[Development of osteoplastic gene-activated 3D matrices based on adenoviral vectors]. / Razrabotka osteoplasticheskikh gen-aktivirovannykh 3D-matriksov na osnove adenovirusnykh vektorov.

Gene therapy is one of the most promising approaches in regenerative medicine for the restoration of extensive bone defects in dentistry and maxillofacial surgery. Matrices obtained using three-dimensional printing from bioresorbable polymers, impregnated with adenoviral constructs with genes for osteoinductive factors, can ensure safe and effective formation of bone tissue. OBJECTIVE: To study the properties of three-dimensional matrices based on polylactic-co-glycolic acid and adenoviral constructs with the GFP gene in vitro. MATERIALS AND METHODS: The matrices were obtained by antisolvent three-dimensional printing. Transduction efficiency was assessed by fluorescence microscopy and flow cytometry. The cytocompatibility of the matrices was assessed by the MTT test and by staining cells with fluorescent dyes. RESULTS: Matrices based on polylactic-co-glycolic acid have high cytocompatibility on adipose tissue-derived mesenchymal stem cells. Impregnation of adenoviral vectors with the green fluorescent protein gene in 3D matrices ensures the release of viral particles within a week, maintaining their high transducing ability. CONCLUSION: The developed method for obtaining gene-activated matrices can serve as the basis for the creation of effective osteoplastic materials for bone regeneration.

Comparative Analysis of Proliferation and Viability of Multipotent Mesenchymal Stromal Cells in 3D Scaffolds with Different Architectonics.

3D biodegradable materials (scaffolds) containing bioactive hydroxyapatite molecules fabricated by foaming in supercritical carbon dioxide and by selective laser sintering were used for culturing of mesenchymal stromal cells from the human adipose tissue. Experiments showed that stromal cells from the human adipose tissue adhered and proliferated on all studied types of structures. Addition of hyproxyapatite to the scaffold stimulated proliferation of stromal adipose tissue cells.

[Kinetics of methyluracil release from bioresorbable polymeric carriers].

Bioresorbable poly(lactic-co-glycolic) matrix-carriers containing 20 wt. % of 6-methyluracil (MU) have been prepared by supercritical fluid monolithization without organic solvents. Raman spectroscopy was used to analyze both the spatial distribution MU over polymer matrices and the MU release kinetics from the carrier into phosphate buffer solution. It was found that, during the first 24 h, the amount of released MU did not exceed 15-20% of its encapsulated content. After that, the MU release kinetics passed to almost linear regime with simultaneous retarding of the process. On the 40th day of observation, the MU content in solution reached up to 80% of its initial content in the carriers. Thus, using 6-methyluracil as a model, it was shown that the proposed bioresorbable and bioactive composites can be used as matrix-carriers for targeted and long-term drug release.

Design of tissue engineering implants for bone tissue regeneration of the basis of new generation polylactoglycolide scaffolds and multipotent mesenchymal stem cells from human exfoliated deciduous teeth (SHED cells).

Cultures of multipotent mesenchymal stromal cells from the pulp of human deciduous teeth (SHED cells) were characterized. The cells were used for population of 3D biodegradable polylactoglycolide scaffolds; their osteogenic potential was preserved under these conditions. Implantation of the scaffolds to mice induced no negative reactions in the recipients. These results suggest that the use of polylactoglycolide scaffolds populated with SHED cells is a promising approach for creation of implants for bone defect replacement.

Biocompatibility of tissue engineering constructions from porous polylactide carriers obtained by the method of selective laser sintering and bone marrow-derived multipotent stromal cells.

We studied the biocompatibility of porous polylactide carrier matrices obtained by means of surface selective laser sintering. Carrier matrices had no cytotoxic activity, but maintained adhesion and proliferation of cells. Subcutaneous transplantation of tissue engineering constructions from these carriers and bone marrow-derived multipotent stromal cells did not cause the inflammatory response and pathological changes in rats. The conditions for organotypic regeneration were provided at the site of transplantation (high degree of blood supply and considerable amount of immature precursor cells).

