Cancer, one of the world's deadliest diseases, is expected to claim an estimated 16 million lives by 2040. Three-dimensional (3D) models of cancer have become invaluable tools for the study of tumor biology and the development of new therapies. The tumor microenvironment (TME) is a determinant of tumor progression and has implications for clinical therapies. Cancer-associated fibroblasts (CAFs) are one of the most important components of the TME. Modeling the interactions between cancer cells and CAFs in vitro can help to create biomimetic tumor equivalents for elucidating the causes of cancer growth and assessing the effectiveness of therapies. Here, we are investigated the effect of the mutual arrangement of tumor cells and fibroblasts on the formation of tumor models and their biomimetic properties. Pancreatic tumor models of three different designs were formed by the bioprinting method. Gelatin-alginate hydrogels with and without PANC-1 (pancreatic cancer) and NIH/3 T3 (mouse fibroblasts) cells, as well as their homo- and heterospheroids, were used as bioink. To enable bioprinting, we have chosen the most suitable compositions of alginate and gelatin that provide both good printability and cell proliferation activity. We also have investigated the kinetics of spheroid formation to identify the optimal cultivation parameters for achieving spheroid sizes suitable for bioprinting. All tumor models remained viable for 3-4 weeks. At the same time, the patterns of model development in the cultivation process and the biomimetic properties of the final tissue-engineered structures depended on the model design.

A new promising approach of creating wound bandages has been proposed. Composite nanofibers based on polycaprolactone (PCL) were obtained by electrospinning using PCL solutions containing 2, 4 and 6 wt% CuO nanoparticles (NPs). To immobilize wound-healing antibiotics on the surface of the PCL nanofiber, a COOH polymer layer was formed by plasma treatment in an Ar/CO2/C2H4 gas mixture. Functional density calculations have shown that the introduction of carboxyl, amide and 1-ethyl-3-(3-dimethylaminopropyl)carbodiimide (EDC) activated carboxyl groups on the PCL surface increases the ability to bind antibiotics due to the formation of hydrogen bond networks. Baneocin, covalently bound to the PCL surface through a COOH polymer layer, increases surface wettability and slows the release of Cu2+ ions, thereby providing long-term and sustainable leaching of bactericidal ions. The modified nanofibers demonstrate excellent bactericidal and fungicidal activity against the bacterial strains Staphylococcus aureus BAA1707 (MW2), Pseudomonas aeruginosa C3945/23, Enterococcus faecium Ya253, Escherichia coli U20 and Acinetobacter baumannii C66627/23, as well as the slightly harmful fungal strain Candida auris KA10. For the first time, Baneocin has been shown to significantly reduce the cytotoxicity of substances and increase fibroblast survival. PCL and PCL-COOH-Ban membranes are non-toxic to cellular fibroblasts, while the addition of CuO results in dose-dependent toxicity. The PCL and PCL-COOH-Ban materials with 2 wt% CuO did not cause hemolysis, indicating excellent blood-related properties. The hemostatic efficacy of the PCL-CuO_2%-COOH-Ban sample, assessed using a mouse tail amputation model, was found to be significantly higher than that of the control group (reduction in blood loss and tail bleeding time by 4.6 and 3.8 times, respectively).

Zinc oxide (ZnO) nanoparticles (NPs) have been investigated for various skin therapies in recent years. These NPs can improve the healing and modulate inflammation in the wounds, but the effective and biosafety concentration in such changes are yet to be known. In this study, we have designed antibacterial, superabsorbent, and hemostatic composite dressings produced through the lyophilization of a curdlan and chitosan polymer combination, incorporating ZnO nanoparticles (NPs) at concentrations of X = 1, 3, 5, and 7 wt% (CUR/CS_ZnO-X%). ZnO nanoparticles are uniformly dispersed within the obtained materials. An increase in ZnO NP concentration correlates with an enhancement in the specific surface area of the composite foams, attributed to the creation of larger pores. Despite all samples demonstrating identical capacity to absorb DMEM cell media (1500 wt%), the swelling rate of the materials escalated with the incorporation of zinc particles and attained a 1500 wt% DMEM media per 9 s. Antibacterial assays demonstrated that CUR/CS_ZnO-3 % entirely inhibited Staphylococcus aureus CSA154, Escherichia coli U20, and Klebsiella pneumoniae C324/23 strains for 24 h, while diminishing the colony-forming units (CFUs) of Clostridium perfringens D46 strain, responsible for gangrene, and the Acinetobacter baumannii C66627/23 strain by 99 %. All samples demonstrated excellent hemostatic properties: the measured blood coagulation indexes of samples were determined in diapason 14-32 %.

