Cell-Permeable Succinate Rescues Mitochondrial Respiration in Cellular Models of Statin Toxicity

Statins are the cornerstone of lipid-lowering therapy. Although generally well tolerated, statin-associated muscle symptoms (SAMS) represent the main reason for treatment discontinuation. Mitochondrial dysfunction of complex I has been implicated in the pathophysiology of SAMS. The present study proposed to assess the concentration-dependent ex vivo effects of three statins on mitochondrial respiration in viable human platelets and to investigate whether a cell-permeable prodrug of succinate (complex II substrate) can compensate for statin-induced mitochondrial dysfunction. Mitochondrial respiration was assessed by high-resolution respirometry in human platelets, acutely exposed to statins in the presence/absence of the prodrug NV118. Statins concentration-dependently inhibited mitochondrial respiration in both intact and permeabilized cells. Further, statins caused an increase in non-ATP generating oxygen consumption (uncoupling), severely limiting the OXPHOS coupling efficiency, a measure of the ATP generating capacity. Cerivastatin (commercially withdrawn due to muscle toxicity) displayed a similar inhibitory capacity compared with the widely prescribed and tolerable atorvastatin, but did not elicit direct complex I inhibition. NV118 increased succinate-supported mitochondrial oxygen consumption in atorvastatin/cerivastatin-exposed platelets leading to normalization of coupled (ATP generating) respiration. The results acquired in isolated human platelets were validated in a limited set of experiments using atorvastatin in HepG2 cells, reinforcing the generalizability of the findings.     

Functionality of chitosan-halloysite nanocomposite films for sustained delivery of antibiotics: The effect of chitosan molar mass

This study was designed to investigate functionality of tetracycline-loaded chitosan-halloysite nanocomposite films, with focus on evaluating the influence of chitosan molar mass on films applicability for sustained local antibiotic delivery. The films were prepared by casting and solvent evaporation using low, medium, and high molar mass chitosan. SEM analysis revealed compact, nonporous and rough surface of the nanocomposite films due to the presence of halloysite agglomerates and tetracycline crystals. Increasing chitosan molar mass led to higher values of elongation at break (from 21.65 ± 2.65 to 34.48 ± 2.34%), tensile strength (from 134.8 ± 13.21 to 246.36 ± 14.69 MPa), and elastic modulus (from 633.79 ± 128.37 to 716.55 ± 60.76 MPa) of the nanocomposite films. FT-IR, XRPD, and thermal analyses confirmed molar mass dependent chitosan-halloysite interactions and improved thermal stability of the nanocomposite films in comparison with chitosan films. The nanocomposite films released tetracycline in a sustained manner, with the slowest release achieved from the films consisting of low molar mass chitosan. Chitosan molar mass was confirmed to be a functionality-related characteristic of chitosan-halloysite nanocomposite films as potential sustained-release carriers for topical delivery of antibiotics. © 2019 Wiley Periodicals, Inc. J. Appl. Polym. Sci. 2020 , 137, 48406.     

Development of hydrophilic gels containing coenzyme Q10-loaded liposomes: characterization,stability and rheology measurements

Objective: The aim of this study was to develop, characterize and evaluate stability of a gel containing coenzyme Q₁₀ (Q₁₀)-loaded liposomes, and enhance the stability of Q₁₀ in the nanocarrier-containing gel compared to the conventional gel.

Methods: Q₁₀-loaded liposome dispersions prepared from unsaturated or saturated lecithin, were characterized for particle size, polydispersity index (PDI), zeta-potential, pH value, oxidation index, Q₁₀-content and morphology, and incorporated into carbomer gel. Liposome gels and liposome-free gel were analyzed for flow properties, pH values, Q₁₀-content, and liposomes size and PDI (liposome gels), 48?h after preparation and in predetermined time intervals during 6?months storage at different temperatures in order to predict their long term stability.

Results: Liposomes were of small particle size, homogeneous, negatively charged, and their incorporation into gel did not significantly change (p?>?.05) their particle size and PDI. All gels revealed non-Newtonian, shear-thinning plastic flow behavior during storage with no marked changes in rheological parameters. Storage of gels did not significantly influence the pH value (p?>?.05), while it significantly decreased Q₁₀-content (p?<?.05). Q₁₀ was significantly more (p?<?.05) stable in liposome gel containing unsaturated lecithin liposomes (G1) than in gel containing saturated lecithin liposomes (G2) and liposome-free gel (G3).

Conclusions: Q₁₀-loaded liposome gel G1 was the optimal formulation, since during storage at different temperatures, it did not show significant increase in liposome size and PDI, it provided significantly higher stability for Q₁₀ than other gels and its pH value was suitable for skin application. Due to limited Q₁₀-stability it should be stored at 4?°C.     

