Angiogenesis outcomes of metformin utilization in diabetes mellitus: a systematic review and meta-analysis.

OBJECTIVE: To review relevant literature regarding the role of metformin in angiogenesis among diabetic patients. METHODS: The systematic review and meta-analysis conducted from May to September 2022, and comprised search on Medline, ScienceDirect, ProQuest, Web of Science, EBSCOhost and Cochrane Library databases. The studies included were published in the English language and were human studies having angiogenesis endothelial markers as the outcomes of interest among patients of type 2 diabetes mellitus undergoing metformin therapy. Endothelial markers, including vascular endothelial growth factor, von-Willebrand-factor, plasminogen activator inhibitor-1, soluble vascular adhesion molecule- 1, intercellular adhesion molecule-1, soluble endothelialselectin, tissue plasminogen activator, urinary albumin excretion, platelet endothelial cell adhesion molecule-1 and thrombin-activatable fibrinolysis inhibitor, were assessed as angiogenesis outcomes. Data was statistically analysed using Review Manager 5.4. RESULTS: Of the 413 studies identified, 8(1.9%) were included; 5(62.5%) randomised control trials, 2(25.0%) cross-sectional, and 1(12.5%) cohort studies, with overall 1199 patients. Among the outcomes, von-Willebrandfactor (p=0.01), soluble vascular adhesion molecule-1 (p<0.00001), intercellular adhesion molecule-1 (p=0.0003), soluble endothelial-selectin (p=0.007), and tissue plasminogen activator (p<0.00001) showed significantly lower levels after metformin treatment using the random effect methods. CONCLUSIONS: Metformin was found to have an additional effect of endothelial function improvement.

Isolation and Culture of Non-adherent Cells for Cell Reprogramming.

Coronary heart disease (CHD) is a leading cause of death globally, while its current management is limited to reducing the myocardial infarction area without actually replacing dead cardiomyocytes. Direct cell reprogramming is a method of cellular cardiomyoplasty which aims for myocardial tissue regeneration, and CD34+ cells are one of the potential sources due to their shared embryonic origin with cardiomyocytes. However, the isolation and culture of non-adherent CD34+ cells is crucial to obtain adequate cells for high-efficiency genetic modification. This study aimed to investigate the optimal method for isolation and culture of CD34+ peripheral blood cells using certain culture media. A peripheral blood sample was obtained from a healthy subject and underwent pre-enrichment, isolation, and expansion. The culture was subsequently observed for their viability, adherence, and confluence. Day 0 observation of the culture showed a healthy CD34+ cell with a round cell shape, without any adherent cells present yet. Day 4 of observation showed that CD34+ cells within the blood plasma medium became adherent, indicated by their transformations into spindle or oval morphologies. Meanwhile, CD34+ cells in vitronectin and fibronectin media showed no adherent cells and many of them died. Day 7 observation revealed more adherent CD34+ cells in blood plasma medium, and which had 75% of confluence. In conclusion, the CD34+ cells that were isolated using a combination of density and magnetic methods may be viable and adequately adhere in culture using blood plasma medium, but not in cultures using fibronectin and vitronectin.

Efficiency Comparison of Direct Reprogramming CD34+ Cells into Cardiomyocytes Using Cardiomyocyte Differentiation Medium vs MicroRNA-1.

The development of a direct reprogramming method to provide cell availability for regenerative therapy has led to a lot of studies. However, the search for appropriate cell sources and methods is still being carried out until now. Direct reprogramming using microRNA-1 (miR-1) is an option to obtain cardiomyocytes from other cells because miR-1 has evidence to play a role in the development of cardiac muscle cells in the embryo. This study aimed to compare the direct reprogramming efficiency of CD34+ cells from peripheral blood into cardiomyocytes between cardiomyocyte differentiation medium and miR-1. CD34+ cells from peripheral blood isolation and expansion process was conducted for 7 days using magnetic-activated cell sorting. Cardiomyocyte differentiation medium added in P1 group and transfection of miR-1 in P2 group of cell culture. Cardiac troponin immunocytochemistry staining and measurement were done on the fifth day after cell culture treatment. Cardiac troponin expression was observed higher in the P2 group (median 31.34) compared to the P1 group (median 21.06) (p = 0.000). The efficiency of direct reprogramming of CD34+ cells into cardiomyocytes with cardiomyocyte differentiation medium was 13%-21% and with miR-1 transfection was 31%-32%. Both the addition of miR-1 and cardiomyocyte differentiation medium could directly reprogram CD34+ cells into cardiomyocytes. The efficiency of miR-1 in reprogramming CD34+ cells into cardiomyocytes is superior to cardiomyocytes differentiation medium.