Local drug delivery systems based on bioceramics ensure safe and effective treatment of bone defects and anticancer therapy. A promising drug delivery scaffold material for bone treatment applications is diopside (CaMgSi2O6) which is bioactive, degradable, and possesses drug-release ability. Currently, in vitro assessment of drug release from biomaterials is performed mostly on a 2D cell monolayer. However, to interpret and integrate biochemical signals, cells need a 3D microenvironment that provides cell-cell and cell-extracellular matrix interactions. In this regard, 3D cell models are gaining popularity. In this work, we proposed the protocol for evaluation of the effect of doxorubicin released from diopside on MG-63 cells and primary human fibroblasts in 3D culture conditions. Tissue spheroids with similar diameters were incubated with doxorubicin-loaded diopside for 72 h, the amount of diopside was calculated in accordance with the required doxorubicin concentration. We demonstrated that doxorubicin is gradually released from diopside and exhibits an activity similar to that of the pure drug at the same total concentration. It is important to note that doxorubicin was more potent on MG-63 spheroids compared to HF spheroids, which confirmed the reliability of spheroids as 3D models of tumor and healthy tissues.

This work presents the results of studies of the features, mechanisms and effectiveness of the photodynamic effect of PS based on polycationic derivatives of long-wave phthalocyanines and synthetic bacteriochlorin on bacteria, bacterial biofilms, tumor cells and experimental animal tumor model.

Superelastic Ti-18Zr-15Nb alloy capable of mimicking the mechanical behavior of a bone tissue is a promising biomaterial but suffers from the lack of antibacterial properties. To address this problem, we developed a combined surface treatment method: formation of a porous sub-surface layer, deposition/precipitation of Au nanoparticles (AuNPs), surface functionalization with cysteine amino acid and grafting of gentamicin. The sizes distribution and AuNPs content were greatly effected by the synthesis methods. The AuNPs with an average size of 3 nm were obtained by precipitation from a AuNP colloidal solution. Larger AuNPs were formed by preliminary alloy functionalization in a NaBH4 solution followed by treatment in a HAuCl4 solution. Successful surface functionalization with L-cysteine and attachment of gentamicin were confirmed by XPS analysis. Due to the formation of stable cysteine-gentamicin complexes attached to AuNPs, the materials showed high antibacterial activity against Escherichia coli and Staphylococcus aureus strains. Better antibacterial properties are ascribed to fine isolated AuNPs as compared to larger ones. Cytocompatibility was assessed for osteoblast cells. In the case of smaller AuNPs, an accelerated restoration of osteoblastic cell proliferation and a more organized actin cytoskeleton were observed. Hemolytic activity of the developed materials was investigated. Functional mechanical properties were not affected by the surface treatment.