[Regeneration processes in bone defects after implantation of composite material of different density of polylactide origin filled with HAP (experimental-morphological study)].

In experimental-morphological study on 6 dogs the dynamics of regenerate formation in ulna and mandible defects after implantation in them composite material of different density (0.46-0.50 and 0.38-0.42 g/cm(3)) of polylactide (PL) origin filled with HAP was followed at the terms of 6 and 9 months. Histologic study and structural determinant distribution analysis in the content of regenerate showed that optimal results according to the bone defect substitution by bone regenerate criterion at 9th month of the experiment were received after composite material from PL and HAP with the density of 0.38-0.42 g/cm(3) implantation. Newly formed trabecular bone tissue was seen in the regenerate and as well as strong tendency for bone matrix maturation. It was confirmed by the appearance of lamellar structures in newly formed bone trabecules situated in peripheral zones of bone defect. In bone mandible defects the substitution process of the implants from PL with HAP by the bone tissue was much slower than in ulna defects.

Atomic force microscopic study of the surface morphology of apatite films deposited by pulsed laser ablation.

Atomic force microscopy (AFM) has been used to study the surface morphology of apatite films deposited on metallic and polyethylene substrates by laser ablation using KrF and transversely excited atmospheric CO2 lasers. The films are found to consist of a smooth apatite coating with macroparticles scattered on the surface. A wide variety of macroparticles, differing in size, shape and roughness, were found and analysed employing the high spatial resolution of AFM (< 1 nm). We have investigated the correlation between the apatite film morphology and the deposition conditions. Of particular importance are laser fluence, gas pressure, the nature of the target and the substrate temperature. We have explained these dependencies on the basis of a theoretical model which includes evaporation and a cluster-type laser ablation mechanism.

Osteoblast growth on titanium foils coated with hydroxyapatite by pulsed laser ablation.

Pulsed laser ablation is a new method for deposition of thin layers of hydroxyapatite (HA) on to biomaterial surfaces. In this paper, we report activity and morphology of osteoblasts grown on HA surfaces fabricated using different laser conditions. Two sets of films were deposited from dense HA targets, at three different laser fluences: 3, 6 and 9 Jcm(-2). One set of the surfaces was annealed at 575 degrees C to increase the crystallinity of the deposited films. Primary human osteoblasts were seeded onto the material surfaces and cytoskeletal actin organisation was examined using confocal laser scanning microscopy. The annealed surfaces supported greater cell attachment and more defined cytoskeletal actin organisation. Cell activity, measured using the alamar Blue assay, was also found to be significantly higher on the annealed samples. In addition, our results show distinct trends that correlate with the laser fluence used for deposition. The cell activity increases with increasing fluence. This pattern was repeated for alkaline phosphatase production by the cells. Differences in cell spreading were apparent which were correlated with the fluence used to deposit the HA. The optimum surface for initial attachment and spreading of osteoblasts was one of the HA films deposited using 9 J cm(-2) laser fluence and subsequently annealed at 575 degrees C.

Biocompatibility of Laser-deposited Hydroxyapatite Coatings on Titanium and Polymer Implant Materials.

We have investigated the biocompatibility of calcium phosphate coatings deposited by pulsed laser ablation from hydroxyapatite (HA) targets onto polyethylene and Teflon substrates. It was found that the cell density, attachment, and morphology of primary rat calvaria osteoblasts were influenced by both the original polymer and by the nature of the apatite coatings. HA coatings on Teflon were found to have higher biocompatibility in terms of cell adhesion and spreading. In vivo studies of bone response to coatings deposited by KrF excimer and CO2 lasers on commercial Ti6A14V alloy implants show that both deposition techniques suppress fibrous tissue formation and promote osteogenesis. © 1998 Society of Photo-Optical Instrumentation Engineers.

