Features of Remyelination after Transplantation of Olfactory Ensheathing Cells with Neurotrophic Factors into Spinal Cord Cysts.

This paper shows for the first time that co-transplantation of human olfactory ensheathing cells with neurotrophin-3 into spinal cord cysts is more effective for activation of remyelination than transplantation of cells with brain-derived neurotrophic factor and a combination of these two factors. The studied neurotrophic factors do not affect proliferation and migration of ensheathing cells in vitro. It can be concluded that the maximum improvement of motor function in rats receiving ensheathing cells with neurotrophin-3 is largely determined by activation of remyelination.

Application of Behavioral Tests for Evaluation of an Experimental Model of Alzheimer's Disease in Female Rats.

Alzheimer's disease was modeled in female Wistar rats aged 4 months by stereotaxic bilateral injection of a synthetic peptide ß-amyloid (Aß1-42) into the hippocampus. Behavioral tests (open field, Y-maze, passive avoidance, and Morris water maze) revealed significant impairment of memory and spatial navigation 8 weeks after ß-amyloid administration. At this term, the cognitive impairments typical of Alzheimer's disease are reproduced. The experimental model of Alzheimer's disease proposed by us can be used in preclinical studies of drugs for the treatment of this pathology.

Modeling of Alzheimer's Disease to Study the Efficacy of Cell Therapy (Review).

We analyzed the main approaches to the modeling of Alzheimer's disease for studying the effectiveness of cell therapy. Recent advances in regenerative medicine in the field of neuroscience create prospects for the use of various cell preparations for the treatment of Alzheimer's disease. Experimental data on the use of neural stem/progenitor cells, mesenchymal stem cells, embryonic stem cells, and induced pluripotent stem cells in various models of Alzheimer's disease are presented. Of particular importance is the standardization of protocols. The use of a standardized protocol in modeling of Alzheimer's disease will allow a comparative analysis of the effectiveness and safety of treatment to identify the optimal cell preparation. The data obtained on experimental animals can form the basis for further preclinical and clinical studies of cell therapy for Alzheimer's disease.

Neurotrophin-3 Enhances the Effectiveness of Cell Therapy in Chronic Spinal Cord Injuries.

Neurotrophin-3 enhances the effectiveness of human olfactory ensheathing cells in improving hind limb mobility in rats with post-traumatic cysts of the spinal cord. Transplantation of olfactory ensheathing cells into spinal cord cysts reduced their size; neurotrophin-3 did not modulate this effect. Combined preparation of human olfactory ensheathing cells and neurotrophin- 3 can be used in neurosurgery for the treatment of patients with spinal cord injuries.

Obtaining a New Gene-Cell Construct Based on Transduced Olfactory Ensheathing Cells for the Treatment of Spinal Cord Injuries.

We developed a viral vector Ad5/35-CAG-mBDNF expressing the mature form of BDNF (mBDNF). On the basis of olfactory ensheathing cells transduced with this adenovector, a new gene-cell construct was obtained. In experiments in vitro, high viability of the transduced olfactory ensheathing cells and enhanced secretion of BDNF by these cells were observed. It is possible that a new gene-cell construct will significantly increase the regenerative effects of transplanted olfactory ensheathing cells.

Effect of Anesthesia on Postoperative Pain in Patients after Septoplasty.

The aim of the study was to assess acute pain syndrome in patients after septoplasty using different tactics of general anesthesia. All patients received local anesthesia with 2% procaine solution. In group 1 (95 patients), premedication with 2% promedol solution and 60 mg of ketorolac in the evening was used; group 2 (72 patients) was administered with fentanyl, propofol, cisatracuria besylate, tranexamic acid, atropine, and metoclopramide; and group 3 (89 patients) received atracuria besylate, sodium thiopental, nitrous oxide, and halothane. In groups 2 and 3, 100 mg of ketoprofen was administered intramuscularly in the evening on the day of surgery. Anterior tamponade was performed with parolon tampons in glove rubber. In groups 1 and 2, the tamponade was removed on day 2, and in group 3 it was removed 1 day after surgery. Pain syndrome was assessed on 1, 3, and 6 h and on days 1 and 2 after surgery. It was found that the scheme of anesthesia in group 2 is the most preferable, and the nasal tamponade must be removed on the 2nd day after the surgery.

