1.Generation of Scalable Hepatic Micro-Tissues as a Platform for Toxicological Studies
Sara DARAKHSHAN ; Ali Bidmeshki POUR ; Reza KOWSARI-ESFAHAN ; Massoud VOSOUGH ; Leila MONTAZERI ; Mohammad Hossein GHANIAN ; Hossein BAHARVAND ; Abbas PIRYAEI
Tissue Engineering and Regenerative Medicine 2020;17(4):459-475
BACKGROUND:
Currently, there is an urgent need for scalable and reliable in vitro models to assess the effects oftherapeutic entities on the human liver. Hepatoma cell lines, including Huh-7, show weakly resemblance to humanhepatocytes, limiting their significance in toxicity studies. Co-culture of hepatic cells with non-parenchymal cells, and thepresence of extracellular matrix have been shown to influence the biological behavior of hepatocytes. The aim of this studywas to generate the scalable and functional hepatic micro-tissues (HMTs).
METHODS:
The size-controllable HMTs were generated through co-culturing of Huh-7 cells by mesenchymal stem cellsand human umbilical vein endothelial cells in a composite hydrogel of liver-derived extracellular matrix and alginate, usingan air-driven droplet generator.
RESULTS:
The generated HMTs were functional throughout a culture period of 28 days, as assessed by monitoringglycogen storage, uptake of low-density lipoprotein and indocyanine green. The HMTs also showed increased secretionlevels of albumin, alpha-1-antitrypsin, and fibrinogen, and production of urea. Evaluating the expression of genes involvedin hepatic-specific and drug metabolism functions indicated a significant improvement in HMTs compared to two-dimensional(2D) culture of Huh-7 cells. Moreover, in drug testing assessments, HMTs showed higher sensitivity tohepatotoxins compared to 2D cultured Huh-7 cells. Furthermore, induction and inhibition potency of cytochrome P450enzymes confirmed that the HMTs can be used for in vitro drug screening.
CONCLUSION
Overall, we developed a simple and scalable method for generation of liver micro-tissues, using Huh-7,with improved hepatic-specific functionality, which may represent a biologically relevant platform for drug studies.
2.Generation of Scalable Hepatic Micro-Tissues as a Platform for Toxicological Studies
Sara DARAKHSHAN ; Ali Bidmeshki POUR ; Reza KOWSARI-ESFAHAN ; Massoud VOSOUGH ; Leila MONTAZERI ; Mohammad Hossein GHANIAN ; Hossein BAHARVAND ; Abbas PIRYAEI
Tissue Engineering and Regenerative Medicine 2020;17(4):459-475
BACKGROUND:
Currently, there is an urgent need for scalable and reliable in vitro models to assess the effects oftherapeutic entities on the human liver. Hepatoma cell lines, including Huh-7, show weakly resemblance to humanhepatocytes, limiting their significance in toxicity studies. Co-culture of hepatic cells with non-parenchymal cells, and thepresence of extracellular matrix have been shown to influence the biological behavior of hepatocytes. The aim of this studywas to generate the scalable and functional hepatic micro-tissues (HMTs).
METHODS:
The size-controllable HMTs were generated through co-culturing of Huh-7 cells by mesenchymal stem cellsand human umbilical vein endothelial cells in a composite hydrogel of liver-derived extracellular matrix and alginate, usingan air-driven droplet generator.
RESULTS:
The generated HMTs were functional throughout a culture period of 28 days, as assessed by monitoringglycogen storage, uptake of low-density lipoprotein and indocyanine green. The HMTs also showed increased secretionlevels of albumin, alpha-1-antitrypsin, and fibrinogen, and production of urea. Evaluating the expression of genes involvedin hepatic-specific and drug metabolism functions indicated a significant improvement in HMTs compared to two-dimensional(2D) culture of Huh-7 cells. Moreover, in drug testing assessments, HMTs showed higher sensitivity tohepatotoxins compared to 2D cultured Huh-7 cells. Furthermore, induction and inhibition potency of cytochrome P450enzymes confirmed that the HMTs can be used for in vitro drug screening.
