Skeletal Muscle Resident Progenitor Cells Coexpress Mesenchymal and Myogenic Markers and Are Not Affected by Chronic Heart Failure-Induced Dysregulations.

BACKGROUND AND PURPOSE: In heart failure (HF), metabolic alterations induce skeletal muscle wasting and decrease of exercise capacity and quality of life. The activation of skeletal muscle regeneration potential is a prospective strategy to reduce muscle wasting; therefore, the aim of this project was to determine if functional properties of skeletal muscle mesenchymal progenitor cells (SM-MPC) were affected by HF-induced functional and metabolic dysregulations. METHODS: Gastrocnemius muscle biopsy samples were obtained from 3 healthy donors (HD) and 12 HF patients to purify mRNA for further analysis and to isolate SM-MPC. Cells were expanded in vitro and characterized by immunocytochemistry and flow cytometry for expression of mesenchymal (CD105/CD73/CD166/CD146/CD140b/CD140a/VIM) and myogenic (Myf5/CD56/MyoG) markers. Cells were induced to differentiate and were then analyzed by immunostaining and Q-PCR to verify the efficiency of differentiation. The expression of genes that control muscle metabolism and development was compared for HD/HF patients in both muscle biopsy and in vitro-differentiated myotubes. RESULTS: The upregulation of MYH3/MYH8/Myf6 detected in HF skeletal muscle along with metabolic alterations indicates chronic pathological activation of the muscle developmental program. SM-MPC isolated from HD and HF patients represented a mixed population that coexpresses both mesenchymal and myogenic markers and differs from AD-MMSC, BM-MMSC, and IMF-MSC. The functional properties of SM-MPC did not differ between HD and HF patients. CONCLUSION: In the present work, we demonstrate that the metabolic and functional alterations we detected in skeletal muscle from HF patients do not dramatically affect the functional properties of purified and expanded in vitro SM-MPC. We speculate that skeletal muscle progenitor cells are protected by their niche and under beneficial circumstances could contribute to muscle restoration and prevention and treatment of muscle wasting. The potential new therapeutic strategies of HF-induced skeletal muscle wasting should be targeted on both activation of SM-MPC regeneration potential and improvement of skeletal muscle metabolic status to provide a favorable environment for SM-MPC-driven muscle restoration.

"Solvent/non-solvent" treatment as a method for non-covalent immobilization of gelatin on the surface of poly(l-lactic acid) electrospun scaffolds.

In the present study, we report a simple and efficient method of gelatin immobilization on the surface of PLA electrospun fibers using pre-treatment with a mixture of toluene and ethanol allowing to form swelled surface layer followed by gelatin adsorption from its solution in PBS. Our results demonstrate that gelatin immobilization leads to a decrease in the water contact angle from 120° to 0°, enhances scaffold strength up to 50%, and doubles the number of adhered cells and their average area. We observed that the maximum amount of gelatin (0.07 ± 0.01 mg per cm3 of the scaffold) was immobilized during the first five minutes of exposure to the gelatin solution. Modified scaffolds demonstrated increased strength.

Flexible intramedullary nails for limb lengthening: a comprehensive comparative study of three nails types.

This study aims to investigate the comparative study of calcium phosphate coatings for flexible intramedullary nails (FINs) used to lengthen long tubular bones. The presence of a calcium phosphate coating deposited by micro-arc oxidation (MAO) or a composite coating based on a co-polymer of vinylidene fluoride with tetrafluoroethylene (VDF-TeFE) and hydroxyapatite (HA) on the surface of the FIN significantly enhanced the regeneration of bone in the area of osteotomy during limb lengthening by combined osteosynthesis. The investigation of the physico-chemical properties of the FIN coated with calcium phosphate via MAO demonstrated that the improved bone tissue formation resulted from favourable conditions for adhesion, proliferation and differentiation of multipotent stem cells into osteoblasts on the coating surface. The composite coatings only stimulated the formation of bone tissue in vivo, primarily because of the piezoelectric properties of the VDF-TeFE co-polymer.

Cellular Mechanisms of Aortic Valve Calcification.