The Relationship between p53-Positive Neurons and Dark Neurons in the Hippocampus of Rats after Surgical Interventions on the Nasal Septum.

The study evaluates the dependence of p53 protein expression on the appearance of dark neurons (DNs) in the hippocampus in rats during experimental modeling of septoplasty. Septoplasty simulation was carried out on 15 sexually mature male Wistar rats. We studied histological sections of the hippocampus stained with Nissl toluidine blue and antibodies to the p53 protein. In the CA1 subfield, the number of p53-positive neurons significantly increased on the 2nd, 4th (p < 0.001) and 6th days (p < 0.05). In the dynamics, the peak of the growth of p53 protein expression in the cytoplasm of CA1 and CA2 neurons fell on the 2-4th day after the operation, and on the 6th day the number of these neurons decreased (p < 0.001). In the cytoplasm of CA3 neurons in all periods after surgery, an increase in the expression of the p53 protein as compared to the control group was noted. In the CA1 pyramidal layer, the number of DNs decreased on the 6th day (p < 0.001). In CA2, after 2 days, a minimum of DNs as compared with the 4th day (p < 0.001) was noted. In CA3, on the 4th day, there was a peak in DNs as compared with the rest of the days (p < 0.001). A positive strong association was found in all periods of assessment and in all subfields of the hippocampus between an increase in the number of dark and p53-positive neurons. The appearance of dark and p53-positive neurons in the hippocampal formation in rats after simulating septoplasty are typical responses of nervous tissue to stress. It is obvious that the expression of the p53 protein is associated with the basophilia of the cytoplasm of neurons, their morpho-functional state. Presumably, the p53 protein can trigger not only the activation of damaged neurons in the hippocampus but also play a neuroprotective role. Upcoming studies should determine the role of the p53 protein in the further fate of damaged neurons in the pyramidal layer and differentiate the mechanisms of its expression.

[Morphogenetic and pathogenetic features of hypertrophic and keloid scars of the head and neck]. / Morfogeneticheskie i patogeneticheskie osobennosti gipertroficheskikh i keloidnykh rubtsov golovy i shei.

OBJECTIVE: Evaluate the morphogenetic and pathogenetic features of hypertrophic and keloid scars of the head and neck. MATERIAL AND METHODS: The study included 286 patients, among them 176 (61.5%) patients with hypertrophic and 110 (38.5%) with keloid scars aged 18 to 65 years with a disease duration from 1 month to 2 years. Material for histological and immunohistochemical (IHC) studies of scar tissue was fixed in 10% buffered formalin. Serial paraffin sections were stained with H&E, according to Van Gieson and Weigert. IHC was performed using monoclonal mouse antibodies to collagen type I (clone 3G3, Santa Cruz, dilution 1:100), collagen type III (clone B-4, Santa Cruz, dilution 1:50), collagen type IV (clone COL-94, Santa Cruz, dilution 1:50), MMP-1 (clone 3B6, Santa Cruz, dilution 1:100), α-SMA1 (clone 1A4, Dako Agilent, dilution 1:100) and rabbit polyclonal anti-TGFß antibodies (clone 3C11, Santa Cruz, 1:100 dilution). RESULTS: Pathogenetic, morphological and immunohistochemical differences in hypertrophic and keloid scars were established depending on their degree of maturity. In the formation of hypertrophic scars, the key factor in sclerotic processes is TGF-b on the background of low MMP1 activity. Keloid scars were distinguished not only by the accumulation of hard-to-degrade collagens, but also by the development of an osteoclast-like reaction with a high content of MMP1. Immature scar tissue was characterized by the presence of myofibroblastic α-SMA1 positive focus and center of inflammatory changes. CONCLUSIONS: The data obtained allow substantiating new approaches to the treatment of patients with hypertrophic and keloid scars.

