Indirect co-culture of islet cells in 3D biocompatible collagen/laminin scaffold with angiomiRs transfected mesenchymal stem cells.

Diabetes is an autoimmune disease in which the pancreatic islets produce insufficient insulin. One of the treatment strategies is islet isolation, which may damage these cells as they lack vasculature. Biocompatible scaffolds are one of the efficient techniques for dealing with this issue. The current study is aimed to determine the effect of transfected BM-MSCS with angiomiR-126 and -210 on the survival and functionality of islets loaded into a 3D scaffold via laminin (LMN). AngiomiRs/Poly Ethylenimine polyplexes were transfected into bone marrow-mesenchymal stem cells (BM-MSCs), followed by 3-day indirect co-culturing with islets laden in collagen (Col)-based hydrogel scaffolds containing LMN. Islet proliferation and viability were significantly increased in LMN-containing scaffolds, particularly in the miRNA-126 treated group. Insulin gene expression was superior in Col scaffolds, especially, in the BM-MSCs/miRNA-126 treated group. VEGF was upregulated in the LMN-containing scaffolds in both miRNA-treated groups, specifically in the miRNA-210, leading to VEGF secretion. MiRNAs' target genes showed no downregulation in LMN-free scaffolds; while a drastic downregulation was seen in the LMN-containing scaffolds. The highest insulin secretion was recorded in the Oxidized dextran (Odex)/ColLMN+ group with miRNA-126. LMN-containing biocompatible scaffolds, once combined with angiomiRs and their downstream effectors, promote islets survival and restore function, leading to enhanced angiogenesis and glycemic status.

Biocompatible scaffolds based on collagen and oxidized dextran for endothelial cell survival and function in tissue engineering.

Angiogenesis is a vital step in tissue regeneration. Hence, the current study aimed to prepare oxidized dextran (Odex)/collagen (Col)-hydrogels with laminin (LMN), as an angiogenic extracellular matrix (ECM) component, for promoting human umbilical vein endothelial cell (HUVEC) proliferation and function. Odex/Col scaffolds were constructed at various concentrations and temperatures. Using oscillatory rheometry, scanning electron microscopy (SEM), and cell viability testing, the scaffolds were characterized, and then HUVEC proliferation and function was compared with or without LMN. The gelation time could be modified by altering the Odex/Col mass ratio as well as the temperature. SEM showed that Odex/Col hydrogels had a more regular three-dimensional (3D) porous structure than the Col hydrogels. Moreover, HUVECs grew faster in the Col scaffold (12 mg/mL), whereas the Odex (30 mg/mL)/Col (6 mg/mL) scaffold exhibited the lowest apoptosis index. Furthermore, the expression level of vascular endothelial growth factor (VEGF) mRNA in the group without LMN was higher than that with LMN, and the Odex (30 mg/mL)/Col (6 mg/mL) scaffold without LMN had the highest VEGF protein secretion, allowing the cells to survive and function effectively. Odex/Col scaffolds, with or without LMN, are proposed as a tissue engineering construct to improve HUVEC survival and function for angiogenesis.

The circulating level of interleukins 6 and 18 in ischemic and idiopathic dilated cardiomyopathy.

Introduction: By aging population, the heart failure and its life-threatening complications have become an enormous issue in public health. Regarding the inflammation as a major contributing pathological factor, the determination of most important inflammatory targets for immunomodulation is a problematic puzzle in the treatment of heart failure patients and the inflammatory pathways primarily involved in different underlying conditions contributing to heart failure can be an area which is worthy of focused research. Considering the dilated cardiomyopathy (DCM) as a relatively high-incident disease leading to heart failure, the aim of this study is to determine the difference in the expression level of interleukin (IL)-6 and IL-18 in patients with ischemic and idiopathic DCM. Methods: 39 non-diabetic patients with ischemic and 37 ones with idiopathic DCM were enrolled in the study. 48 healthy individuals were also considered as control group. For quantitative determination of the mRNA expression level of IL-6 and IL-18 genes, an in-house- SYBR Green real-time PCR was used and Glyceraldehyde 3-phosphate dehydrogenase (GAPDH) was considered as internal control gene. The left ventricular end-diastolic volume (LVEDV) and left ventricular ejection fraction (LVEF) was calculated by 2D echocardiographic assessment. Data were finally analyzed via SPSS statistical software version 19.0 using independent t test and 2-∆∆Ct method and P<0.05 were considered statistically significant. Results: The IL-6 was significantly higher expressed in patients with ischemic and idiopathic DCM than in healthy controls (274.3 and 168.8 times, respectively, both P values <0.001). The same higher expression of IL-18 was observed in ischemic DCM (48.5 times) and idiopathic DCM (45.2 times) compared with healthy individuals (both P values <0.001). Conclusion: Both ischemic and idiopathic DCM associates with IL-6 and IL-18 overexpression. However, no significant difference was observed between these two subtypes of DCM in either interleukin expression level. There is certainly need to further studies for evaluating the uniformity of results and also assessing other molecules in determining their roles in pathophysiology and probable utility for management.

A nested PCR method for the identification of hepatitis B virus genotype in paraffin blocks of formalin-fixed liver biopsies.

Hepatitis B virus is a hepatotropic virus that causes acute and chronic hepatitis, liver cirrhosis, and hepatocellular carcinoma; it is responsible for more than one million deaths annually worldwide despite hepatitis B vaccination. Until now, there are eight known genotypes (A-H).Clinical course of chronic hepatitis B varies according to the genotype of Hepatitis B virus. Liver biopsy becomes necessary to judge the degree of liver lesions and to make the final diagnosis, especially to make the diagnosis for latent liver damage and early-compensated cirrhosis. Genotype is very important for prognostication, but it has not yet been reported on liver tissues. Sometimes, it can be helpful to do genotyping of Hepatitis B virus of the liver tissue; such conditions include research programs, when serum is not available or when serum is negative for Hepatitis B virus DNA. In this study, we wanted to evaluate the feasibility of a simple method for genotyping of liver tissue samples. We performed genotyping of the liver biopsies and intended to find out a simple and reliable method for genotyping in the paraffin- embedded formalin- fixed liver tissue.Genotype D was the only isolated genotype in all the liver biopsies. The tissue genotype was just the same as that found in serum. The procedure was easy and good for large scale studies.Genotyping in the paraffin-embedded formalin-fixed stored liver tissue can be done with the same accuracy of the serum.

Value of serum cystatin C as a marker of renal function in the early post kidney transplant period.

Management of renal transplant patients requires periodic measurement of renal function especially in early post transplant period. This is usually assessed by measuring the creatinine clearance, but because of its limitations, it is not an ideal marker for assessing the renal function. Serum cystatin C (sCyC) appears to be an endogenous marker of glomerular filtration rate (GFR). To assess the use of sCyC as a marker of renal function in kidney transplant patients, we compared it with serum creatinine (sCr) and 24-hour urine creatinine clearance (CrCl) in the first week post-transplantation. Among 60 patients (62.8% men, 37.2% women) undergoing kidney transplantation (average age: 44.87 +/- 13.37 years), we determined renal function at 1, 3, 5, and 7 days after kidney transplantation using: sCr, sCyC and CrCl in a 24-hours urine specimen. During the first 5 days following transplantation, there was a progressive decline in sCr levels. In the first 5 days, post transplantation we could not find good correlation between CrC and sCyC, and the sCyC increased during these 5 days, but after that in day 7, there was a good correlation between CrC and sCyC which is coinciding with decreasing the dose of steroid (r= .625). Therefore, we recommend using sCyC may be used as a marker of renal function after one-week post kidney transplantation.