Comparison of the efficiency of ultrafiltration, precipitation, and ultracentrifugation methods for exosome isolation.

Extracellular vesicles (EVs) are enclosed by a lipid-bilayer membrane and secreted by all types of cells. They are classified into three groups: apoptotic bodies, microvesicles, and exosomes. Exosomes play a number of important roles in the intercellular communication and crosstalk between tissues in the body. In this study, we use three common methods based on different principles for exosome isolation, namely ultrafiltration, precipitation, and ultracentrifugation. We use field emission scanning electron microscopy (FESEM) and dynamic light scattering (DLS) analyses for characterization of exosomes. The functionality and effect of isolated exosomes on the viability of hypoxic cells was investigated by alamarBlue and Flow-cytometry. The results of the FESEM study show that the ultrafiltration method isolates vesicles with higher variability of shapes and sizes when compared to the precipitation and ultracentrifugation methods. DLS results show that mean size of exosomes isolated by ultrafiltration, precipitation, and ultracentrifugation methods are 122, 89, and 60 nm respectively. AlamarBlue analysis show that isolated exosomes increase the viability of damaged cells by 11%, 15%, and 22%, respectively. Flow-cytometry analysis of damaged cells also show that these vesicles increase the content of live cells by 9%, 15%, and 20%, respectively. This study shows that exosomes isolated by the ultracentrifugation method are characterized by smaller size and narrow size distribution. Furthermore, more homogenous particles isolated by this method show increased efficiency of the protection of hypoxic cells in comparison with the exosomes isolated by the two other methods.

Factors Affecting Late Atrial Fibrillation and Its Association With Coronary Artery Bypass Outcomes.

BACKGROUND: Although predictors and outcomes of postoperative atrial fibrillation (POAF) are well studied, evidence is lacking concerning postdischarge late/recurrent atrial fibrillation (AF). This study evaluated factors affecting late/recurrent AF and its association with coronary artery bypass grafting (CABG) outcomes in a real-world setting. METHODS: From 2012 through 2016, 5175 patients were included. Independent factors associated with late/recurrent AF were identified in a competing risk setting. Cox proportional hazard regression was used to evaluate the association between late/recurrent AF and study outcomes, consisting of all-cause mortality, major adverse cardio-cerebrovascular events, acute coronary syndrome, cerebrovascular events, and heart failure admissions. RESULTS: During a median follow-up of 60 months (quartile 1-quartile 3, 59.3-60.7 months), late/recurrent AF developed in 85 patients (1.64%). Independent factors associated with late/recurrent AF were age (subdistribution hazard ratio [sHR], 1.04; 95% CI, 1.02-1.07), left-ventricular ejection fraction (sHR, 0.97; 95% CI, 0.95-0.99), length of stay (sHR, 1.02; 95% CI, 1.01-1.04), and POAF (sHR, 4.02; 95% CI, 2.50-6.45). Late/recurrent AF was not significantly associated with all-cause mortality and major adverse cardio-cerebrovascular events at unadjusted or adjusted levels (adjusted hazard ratio, 0.80 [95% CI, 0.50-1.28] and 0.74 [95% CI, 0.48-1.13], respectively). Nevertheless, it significantly increased the unadjusted risk of cerebrovascular events (hazard ratio, 2.28; 95% CI, 01.07-4.87), which disappeared after adjustments. CONCLUSIONS: Patients with advanced age, a lower left-ventricular ejection fraction, and POAF are more likely to have late/recurrent clinical AF. Albeit counterintuitive, late/recurrent AF was not independently associated with worse midterm post-CABG outcomes. These observations need to be further elucidated in larger-scale studies and interpreted in the context of a developing country with limited resources for late AF surveillance.

Ultrasound-Triggered Microbubbles: Novel Targeted Core-Shell for the Treatment of Myocardial Infarction Disease.