The Effect of Transplantation of Olfactory Ensheathing Cells on the Size of Posttraumatic Spinal Cord Cysts.

We studied the effect of transplantation of ensheathing cells obtained from the olfactory mucosa of rats and humans on the size of posttraumatic spinal cord cysts. MRI examination showed that transplantation of these cells into experimental posttraumatic cysts of the spinal cord led to a significant decrease in cyst volume and even their complete disappearance in two animals receiving transplantation of rat or human cells. These findings attested to regenerative processes developing as a result of ensheathing cell transplantation. Further studies in this field will be aimed at elucidation of the mechanisms underlying spinal cord regeneration in the area of posttraumatic cysts after transplantation of ensheathing cells.

Low-Intensity Laser Therapy As a Method to Reduce Stress Responses after Septoplasty.

The aim of the study was to evaluate the effectiveness of the use of photobiomodulation therapy (PBMT) to minimize acute pain in the early postoperative period in patients after septoplasty. In total, 62 patients underwent septoplasty under general anesthesia (40 men and 22 women, 18-44 years old) followed by nasal tamponade. Patients of the 1st group did not undergo PBMT, and patients of the 2nd group received PBMT 3, 6, and 24 h after septoplasty (infrared pulsed laser radiation, λ = 0.890 µm, P = 10 W, 2 min in the projection of the wings of the nose). After 48 h, after the removal of tampons, intranasal PBMT with a nozzle in the red range, with λ = 0.63 µm, P = 8 mW, 2 min. ULF, HF, LF, and total power of heart rate variability (HRV), pain syndrome was assessed. ULF, LF, HF, total HRV power were significantly lower in group 2, compared to group 1. In the period from 6 to 24 h after septoplasty, patients of group 1 experienced higher pain than patients with PBMT (p < 0.001). The use of PBMT after septoplasty against the background of nasal tamponade helps to reduce the severity of pain syndrome and the inflammatory response to surgical stress and, consequently, leads to less pronounced changes in the autonomic nervous system in response to surgical stress.

Study of the Role of Hearing Aid on the Area of the Acoustic Field of Vowels.

The method of transformation of acoustic vowel triangles (AVT) /a/, /i/, /u/ was used for an objective assessment of the acoustic features of vowels in the speech production of 20 persons with long-term hearing impairment (LHI). The logarithm of the values ​​of the first two formants of each vowel (logF1, logF2) was determined for each subject. AVTs were transformed into the right-angled triangles, the vertices of the sound /u/ of which were moved to the origin of coordinates and the legs were aligned with the coordinate axes. In patients with LHI, the size of the triangles usually decreased, and they were stretched along one of the axes, which probably depends not only on the hearing loss severity but also on the duration of hearing aid use. The presented approach to the normalization of AVTs makes it possible to distinguish at least three groups of persons with LHI: in the first group, vowel triangles are stretched along the logF1 axis; in the second group, vowel triangles are stretched along the logF2 axis; and in the third group, AVT are symmetric.

Effectiveness of the Sequential Use of Plastic and Titanium Implants for Experimental Replacement of the Mandibular Defect in Animals Using Preliminary Digital Design.

Using the templates preliminarily made by 3D design and prototyping methods, defects in the lateral area of the lower jaw of sheep were created using the piezosurgical technique. The defects were replaced by plastic implants obtained by the method of layer-by-layer fusion of the FDM printing-fusing deposition modeling and fixation with titanium screws to the jaw body. In the time interval, plastic implants are replaced by titanium implants obtained by selective laser sintering (SLS) using a 3D printer. To study the processes of reparative osteogenesis, microsamples of tissues of the preimplantation zone were analyzed. As a result, signs of osteo- and fibro-osseointegration were identified. The obtained data are regarded as a prerequisite for further clinical trials of the developed protocols for the sequential replacement of jaw defects using 3D printing.

