Primary 3D Culture of Human Thyroid Follicle-Like Structures in Platelet Lysate-Based Gel.

The results of 3D culturing of human thyroid follicle-like structures in a gel based on platelet lysate at the gel-air interface are presented. During culturing up to 4 months, no new follicle-like structures were formed and none were destroyed. During the first 2 months, most follicle-like structures increased in size; then, their grown decelerated, but they retained viability. Ki-67+ cells were observed in the majority of follicle-like structures. Most of them produced thyroglobulin. Follicle-like structures get closer, the number of contacts between them increased, and cluster appeared. Thus, the developed 3D culturing system in a gel based on platelet lysate is an adequate approach for maintaining structure and functional activity of human follicle-like structures in vitro for at least 2 months.

Physicochemical and biological characteristics of chitosan/κ-carrageenan thin layer-by-layer films for surface modification of nitinol.

Thin films based on the natural polysaccharides κ-carrageenan (Carr) and chitosan (Chit) were formed by layer-by-layer deposition technique. Surface topography and mechanical characteristics (Young's modulus, adhesion strength) of the polymer films with different number of layers were determined using various modes of atomic force microscopy (AFM). Polymer films were used to deposit on the surface of nitinol (NiTi) plates. The creation of polysaccharide coatings on nitinol led to a change in surface properties, such as hydrophilicity and root mean square roughness. in vitro cytotoxicity assay for nitinol plates with and without polymer coating by the MG-63 osteoblast-like cell line was conducted and was shown that all the studied samples are not toxic. A decrease in cytotoxicity for samples with a polymer film consisting of 4 layers of chitosan and carrageenan was shown as compared to an uncoated nitinol plates.

Analysis of Reparative Activity of Platelet Lysate: Effect on Cell Monolayer Recovery In Vitro and Skin Wound Healing In Vivo.

Platelet lysate prepared from donor platelet concentrate and pooled according to a developed technique stimulates migration of multipotent mesenchymal stromal cells of the human adipose tissue and promotes healing of the monolayer defect in cultures of human fibroblasts and multipotent mesenchymal stromal cells in vitro in concentrations close those of fetal calf serum (5-10%). Lysate of platelets from platelet-rich rat blood plasma stimulated healing of the skin defect by promoting epithelialization and granulation tissue formation. The regenerative properties of platelet lysate in vivo increased with increasing its concentration.

Some Physical, Chemical, and Biological Parameters of Samples of Scleractinium Coral Aquaculture Skeleton Used for Reconstruction/Engineering of the Bone Tissue.

Physical and chemical (phase and chemical composition, dynamics of resorption, and strength properties), and biological (cytological compatibility and scaffold properties of the surface) properties of samples of scleractinium coral skeletons from aquacultures of three types and corresponding samples of natural coral skeletons (Pocillopora verrucosa, Acropora formosa, and Acropora nobilis) were studied. Samples of scleractinium coral aquaculture skeleton of A. nobilis, A. formosa, and P. verrucosa met the requirements (all study parameters) to materials for osteoplasty and 3D-scaffolds for engineering of bone tissue.

Criteria of biocompatibility of materials for bone defect repair.

We analyzed and systematized the criteria of biocompatibility of materials intended for reconstruction of bone defects on the basis of previously studied synthetic, natural, and composite biomaterials (carbonate hydroxyapatite, calcium phosphate cement, silica, natural corals, freshwater pearls, chitosan/carbonate hydroxyapatite) in the model of subcutaneous implantation to small laboratory animals.

Scleractinium coral aquaculture skeleton: a possible 3D scaffold for cell cultures and bone tissue engineering.

Cytocompatibility of 5 coral aquaculture skeleton species derived from two families (Acroporidae and Pocilloporidae) was studied over the course of in vitro culturing in continuous human fibroblast culture by the MMT test. Biocompatibility and capacity of scaffold to "transfer" cell cultures (specifically, multipotent mesenchymal stromal cells) to sites of implantation were studied in vivo by subcutaneous implantation of skeletal fragments to rats. All coral skeleton aquaculture specimens were cytocompatible (nontoxic and with surface matrix characteristics satisfactory for cells), biocompatible, and could be tried as 3D matrices for bone tissue engineering.