Comparative in vitro study examined the osteogenic potential of interstitial cells of aortic valve obtained from the patients with aortic stenosis and from control recipients of orthotopic heart transplantation with intact aortic valve. The osteogenic inductors augmented mineralization of aortic valve interstitial cells (AVIC) in patients with aortic stenosis in comparison with the control level. Native AVIC culture of aortic stenosis patients demonstrated overexpression of osteopontin gene (OPN) and underexpression of osteoprotegerin gene (OPG) in comparison with control levels. In both groups, AVIC differentiation was associated with overexpression of RUNX2 and SPRY1 genes. In AVIC of aortic stenosis patients, expression of BMP2 gene was significantly greater than the control level. The study revealed an enhanced sensitivity of AVIC to osteogenic inductors in aortic stenosis patients, which indicates probable implication of OPN, OPG, and BMP2 genes in pathogenesis of aortic valve calcification.

Osteoinductive composite coatings for flexible intramedullary nails.

This work presents composite coatings based on a copolymer of vinylidene fluoride with tetrafluoroethylene (VDF-TeFE) and hydroxyapatite (HA) for flexible intramedullary nails (FIN). The effect of the proportion of VDF-TeFE (100-25% wt.) on physicochemical and biological properties of the composite coatings was investigated. It was shown that a decrease of VDF-TeFE in the coating hinders its crystallization in ß and γ forms which have piezoelectric properties. The decrease also reduces an adhesive strength to 9.9±2.4MPa and a relative elongation to 5.9±1.2%, but results in increased osteogenesis. It was demonstrated that the composite coatings with 35% VDF-TeFE has the required combination of physicochemical properties and osteogenic activity. Comparative studies of composite coatings (35% VDF-TeFE) and calcium phosphate coatings produced using micro-arc oxidation, demonstrated comparable results for strength of bonding of these FINs with trabecular bones (~530MPa). It was hypothesized that the high osteoinductive properties of the composite coatings are due to their piezoelectric properties.

[Effect of radio frequency discharge plasma on surface properties and biocompatibility of polycaprolactone matrices].

Surface modification of bioresorbable polymer material (polycaprolactone, PCL) with abnormal glow discharge, initiated during radio-frequency magnetron sputtering of a hydroxyapatite target was investigated. Plasma treatment resulted in an increase of surface roughness of PCL, crystallite size, the surface free energy and hydrophilicity. Increased treatment time (30, 60, 150 seconds) provoked the polymer surface saturation with the sputtering target ions (calcium, phosphorus). The assessment of plasma exposure of PCL surface on bone marrow multipotent mesenchymal stromal cells behavior (BM MSCs) has been performed. Modification of the polymer surface with the abnormal glow discharge stimulated adhesion and subsequent proliferation of BM MSCs; thus, maximum values were achieved with the surface treatment for 60 s. This type of plasma modification did not affect cell viability (apoptosis, necrosis). Thus, the surface modification with abnormal glow discharge, initiated during radio-frequency magnetron sputtering of a hydroxyapatite target, appear to be a promising method of surface modification of bioresorbable polymer material (PCL) for tissue engineering.

[PHARMACOLOGICAL CARDIOPROTECTION DURING REPERFUSION OF ISOLATED HEART].

PURPOSE: To study the contractile function, the degree of damage and regional myocardial metabolism in the isolated rat heart model subjected to cardioplegic stop and reperfusion under the protection of levosimendan. MATERIALS AND METHODS: The study was performed on isolated rat hearts Wistar (group using "Custodiol" vs group using "Custodiol" + "Levosimendan". We assessed the extent of myocardial damage (in terms of markers of myocardial necrosis), the contractile function of the myocardium (coronary flow, heart rate, left ventricular pressure), the dynamics of redox processes during reperfusion with a parallel study of histology of the myocardium. RESULTS: We found a presence of cardioprotective effect of levosimendan in respect of the isolated heart in the reperfusion period of cardioplegic ischemia. The effect related to reducing the emission of reperfusion enzyme markers of myocardial damage, reducing the severity of pathological changes in the myocardium and reducing the intensity of free radical reactions in the myocardium. CONCLUSIONS: Cardioprotection with levosimendan reduces the severity of free radical attack the isolated heart, reduces the severity of damage to cardiomyocytes and preserves the contractile activity of the myocardium during reperfusion due to the effect of postconditioning.

Use of polycaprolactone grafts for small-diameter blood vessels.

