The Pattern of Granuloma Formation in the Liver of Rats as a Reflection of the Mechanism of Internalization of Submicron Silicon Particles.

A morphological analysis of the liver of Wistar rats was performed 2 months after a single intravenous injection of porous silicon particles of different sizes (60-80, 250-300, and 500-600 nm; 2 mg/ml, 1 ml). Histological, immunohistochemical, and electron microscopic methods showed the development of CD68+ granulomas in all experimental groups. Injection of 60-80-nm porous silicon particles led to the formation of single large granulomas (>2000 µm2), while 500-600-nm nanoparticles caused the formation of numerous smaller granulomas. The mechanism of involution of granulomas by apoptosis of Kupffer cells and the absence of subsequent connective tissue remodeling of the organ tissue is shown.

Morphological Analysis of the Respiratory Tract of Rats after Parenteral Administration of Silicon Dioxide Nanoparticles.

Morphological analysis of the respiratory tract of Wistar rats was performed after a single parenteral administration of 12-nm silicon dioxide nanoparticles (1 ml, 2 mg/ml, intravenously) was performed. On day 21 and in 2, 4, and 6 months after the administration of nanoparticles, the development of macrophage infiltration in the interstitium of the respiratory tract was demonstrated by histological and immunohistochemical methods. The pool of alveolar macrophages increased in 4 months after administration (p=0.004) and returned to the control values in 6 months. The number of mast cells did not significantly change at all stages of the experiment. Connective tissue remodeling in the interstitium of the respiratory tract was not observed throughout the observation period.

Reaction of the Palatine Tonsillar Immunocompetent Cells to Irrigation of Crypts by Suspension of Silica Nanoparticles under Conditions of Chronic Tonsillitis.

We studied the response of neutrophils, macrophages, and mast cells to local application of silica nanoparticles (10-20 nm). Histological examination of tonsillar postoperative material from 6 patients aged 24-44 years with recurrent tonsillitis was carried out. Irrigation of the tonsillar lacunae was carried out over 5 days before bilateral tonsillectomy: on the right by Polysorb MP suspension (1 g/liter), on the left by saline. The contact of nanoparticles with the mucosa led to a decrease in the number of cells expressing myeloperoxidase (p=0.02) and an increase in the count of CD68+ cells (p=0.04); the count of mast cells remained unchanged. Local use of medical adsorbent based on silica nanoparticles induced changes in cells due to their resorption by the tissue. Positive chemotaxis of CD68+ macrophages revealed in the tonsillar lymphoid tissue attested to stimulation of non-specific immunity and inductive phase of specific immunity. The authors hypothesized that internalization of medical nanoparticles by resident phagocytes of the mucosa could support targeted biodistribution of drugs in the palatine tonsils.

Tissue-Engineered Vascular Graft of Small Diameter Based on Electrospun Polylactide Microfibers.

Tubular vascular grafts 1.1 mm in diameter based on poly(L-lactide) microfibers were obtained by electrospinning. X-ray diffraction and scanning electron microscopy data demonstrated that the samples treated at T = 70°C for 1 h in the fixed state on a cylindrical mandrel possessed dense fibrous structure; their degree of crystallinity was approximately 44%. Strength and deformation stability of these samples were higher than those of the native blood vessels; thus, it was possible to use them in tissue engineering as bioresorbable vascular grafts. The experiments on including implantation into rat abdominal aorta demonstrated that the obtained vascular grafts did not cause pathological reactions in the rats; in four weeks, inner side of the grafts became completely covered with endothelial cells, and fibroblasts grew throughout the wall. After exposure for 12 weeks, resorption of PLLA fibers started, and this process was completed in 64 weeks. Resorbed synthetic fibers were replaced by collagen and fibroblasts. At that time, the blood vessel was formed; its neointima and neoadventitia were close to those of the native vessel in structure and composition.

Vascular Prostheses Based on Nanofibers from Aliphatic Copolyamide.

Tubular grafts based on nanofibers of copolymer of ε-caprolactam and hexamethylendiaminadipate were obtained by the electrospinning method. The strength of materials based on the dry nanofibers was 6.2 MPa with elongation at break of 133%, or 7.5 MPa and 299% in saline, respectively. The pressure value at which liquid started seeping through the tube wall was P = 10 kPa. Absence of cytotoxicity was proved, as well as adhesion and proliferation of mesenchymal stem cells on the surface. Tubes with inner diameter of 1 mm were tested in vivo in rat abdominal aorta. A layer of endothelial cells was shown to form on the inner side of the prosthesis after 30 days. There was no evidence of stenosis or dilatation of the prosthesis after 14 months with observation of endothelial and subendothelial layers.