Myocardial infarction (MI) is known as a main cardiovascular disease that leads to extensive cell death by destroying vasculature in the affected cardiac muscle. The development of ultrasound-mediated microbubble destruction has inspired extensive interest in myocardial infarction therapeutics, targeted delivery of drugs, and biomedical imaging. In this work, we describe a novel therapeutic ultrasound system for the targeted delivery of biocompatible microstructures containing basic fibroblast growth factor (bFGF) to the MI region. The microspheres were fabricated using poly(lactic-co-glycolic acid)-heparin-polyethylene glycol- cyclic arginine-glycine-aspartate-platelet (PLGA-HP-PEG-cRGD-platelet). The micrometer-sized core-shell particles consisting of a perfluorohexane (PFH)-core and a PLGA-HP-PEG-cRGD-platelet-shell were prepared using microfluidics. These particles responded adequately to ultrasound irradiation by triggering the vaporization and phase transition of PFH from liquid to gas in order to achieve microbubbles. Ultrasound imaging, encapsulation efficiency cytotoxicity, and cellular uptake of bFGF-MSs were evaluated using human umbilical vein endothelial cells (HUVECs) in vitro. In vivo imaging demonstrated effective accumulation of platelet- microspheres injected into the ischemic myocardium region. The results revealed the potential use of bFGF-loaded microbubbles as a noninvasive and effective carrier for MI therapy.

BMI modifies HDL-C effects on coronary artery bypass grafting outcomes.

BACKGROUND: Despite the recognized implications of high-density lipoprotein cholesterol (HDL-C) in cardiovascular diseases, the role of body mass index (BMI) in HDL-C association with cardiovascular outcomes remains unclear. This study investigated the possible modifying implications of BMI on the correlation between HDL-C and coronary artery bypass grafting (CABG) outcomes. METHODS: The present cohort included isolated CABG patients (median follow-up: 76.58 [75.79-77.38] months). The participants were classified into three groups: 18.5 ≤ BMI < 25 (normal), 25 ≤ BMI < 30 (overweight), and 30 ≤ BMI < 35 (obese) kg/m2. Cox proportional hazard models (CPHs) and restricted cubic splines (RCSs) were applied to evaluate the relationship between HDL-C and all-cause mortality as well as major adverse cardio-cerebrovascular events (MACCEs) in different BMI categories. RESULTS: This study enrolled a total of 15,639 patients. Considering the final Cox analysis among the normal and overweight groups, HDL-C ≥ 60 was a significant protective factor compared to 40 < HDL-C < 60 for all-cause mortality (adjusted hazard ratio (aHR): 0.47, P: 0.027; and aHR: 0.64, P: 0.007, respectively). However, the protective effect of HDL-C ≥ 60 was no longer observed among patients with 30 ≤ BMI < 35 (aHR: 1.16, P = 0.668). RCS trend analyses recapitulated these findings; among 30 ≤ BMI < 35, no uniform inverse linear association was observed; after approximately HDL-C≈55, its increase was no longer associated with reduced mortality risk. RCS analyses on MACCE revealed a plateau effect followed by a modest rise in overweight and obese patients from HDL-C = 40 onward (nonlinear association). CONCLUSIONS: Very high HDL-C (≥ 60 mg/dL) was not related to better outcomes among obese CABG patients. Furthermore, HDL-C was related to the post-CABG outcomes in a nonlinear manner, and the magnitude of its effects also differed across BMI subgroups.

Synergic role of zinc and gallium doping in hydroxyapatite nanoparticles to improve osteogenesis and antibacterial activity.

