Immunosuppressive properties of cytochalasin B-induced membrane vesicles of mesenchymal stem cells: comparing with extracellular vesicles derived from mesenchymal stem cells.

Extracellular vesicles derived from mesenchymal stem cells (MSCs) represent a novel approach for regenerative and immunosuppressive therapy. Recently, cytochalasin B-induced microvesicles (CIMVs) were shown to be effective drug delivery mediators. However, little is known about their immunological properties. We propose that the immunophenotype and molecular composition of these vesicles could contribute to the therapeutic efficacy of CIMVs. To address this issue, CIMVs were generated from murine MSC (CIMVs-MSCs) and their cytokine content and surface marker expression determined. For the first time, we show that CIMVs-MSCs retain parental MSCs phenotype (Sca-1+, CD49e+, CD44+, CD45-). Also, CIMVs-MSCs contained a cytokine repertoire reflective of the parental MSCs, including IL-1ß, IL-2, IL-3, IL-4, IL-5, IL-6, IL-9, IL-10, IL-12(p40), IL-13, IL-17, CCL2, CCL3, CCL4, CCL5, CCL11, G-CSF, GM-CSF and TNF-α. Next, we evaluated the immune-modulating properties of CIMVs-MSCs in vivo using standard preclinical tests. MSCs and CIMVs-MSCs reduced serum levels of anti-sheep red blood cell antibody and have limited effects on neutrophil and peritoneal macrophage activity. We compared the immunomodulatory effect of MSCs, CIMVs and EVs. We observed no immunosuppression in mice pretreated with natural EVs, whereas MSCs and CIMVs-MSCs suppressed antibody production in vivo. Additionally, we have investigated the biodistribution of CIMVs-MSCs in vivo and demonstrated that CIMVs-MSCs localized in liver, lung, brain, heart, spleen and kidneys 48 h after intravenous injection and can be detected 14 days after subcutaneous and intramuscular injection. Collectively our data demonstrates immunomodulatory efficacy of CIMVs and supports their further preclinical testing as an effective therapeutic delivery modality.

Synthesis, characterization, in vitro biocompatibility and antibacterial properties study of nanocomposite materials based on hydroxyapatite-biphasic ZnO micro- and nanoparticles embedded in Alginate matrix.

The paper presents the results of studies of biocompatibility and antibacterial properties of multiphase nanocomposite materials based on HA-Alg-ZnO (hydroxyapatite­sodium alginate-biphasic zinc oxide) and HA-ZnO (hydroxyapatite­zinc oxide), which were synthesized from the analytically pure calcium nitrate tetrahydrate, ammonium hydrophosphate, hydrous ammonia, zinc nitrate hexahydrate and calcium chloride. The samples' antimicrobial activity assessment was carried out on Gram-negative (E. coli, P. aeruginosa) and Gram-positive bacteria (S. aureus and S. epidermidis) test cultures by the co-incubation and modified "agar diffusion" methods. The murine fibroblast cells were used for the biocompatibility tests and cytotoxicity evaluation. It was shown that synthesized nanocomposite material has a multiphase nanoscale architecture, where ZnO nanocrystals are represented by two lattices: cubic and hexagonal. The possible explanation of ZnO nanocrystals' phase transition is given. At the same time, a partial replacement of Ca2+ ions by Zn2+ ions in the HA lattice possibly occurs due to processing of composite by US radiation. The replacement was evidenced by the non-stoichiometric Ca/P ratio < 2.16, OPO lines' shifting on FTIR spectrum and TEM analysis. The studied composite demonstrate a pronounced antibacterial activity due to the incorporation of ZnO particles into sodium alginate and moistened powder of hydroxyapatite. Both forms of HA-ZnO (suspension) and HA-Alg-ZnO (beads) are biocompatible. An interpretation of the process of Zn ions' embedding into hydroxyapatite and alginate matrix is given, as well as their influence on the biomimetic composite properties is discussed in details. STATEMENT OF SIGNIFICANCE: A number of studies have shown that Zn effectively inhibits the growth and development of bacteria and yeast fungi. Zinc plays an important role in the creation of new antimicrobial agents, and zinc-doped hydroxyapatite will find further application in biomedicine. In this regard, the phase states of zinc oxide, as well as the processes of calcium replacement by zinc in calcium apatite and in alginate should be explored fully. Nowadays we have lack of information and the study's results about those interactions. The present study provides data of the multiphase morphology, antimicrobial activity, biocompatibility and cytotoxicity of the biomimetic nanostructured composite materials, such as sodium alginate/hydroxyapatite/ZnO based granules and hydroxyapatite/ZnO based hydrogel, and the establishing Zn ions' behavior patterns with another composite components.