Changes in the Time-Domain of Heart Rate Variability and Corticosterone after Surgical Trauma to the Nasal Septum in Rats.

The aim of the work was to study changes in the time range of heart rate variability (HRV) against the background of changes in the concentration of corticosterone in blood plasma in rats after surgical trauma to the nasal septum. Septoplasty was simulated in 30 mature male Wistar rats weighing 210-290 g. ECG was recorded with subsequent analysis of the time domain of HRV, as well as blood sampling to estimate changes in the concentration of corticosterone in the blood plasma was performed. As a result, SDNN significantly increased in comparison with the control on days 2 and 3 (p < 0.001) but decreased on days 4-5 (p < 0.001) and 6 (p < 0.01). rMSSD changed in waves with two irregular peaks on days 1 and 6. SDNN/rMSSD, in comparison with the 1st day of the postoperative period, increased on the 2nd day, continued to increase (p < 0.05), and then began to decrease on day 4 (p < 0.01). The total HRV power of was low throughout the postoperative period (p < 0.001), except for day 3, when it was equal to the control data. The increase in the total power index fell on day 3 after the operation (p <0.01), after which its decline was observed again. The concentration of corticosterone in the blood plasma in rats was significantly higher than before (p < 0.001). On postoperative day 2 to 4, its plateau was determined. Simulation of septoplasty leads to changes in the time range of HRV, an increase in the concentration of corticosterone in the blood plasma in rats with its maximum at the time of surgery and 24 h later, and the formation of a "plateau" on postoperative days 2 to 4, which coincides with the changes in HRV.

Influence of Surgical Trauma in the Nasal Cavity on the Expression of p53 Protein in the Hippocampus of Rats.

The aim of the study was to determine the role of the stress effect of septoplasty modeling on p53 protein expression in the hippocampus of rats under conditions of sensory olfactory deprivation. Simulation of septoplasty was carried out on 30 sexually mature male rats. A quantitative assessment of the apoptosis of neurons in the pyramidal layer of the hippocampus in the subfields CA1, CA2, CA3, and dentate gyrus (DG) on days 2, 4, and 6 after surgery was carried out. Histological sections were stained by the immunohistochemical method with antibodies to the p53 protein. An increase in the number of p53-positive neurons was noted in all subfields; the maximum increase in the number of apoptotic neurons was noted on day 4 after surgery. The stress effect of modeling septoplasty in rats, accompanied by sensory deprivation of the peripheral part of the olfactory analyzer, provoked the expression of p53 and the initiation of apoptosis mechanisms in various subfields of the hippocampus.

Comparison of the Effects of Septoplasty and Sinus Lifting Simulation in Rats on Changes in Heart Rate Variability.

The effects of septoplasty and sinus lifting simulation in rats on changes in the frequency domain of heart rate variability were compared. In the early postoperative period, the low-frequency component increases, and the high-frequency component decreases. The VLF range also increased. This indicates an increase in the activity of the sympathetic nervous system and a shift in metabolism under the influence of post-surgical inflammation.

[Methods of decreasing the risk of repeat interventions in patients after arterial revascularization]. / Sposoby snizheniia riska povtornogo vmeshatel'stva u patsientov posle arterial'noi revaskuliarizatsii.

Discussed in the article are the main problems related to surgical treatment of patients with peripheral artery disease, particularly taking into consideration that in the world there are from 160 to 202 million people suffering from this disease, with two thirds of such patients having signs of lesions of coronary or cerebral arteries. Vascular reconstructive interventions cannot completely eliminate the problem, since in the postoperative period there may develop cardiovascular complications related to both the limb involved as either acute or progressing chronic ischaemia and arteries of other localization (coronary, cerebral). The risk of serious cardiovascular complications in patients with a history of endured adverse ischaemic events on the part of limbs is severalfold higher. To solve these problems and decrease complications, salicylic acid is used as basic therapy. Attempts at replacing it by another drug or combined therapy with an alternative antiaggregant showed no advantages in increased risk of massive haemorrhage. On the other hand, a combination of salicylic acid with an anticoagulant at a low dose, i. e., rivaroxaban 2.5 mg twice daily as compared with acetylsalicylic acid monotherapy made it possible to significantly decrease the incidence of various cardiovascular complications in the form of myocardial infarction, stroke, adverse ischaemic events on the part of the extremity, limb amputation.