Background: One of the tasks of anticancer photodynamic therapy is increasing the efficacy of treatment of cancer nodes with large (clinically relevant) sizes using near-infrared photosensitizers (PS). Methods: The anticancer efficacy and mechanisms of the photodynamic action of PS based on polycationic de-rivatives of synthetic bacteriochlorin against Lewis lung carcinoma were studied in vitro and in vivo. Results: It was found that studied PS have high phototoxicity against Lewis lung carcinoma cells: the IC50 values were about 0.8 & mu;M for tetracationic PS and 0.5 & mu;M for octacationic PS. In vivo studies have shown that these PS provide effective inhibition of the tumor growth with an increase in the lifespan of mice in the group by more than 130%, and more than 50% survival of mice in the group. Conclusions: Photosensitizers based on polycationic derivatives of synthetic bacteriochlorin have high photody-namic efficacy caused by the induction of necrosis and apoptosis of cancer cells, including cancer stem cells, and a sharp decrease of mitotic and proliferative activity. Studied polycationic photosensitizers are much more effective at destroying cancer stem cells and newly formed cancer vessels in comparison with anionic photo-sensitizers, and ensure the cessation of tumor blood flow without hemorrhages and thrombosis.

Efficient screening of photosensitizers (PS) as well as studying their photodynamic activity, especially PS excited in the near-infrared region, require informative in vitro models to adequately reflect the architecture, thickness, and intercellular interactions in tumors. In our study, we used spheroids formed from human colon cancer HCT-116 cells and liver cancer Huh7 cells to assess the phototoxicity of a new PS based on tetracationic derivative of synthetic bacteriochlorin (BC4). We optimized conditions for the irradiation regime based on the kinetics of BC4 accumulation in spheroids and kinetics of spheroid growth. Although PS accumulated more efficiently in HCT-116 cells, characterized by more aggressive growth and high proliferative potential, they were less susceptible to the photodynamic therapy (PDT) compared to the slower growing Huh7 cells. We also showed that 3D models of spheroids were less sensitive to BC4 than conventional 2D cultures with relatively identical kinetics of drug accumulation. Our findings suggest that BC4 is a perspective agent for photodynamic therapy against cancer cells.

In view of their potential in the treatment of oncological diseases, photosensitizers (PS) have been the object of active research for the last years. The applied therapeutic photodynamic method involves the activation of a PS by light at a specific wavelength. PS interacts with oxygen and catalyzes the production of singlet oxygen molecules or radical oxygen species which leads to tumor cell death. This work is devoted to the study of the effectiveness of the photodynamic action of new PSs based on polycationic derivatives of synthetic bacteriochlorin on human lung cancer cells A549. The results obtained show that these PS effectively bind to these cells, have high phototoxicity and low ‘dark’ cytotoxicity for them. Morphological and immunohistochemical studies show that photodynamic effect leads to induction of cell necrosis and apoptosis, as well as a sharp decrease in mitotic activity.

Recently, spheroids have attracted the wide attention of researchers as 3D models for drug testing. One of the key features of spheroids is that they can be composed of one or more cell types. There is a hypothesis that the use of heterospheroids from several cell types when testing anticancer drugs can better reproduce the structure of the tumor and, as a result, increase the predictive value of the study. To verify this hypothesis, we tested the activity of eight antitumor drugs on homospheroids consisting of PANC-1 pancreatic cancer cells and heterospheroids from three types of cells: PANC-1 pancreatic cancer cells, primary human fibroblasts, and human umbilical vein endothelial cells. It has been found that the use of heterospheroids from several cell types, which more accurately reflect the tumor microenvironment, does not lead to a noticeable change in the activity of drugs. Subcutaneous transplantation was carried out of spheroids from PANC-1 to immunodeficient mice which showed that the obtained model of pancreatic cancer more effectively reproduces the aggressive-tumor phenotype compared to transplantation of cells in suspension.

In situ 3D bioprinting is a new emerging therapeutic modality for treating human skin diseases. The tissue spheroids have been previously suggested as a powerful tool in rapidly expanding bioprinting technology. It has been demonstrated that the regenerative potential of human dermal fibroblasts could be quantitatively evaluated in 2D cell culture and confirmed after implantation in vivo. However, the development of unbiassed quantitative criteria of the regenerative potential of 3D tissue spheroids in vitro before their in situ bioprinting remains to be investigated. Here it has been demonstrated for the first time that specific correlations exist between the regenerative potential of human dermal fibroblasts cultured in vitro as 2D cell monolayer with biological properties of 3D tissue spheroids fabricated from these fibroblasts. In vitro assessment of biological properties included diameter, spreading and fusion kinetics, and biomechanical properties of 3D tissue spheroids. This comprehensive characterization could be used to predict tissue spheroids’ regenerative potential in vivo.