The Kі-67 marker for assessing the effectiveness of systemic or regional neoadjuvant polychemotherapy in patients with locally advanced breast cancer.

Over the past decades, breast cancer (BC) is the most common cancer and one of the key causes of mortality and disability among women in developed countries. AIM: Determination of the role of Ki-67 index in assessing the quality of neoadjuvant polychemotherapy treatment using regional or systemic delivery routes of pharmacological agents in patients with locally advanced breast cancer (LABC). MATERIALS AND METHODS: The retrospective analysis of 30 clinical trials of LABC treatment based on selective intra-arterial therapy in patients with BC (T4A-DN0-3M0) was used. RESULTS: The decrease in Ki-67 level in LABC after selective intra-arterial polychemotherapy was more pronounced than after systemic polychemotherapy. No correlation of the tumor metastatic potential with a Ki-67 level was detected. CONCLUSION: Assessment of Ki-67 expression allows to evaluate effectively the biological properties of the tumor, predict the course of the disease and choose the optimal tactics of neoadjuvant polychemotherapy (regional or systemic variant) as part of integrated antitumor treatment.

Conjugation of succinic acid to non-ionogenic amphiphilic polymers modulates their interaction with cell plasma membrane and reduces cytotoxic activity.

Pluronic block copolymers L61 and L121 were reacted with succinic anhydride to produce, respectively, their mono- and bisderivatives with succinic acid. The critical micelle concentration of Pluronics decreased after modification. The modification of Pluronic L61 promoted its association with the plasma membrane of human cells and increased membrane damage, while the membranotropic activity of modified Pluronic L121 reduced compared to the initial copolymer. Modified Pluronics interfered with the viability, apoptosis induction and metabolism of A549 cells and skin fibroblasts to a much lesser extent presumably due to the introduction of succinic acid residue inhibited intracellular penetration of copolymers. Modified Pluronic L121 promoted the cellular uptake of doxorubicin and rhodamine 123 in A549 cells attributed to the inhibition of membrane P-glycoprotein. Our study provides an approach to assessing the mechanism of interaction of amphiphilic polymers with living cells and demonstrates that Pluronic-succinic acid conjugates can be used as safe and efficient modulators of intracellular drug delivery.

Monitoring of the Zeta Potential of Human Cells upon Reduction in Their Viability and Interaction with Polymers.

The dynamic light scattering (DLS) technique was applied in order to assess the zeta potential of the plasma membrane of human cells. At pH 7.4, the cell zeta potential for different types of cells showed variations over a wide range and was equal to -19.4 ± 0.8 mV for HeLa cells and -31.8 ± 1.1 mV for erythrocytes. The difference could presumably be attributed to the differences in the biochemical composition of the cell plasma membrane. As a result of the heating of HeLa cells, the zeta potential shifted towards more negative voltages by 4.2 mV. An increase in the zeta potential correlated with an increase in the content of phosphatidylserine on the cell surface, which is considered to be an early marker of apoptosis. The DLS technique was also used to study the interactions between the cells and membranotropic polymers, such as polycations and nonionogenic Pluronic L121.

[Carbon nanotube-based biosensors for DNA structure characterization].

The possibility of DNA detection using electrodes modified with carbon nanotubes (CNTs) was studied. CNTs facilitate the electrochemical oxidation of DNA guanine nucleotide, which allows direct detection of DNA on a modified electrode. Electrochemical properties of DNA depend on its secondary structure and molecular weight. Denaturation of native DNA improves the adsorption of biopolymer on CNTs and results in an increase in DNA oxidation current on the modified electrode. A similar effect is observed after ultrasonic shearing of DNA or its treatment with Fenton's reagent due to the fragmentation of biopolymer. Our results demonstrate the feasibility of biosensors based on CNT-modified electrodes for the direct detection and characterization of DNA and DNA damaging factors.