Proof of concept for continuously-tunable terahertz bandpass filter based on a gradient metal-hole array.

A continuously-tunable terahertz (THz) bandpass filter based on the resonant electromagnetic-wave transmission through a metal-hole array featuring a gradually changing period was developed and fabricated on a silicon substrate using optical lithography. A gradient geometry of the metal-hole array yields a wide tunability of the filter transmission, when operating with a focussed THz beam. The filter was studied numerically, using the finite element method, and experimentally, using the THz pulsed spectroscopy. We find that the central wavelength of the filter transmission band can be tuned in the wide range of λc = 400-800 µm with the relative bandwidth of Δλ/λc ≃ ~0.4. Finally, Kapton-based anti-reflection coating was applied to the filter flat side, in order to suppress an interference pattern in the filter transmission spectrum. We believe that the developed filter holds strong potential for multispectral THz imaging and sensing due to its conceptual simplicity and case of operation. Moreover, the presented filter concept can be translated to other spectral ranges, where appropriate technologies are available for the fabrication of gradient sub-wavelength metal-hole arrays.

Combined Preparation of Human Olfactory Ensheathing Cells in the Therapy of Post-Traumatic Cysts of the Spinal Cord.

In experiments on rats, co-transplantation of olfactory ensheathing cells of the human olfactory mucosa and neural stem/progenitor cells from the same source into post-traumatic cysts of the spinal cord led to improvement of the motor activity of the hind limbs and reduced the size of the cysts in some animals by 4-12%. The transplantation of a combination of the olfactory mucosa cells is effective and can be used in preclinical trials for the treatment of spinal cord injuries.

Preparation of Adhesion Culture of Neural Stem/Progenitor Cells of the Olfactory Mucosa for the Treatment of Spinal Cord Injuries.

In this work, an optimal protocol was developed for obtaining adhesion culture of neural stem/progenitor cells (NSPC) of rat olfactory mucosa. During the development of the protocol, the conditions for cell culturing on adhesion substrates fibronectin and laminin in DMEM/F-12 and neurobasal media with the same culture additives were compared. Cell proliferation was maximum during culturing on both substrates in the neurobasal medium. Using the immunofluorescence method, we found that culturing on fibronectin in the neurobasal medium ensured maximum (52.22%) content of nestin-positive cells in comparison with other culturing conditions. The highest percentage of ßIII-tubulin-positive cells was detected in cultures growing on fibronectin in the neurobasal medium and in DMEM/F-12 (79.11 and 83.52%, respectively). Culturing in adhesion cultures in the neurobasal medium on fibronectin allowed obtaining cultures enriched with NSPC and neurons differentiating from them in a quantity sufficient for further transplantation. The developed protocol can be recommended for obtaining NPSC from human olfactory mucosa for the treatment of spinal cord injuries.

Energy Metabolism in Cellular Regenerative Processes: Focus on PPARα.

Reduced expression of the key regulator of cardiac metabolism, transcription factor PPARα, in surgical samples of the auricles from patients with coronary heart disease and heart failure was detected by real-time quantitative PCR. These changes indicate reduced activity of this factor and a shift of energy metabolism from oxidative phosphorylation to glycolysis typical of dedifferentiated cells. Electron microscopy revealed dedifferentiated cardiomyocytes with disassembled contractile apparatus and disorganized sarcomeres. In the examined specimens from patients with heart failure, severe myocardial fibrosis was revealed.