Case Report: Compound Heterozygous Variants of the MAN1B1 Gene in a Russian Patient with Rafiq Syndrome

Rafiq syndrome (RAFQS) is a congenital disorder of glycosylation (CDG) that is caused by mutations in the MAN1B1 gene and characterized by impaired protein and lipid glycosylation. RAFQS is characterized by a delay in intellectual and motor development, facial and other dysmorphism, truncal obesity, behavior problems, and hypotonia. We describe a Russian patient with delayed intellectual and motor development, a lack of speech, disorientation in space and time, impaired attention and memory, and episodes of aggression. Screening for lysosomal, amino acid, organic acid, and mitochondrial disorders was normal. The patient was referred for the targeted sequencing of the “Hereditary Metabolic Disorders” panel. The genetic testing revealed two heterozygous pathogenic variants in the MAN1B1 gene: the previously reported c.1000C > T (p.Arg334Cys) and the novel c.1065 + 1 G > C. Thus, the patient’s clinical picture and genetic analysis confirmed RAFQS in the patient.     

Assessment of Betulinic Acid Cytotoxicity and Mitochondrial Metabolism Impairment in a Human Melanoma Cell Line

Melanoma represents one of the most aggressive and drug resistant skin cancers with poor prognosis in its advanced stages. Despite the increasing number of targeted therapies, novel approaches are needed to counteract both therapeutic resistance and the side effects of classic therapy. Betulinic acid (BA) is a bioactive phytocompound that has been reported to induce apoptosis in several types of cancers including melanomas; however, its effects on mitochondrial bioenergetics are less investigated. The present study performed in A375 human melanoma cells was aimed to characterize the effects of BA on mitochondrial bioenergetics and cellular behavior. BA demonstrated a dose-dependent inhibitory effect in both mitochondrial respiration and glycolysis in A375 melanoma cells and at sub-toxic concentrations (10 μM) induced mitochondrial dysfunction by eliciting a decrease in the mitochondrial membrane potential and changes in mitochondria morphology and localization. In addition, BA triggered a dose-dependent cytotoxic effect characterized by apoptotic features: morphological alterations (nuclear fragmentation, apoptotic bodies) and the upregulation of pro-apoptotic markers mRNA expression (Bax, Bad and Bak). BA represents a viable therapeutic option via a complex modulatory effect on mitochondrial metabolism that might be useful in advanced melanoma or as reliable strategy to counteract resistance to standard therapy.     

pH‐sensitive polyelectrolyte films derived from submicron chitosan/Eudragit® L 100‐55 complexes: Physicochemical characterization and in vitro drug release

The objectives of this study were to prepare films from submicron chitosan/Eudragit^® L100‐55 polyelectrolyte complexes (CH/EL PEC) and to assess the influence of CH molecular weight and CH/EL mass ratio on their structure and drug‐release properties. The films were obtained by a simple, environmentally friendly, casting/solvent evaporation method and the verapamil hydrochloride (VH) was used as model drug. Submicron size, narrow size distribution, and acceptable stability of CH/EL PECs were confirmed by DLS and laser Doppler microelectrophoresis. SEM analysis revealed nonporous inner structure and flat surface of the films. Interactions between comprising polymers and formation of CH/EL PEC were established by DSC and FT‐IR spectroscopy. In vitro swelling and drug release studies revealed the pH sensitivity of the films, with burst drug release in acidic conditions (pH 1.2) and sustained release in phosphate buffers pH 5.8, 6.8, and 7.4. The slowest VH release was achieved from the films prepared from equal amounts of EL and CH of higher molecular weight, confirming the significance of the CH/EL ratio and CH molecular weight on their ability to sustain drug release. The obtained results suggested that presented, simple, and eco‐friendly preparation procedure can be used to obtain pH‐sensitive CH/EL PEC films with a promising potential as drug carriers. © 2015 Wiley Periodicals, Inc. J. Appl. Polym. Sci. 2015 , 132, 42583.     

Polymer-stabilized emulsion systems: structural characteristics and physical stability evaluation

In the course of investigation of the effects of polymeric emulsifier (Simulgel®A) on the structural characteristics and physical stability (during storage time) of semisolid o/w emulsion systems with different emulsifier concentrations, dispersed droplets of an internal oil phase (4.5-11.5 µm size) were discovered. The degree of dispersity was increased along with increasing polymeric emulsifier concentration (from 1 to 5% m/m), which was shown by microscopic analysis. Experiments were also carried out using pH, electrical conductivity, and rheological (rotational and oscillatory) measurements. Emulsion samples expressed a mild acid value acceptable for topical preparations during a 6-month period. Electrical conductivity of fresh samples ranged from 0.423 to 2.009 mS/cm and after a 6-month period this slightly increased to 0.663-2.890 mS/cm. All emulsion samples revealed plastic flow behavior in fresh state, with changes in flow pattern during storage time. Viscoelastic properties defined by the oscillatory rheometry results were in good correlation with parameters of rotational rheometry and degree of internal phase dispersity. High yield stress values (flow tests) and favorable low values of damping factor (oscillatory studies) both indicated good internal structuring and long-term stability of sample containing 3% m/m of polymeric emulsifier.