Current trends are toward actively developing approaches of tissue engineering, aimed at creating vascular grafts of small diameter. This is due to the existing in cardiovascular surgery demand for prostheses to be used in coronary artery bypass grafting. The present work was undertaken in order to assess possibilities of using smalldiameter vascular grafts made of biodegradable polymer polycaprolactone by means of electrospinning. The authors studied physico-mechanical properties and structure of polycaprolactone grafts, as well as their thromboresistance and patency after implantation into the vascular bed of rats. The obtained results demonstrated optimal physicomechanical properties of the vascular grafts, their biocompatibility, endothelialisation of the internal surface, and infiltration of the graft's wall by cells with the formation of new tissue, accompanied and followed by the development of an extensive intimal layer in the zones of the anastomoses. Hence, the study showed possibilities of using polycaprolactone grafts as vascular prostheses, however requiring their further modification which would promote and contribute to a decrease in hyperplasia of connective tissue in the graft's lumen.

[Proliferative and secretory activity of human umbilical vein endothelial cells cultured under varying degrees of hypoxia].

In this study we examined the impact of 3-day hypoxia of varying degrees on the viability, proliferative and secretory activity of endothelial cells in human umbilical vein (HUVEC). The gas mixture of the three components (%) was used: 1) 10 O2, 5 CO2, 85 Ar; 2) 5 O2, 5 CO2, 90 Ar and 3) 1 O2, 5 CO2, 94 Ar. The HUVEC, cultivated in CO2-incubator under conditions of atmospheric oxygen (21% O2) were the controls. Comprehensive assessment of the results after has shown that 3-day HUVEC cultivating in the presence of 1% O2 led to pathological activation of endotheliocytes: increased NO synthesis combined with the marked secretion of endothelin-1, IL-6, IL-8 and TNF-alpha, sVCAM-1, sE-cadherin and of sE-selectin, VEGF-A and bFGF, and slow proliferation. When HUVEC were cultivated at 10% O2 and 5% O2, the level of basal secretion of the substances listed above was the least against the background of increased proliferative activity. The results showing the changes in the secretory activity of endothelial cells when cultivated under the conditions of atmospheric oxygen levels have demonstrate HUVEC activation, because the secretion of NO, IL-6, IL-8 and von Willebrand factor after 3 days of cultivation in 21% 02 exceeded that in the case of 10 and 5% O2. Thus, a gaseous medium with reduced oxygen content of up to 5% provides more physiological conditions for HUVEC cultivation. The maximum proliferative activity of HUVEC with minimal basal secretion proved such a composition to be comfortable. Increasing the oxygen content to the atmospheric level leads to the activation of endotheliocytes with signs of endothelial dysfunction, and the critical reduction in oxygen to 1% causes the development of endothelial dysfunction and reduces the proliferative potential.

[Inhalation anaesthesia and total intravenous anaesthesia for hepatic resections].

Recently number of surgeries in patients with pathology of the liver increases. Therefore optimal and safe anaesthetic management for these surgeries is very topical. The article deals with analysis of 51 anaesthesia cases: 26 cases (51%) of multimodal balanced anaesthesia with sevoflurane inhalation and 25 cases (49%) of multimodal balanced anaesthesia with continuous propofol infusion. Both techniques provide optimal anaesthetic protection, however sevoflurane use influences on the liver metabolism more significantly.

Perioperative Dynamics of TLR2, TLR4, and TREM-1 Expression in Monocyte Subpopulations in the Setting of On-Pump Coronary Artery Bypass Surgery.

Hypercytokinemia plays a key role in the pathogenesis of systemic inflammatory response syndrome (SIRS). Monocytes are the main source of cytokines in the early inflammatory phase. Simultaneous stimulation of toll-like receptors (TLRs) and triggering receptor expressed on myeloid cells (TREM-1) activating receptor on monocytes results in the amplification of the inflammatory signal and multiple increase in proinflammatory cytokine production. The dynamics of those receptors expression on monocyte surface of patients with uncomplicated SIRS course followed coronary artery bypass surgery (CABG) was studied. The increase in TLR2 and TREM-1 expression on the first day after CABG induces proinflammatory and amplification potentials of monocytes in that period. The decrease in TLR2 surface expression on the seventh day compared to the preoperative values can be regarded as a mechanism limiting inflammatory response. The highest level of TLR2, TLR4, and TREM-1 surface expression was observed in CD14(hi)CD16(+) monocyte subpopulation, confirming its proinflammatory profile.

[The role of immune monitoring in major liver resections from the position of the operative trauma and anaesthesia protection level].