CONCLUSION
Overall, we developed a simple and scalable method for generation of liver micro-tissues, using Huh-7,with improved hepatic-specific functionality, which may represent a biologically relevant platform for drug studies.
3.The effect of embryo catheter loading technique on the live birth rate.
Marjan OMIDI ; Iman HALVAEI ; Esmat MANGOLI ; Mohammad Ali KHALILI ; Mohammad Hossein RAZI
Clinical and Experimental Reproductive Medicine 2015;42(4):175-180
OBJECTIVE: Embryo loading (EL) is a major step in embryo transfer (ET) and affect on the success of in vitro fertilization (IVF). This study aimed to compare the effect of two different EL techniques on the rates of pregnancy and delivery in IVF/ET cycles. METHODS: 207 fresh ET and 194 Frozen-thawed ET (FET) cycles were included in this retrospective study. Two groups (A and B) were defined based on the EL technique used. In group A, the entire catheter was flushed with Ham's F-10 medium. The embryos were then drawn into the catheter using one air bracket. In group B, 70 microL of air was aspirated into the syringe and the catheter was flushed using Ham's F10 medium. The medium, air, embryos, air, and finally another layer of medium were then sequentially drawn into the catheter. The main outcome measures were the pregnancy and delivery rates. RESULTS: The groups did not differ with respect to the etiology of infertility, the source of spermatozoa, the quality of the embryos, the type of EL catheter, and the ease of transfer. The pregnancy rate was similar between two groups. In fresh ET cycles, a higher delivery rate was observed in group B than it group A (78.1% vs. 60%, p=0.1). In FET cycles, the rate of delivery was significantly higher in group B than in group A to a nonsignificant extent (88.9% vs. 58.8%, p=0.06). CONCLUSION: EL techniques did not have a significant impact on the delivery rate in either fresh or FET cycles.
Catheters*
;
Embryo Transfer
;
Embryonic Structures*
;
Fertilization in Vitro
;
Infertility
;
Live Birth*
;
Outcome Assessment (Health Care)
;
Pregnancy
;
Pregnancy Rate
;
Retrospective Studies
;
Spermatozoa
;
Syringes
4. Epidemiological, molecular characterization and risk factors of human brucellosis in Iran
Afshar ETEMADI ; Rezvan MONIRI ; Mahmood SAFFARI ; Hossein AKBARI ; Saeed ALAMIAN ; Ali Mohammad BEHROZIKHAH
Asian Pacific Journal of Tropical Medicine 2020;13(4):169-175
Objective: To determine epidemiological, molecular characterization, and potential risk factors of human brucellosis. Methods: This descriptive study was carried out in the clinical setting in Iran between 2017 and 2018. A total of 297 participants enrolled in the study. The sample size was calculated based on the occurrence rate of brucellosis in different areas. Patients were assessed using serological tests and conventional culture methods. Phage and multiplex PCR methods typed all of Brucella isolates. Potential risk factors of disease were determined. Results: A total of 141 of 297 (47.5%) Brucella strains were isolated and all of them were detected as Brucella melitensis biovar 1. Based on serologic titers, high culture positivity was recorded at 1/640 titer (P< 0.006). The risk factors for brucellosis were patients older than 40 years (OR=2.23, 95%CI: 1.4-3.55, P=0.001), animal keeper (OR=7, 95%CI: 1.51-32.41, P=0.005), housewife (OR=8.76, 95%CI: 1.85-41.37, P=0.002), farmer (OR=6.42, 95%CI: 1.21-33.97, P=0.019), and contact with animal (OR=1.31, 95%CI: 0.60-2.85, P=0.005). Conclusions: To the best of our knowledge, this is the first comprehensive report from Iran presenting the detection of Brucella species by the multiplex PCR. Brucella melitensis biovar 1 is still the dominant causative agent in Iran. The consumption of unpasteurized dairy products, living in rural areas, and animal contact were risk factors of brucellosis.