Laser Thermo-Photobiomodulation of Bone Marrow Mesenchymal Stem Cells.

We studied the effect of laser radiation of moderate intensity with a wavelength of 970 nm on the efficiency of colony formation of rat bone marrow mesenchymal stem cells (MSC) in vitro. In this case, photobimodulation and thermal heating of MSC occur simultaneously. This combined laser treatment allows increasing the number of colonies by 6 times in comparison with the control and by more than 3 times in comparison with thermal heating alone. The mechanism of such an increase is associated with combined thermal and light effects of laser radiation of moderate intensity, which stimulates cell proliferation. This phenomenon can be used as the basis for solving the most important task of cell transplantation, associated with the expansion of autologous stem cells and activation of their proliferative potential.

Changes in the Number of Mesenchymal Stem Cells in Bone Marrow at Different Periods after In Vivo Exposure of the Bone Marrow to Local Infrared Laser Radiation.

We determined optimal parameters of bone marrow (BM) irradiation in vivo for rapid increase in the number of mesenchymal stem cells (MSC) at the initial stages of the culturing without changing the karyotype, polyploidy, which are observed at higher passages. Such an increase is necessary to achieve the required number of cells at the initial passages for subsequent transplantation into the body. It was shown that after irradiation with λ=0.97 µm, the maximum and similar increase in the content of BM MSC in comparison with the control (by 2.4 times) was observed on day 2 in the irradiated and contralateral tibia. An insignificant difference in the number of BM MSC for the irradiated and contralateral tibia remained at all terms after irradiation, with a general decrease in the number of BM MSC up to 21 days. After laser irradiation with λ=1.56 µm, the maximum number of BM MSC was also observed on day 2. In this case, the concentration of these cells in the irradiated and contralateral limbs exceeded the control by 3.1 and 1.7 times, respectively. With increasing the time after exposure, the number of BM MSC in both limbs showed the same tendency to a decrease as after irradiation at λ=0.97 µm.

Cytotoxicity control of silicon nanoparticles by biopolymer coating and ultrasound irradiation for cancer theranostic applications.

Silicon nanoparticles (SiNPs) prepared by mechanical grinding of luminescent porous silicon were coated with a biopolymer (dextran) and investigated as a potential theranostic agent for bioimaging and sonodynamic therapy. Transmission electron microscopy, photoluminescence and Raman scattering measurements of dextran-coated SiNPs gave evidence of their enhanced stability in water. In vitro experiments confirmed the lower cytotoxicity of the dextran-coated NPs in comparison with uncoated ones, especially for high concentrations of about 2 mg ml-1. Efficient uptake of the NPs by cancer cells was found using bioimaging in the optical transmittance and photoluminescence modes. Treatment of the cells with uptaken SiNPs by therapeutic ultrasound for 5-20 min resulted in a strong decrease in the number of living cells, while the total number of cells remained nearly unchanged. The obtained data indicate a 'mild' effect of the combined action of ultrasonic irradiation and SiNPs on cancer cells. The observed results reveal new opportunities for controlling the photoluminescent and sonosensitizing properties of silicon-based NPs for applications in the diagnostics and mild therapy of cancer.

Silicon Nanoparticles as Amplifiers of the Ultrasonic Effect in Sonodynamic Therapy.

The possibility of using mesoporous silicon nanoparticles as amplifiers (sensitizers) of therapeutic ultrasonic exposure were studied experimentally in vitro and in vivo. The combination of nanoparticles and ultrasound led to a significant inhibition of Hep-2 cancer cell proliferation and Lewis lung carcinoma growth in mice. These results indicated good prospects of using silicon nanoparticles as sensitizers for sonodynamic therapy of tumors.

Effects of acoustic and EHF impulses on multipotent stromal cells during formation of bone marrow containing heterotopic organs in tissue engineered constructions.

We studied the effects of physical factors (acoustic impulses of laser-induced hydrodynamics, AILIH, and EHF-radiation) on the formation of heterotopic bone marrow organs. Suspension of precipitated mouse bone marrow cells was exposed to AILIH and EHF or their combinations (AILIH+EHF, EHF+AILIH). The developed tissue engineering constructions (gelatin sponges containing 107 nucleated bone marrow cells exposed to physical factors) were transplanted under the renal capsule of syngeneic mice. Analysis of newly formed hemopoietic organs was performed after 3 and 5 months. The total amount of hemopoietic cells, number of multipotent stromal cells, efficiency of colony formation from these cells, and weight of bone capsule of the transplants were measured. Microscopic study showed that 5-month transplants were significantly larger than 3-month transplants and contained 3-fold more hemopoietic cells (20-fold in the AILIH+EHF group). The number of multipotent stromal cells was maximum in EHF+AILIH group (by 2.2 times higher than in the control) and minimum in AILIH+EHF group. Exposure to EHF+AILIH had most pronounced effect on the formation of the bone marrow transplants. The weight of bone capsules more rapidly increased in gelatin sponges of 3-month transplants of EHF+AILIH and AILIH groups. These data suggest that the studied physical factors can be used for acceleration of rehabilitation process.

