Background: Endometrium is recently introduced as an available source of mesenchymal stem cells (EnMSCs), which can be obtained without anesthesia and side effects. Regarding the issues and complexities of cell-based therapies, exosomes gain tremendous attention as a novel tool for cell-free therapies. Although several clinical trials are recently established based on therapeutic potential of EnMSCs, biological roles of EnMSC-derived exosomes are still unclear. Methods: The current study was conducted to investigate the potential effects of EnMSC-derived exosomes on proliferation, migration, and angiogenesis of human umbilical cord vein endothelial cells (HUVECs). For this purpose, EnMSCs and then EnMSC-derived exosomes were isolated and characterized. MTT assay and wound healing assay as well as tube formation assay were applied. Results: The collected data showed that EnMSC-derived exosomes significantly increased proliferation, migration, and angiogenesis of HUVECs. It was observed that the effects of exosomes were applied in a dose dependent manner. In addition, expression analysis by quantitative real-time PCR showed that increased expression of proliferation and angiogenesis genes in HUVECs were treated with EnMSC-derived exosomes in a dose dependent manner. Conclusions: The current study results showed that EnMSC-derived exosomes can exert biological effects such as their source cells and become new candidates for cell-free therapies. Taken together, increased angiogenesis makes EnMSC-derived exosomes a promising tool in regenerative medicine, especially wound healing and treatment of vascular disease.

Background Hepatocellular carcinoma (HCC) can lead to liver failure which renders to liver transplant. miRNAs might be detected as biomarkers in subclinical stage of several hepatobiliary disorders like HCC. Therefore, in the present study, alterations in miRNAs as biomarkers were detected in LT patients with HCC. Methods Fourteen tissue samples composed of 5 rejected and 9 non-rejected ones were used for studying the miRNAs expression pattern using LNA-array probe assay and the result was evaluated by in house SYBR Green Real-time PCR protocols on 30 other tissue samples composed of 10 rejected and 20 non-rejected ones for the selected miRNAs. All samples were collected from liver transplanted patients with HCC. Results The study results revealed that in rejected patients compared to non-rejected ones, hsa-miR-3158-5p, -4449, -4511, and -4633-5p were up-regulated and hsa-miR-122-3p, -194-5p, 548as-3p, and -4284 were down-regulated. ROC curve analysis also confirmed that miR194-5p and -548as-3p in up-regulated and also, miR-3158-5p, -4449 in down-regulated microRNAs are significantly important molecules in rejection. Conclusion Finally, the tissue levels of specific miRNAs (especially hsa-miR-3158-5p, -4449, -194-5p and -548as-3p) significantly correlated with the development of HCC, which can be present as biomarkers after further completing studies.

Background RADA16I represents one of promising hydrogel forming peptides. Several implementations of RADA16I hydrogels have proven successful in the field of regenerative medicine and tissue engineering. However, RADA16I peptides used in various studies utilize synthetic peptides and so far, only two research articles have been published on RADA16I peptide recombinant production. Moreover, previous studies utilized non- or less routine expression and purification methods to produce RADA16I peptide recombinantly. Objectives The main goal was to produce the self-assembling peptide, RADA16I, in Escherichia coli by exploiting routine and widely used vectors and purification methods, in shake flask. Material and Methods RADA16I coding sequence was inserted in pET31b+, and the construct was transformed into E. coli. Purified fusion constructs were purified using Nickel Sepharose. RADA16I unimers were released using CNBr cleavage. CD and FTIR spectroscopy were used to study recombinant RADA16I’s confirmation. TEM was used to confirm fibril formation of recombinant RADA16I. Furthermore, MTT assay was implemented to assess cytocompatibility of recombinant RADA16I. Results The biochemical, biophysical and structural analysis proved the ability of the recombinant RADA16I to form self-assembling peptide nanofibers. Furthermore, the nanofibers exhibited no cytotoxicity and retained their cell adhesive activity. Conclusions We successfully produced RADA16I in acceptable levels and established a basis for future investigation for the production of RADA16I under fermentation conditions.

Studies on the blood of patients with prostate cancer using Dynamic Light Scattering (DLS) and corona protein size changes have shown that this test is highly specific and sensitive, but this method has not been studied in Iran, and therefore this study intends to perform this procedure using gold nanoparticles in prostate cancer detection. Blood samples of 60 male subjects aged 40–90 years were collected from 20 healthy, 20 benign and 20 prostate cancer patients. Optical scattering changes were measured by the level of gold nanoparticles mixed with these sera, and the responses were compared with the PSA index (Prostate Specific Antigen) of the subjects. Results of D2/D1 ratio analysis were performed using SPSS statistical software R. No significant differences were found in the size of the corona protein structure between the three groups of males with cancer, males with benign tumor, and healthy males. No correlation was found between the light scattering concentration and PSA serum level Due to changes in ambient temperature, prolonged test duration or high IgG levels in apparently healthy individuals, this test is not feasible in Iran. Performing this test requires advanced equipment to maintain the same temperature that do not exist in Iran. DLS also has major limitations for prostate cancer detection, so it cannot be a simple and accurate method for the early detection of prostate cancer, and it is suggested that other methods be used to diagnose.

Poster session 3, September 23, 2022, 12:30 PM - 1:30 PM Objectives Rhizopus oryzae is one of the most common causes of mucormycosis. Among the virulence factors of the Mucorales, CotH protein has recently been identified, which causes the invasion of R. oryzae into endothelial cells. In this study, we aimed to examine the reaction between GRP78 at the level of human cells and different groups of mice and CotH3 at the surface of the R. oryzae hyphae. We evaluated the relative expression of GRP78 and CotH3 genes and changes in the expression of some miRNAs that target the human GRP78 gene. Methods In this study, the relative changes in gene expression were studied. In three groups (1) Macrophages derived from human monocytes: monocytes from the blood of healthy donors were isolated using Ficoll and in RPMI 1640 medium containing FCS 10% and with penicillin-streptomycin after 2 weeks were differentiated into macrophages. Two groups were investigated, including control and infected with R. oryzae hyphae, for 6 and 16 hours after infection. (2) Hematogenous dissemination mucormycosis model: Seven groups of male BALb/c mice were examined in control, infected, and treated groups with Liposomal amphotericin B. (3) Human mucormycosis: in this study, two samples of patients with rhinocerebral mucormycosis with diabetes mellitus treated and untreated with liposomal amphotericin B were examined. Total RNA extraction and cDNA synthesis were performed from the studied samples. The relative expression changes of the target genes and miRNAs were evaluated using real-time PCR carried out using Sybrgreen-based detection methods. Results Monocyte-derived macrophages had a steady pattern in relative changes in gene expression. An increase in expression of two genes, GRP78 and CotH3, was observed in the samples, and all miRNAs targeted by the GRP78 gene included hsa - miR-16-5p; hsa -miR-335-5p and hsa -miR-93-3p showed a decreasing pattern. In the mice mucormycosis model, relative gene expression changes were observed, and mmu-miR-181b-5p showed increased expression deviation in all groups. The clinical sample of diabetic patients with untreated rhinocerebral mucormycosis also had a consistent pattern of GRP78 and CotH3 increased gene expression. The hsa-miR-16-5p and hsa-miR-335-5p have increased expression, while hsa-miR-93-3p decreased expression. Conclusion After validation, these micro RNAs can be used as valuable markers in mucormycosis detection and treatment processes.