleukin-10 (IL-10) and tumor necrosis factor α (TNF-α). Additionally, in FRH-treated cells, we analyzed an expression of Toll-like receptor 4 (TLR-4) and its role in macrophage polarization. We also checked whether FRH can switch the polarization of macrophages in pro-inflammatory condition triggered by lipopolysaccharide (LPS). FRH induced M2-like polarization, evident in increased CD163, IL-10, and Arg-1 expression. Notably, elevated COX-2, TNF-α, and TLR-4 indicated potential pro-inflammatory properties, suggesting polarization towards the M2b phenotype. Additionally, FRH shifted lipopolysaccharide (LPS)-induced M1 polarization to an M2-like phenotype, reducing antimicrobial molecules (ROS and NO). In summary, FRH emerged as a modulator favoring M2-like macrophage polarization, even under pro-inflammatory conditions, showcasing its potential therapeutic relevance.

The Impact of Liquid Components on Alteration of the Adhesion of Polyacrylate and Silicone Patches.

Polyacrylates and polysiloxanes are polymers used in pressure-sensitive adhesive (PSA) patches. Liquid additives are co-solvents of the active substances or permeation enhancers, and their compatibility with the polymeric matrix and the effect on adhesive properties should be considered. The patches were prepared from commercial polyacrylates (three types of Duro-Tak®) and siloxanes (Bio-PSA® and Soft Skin Adhesive®). Propylene glycol, polyoxyethylene glycol, isopropyl myristate, triacetin, triethyl citrate and silicone oil were added (10% w/w). Formulations were evaluated microscopically and with a texture analyzer in terms of in vitro adhesiveness and hardness. Only silicone oil was compatible with the silicone matrices. The best compatibility of acrylic PSA was observed with triethyl citrate; one out of three Duro-Tak matrices was incompatible with every additive. In all compositions, the adhesiveness was impaired by the liquid additives. A significant drop in adhesiveness was noted after immersion of the patches in buffer and drying. The probe tack test was considered as the most useful for evaluation of the effect of the liquid additive on adhesiveness, but the results obtained with a spherical and cylindrical probe were contradictory. The structural changes caused by the additives were also demonstrated by a 90° peel test, considered as complementary to the tack test.

Endotoxin Tolerance Creates Favourable Conditions for Cancer Development.

Endotoxin tolerance (ET) is an adaptive phenomenon of the immune system that protects the host from clinical complications due to repeated exposure of the body to endotoxins such as lipopolysaccharide (LPS). Since ET is an immunosuppressive mechanism in which a significant reprogramming of macrophages is observed, we hypothesized that it could influence cancer development by modifying the tumour environment. This study aimed to explore whether ET influences cancer progression by altering the tumour microenvironment. Endotoxin-tolerant macrophages (MoET) were examined for their impact on breast and colon cancer cells via direct interaction and conditioned media exposure. We characterized cancer cell behaviour by viability, clonogenic potential, motility, scratch assays, and 3D spheroidal assays. MoET-derived factors increased cancer cell viability, motility, and clonogenicity, suggesting a conducive environment for cancer development. Remarkably, despite reduced TNFα and IL-6 levels, MoET exhibited M1 polarization. These findings uncover an ET-associated macrophage reprogramming that fosters a favourable context for cancer progression across diverse tumours. Targeting ET could emerge as a promising avenue for cancer therapy and prevention.

Mucoadhesive Alginate/Pectin Films Crosslinked by Calcium Carbonate as Carriers of a Model Antifungal Drug-Posaconazole.

The mucosal membrane of the oral cavity, due to its unique structure and availability, constitutes an appropriate site for the delivery of drugs, both with local and systemic effects. Mucoadhesive buccal films are drug dosage forms that due to their convenience of application, flexibility and size, are characterized by patients' compliance. Sodium alginate and pectin are natural polymers from the polysaccharides group, with mucoadhesive properties, that are widely applied to obtain buccal films. However, their hydrophilic nature and poor water resistance limit their application in sustained drug release formulations. Hence, the aim of this investigation was to design alginate/pectin buccal films by a one-step crosslinking technique-with the application of calcium carbonate. This technique was applied to prepare crosslinked alginate and alginate/pectin mucoadhesive films with a model antifungal drug-posaconazole. The obtained formulations were evaluated for the impact of crosslinking and pectin's presence on their pharmaceutical, mucoadhesive, mechanical and physicochemical properties. Additionally, the antifungal activity of the prepared films against Candida spp. was evaluated. It was shown that pectin's presence in the formulations improved flexibility, mucoadhesion and antifungal activity. The crosslinking process reduced mucoadhesiveness and antifungal activity but significantly enhanced the mechanical properties and stability and enabled prolonged drug release.

Fever-range whole body hyperthermia leads to changes in immune-related genes and miRNA machinery in Wistar rats.