Recently, postoperative bone infections have been one of the most crucial challenges for surgeons. This study aims to synergistically promote antibacterial and osteoconductive properties of hydroxyapatite (HAp) nanoparticles through binary doping of Zn2+ and Ga3+ ions (Zn-Ga:HAp). Zn-Ga:HAp nanopowders with spherical morphology and homogeneous size are synthesized using a simple sol-gel method. Substitution of both zinc and gallium in the structure of HAp results in a gradual decrease in the lattice parameters as doping level increases, limits the growth of HAp particles and reduces its crystallinity. Noticeably, the crystallinity of HAp (85%) reduces to less than 73% (for XZn = 0.1), 78% (for XGa = 0.4) and 75% (for XZn = 0.1 and XGa = 0.4). Ion doping also significantly modulate the release of bioactive ions (Ca2+, PO43-, Zn2+, Ga3+) from the Zn-Ga:HAp depended on the overall amount of Ga and Zn in the HAp, which could mediate the biological responses. Incorporating both Zn2+ and Ga3+ ions in HAp structure could significantly improve the antibacterial activity of HAp nanopowders against Staphylococcus aureus (S. aureus) and Escherichia coli (E. coli) with a concentration-dependent effect. Noticeably, Zn-Ga:HAp (XZn = 0.1 and XGa = 0.4) powder shows the antibacterial activity of more than 68% and 84% against E. coli and S. aureus, respectively, at the concentration of 500 µg/ml, thereby showing excellent antibacterial properties. In addition, Zn-Ga:HAp nanopowders not only do not exhibit any cytotoxicity towards hMSCs, but also show significantly superior osteogenic properties. For instance, Zn-Ga:HAp (XZn = 0.1 and XGa = 0.4) nanopowders significantly enhance the alkaline phosphatase activity (approximately 2-fold) and mineralization (approximately 3-fold) of hMSCs after 14 days of culture, compared to pure HAp. Overall, Zn-Ga:HAp (XZn = 0.1 and XGa = 0.4) with desired osteogenesis and antibacterial activity compared to pure HAp, Zn:HAp and Ga:HAp shows promising opportunities for the implant-associated infections and the efficient healing of bone defects.

Patient-specific finite element analysis of heart failure and the impact of surgical intervention in pulmonary hypertension secondary to mitral valve disease.

Pulmonary hypertension (PH), a chronic and complex medical condition affecting 1% of the global population, requires clinical evaluation of right ventricular maladaptation patterns under various conditions. A particular challenge for clinicians is a proper quantitative assessment of the right ventricle (RV) owing to its intimate coupling to the left ventricle (LV). We, thus, proposed a patient-specific computational approach to simulate PH caused by left heart disease and its main adverse functional and structural effects on the whole heart. Information obtained from both prospective and retrospective studies of two patients with severe PH, a 72-year-old female and a 61-year-old male, is used to present patient-specific versions of the Living Heart Human Model (LHHM) for the pre-operative and post-operative cardiac surgery. Our findings suggest that before mitral and tricuspid valve repair, the patients were at risk of right ventricular dilatation which may progress to right ventricular failure secondary to their mitral valve disease and left ventricular dysfunction. Our analysis provides detailed evidence that mitral valve replacement and subsequent chamber pressure unloading are associated with a significant decrease in failure risk post-operatively in the context of pulmonary hypertension. In particular, right-sided strain markers, such as tricuspid annular plane systolic excursion (TAPSE) and circumferential and longitudinal strains, indicate a transition from a range representative of disease to within typical values after surgery. Furthermore, the wall stresses across the RV and the interventricular septum showed a notable decrease during the systolic phase after surgery, lessening the drive for further RV maladaptation and significantly reducing the risk of RV failure.

Targeted and Controlled Drug Delivery to a Rat Model of Heart Failure Through a Magnetic Nanocomposite.

As a novel cardiac myosin activator, Omecamtive Mecarbil (OM) has shown promising results in the management of systolic heart failure in clinical examinations. However, the need for repeated administration along with dose-dependent side effects made its use elusive as a standard treatment for heart failure (HF). We hypothesized that improved cardiac function in systolic HF models would be achieved in lower doses by targeted delivery of OM to the heart. To test this hypothesis, a nanocomposite system was developed by composing chitosan and a magnetic core (Fe3O4), loaded with OM, and directed toward the rats' heart via a 0.3 T magnet. HF-induced rats were injected with saline, OM, and OM-loaded nanocomposite (n = 8 in each group) and compared with a group of healthy animals (saline injected, n = 8). Knowing the ejection fraction (EF) of healthy (93.68 ± 1.37%) and HF (71.7 ± 1.41%) rats, injection of nanocomposites was associated with improved EF (EF = 89.6 ± 1.40%). Due to increased heart targeting of nanocomposite (2.5 folds), improved cardiac function was seen with only 4% of the OM dose required for infusion, while injecting the same dose of OM without targeting was unable to stop HF progression (EF = 55.33 ± 3.16%) during 7 days. In conclusion, heart nanocomposites targeting improves the EF by up to 18% by only using 4% of the doses traditionally used in treating the HF.