L-lysine α-oxidase (LO) is an L-amino acid oxidase with antitumor, antimicrobial and antiviral properties. Pharmacokinetic (PK) studies were carried out by measuring LO concentration in plasma and tissue samples by enzyme immunoassay. L-lysine concentration in samples was measured spectrophotometrically using LO. After single i.v. injection of 1.0, 1.5, 3.0 mg/kg the circulating T1/2 of enzyme in mice varied from 51 to 74 min and the AUC0–inf values were 6.54 ± 0.46, 8.66 ± 0.59, 9.47 ± 1.45 μg/ml × h, respectively. LO was distributed in tissues and determined within 48 h after administration with maximal accumulation in liver and heart tissues. Mean time to reach the maximum concentration was highest for the liver—9 h, kidney—1 h and 15 min for the tissues of heart, spleen and brain. T1/2 of LO in tissues ranged from 7.75 ± 0.73 to 26.10 ± 2.60 h. In mice, plasma L-lysine decreased by 79% 15 min after LO administration in dose 1.6 mg/kg. The serum L-lysine levels remained very low from 1 to 9 h (< 25 μM, 17%), indicating an acute lack of L-lysine in animals for at least 9 h. Concentration of L-lysine in serum restored only 24 h after LO administration. The results of LO PK study show that it might be considered as a promising enzyme for further investigation as a potential anticancer agent.

Magnetic tissue engineering is one of the rapidly emerging and promising directions of tissue engineering and biofabrication where the magnetic field is employed as temporal removal support or scaffold. Iron oxide nanoparticles are used to label living cells and provide the desired magnetic properties. Recently, polymer microcapsules loaded with iron oxide nanoparticles have been proposed as a novel approach to designing magnetic materials with high local concentrations. These microcapsules can be readily internalized and retained intracellularly for a long time in various types of cells. The low cytotoxicity of these microcapsules was previously shown in 2D cell culture. This paper has demonstrated that cells containing these nontoxic nanomaterials can form viable 3D tissue spheroids for the first time. The spheroids retained labeled fluorescent microcapsules with magnetic nanoparticles without a detectable cytotoxic effect. The high concentration of packed nanoparticles inside the microcapsules enables the evident magnetic properties of the labeled spheroids to be maintained. Finally, magnetic spheroids can be effectively used for magnetic patterning and biofabrication of tissue-engineering constructs.

Introduction. Transformed cells are selectively vulnerable to a deficiency of certain amino acids. L-Lysine α-oxidase (LO) catalyzes the oxidative deamination of L-lysine, resulting in lysine depletion and hydrogen peroxide production. In preclinical models with intravenous and intraperitoneal administration LO shows a wide spectrum of antitumor activity. The purpose of this study was to determine the antitumor effect of orally administered to mice LO under tolerance control. Materials and methods. LO efficiency was studied on models Ca755 and SW620 when administered orally, evaluated using adequate criteria and the method of variation statistics and statistically significant differences at p≤0.05. Results. The antitumor activity and tolerance of LO from Trichoderma cf. aureoviride Rifai BKM F-4268D when administered orally was studied for the first time on murine and human tumor models. It was shown that LO in total doses of 6000-8000 U/kg (discrete mode of administration) reliably and significantly inhibits the growth of mouse breast adenocarcinoma Ca755, Tumor Growth Inhibition, TGImax=80-89% (p<0.05) (criterion of TGI≥50%). A smaller, but reliable effect was obtained on SW620 human colon cancer xenografts at a total dose of 6000 U/kg, T/C=75% (p<0.05) (criterion of T/C≤42%). By oral administration LO did not cause any side effects or death of mice from toxicity in the range of studied doses. Conclusion. This study makes it possible to consider LO promising for preclinical study with oral administration.