[THE DETECTION OF CONTENT OF DIAGNOSTICALLY SIGNIFICANT FATTY ACIDS AND INDIVIDUAL TRIGLYCERIDES IN BIOLOGICAL MEDIUMS BASED ON INFRARED SPECTROMETRY].

The content of clinically important fatty acids and individual triglycerides in food and biological mediums are traditionally detected by gas and fluid chromatography in various methodical modifications. The techniques are hard-to-get in laboratories of clinical biochemistry. The study was carried out to develop procedures and equipment for operative quantitative detection of concentration of fatty acids, primarily palmitic saturated fatty acid and oleic mono unsaturated fatty acid. Also detection was applied to sums ofpolyenoic (eicosapentaenoic and docosahexaenoic acid) fatty acids in biological mediums (cod-liver oil, tissues, blood plasma) using spectrometers of short-range infrared band of different types: with Fourier transform, diffraction and combined scattering. The evidences of reliable and reproducible quantitative detection offatty acids were received on the basis of technique of calibration (regression) by projection on latent structures using standard samples of mixtures of oils and fats. The evaluation is implemented concerning possibility of separate detection of content of palmitic and oleic triglycerides in mediums with presence of water The choice of technical conditions and mode of application of certain types of infrared spectrometers and techniques of their calibration is substantiated

Express Analysis of Cartilage Tissue Using Multivariate Analysis of IR Spectra.

The aim of the study was to develop a diagnostic method for the quantitative determination of the main components of cartilage tissue of various types based on multivariate IR spectral analysis and verification of data using classical chemical analysis. Materials and Methods: Cartilages of the nasal septum, knee joint, rib, and nucleus pulposus of the intervertebral disc, as well as trypsinized and defective cartilage samples, were examined as samples. The IR spectra of the cartilage samples, as well as calibration mixtures of collagen and chondroitin sulfate, were obtained. The IR spectra were collected using the attenuated total reflectance techniques, and their processing was performed using the TQ Analyst software and the principal component regression calibration technique. Based on calibration dependence, the Ksp coefficient was determined as the ratio of the mass fractions of collagen and chondroitin sulfate. Its value was compared with the value of Kchem, equal to the ratio of the mass fractions of collagen and chondroitin sulfate, obtained using the classical chemical analysis of these substances. Results: The IR spectra of cartilage tissues are a superposition of the IR spectra of collagen and chondroitin sulfate and qualitatively reflect their composition. A change in the ratio between the relative intensities of the characteristic bands of compounds in the IR spectrum is obvious only with a significant change in the content of these compounds in cartilage. This change occurs after trypsinization, when Ksp increases from 0.88±0.05 (Kchem~0.8) to 4.55. The use of a calibration model with a complete analysis of the cartilage IR spectrum made it possible to determine the difference in the ratio of the main components in the matrix of different samples in the absence of obvious changes in the IR spectra. Thus, a statistically significant decrease in the content of chondroitin sulfate in degraded articular cartilage (Ksp=4.4±1.8; Kchem~5.5) was shown compared with intact samples (Ksp=2.8±1.1; Kchem~2.6). Conclusion: IR spectrometric express analysis of cartilage tissue employing the principal component regression method allows a correct determination of the ratio of the main components in the cartilage matrix, those of collagen and glycosaminoglycans. The proposed technique includes one measurement, does not require prolonged and laborious sample preparation, does not require long, multi-stage and laborious chemical manipulations to determine each of the components, and makes it possible to determine the features and changes in the composition for a large set of samples of cartilage tissue of different types. In future, this approach can be used for non-invasive diagnostics of cartilage tissue.

Biodegradation model of porous silicon nanoparticles.

Development of porous silicon-based drug delivery systems for theranostics requires a precise control of their biodegradation. Thus, we propose a model for the biodegradation of porous silicon nanoparticles (PSi NPs) based on a diffusion equation combined with Nernst-Brunner mass transfer equation describing the dissolution of silicon and formation of silicic acid (SA). The spatiotemporal distributions of PSi NP porosity and SA concentration were calculated. The model was successfully applied to fitting a great variety of experimental data on more than 10 factors influencing the PSi NP biodegradation kinetics, such as the morphology of PSi NPs, surface composition, properties of surrounding media and protective coating layer. Two principal regimes were found out for systems with either diffusion or dissolution dominating over each other. The results of simulations revealed the values of several important parameters, which are hard to be measured experimentally.