Recently, the number of liver resection has increased, which requires adequate and safe anesthesia provision in this surgery area. Cytokine plasma spectrum is the one of the most important indicators characterizing inflammatory reaction intensity during and after surgery and postoperative period flow prognosis. Immune monitoring gives a notion about operative rauma features, liver damage severity, anaesthesia adequacy. Interleukin dynamics evaluation during liver resections is a topical theoretical and practical problem. The aim of this research was to evaluate the interleukins intraoperative dynamics in liver resection patients. Anaesthesia protocols were analyzed in 51 patients, 26 (51%) of which was a Sevorane multi-component balanced anaesthesia, and 25 (49%) - Propofol multi-component balanced anaesthesia. Intraoperative haemodynamics, metabolism, General and biochemical blood tests, coagulogramm, C3 and C4 complement factors dynamics, cytokines data were evaluated The obtained data testified systemic inflammatory response with a marked pro - and anti-inflammatory cytokines imbalance formation development in patients underwent organ-conserving and major liver resection. Sevorane use leads to a more pronounced anti-inflammatory cytokines synthesis in comparison with TIVA-group, the imbalance of pro - and anti-inflammatory cytokines is also more pronounced in Sevorane-group.

[Effect of different concentrations liposomal emoxipine on coronary flow, contractive and pump function of the isolated rat heart after normotermic ischema and further reperfusion].

In the experiments on the isolated perfused rat heart we studied the effects of liposomes containing different concentrations (0.25 and 0.1 mg/ml) of emoxipine on coronary flow, contractive and pump function of the isolated heart, which was effected by total normothermical ischemia and reperfusion. The parameters of the contractile activity of hearts, coronary flow and pump function of the hearts were assessed. It was detected that the introduction of the liposomal emoxipine during ischemia provides a protective effect against ischemic and reperfusion myocardial damage and smaller concentration of the emoxipin (0.1 mg/ml) in composition with the liposomes promote the best recovery of contractile activity and the pumping function of the ischemic heart in the period of the reperfusion.

[Mechanisms of coronary flow recovery in the isolated heart during reperfusion with cardioprotective liposomal emoxipin form].

In the experiments on the isolated perfused rat heart the effects of liposomes, containing different concentrations (0.25 and 0.1 mg/mL) of emoxipine, on coronary flow restoration after total normothermic ischemia and reperfusion were studied. The coronary flow, levels of nitrates and nitrites in the outflowing perfusate from heart and level of free radical processes were assessed, The obtained results showed that 0.1 mg/mL liposomal emoxipine provide with stronger increase coronary flow during reperfusion mostly due to the increase concentration of endothelial nitric oxide compare with treatments at 0.25 mg/mL.

[Assessment of anaesthesia sufficiency and stress response during liver resections].

Recently number of patients undergoing a surgery for primary and secondary liver damages is increased. Thus an adequate and safe anaesthesiological care for the surgeries is a very actual problem. The article deals with a study of anaesthesiological care in 51 patients. 26 patients (51%) received multimodal balanced anaesthesia based on sevoflurane and 25 patients (49%) received multimodal balanced anaesthesia based on continuous propofol infusion. Monitoring of haemodynamics, acid-base balance, common liquid volume, intracellular and extracellular liquid, stress hormones (cortisol and prolactin) was carried out during the surgeries. Haemodynamics and infusion and transfusion therapy were adequate during both methods of anaesthesiological care for liver resections. Strongly marked tissue injury during surgery causes neuroendocrine stress. Cortisol activity during anaesthesia based on continuous propofol infusion was less than during anaesthesia based on sevoflurane. This fact shows that propofol provides stronger protection than sevoflurane. Adequate level of anaesthesia does not cause outoregulative mechanisms suppression which is important during strongly traumatic surgery. Anaesthesia based on sevoflurane both to anaesthesia based on continuous propofol infusion is a method of choice for liver resection.

[Postoperative dynamic changes in matrix metalloproteinase levels in patients with coronary artery bypass graft procedure complications].