The Therapeutic Potential of Mesenchymal Stem Cell-Derived Exosomes in Treatment of Neurodegenerative Diseases.

Neurologic complications are commonly regarded as irreversible impairments that stem from limited potential of regeneration of the central nervous system (CNS). On the other side, the regenerative potential of stem cells has been evaluated in basic research, as well as in preclinical studies. Mesenchymal stem cells (MSCs) have been regarded as candidate cell sources for therapeutic purposes of various neurological disorders, because of their self-renewal ability, plasticity in differentiation, neurotrophic characteristics, and immunomodulatory properties. Exosomes are extracellular vesicles which can deliver biological information over long distances and thereby influencing normal and abnormal processes in cells and tissues. The therapeutic capacity of exosomes relies on the type of cell, as well as on the physiological condition of a given cell. Therefore, based on tissue type and physiological condition of CNS, exosomes may function as contributors or suppressors of pathological conditions in this tissue. When it comes to the therapeutic viewpoint, the most promising cellular source of exosomes is considered to be MSCs. The aim of this review article is to discuss the current knowledge around the potential of stem cells and MSC-derived exosomes in the treatment of neurodegenerative diseases.

Evaluation of quality of life after cardiac surgery in high-risk patients.

BACKGROUND: Conventionally, there is controversy over subjecting high-risk patients to cardiac operations, due to major postoperative complications. Higher survival rates and less morbidity as well as better quality of life can be good predictors of the outcome of surgery. This study evaluates the quality of life before and 12 months after cardiac operations on high-risk patients. METHODS: In this study, the European System for Cardiac Operative Risk Evaluation (EuroSCORE) II was used to separate high-risk patients from others. The quality of life was assessed using the Medical Outcomes Study 36-item Short Form Health Survey (SF-36) before surgery and one year afterward. Based on SF-36, the score for each of the eight different dimensions of the quality of life was quantified; and, their differences between pre-surgery and post-follow up period were analyzed. RESULTS: 126 high-risk patients were included in this study. The mean age of the patients was 64.29 ± 12.35 years. The median of EuroSCORE II score in these cases was 6.83 (6.04-25.98). The results reveal that the majority of the quality of life dimensions, except mental health, improved significantly after the follow-up period. CONCLUSION: Cardiac surgery on high-risk patients can noticeably promote the different aspects of their quality of life; although, such improvements should be considered against surgical complications.

Comparison between mechanical properties of human saphenous vein and umbilical vein.

BACKGROUND: As a main cause of mortality in developed countries, Coronary Artery Disease (CAD) is known as silent killer with a considerable cost to be dedicated for its treatment. Coronary Artery Bypass Graft (CABG) is a common remedy for CAD for which different blood vessels are used as a detour. There is a lack of knowledge about mechanical properties of human blood vessels used for CABG, and while these properties have a great impact on long-term patency of a CABG. Thus, studying these properties, especially those of human umbilical veins which have not been considered yet, looks utterly necessary. METHODS: Umbilical vein, as well as human Saphenous vein, are respectively obtained after cesarean and CABG. First, histological tests were performed to investigate different fiber contents of the samples. Having prepared samples carefully, force-displacement results of samples were rendered to real stress-strain measurements and then a fourth-order polynomial was used to prove the non-linear behavior of these two vessels. RESULTS: Results were analyzed in two directions, i.e. circumferentially and longitudinally, which then were compared with each other. The comparison between stiffness and elasticity of these veins showed that Saphenous vein's stiffness is much higher than that of umbilical vein and also, it is less stretchable. Furthermore, for both vessels, longitudinal stiffness was higher than that of circumferential and in stark contrast, stretch ratio in circumferential direction came much higher than longitudinal orientation. CONCLUSION: Blood pressure is very high in the region of aorta, so there should be a stiff blood vessel in this area and previous investigations showed that stiffer vessels would have a better influence on the flow of bypass. To this end, the current study has made an attempt to compare these two blood vessels' stiffness, finding that Saphenous vein is stiffer than umbilical vein which is somehow as stiff as rat aortic vessels. As blood vessel's stiffness is directly related to elastin and mainly collagen content, results showed the lower amount of these two contents in umbilical vein regarding Saphenous vein.