Oncolytic viral therapy is a promising approach to targeted therapy of malignant tumors. In this article we consider the therapeutic potential of a non-pathogenic Coxsackie A7 virus (CA7V) with neurotropic properties on a model of human neuroblastoma. Purpose to study in vitro/in vivo sensitivity of human neuroblastoma HNB (from cell line JMR-32) to Coxsackie virus A7 (CA7V). Objectives: еvaluation of cytolytic activity in vitro on NB cells verified by cytomorphology and assessment of dynamics of the growth of subcutaneous neuroblastoma xenografts in Balb/c nude male mice exposed to CA7V multiple i.v. injections. Material and methods. CA7V was produced in the cells of line-producer С-33А. Cell culture and the strain of transplanted NB (JMR-32) were obtained from the Collection of N.N. Blokhin Russian Cancer Research Center. Cytomorphologic verification of neuroblastoma and CA7V cytolytic activity were executed with the use of standard cultural methods, TCID50 and IC50 criteria. Experiments «in vivo» were performed on immunodeficient Balb/c nude male mice bred and reared in the N.N. Blokhin Russian Cancer Research Center. The experiments were made at day 6 when neuroblastoma subcutaneous xenografts developed to the Vmean = 79-82 mm3 by day 6. The treatment with CA7V at the i.v. single dose of 1×108 cells per mouse was performed 3 times with 72-hours intervals; evaluation of the efficacy was made according to standard criterion Т/С ≤ 42%; and control of the tumor growth rate (Vt/V0) in the dynamics. Statistical assessment was made with the software Excel for Windows 2007 with the use of T-test under p ≤ 0.05. Results. Cytolytic effect of CA7V on neuroblastoma cells was registered similar to basic parameters of the original line-producer С-33А: TCID50 = 0.99×10-4 pfu/cell, and IC50 = 1.11×10-4 pfu/cell; 48 and 72 hours after virus reproduction in NB cells the rate was 2.0 and 1.5-fold higher than in the line-producer cells. СA7V inhibiting effect on the growth of large subcutaneous neuroblstoma xenografts is registered after the first i.v. injection at the minimal level of T/C = 67% (criterion ≤ 42%) with the 1.5-fold decrease of the tumor growth rate and cancellation of early mice death by day 22 vs day 15 in the control group of untreated mice (n = 8). Conclusion. The obtained results allow to consider human neuroblastoma (JMR-32) to possess the low sensitivity to oncolytic effect of in vitro/in vivo. In order to obtain significant effect in vivo the treatment should be started in mice with 2-fold smaller tumors and a higher initial dose of the oncolytic agent.

The aim of this study was to estimate the efficacy of mesenchymal stem cell-based suicide gene therapy in mice bearing murine melanoma B16F10. Adipose mesenchymal stem cells (MSCs) were transfected with plasmid constructs expressing cytosine deaminase fused with uracil phosphoribosyltransferase (CDA/UPRT) or CDA/UPRT fused with HSV-1 tegument protein VP22 (CDA/UPRT/VP22). In this study, we demonstrate that direct intratumoral transplantation of MSCs expressing CDA/UPRT or CDA/UPRT/VP22 followed by systemic administration of 5-fluorocytosine (5-FC) results in a significant inhibition of tumor growth. There was a 53% reduction in tumor volume in mice treated with CDA/UPRT-MSCs and 58% reduction in tumor volume in mice treated with CDA/UPRT/VP22-MSCs as compared with control animals transplanted with B16F10 melanoma alone. Injection of CDA/UPRT-MSC and CDA/UPRT/VP22-MSC prolonged the life span of mice bearing B16F10 melanoma by 15 and 26%, respectively. The data indicate that in murine B16F10 melanoma model, MSCs encoding CDA/UPRT suicide gene have a significant antitumor effect.