BIORESORPTION OF POROUS 3D-MATERIALS BASED ON CHITOSAN.

3D-materials with high porosity were prepared by the method of lyophilization of chitosan solution. In vivo investigation of the mechanism and resorption rate of the resulting material in the muscle tissue showed complete resorption occuring in 12 months after implantation in an animal. The formation of scar tissue was not observed; there was no change and damage of the surrounding tissue. Histological analysis showed that chitosan resorption occurred simultaneously with the formation of collagen fibers and blood vessels. This allows us to recommend such porous material based on chitosan as a matrix for tissue engineering.

[CHANGE IN BCL-2 PROTEIN EXPRESSION IN NEURONS OF THE HIPPOCAMPAL AREAS AFTER THE APPLICATION OF ISCHEMIC CEREBRAL POSTCONDITIONING].

Bcl-2 protein expression was studied in hippocampal CA1, CA2, CA3 and CA4 pyramidal neurons in Mongolian gerbils (Meriones unguiculatus), of the in in the early (Day 2) and late (Day 7) reperfusion period after a 7-minute forebrain ischemia and following ischemic postconditioning (IPostC), as well as in sham-operated animals (n=60). In the latter, the highest level, of Bcl-2-expression was found in CA4 neurons, while the lowest--in-CA1 neurons (P<0.01). Reversible ischemic brain damage led to the increasing deficit of morphologically unchanged hippocampal neurons with the increasing duration of reperfusion period. This was accompanied by a significant decrease in expression of Bcl-2 in the early reperfusion period, but in the late reperfu- sion period this decrease largely disappeared. IPostC, applied as three episodes of ischemia-reperfusion lasting 15/15 seconds, contributed to significant increase in the number of morphologically unchanged CA1 and CA3 neurons in the early reperfusion period, while the expression of Bel-2 was increased in morphologically unchanged neurons in all the hippocampal areas. In the late reperfusion period after IPostC, the number of unchanged neurons was increased in hippocampal areas CA1, CA3 and CA4 (P<0.05), while a significant increase in Bcl-2 expression (by 12.7%, P<0.01) was detected only in CA1 neurons. The results suggest that the cytoprotective effect of IPostC in hippocampal CA1 area is realized through a mechanism leading to increased expression of Bcl-2 protein, i.e., by blocking apoptosis.

[Morpho-functional changes of the pyramidal neurons in various hippocampal areas after the ischemic postconditioning].

The aim of this study was to determine the effect of ischemic postconditioning (IP) on the viability of neurons in various hippocampal areas as well as on cytoplasmic activity of succinatedehydrogenase (SDH) in these cells in 30 male Mongolian gerbils (Meriones unguiculatus). Ischemic brain injury was induced by bilateral common carotid artery occlusion for 7 min. IP protocol comprised 3 cycles of 15 s of reperfusion/15 s of ischemia. After reperfusion for 48 h, the morphometric analysis was conducted, and SDH cytoplasmic activity was assessed using quantitative histochemistry in the pyramidal neurons of the hippocampal areas CA1, CA2, CA3, CA4. The experiment has demonstrated that 7-minute-long ischemia resulted in a significant decrease in the number of viable neurons in CA1 area (up to 24%) and in the CA3 (to 56%) of the hippocampus; besides, it lead to the elevation of SDH activity in the cytoplasm of the neurons in all the hippocampal areas as compared to that in sham-operated animals. The application of IP significantly increased the number of viable neurons in CA1 (up to 52.9%, P<0,01) and in CA3 areas of the hippocampus(up to 88%, P<0,05), and it was accompanied by reduction of SDH activity in surviving neurons in all the hippocampal areas.

Activity of succinate dehydrogenase in the neocortex and hippocampus of Mongolian gerbils with ischemic and reperfusion brain injury.

We analyzed changes in activity of SDH, one of the most important enzymes of the Krebs cycle, in the cytoplasm of hippocampal and cortical neurons of Mongolian gerbils (Meriones unguiculatus) at the early and delayed reperfusion period after global brain ischemia. The data indicate that SDH activity in pyramidal neurons of various hippocampal areas and in neurons of II, III and V layers of cerebral cortex after 7-min forebrain ischemia depends on both the localization of these neurons and duration of the postischemic reperfusion. SDH activity in neurons significantly increased on days 2 and 7 after reperfusion.