Thermal and diffusion processes in laser-induced stress relaxation and reshaping of cartilage.

The thermodynamic characteristics of the 'bound-to-free' phase transformation of water are studied by means of differential microcalorimetry and FTIR spectroscopy. This phase transition controls the stress relaxation and reshaping of cartilage which we have observed previously under moderate laser heating. It is shown that the FTIR spectrum of bound water in cartilage differs from that of free water in cartilage, and that both show differences to the FTIR spectrum of pure water. The proportion of bound water in cartilage is found to be of the order 4%. We have examined water liberation and absorption kinetics and found them to be controlled by diffusion through the tissue and also by the bound-to-free water transformation. The theoretical modelling and experimental data allowed calculation of diffusion coefficients and the activation energy for water transfer. The latter was found to be close to the heat of phase transformation of water. We have established that the drying and wetting processes in cartilage are reversible (fully or in a part, depending on the temperature of preliminary drying) and can be described by identical solutions of the diffusion problem, with coefficients of water diffusion being the same. The mechanism of water diffusion in cartilaginous tissue is also discussed.

Daily copper supplement effects on copper balance in trained subjects during prolonged restriction of muscular activity.

The aim of this study was to assess the effect of a daily intake of copper supplements on negative copper balance during prolonged exposure to hypokinesia (decreased number of kilometers per day). During hypokinesia (HK), negative copper balance is shown by increased, not by decreased, serum copper concentration, as it happens in other situations. Studies were done during a 30-d prehypokinetic period and a 364-d hypokinetic period. Forty male trained volunteers aged 22-26 yr with a peak oxygen uptake of 66.4 mL/min/kg and with an average of 13.7 km/d running distance were chosen as subjects. They were equally divided into four groups: unsupplemented ambulatory control subjects (UACS), unsupplemented hypokinetic subjects (UHKS), supplemented hypokinetic subjects (SHKS), and supplemented ambulatory control subjects (SACS). The SACS and SHKS groups took 0.09 mg copper carbonate/kg body weight daily. The SHKS and UHKS groups were maintained under an average running distance of 1.7 km/d, whereas the SACS and UACS groups did not experience any modifications in their normal training routines. During the 30-d prehypokinetic period and the 346-d hypokinetic period, urinary excretion of copper, calcium, and magnesium and serum concentrations of copper, calcium, and magnesium were measured. Copper loss in feces and copper balance was also determined. In both UHKS and SHKS groups, urinary excretion of copper, calcium, and magnesium and concentrations of copper, magnesium, and calcium in serum increased significantly when compared with the SACS and UACS groups. Loss of copper in feces was also increased significantly in the SHKS and UHKS groups when compared with the UACS and SACS groups. Throughout the study, the copper balance was negative in the SHKS and UHKS groups, whereas in the SACS and UACS groups, the copper balance was positive. It was concluded that a daily intake of copper supplements cannot be used to prevent copper deficiency shown by increased copper concentration. Copper supplements also failed to prevent negative copper balance and copper losses in feces and urine in endurance-trained subjects during prolonged exposure to HK.

Changes in thyroid hormones and lipids in endurance trained volunteers during acute and rigorous bed rest conditions.