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.

[Photochemical reactions in bone tissue induced by excimer laser ultraviolet]. / O proiavleniiakh fotokhimicheskikh reaktsi'i v kostno'i tkani pod de'istviem ul'trafioletovogo izlucheniia eksimernogo lasera.

A phenomenological model of an increase in ultraviolet absorption in bone tissue induced by ultraviolet radiation of excimer laser has been developed. It is assumed that the increase in absorption is related to photochemical reactions in collagen. The model accounts for changes in the intensity of laser radiation due to its absorption inside the specimen. From the comparison of experimental and calculated results the parameters of the photochemical model were estimated. The temperature fields in the specimen were calculated with regard to laser-induced changes in absorption coefficient. The limits of applicability of the model are discussed.

[Holmium laser in chondrology: potentials and prospects]. / Vozmozhnosti i perspektivy primeneniia gol'mievogo lazera v khondrologii.

To study the mechanism of simulation of a cartilage, to define the optimum parameters and laser radiation regimens, the authors made a complex of in vitro experiments on the cartilages of the nasal septum in man and domestic animals and in vivo experiments on those of the concha in rabbits and pigs. Holmium laser radiation is shown to be the most suitable tool for formation of a cartilage. Evidence is provided for the possibility of irreversibly altering the shape of a cartilage during laser radiation. A procedure has been developed for non-invasive correction of the septum of the nose for its curvature. Restored or improved nasal respiration was observed in more than two thirds of the patients. Laser surgery is not of age-limited application, it is noninvasive, can be performed in the outpatient settings and requires no drug treatment in the postoperative treatment.

[Mathematical analysis of the causes of unsatisfactory results of scleroplasty in progressive myopia]. / Matematicheskii analiz prichin neudovletvoritel'nykh rezul'tatov skleroplastiki pri progressiruiushchei blizorukosti.

Mathematical statistics-based analysis of the results of 256 surgeries, carried out in 219 patients with high progressive myopia, has revealed a number of factors significant for effective outcomes and failures of the operations. Among prognostically favorable factors are the school and congenital forms of myopia, young or adult age of the patients, a moderate progress of myopia (up to 1 diopter yearly), a noninherited pattern of myopia or a dominant type of inheritance, absence of somatic diseases. The factors that predict a possible unfavorable outcome are early acquired myopia, patients' childhood age, high rate of myopia progress, the recessive type of inheritance, concomitant diseases. The contribution of every factor to the result has been determined. The analysis has helped single out the group of patients at risk of an unfavorable result of surgery.

[Possibilities of the use of surgical laser irradiation in spontaneous formation of cartilage tissue in ENT plastic surgery]. / Vozmozhnosti primeneniia izlucheniia khirurgicheskikh lazerov s tsel'iu proizvol'nogo formirovaniia khriashchevoi tkani v plasticheskoi LOR-khirurgii.

The experiments were made to study CO2-laser ability to promote spontaneous formation of nasal septum cartilage. Laser-induced changes in physical and histological characteristics of the cartilage tissue were followed up. Optimal regimens of radiation were selected which allowed to modify cartilage shape without impairment of its morphological and histological structure. In vivo tests were performed on rabbit auricular floor cartilage. The present shape of the cartilage remained stable. It is inferred that laser energy may be a promising tool in plastic ENT surgery.

[Changes in the cartilage shape under the action of laser irradiation]. / Proizvol'noe izmenenie formy khriashcha pod vliianiem lazernogo izlucheniia.

Investigations of structural and mechanical properties of cartilage tissue exposed to laser radiation have shown the effect of laser relaxation of the tension existing in the cartilage and changes in the latter form to be due to phasic transition of water from bound into free state. Laser radiation does not induce macroscopic transformations of the matrix structure. New form of the cartilage proved stable this offering new prospects for wide application of this technique in otorhinolaryngology and plastic surgery.

[Reestablishing of nasal respiration with Holmium laser in distortion of nasal septum in cartilaginous part]. / Vosstanovlenie nosovogo dykhaniia s pomoshch'iu izlucheniia gol'mievogo lazera pri iskrevlenii peregorodki nosa v khriashchevom otdele.

To study mechanisms of cartilage modelling, estimation of optimal parameters and regimens of laser radiation, the authors made in vitro and human experiments on nasal septum cartilages and in vivo experiments on cartilages of the floor of the auricle in rabbits and pig. Ho-laser radiation is thought the most appropriate tool for cartilage shaping. A method of non-invasive correction of the nasal septum cartilage in its distortion has been designed. 2/3 of the patients recovered or improved nasal respiration. Laser operation has no age restrictions, is non-invasive, can be made outpatiently, is not followed by pharmacological treatment.