Human platelet lysate as a promising growth-stimulating additive for culturing of stem cells and other cell types.

We compared the composition and biological activity of fetal calf serum and platelet lysate from donor platelet concentrate. In platelet lysate, the concentrations of alkaline phosphatase, lactate dehydrogenase, creatinine, and mineral metabolism parameters were lower, while parameters of lipid and protein metabolism were higher than in fetal calf serum. The concentrations of growth factors (platelet-derived (AA, AB, BB), vascular endothelial, insulin-like, and transforming growth factor ß) in platelet lysate 1.7-148.7-fold surpassed the corresponding parameters in fetal calf serum. After replacement of fetal calf serum with platelet lysate in the culture medium (0, 25, 50, 75, and 100%), the count of multipotent mesenchymal stromal cells on day 7 (in comparison with day 1) increased by 154.8, 206.6, 228.2, 367.7, and 396.5%, respectively. Thus, platelet lysate can be an adequate non-xenogenic alternative for fetal calf serum.

In vitro study of matrix surface properties of porous granulated calcium phosphate ceramic materials made in Russia.

We performed in vitro screening of monophasic (hydroxyapatite, beta-tricalcium phosphate, carbonate-substituted hydroxyapatite with 0.59 and 5.9 wt% substitution with CO(3)(2-)) and biphasic (hydroxyapatite-tricalcium phosphate with various percentage of the components 80/20, 60/40, 20/80, silicon-substituted hydroxyapatite with 0.79 wt% SiO(2)) porous granulated ceramics composed of calcium phosphate powders synthesized by methods of heterophasic interaction of reagents and precipitation from aqueous solutions using MTT test and cultured human fibroblasts. Acute toxicity of materials (24-h incubation with cell culture) and matrix properties (3, 5, 7, 14, 18, 21, 28 days in culture) were evaluated. We selected a batch of materials obtained by precipitation from aqueous solutions, which were non-toxic and were characterized by good matrix properties (for cells). Biphasic ceramics with hydroxyapatite-tricalcium phosphate ratio of 80/20 exhibited best characteristics, and ceramics on the basis of silicon-substituted hydroxyapatite showed moderate characteristics.

Study of in vivo biocompatibility and dynamics of replacement of rat shin defect with porous granulated bioceramic materials.

Biocompatibility of porous granulated bioceramic materials (hydroxyapatite, beta-tricalcium phosphate, hydroxyapatite-b-tricalcium phosphate complex (80:20 wt%), carbonate-containing hydroxyapatite, and silicon-containing hydroxyapatite) was shown in a subcutaneous test on BDF1 mice. Dynamic (up to 8 months) observation showed gradual replacement of the granular substance with de novo forming bone tissue with hemopoiesis foci on a model of fenestral defect in the shin bone in Wistar rats. By the rate of resorption, the materials rank as follows: silicon-containing hydroxyapatite

Study of growth parameters and potentialities of differentiation of multipotent mesenchymal stromal cells from rat bone marrow in vitro.

The dynamics of growth and proliferative activity of the population of multipotent mesenchymal stromal cells from rat bone marrow was studied during 7 passages. The efficiency of colony formation, the morphology of multipotent mesenchymal stromal cells, and the possibility of spontaneous and induced differentiation were studied. The rat bone marrow fibroblast-like multipotent mesenchymal stromal cells are capable of clonal growth; their proliferative activity and the yield remained high until passage 4, but then decreased. Induction of osteo- or adipogenic differentiation of bone marrow multipotent mesenchymal stromal cells increased the percentage of morphologically modified cells carrying specific markers.

[Application of cell technologies to tissue defect closure in oncology].