The purpose of this study was to evaluate diagnostic and prognostic value of matrix metalloproteinases in postoperative complication development after on-pump coronary artery bypass grafting. 29 coronary artery disease patients who had undergone on-pump coronary artery bypass grafting were examined, 4 of those had complicated systemic inflammatory response and 5 of those had isolated renal failure. Serum matrix metalloproteinase/tissue inhibitor of matrix metalloproteinase concentrations were measured before surgery, at day 1 and day 7 after surgery. Postoperative period was found to be characterized by higher levels of serum matrix metalloproteinases (MMP-9, proMMP-1) and lower levels of matrix metalloproteinase 1 tissue inhibitor. Complicated systemic inflammatory response was associated with higher levels of MMP-9, MMP-3 and proMMP-1, on particular, at day 7.

sTREM-1 as a Prognostic Marker of Postoperative Complications in Cardiac Surgery.

Cell-activating receptor TREM-1 (triggering receptor expressed on myeloid cells 1) regulates congenital immune response and contributes to systemic inflammatory response syndrome (SIRS) development. It is able to multiply cytokine production while stimulated together with the main receptors of the congenital immune system. The purpose of the paper is to study the potential use of soluble TREM-1 (sTREM-1) as a marker of intensive SIRS and a criterion for postoperative complications prediction following on-pump coronary artery bypass surgery (CABG). Results show that early postoperative sTREM-1 concentrations demonstrate their potential prognostic value regarding SIRS-associated complications.

[The autonomic nervous system and blood circulation system during different modes of anesthetic coinduction].

The autonomic nervous system (ANS) at the stage of induction was studied in 128 spinal surgical patients with chronic pain syndrome (CPS) (ASA 2-3). In all groups premedication was similar (diazepam 10 mg i.m.), intramuscular and intravenous atropine 0.5 was additionally injected in Groups 3 and 4, respectively. Introductory anesthesia used 4 coinduction modes: thiopental, fentanyl, and propofol (Group 1); propofol and fentanyl (Group 2); propofol, medazolam, and fentanyl (Groups 3 and 4). BIS monitoring was used to evaluate the impact of anesthesia and sedation on the central nervous system; autonomic screening and the Kerdo index were employed for the ANS. Intraoperative circulatory function was controlled by the values of blood pressure, heart rate, stroke index, cardiac index (CI), total peripheral resistance, and double product. Assessment of the time course of changes in ANS tone in patients with CPS indicated that after premedication, the number of parasympathotonics ranged up to 82% and that of normo- and sympathotonics was reduced to 13 and 5%, respectively. During induction with vagotonic agents, ignoring the baseline ANS tone was followed by a progression of the parasympathetic reaction of the circulatory system as a drop of CI to the low values of relevant dissadjustment in the hypokinetic hemodynamic type. There was a transition of ANS tone to physiological sympathicotonia and, accordingly, to the eukinetic circulatory type during coinduction mode 4 that suppressed vagus reflex.

[Use of average-acting nondepolarizing myorelaxants in abdominal surgery].

The efficiency and safety of use of average-acting nondepolarizing myorelaxants, such as atracurium, vecuronuim, rocuronium, and cisatracurum, during abdominal surgical interventions under general anesthesia (combined or total intravenous anesthesia) were investigated in 162 patients. There is evidence that the use of the above agents ensures a qualitative myorelaxation during different types of operations (laparoscopic and laparotomic) and no impact on hemodynamics (according to the data of continuous non-invasive monitoring) allows these myorelaxants to be used in patients with concomitant cardiovascular diseases. Rocuronium is the myorelaxant of choice for rapid tracheal intubation. The high degree of predictability of the duration of an effect produced by atracurium and cisatracurium enables these myorelaxants to be used without monitoring neuromuscular conduction. With the equally good quality of myoplegia and with virtually comparable consumed diseases, the infusion procedure of relaxation maintenance is preferable for early activation of patients. Insignificant effects on histamine release make it possible to use vecuronium, rocuronium, and cisatracurium in patients with the compromised immune system.

[Epidural block as a component of anesthesiological provision during abdominal operations].

Two modes of general anesthesia with artificial ventilation in combination with epidural block differing in a hypnotic component (inhalational or intravenous) were studied in 27 patients. A combination of general and epidural anesthesia afforded the best protection from surgical stress during major abdominal operations. Inhalational or intravenous anesthetics may be equally used to induce unconsciousness. The fact that there may be blood loss is not an absolute contraindication to epidural anesthesia. The safety and controllability of combined anesthesia increase when its individual components are monitored and the protocol is strictly observed. There is a need for further development of objective criteria for efficient regional block under general anesthesia.