The aim of this study was to evaluate the effect of an acute rigorous bed rest (ABR) (abrupt confinement to a rigorous bed rest regimen) and a rigorous bed rest (RBR) regimen on serum concentrations of thyroid hormones and lipids. The studies were done during seven days of a pre bed rest period and during seven days of an ABR and RBR period. Thirty endurance trained male volunteers aged 22 to 26 years with a peak oxygen uptake of 66.0 mL.min-1.kg-1 and running an average of 14.0 km.day-1 were chosen as subjects. They were divided equally into three groups: (one) ten athletes placed under ambulatory conditions served as ambulatory control subjects (ACS), (two) ten athletes subjected to an acute rigorous bed rest regimen served as acute bed rested subjects (ABRS) and (three) ten athletes submitted to a rigorous bed rest regimen served as rigorous bed rested subjects (RBRS). For the simulation of the effect of ABR the ABRS group was submitted abruptly to a rigorous bed rest regimen. They did not have any prior knowledge of the exact date and time when they would be asked to submit to the RBR. For the simulation of the effect of RBR the RBRS group was subjected to a rigorous bed rest regimen on a predetermined date and time known to them right from the start of the study. During the pre bed rest period and during the ABR and RBR periods serum concentrations of thyroxine (T3), triiodothyronine (T4), thyroid-stimulating hormone (TSH), cholesterol, triglycerides, total protein and albumin were measured. In the ABRS and RBRS groups serum concentrations of T3, T4, TSH, cholesterol, triglycerides, total protein and albumin increased significantly when compared with the ACS group. Serum concentrations of thyroid hormones and lipids in the ABRS group increased much faster and were more pronounced than in the RBRS group. It is concluded that exposure to RBR and, especially, to ABR conditions induces significant increases in serum concentrations of thyroid hormones and lipids. These changes occurred much earlier and were much greater in the ABRS group than in the RBRS group.

Evolution of the regulation of the environmental radiation protection and monitoring in the Russian Federation.

The evolution of the Russian Federation legislation and regulation concerning the radiation protection of the population, the environment and the monitoring is presented. The main federal laws--"On the Environmental Protection", "On the Atomic Energy Use", "On the Radiation Safety of Population", etc. as well as standards of radiation safety, sanitary and hygienic norms, international agreements and conventions are briefly summarized. Main directions towards further improvement of the environmental radioprotection regulation in Russia are pointed out.

Morphometric examination of glomerulus and juxta glomerural system of rat kidney during prolonged restriction of motor activity.

The aim of this study was to examine the structural changes of glomerular and juxta glomerular system (JGS) of the kidney of rats during prolonged restriction of motor activity (hypokinesia). The studies were performed during 90 days of hypokinesia (HK) on 144 male Wistar rats divided into two groups: Group one placed under ordinary vivarium conditions and serving as vivarium control rats (VCR) and Group two subjected to HK and serving as hypokinetic rats (HKR). For the simulation of the hypokinetic effect the HKR group was kept in small individual cages made of wood that restricted the movements of rats in all directions without hindering food and water intake. During a prehypokinetic period of 15 days and the hypokinetic period of 90 days, body weight and food intake were measured and morphometric examinations were done to measure the cortical and juxta medullary glomerulus on kidney sections from the VCR and HKR groups (eight rats from each group). Body weight and food intake decreased significantly in the HKR groups when compared with the VCR group. Kidney weight of rats increased, the superficial volume decreased and that of the juxta medullary glomerules increased, whereas juxta glomerular granularity indexes decreased significantly in the HKR group when compared with the VCR group. The measured parameters did not change significantly in the VCR group when compared with the baseline control values. It was concluded that prolonged exposure to HK induces a significant increase in the kidney weight and a relationship appeared between variations of the volume of cortical and juxta medullary glomerulus and the function of the juxtaglomerular apparatus.

[Relation of optokinetic nystagmus and the width of the visual field]. / Zavisimos't optokineticheskogo nistagma ot shiriny polia zreniia.

Optokinetic nystagmus (OKN) was elicited in humans by a moving vertical grating wide 30-170 degrees. A large range of velocities (10-120 degrees/sec) and a constant spatial stimulus frequency (0.13 cycle/degree) were used. It has been shown that the qualitative and quantitative OKN parameters can be controlled by stimulus area width. The optimal OKN was found under conditions of the whole field stimulation. Progressive masking of the periphery caused a OKN reduction whose magnitude was fixed in different stimulus ranges. The elimination of central stimulation resulted in a complete OKN suppression. The role of the stimulation eccentricity and of the stationary edges in the observed OKN changes is discussed. The findings offer a possibility for a more precise evaluation of OK responses.