Relationship between serum magnesium level and arrythmias following post-coronary artery bypass grafting.

INTRODUCTION: Atrial and ventricular arrhythmias are among the most common complications after coronary artery bypass graft (CABG) surgery. It is known that cardiopulmonary bypass reduces serum magnesium level. In this study, we evaluated the relationship between total blood magnesium level (TMG) and the incidence of perioperative arrhythmias. METHODS: TMG was measured in patients who were scheduled for CABG on three occasions: just before anesthesia, on intensive care unit (ICU) arrival and on the first morning after operation. Patients were evaluated for primary cardiac rhythm, serum creatinine, urine output in operating room and diuretic therapy. Supplemental magnesium (SMG) was also recorded in operating room and ICU. Patients were then evaluated for the rate and kind of arrhythmia occurring during the next 3 days. RESULTS: Mean TMG level in 170 cases was 2.2 (0.5), 2.6 (0.6) and 2.4 (0.6) mg/dl on three occasions respectively. 53 patients developed post-operative arrhythmia (31%) [Atrial Fibrillation (AF) (7.1%), Non-AF Supraventricular arrhythmia (14.7%) and Ventricular arrhythmia (16.5%)]. Although there was a significant difference between TMG on three occasions (P <0.001), all values were within normal range. Although TMG was higher in arrhythmic patients compared to non- arrhythmics (2.26 vs. 2.14), both values were in normal range and there was no significant difference between two groups. DISCUSSION: This study shows that routine magnesium administration has no significant effect on serum magnesium level. We conclude that though routine regimen of magnesium administration has no effect on incidence of perioperative arrhythmia, it is probably necessary for maintaining normal magnesium level.

Magnesium infusion and postoperative atrial fibrillation: a randomized clinical trial.

BACKGROUND: Postoperative arrhythmias are among the most common complications of cardiac surgery. Total serum magnesium concentration will change after coronary bypass surgery but compensatory prophylactic administration of magnesium has remained a controversial issue. We studied whether prophylactic administration of magnesium could prevent post-coronary artery bypass grafting (CABG) arrhythmias and evaluated the effects of diabetes mellitus on prophylactic magnesium administration. METHODS: In a clinical trial, 345 consecutive CABG candidates were randomly assigned to study (n = 166, 48.1%) and control groups. Patients in study group received supplemental magnesium infusion as following: 2 g [corrected] after induction of anesthesia until cardio-pulmonary bypass and then 8 g upon arrival in Intensive Care Unit (ICU) until 24 hr. Total serum magnesium concentration was measured at four designated time points: onset of induction, and 0, 24 and 48 hr after ICU admission. Cardiac arrhythmias were sought with a 12-lead electrocardiogram (ECG) from the end of surgery up till discharge. RESULTS: Atrial Fibrillation (Af) occurred in 34 patients (9.9%). Total serum magnesium concentration was significantly higher in patients who received supplemental magnesium (P < 0.001) and significantly lower in Af patients (P= 0.02). Among non-diabetics, Af incidence was significantly lower in study group compared with control group. CONCLUSIONS: The occurrence of atrial fibrillation correlates with serum magnesium level. Diabetes mellitus probably hampers prophylactic effect of supplemental magnesium in preventing the occurrence of Af.