In vivo and in vitro experiments on the application of cell technologies to tissue defect closure were conducted; autologic mesenchymal stem cells on 3-demensional matrices were used. The authors analyze the results of the application of bioengineering tissue equivalents for the closure of soft tissue and upper airway defects after extensive resections performed in 52 oncological patients. Tissue equivalents with stem cells provide engraftment and long-term graft functioning; they also modify wound surface, thus stimulating wound epithelization. In this study the application of tissue equivalents led to wound healing and functional recovery in 87% of patients.

Effects of various modes of sonication with low frequency ultrasound on in vitro survival of human tumor cells.

The effect of low frequency ultrasound in the cavitation (5-15 sec exposure) and subcavitation (5-210 sec exposure) modes on in vitro survival of cultured human tumor cells was studied. Analysis of the dose-effect curves and mathematical estimation of the effects low-frequency ultrasonication in the subcavitation mode on cell membranes helped to choose time intervals of tumor cell sonication ensuring potentiation of the cytostatic effects.

[The prediction of the individual chemosensitivity of human malignant tumors: the biological patterns and clinical results]. / Prognozirovanie individual'noi khimiochuvstvitel'nosti zlokachestvennykh opukholei cheloveka: biologicheskie zakonomernosti i klinicheskie rezul'taty.

Individual laboratory prediction of the sensitivity of human neoplasms to chemicals is justified on the basis of the authors' own data and those available in the literature. The potentialities and limitations of methods for individual prediction, as well as clinical results are discussed. Data are given on some biological mechanisms responsible for tumor growth which have been obtained in in vitro and in vivo tests with human tumor tissue and carcinoma cells. Further perspectives of this line of studies are discussed.

[The development and testing of an MTT method for assessing the reaction of peripheral blood lymphocytes to irradiation]. / Razrabotka i aprobatsiia MTT-metoda dlia otsenki reaktsii limfotsitov perifericheskoi krovi na obluchenie.

The semiautomatic MTT assay for evaluation of differences in the reaction of human donors' peripheral blood lymphocytes to ionizing radiation was developed. The stimulation conditions and period of cultivation were determined, and redox activity of lymphocytes and their coefficient of stimulation after irradiation with different doses were defined.

[A comparative evaluation of 2 methods for cloning human lung tumors]. / Sravnitel'naia otsenka dvukh sposobov klonirovaniia opukholei legkogo cheloveka.

Clonogenic capacity of tumor cells from 59 human lung cancer specimens was investigated in Petri dishes (24 specimens) and in capillaries (40 specimens), and parallel cloning was done in 5 cases. The results were as follows: (a) bacterial contamination in capillaries was noted less frequently than in Petri dishes (12.5 vs. 25%); (b) cells did not form colonies in capillaries in 42.5% and in Petri dishes in 25%; (c) colony-forming effectiveness in capillaries was 3.46-fold higher than that in Petri dishes; (d) the amount of tumor cells for analysis in capillaries is 10-fold less than that for analysis in Petri dishes. However, the absolute number of colonies, indispensable for assessment of tumor sensitivity to antitumor action (not less than 15 colonies per capillary or Petri dish) was obtained in 67% in Petri dishes and in 33% only in capillaries.

[Preparation of tumor cells from human lung cancer tissue for the purpose of cloning]. / Poluchenie opukholevykh kletok iz tkanei raka legkogo cheloveka s tsel'iu klonirovaniia.

Various approaches to isolation of tumor cells are analyzed on the basis of examination of 46 human pulmonary tumors of different histologic types. Mechanical treatment alone resulted in the death of the majority of tumor cells. Short trypsin treatment (for 10-15 min in several stages) proved to be the most suitable for disaggregation of small-cell carcinoma, adenocarcinoma, and carcinoid samples. A specific approach has been developed for isolation of cells from squamous-cell carcinoma, consisting in tumor treatment with 0.25 percent trypsin at 4 degrees C for 16-18 h, followed by DNAase, collagenase, and trypsin treatment, and then again trypsin treatment at 37 degrees C for 10-15 min. The suggested approaches permit a harvest of at least 6 x 10(6) tumor cells from 1 g of tissue, with more than 70 percent of these cells viable.