OBJECTIVE: Fever is defined as a rise in body temperature upon disease. Fever-range hyperthermia (FRH) is a simplified model of fever and a well-established medical procedure. Despite its beneficial effects, the molecular changes induced by FRH remain poorly characterized. The aim of this study was to investigate the influence of FRH on regulatory molecules such as cytokines and miRNAs involved in inflammatory processes. METHODS: We developed a novel, fast rat model of infrared-induced FRH. The body temperature of animals was monitored using biotelemetry. FRH was induced by the infrared lamp and heating pad. White blood cell counts were monitored using Auto Hematology Analyzer. In peripheral blood mononuclear cells, spleen and liver expression of immune-related genes (IL-10, MIF and G-CSF, IFN-γ) and miRNA machinery (DICER1, TARBP2) was analyzed with RT-qPCR. Furthermore, RT-qPCR was used to explore miRNA-155 levels in the plasma of rats. RESULTS: We observed a decrease in the total number of leukocytes due to lower number of lymphocytes, and an increase in the number of granulocytes. Furthermore, we observed elevated expressions of DICER1, TARBP2 and granulocyte colony-stimulating factor (G-CSF) in the spleen, liver and PBMCs immediately following FRH. FRH treatment also had anti-inflammatory effects, evidenced by the downregulation of pro-inflammatory macrophage migration inhibitor factor (MIF) and miR-155, and the increased expression of anti-inflammatory IL-10. CONCLUSION: FRH affects the expression of molecules involved in inflammatory processes leading to alleviated inflammation. We suppose these effects may be miRNAs-dependent and FRH can be involved in therapies where anti-inflammatory action is needed.

Orodispersible Films with Rupatadine Fumarate Enclosed in Ethylcellulose Microparticles as Drug Delivery Platform with Taste-Masking Effect.

Orally disintegrating (orodispersible) films provide a versatile tool for drug administration, especially in the pediatric and geriatric population, since they reduce the risk of choking and do not necessitate drinking water during application. By considering their direct contact with the taste buds, palatability is an influential aspect related to patient compliance. The microparticles based on taste-masking polymers containing drugs enclosed inside effectively mask the unpleasant taste of medicines. Ethylcellulose is a hydrophobic polymer widely used as a taste-masking material. Rupatadine fumarate, a second-generation antihistamine drug, is characterised by an intense bitter taste; therefore, it is crucial to achieve a tolerable taste whilst developing orodispersible formulations with its content. The objective of this study was to develop orally disintegrating films with rupatadine fumarate in the form of ethylcellulose-based microparticles obtained from aqueous dispersions of ethylcellulose-Surelease® or Aquacoat® ECD. It was a technological challenge to achieve homogenous drug content per dosage unit and sufficient mechanical properties for film operating due to the necessity to suspend the microparticles in the casting solution. Although the process of obtaining films consisted of several steps (mixing, pouring, drying), the particles were homogeneously dispersed, and each film of the desired size contained the proper dose of the drug. The taste-masking effect was also maintained. This parameter was confirmed by three independent methods: in vivo by healthy volunteers, an electronic tongue and a dissolution test. The applied taste-evaluation techniques showed that the films containing Aquacoat® ECD microparticles have the highest degree of bitter taste reduction, which confirms the results obtained in our previous studies.

Prototype Gastro-Resistant Soft Gelatin Films and Capsules-Imaging and Performance In Vitro.

The following study is a continuation of the previous work on preparation of gastro-resistant films by incorporation of cellulose acetate phthalate (CAP) into the soft gelatin film. An extended investigation on the previously described binary Gelatin-CAP and ternary Gelatin-CAP-carrageenan polymer films was performed. The results suggest that the critical feature behind formation of the acid-resistant films is a spinodal decomposition in the film-forming mixture. In the obtained films, upon submersion in an acidic medium, gelatin swells and dissolves, exposing a CAP-based acid-insoluble skeleton, partially coated by a residue of other ingredients. The dissolution-hindering effect appears to be stronger when iota-carrageenan is added to the film-forming mixture. The drug release study performed in enhancer cells confirmed that diclofenac sodium is not released in the acidic medium, however, at pH 6.8 the drug release occurs. The capsules prepared with a simple lab-scale process appear to be resistant to disintegration of the shell structure in acid, although imperfections of the sealing have been noticed.

Gelatin Films Modified with Acidic and Polyelectrolyte Polymers-Material Selection for Soft Gastroresistant Capsules.

The following investigation comprised the formation of acid-resistant gelatin-based films, intended for future use in soft-capsule technology. Such film compositions were obtained by including nonionized forms of acid-insoluble polymers in a gelatin-based film-forming mixture. The selected films were additionally modified with small amounts of anionic polysaccharides that have potential to interact with gelatin, forming polyelectrolyte complexes. The obtained film compositions were subjected to, e.g., disintegration tests, adhesiveness tests, differential scanning calorimetry (DSC), and a transparency study. As a result of the performed study, some commercial enteric polymers (acrylates), as well as cellulose acetate phthalate, were selected as components that have the ability to coalesce and form a continuous phase within a gelatin film. The use of a small amount (1.5%) of additional gelling polymers improved the rheological characteristics and adhesive properties of the obtained films, with ί-carrageenan and gellan gum appearing to be the most beneficial.

Soft Gelatin Films Modified with Cellulose Acetate Phthalate Pseudolatex Dispersion-Structure and Permeability.

Gastroresistant material, based on gelatin and intended to form capsule shells, was characterized. The films were obtained by mixing a gelatin solution with cellulose acetate phthalate (CAP) pseudolatex at an elevated temperature. Microscopic and spectroscopic analyses of the films-intact or subjected to the acidic treatment-were performed, along with a permeability study of tritium-labeled water. A uniform porous structure formed by CAP within the gelatin gel was observed. The results demonstrated that no interaction of a chemical nature occurred between the components. Additionally, the performed permeability and solubility studies proved that the diffusion of water through the membranes at an acidic pH can be noticeably reduced by adding carrageenan as a secondary gelling/thickening agent.