[Enhancing the biocompatibility of dentures made from polymethylmethacrylate by using hydroxyapatite]. / Povyshenie biologicheskoi sovmestimosti zubnykh protezov iz polimetilmetakrilata s pomoshch'iu gidroksiapatita.

Twenty or 50 weight-percent of hydroxyapatite (Polystome) was added in suspensions of AKR-15 copolymer and acid monomers, then the mixtures were molded and subjected to thermal treatment. Admixtures (supercritical CO2) were extracted from the composites in parallel with infrared spectroscopic measurement of the concentration of methylmetacrylate toxic monomers. Hydroxyapatite in 20% concentration decreased the release of toxic substances from the composite, whereas 50% concentration of the agent did not produce this effect. Three patients with intolerance of acrylic plastic were effectively treated using this method.

[Bone marrow hematopoiesis in polymer composite implants in long-term bone marrow cultures]. / Issledovanie protsessov kostnomosgovogo krovetvoreniia na implantatakh iz polimernykh kompozitov v dlitel'nykh kul'turakh kostnogo mozga.

The morphological composition of cells on implants made of different composition materials was studied on the model of long-term bone marrow cultures at cultivation for 28 days. Three samples of materials were investigated: polymethyl methacrylate (PMMC), PMMC with hydroxyapatite (HA) and PMMC with HA and subsequent processing with supercritical carbon dioxide. A gradual rise in total number of cells was observed on the implants for 4 weeks as well as increasing number of stromal and hemopoietic cells. With increasing time of cultivation the share of stromal cells was growing while the percent of hemopoietic cells fell. The greater number of hemopoietic cells was observed on the samples of the implants made from composites containing HA vs implants made from the initial PMMC.

[The tissue reaction to acrylic plastics modified by supercritical extraction with carbon dioxide]. / Tkanevaia reaktsiia na akrilovye plastmassy, modifitsirovannye sverkhkriticheskoi ékstraktsiei dvuokis'iu ugleroda.

The process of extraction of admixtures from acryl plastic widely used in dentistry by means of supercritical carbon dioxide (sc-CO2) was studied and effects of extraction conditions on biocompatibility and toxicity of resultant materials assessed, sc-CO2 effectively purified the specimens from toxic compounds (monomers and low-molecular oligomers, methylmethacrylate, dichloroethane) and notably improved the biocompatability of polymer implants. Tissue reaction to ethacryl and protacryl depends on the degree of implant polymerization and duration of extraction of toxic substances from polymer.

[Biocompatible coatings for metallic implants obtained by laser spraying]. / Biosovmestimye pokrytiia dlia metallicheskikh implantov, poluchennye lazernym napyleniem.

Use of various lasers and regimens for spraying the coatings and for additional thermal processing makes it possible to purposefully alter the morphology, composition, and crystallinity of the coatings, and hence, the optimal coatings, as regards their biological activity, may be created. Replacement of eximer lasers with CO2 lasers makes the spraying process cheaper and comparable in efficacy with the plasma process, with the quality of coatings and reproducibility of their parameters appreciably improved. Experiments with coatings of three types, sprayed by CO2 lasers, demonstrated that they met the mechanical and physico-chemical requirements of biocompatibility.

[The structural characteristics of the bony tissue at the surface of an implant with hydroxyapatite spray-coated with excimer and CO2 lasers]. / Osobennosti postroeniia kostnoi tkani u poverkhnosti implanta s pokrytiiami iz gidroksiapatita, napylennymi éksimernym i CO2-lazerami.

Rat experiments were performed to study osteogenesis and osseointegration in implanting fragments of dental titanic implants into the spongy bone. The implant spray-coating with hydroxyapatite produced by excimer and CO2 lasers stimulates osteogenesis. Bone tissue integration with implant proceeds more actively in response to CO2 laser radiation. The weakest integration was registered between the metal and bone tissues.