Acute effects of cigarette smoke on Endothelial Nitric Oxide synthase, vascular cell adhesion molecule 1 and aortic intima media thickness.

Background. Cigarette smoking could induce endothelial dysfunction and the increase of circulating markers of inflammation by activation of monocytes. This can lead to increased intima media thickness (IMT) of entire blood vessels and result in acceleration of the atherosclerosis process. However, to our knowledge, little is known about the role of cigarette smoking in this atherosclerotic inflammatory process. The aim of this study is to explore the link between cigarette smoking and its effect on endothelial nitric oxide synthase (e-NOS) and vascular cell adhesion molecule 1 (VCAM-1). Methods. An experimental study with a post-test only controlled group design was used. We used 18 Wistar rats ( Rattus norvegicus) randomly subdivided into two groups: group K (-) were not exposed to tobacco smoke, whereas group K (+) were exposed to smoke equivalent of more than 40 cigarettes for 28 days daily. After 28 days, samples were analyzed for e-NOS, VCAM-1 and aortic IMT. Results . Our results indicate that tobacco smoke can enhance the expression of VCAM-1 on rat cardiac vascular endothelial cells, resulting in a decreased expression of e-NOS level and increase of aortic IMT. Linear regression model found that eNOS level negatively correlated wiith aortic IMT ( r 2 = 0.584, ß = -0.764, p < 0.001), whereas VCAM-1 expression did not correlate with aortic IMT ( r 2 = 0.197, p = 0.065). Conclusion. Low e-NOS level and high VCAM-1 level observed after cigarette smoke exposure which may increase aortic IMT.

Effect of active immunization with IL-17A on B cell function and infection risk in pristane-induced lupus model.

PURPOSE: The aim of this study was to determine the effect of active immunization of interleukin (IL)-17A to inhibit B cell functions and monitor the risk of infection in a pristane-induced lupus mice model. METHODS: Female Balb/c mice were given a single intraperitoneal injection of 0.5 mL pristane. IL-17A was coupled to keyhole limpet hemocyanin (KLH) and given to mice in three different doses: D0 (0 µg/mL), D1 (1 µg/mL), and D2 (10 µg/mL). The vaccine was given three times with 3-week intervals. At day 42, mice were injected intraperitoneally with methicillin-resistant Staphylococcus aureus (MRSA) and monitored for 3 weeks. Plasma cells proliferation, Th17 and plasma cell percentages were measured by flow cytometry; anti-IL-17A antibody titers, IL-17A, and anti-double-stranded DNA (anti-dsDNA) levels were measured by enzyme-linked immunosorbent assay; and MRSA colonization was measured by bacterial counter. RESULTS: Anti-IL-17A antibody titers were significantly higher in D2 compared to D0 (P = 0.012). Serum IL-17A levels were also significantly lower in D2 compared to D0 (P = 0.000) while Th17 percentages were not significantly different between groups. D2 was also had significantly lower anti-dsDNA (P = 0.021), lower plasma cell percentages (P = 0.000) and lower B cell proliferation rate (P = 0.001) compared to D0. Analysis for the risk of infection also revealed that D2 did not increase the risk of infection compared to D0 (P = 0.504). CONCLUSION: Active immunization with IL-17A coupled to KLH was able to induce a high titer of neutralizing antibodies against IL-17A and inhibit B cell functions without increasing the risk of infection in a pristane-induced lupus mice model.