Increased Sensitivity of Computed Tomography Scan for Neoplastic Tissues Using the Extracellular Vesicle Formulation of the Contrast Agent Iohexol.

Computed tomography (CT) is a diagnostic medical imaging modality commonly used to detect disease and injury. Contrast agents containing iodine, such as iohexol, are frequently used in CT examinations to more clearly differentiate anatomic structures and to detect and characterize abnormalities, including tumors. However, these contrast agents do not have a specific tropism for cancer cells, so the ability to detect tumors is severely limited by the degree of vascularization of the tumor itself. Identifying delivery systems allowing enrichment of contrast agents at the tumor site would increase the sensitivity of detection of tumors and metastases, potentially in organs that are normally inaccessible to contrast agents, such as the CNS. Recent work from our laboratory has identified cancer patient-derived extracellular vesicles (PDEVs) as effective delivery vehicles for targeting diagnostic drugs to patients' tumors. Based on this premise, we explored the possibility of introducing iohexol into PDEVs for targeted delivery to neoplastic tissue. Here, we provide preclinical proof-of-principle for the tumor-targeting ability of iohexol-loaded PDEVs, which resulted in an impressive accumulation of the contrast agent selectively into the neoplastic tissue, significantly improving the ability of the contrast agent to delineate tumor boundaries

Inhibition of miR-155-5p in non-small cell lung cancer, a potential target for induction of autophagy

Objectives: The function of autophagy appears to be diverse in cancer. Autophagy also has a complex relationship with apoptosis under different cell conditions. Considering the high mortality and morbidity of Non-small Cell Lung Cancer (NSCLC) around the world, and the potential role of miR-155-5p in cancer progression, we aimed to investigate the autophagy and apoptosis of NSCLC cell line after inhibition of miR-155-5p. Methods and results: The antimiR-155-5p and Scramble oligonucleotides were transfected into the A549 human adenocarcinoma lung cancer. After 24 h, the expression level of SIRT1, SIRT2, TP53INP1, and mTOR were evaluated for both groups performing qRT-PCR. After 48 h, the apoptosis assay was also performed using Annexin V-FITC/PI-based flow cytometry. The transfection of antimiR-155-5p decreased the miR-155-5p expression significantly, which increased the expression levels of SIRT1and TP53INP1 as miR-155-5p target genes and expression of SIRT2 as an indirect target. The high expression of SIRT1 inhibited the mTOR pathway leading to enhanced induction of autophagy. The excess autophagy increased the apoptosis rate compared to the scramble group. Conclusion: The exact control of autophagy is an important issue for normal and cancer cells. Here, inhibition of miR-155-5p induced autophagy in a substantial way leading to autophagic cell death. Therefore, targeting miR-155-mediated autophagy might be a potential therapeutic method for NSCLC

Does survivin overexpression enhance the efficiency of fibroblast cell-based wound therapy?

Cell-based wound therapy is faced with some limiting factors that decrease the therapeutic efficacy of transplanted cells. In this study, we aimed to genetically modify fibroblast cells with anti-apoptotic Survivin gene (Birc5) before cell transplantation. In vitro, pIRES2-eGFP-Survivin plasmid was transfected into the fibroblast cells and the growth curve was evaluated for transfected and normal cells performing MTT assay. In vivo, two 6-diameter cutaneous wounds were created at mice dorsal skin. Fibrin clot was used as a delivery vehicle to transfer cells into the wound bed. The effects of four treatment groups including (a) Cell-SVV-Clot (b) Cell-GFP-Clot, (c) Normal cell-Clot and, (d) Clot alone were evaluated. After 1,2,3,7 and 14 days post-transplantation, the wounds were photographed for evaluating the wound closure rate and wound samples were obtained. Angiogenesis and formation of granulated tissue were assessed via H&E staining for wound samples. The expression levels of Survivin, VEGF, and bFGF genes were also determined using qRT-PCR. The MTT assay showed similar proliferation potential of transfected cells with normal cells verifying that Survivin had no detrimental effect. Compared to the Normal cell-Clot group, the Survivin overexpression was seen for 3 days in the Cell-SVV-Clot group verifying the cell survival during the early stage of wound healing. The Survivin further upregulated VEGF and bFGF expressions resulting in more angiogenesis and formation of granulated tissue by day 3 and 14. The treated wounds with Cell-SVV-Clot were regenerated with a higher wound closure rate by day 7 compared to Normal cell-Clot and Clot groups. Survivin enhanced wound healing through induction of VEGF and bFGF at particular times post-wounding that led to a more structured-epidermis with higher angiogenesis and granulation tissue formation rate

The Effect of Encapsulated Umbilical Cord-derived Mesenchymal Stem Cells in PRPCryogel on Regeneration of Grade-II Burn Wounds

Purpose: To achieve an efficient approach for burn regeneration, we aimed to investigate the effects of encapsulated human umbilical cord-derived mesenchymal stem cells (hUMSCs) in platelet-rich plasma cryogel (PRPCryogel) on regeneration of grade-II burn wounds in a rat model. Methods: MSCs were extracted from the umbilical cord and characterized by flow cytometry. Forty rats were divided into 4 groups including PRPCryogel-MSCs, PRPCryogel, MSCs, and control treatment. After 7, 14, 21, and 28 days post-treatment, the wound sites were photographed and wound samples were taken. H&E staining for wound samples was done to assess re-epithelialization, angiogenesis, and formation of granulated tissue. The gene expression rates of Ang-1 and VEGF growth factors were also determined by qRT-PCR. Results: The higher wound closure rate was seen for the PRPCryogel-MSCs group on day 21 and 28 compared to other groups. Based on H&E results, an ascending rate of re-epithelialization was seen for the PRPCryogel-MSCs group on day 28. The most angiogenesis rate was detected for MSCs and PRPCryogel-MSCs groups on day 14. The qRT-PCR results showed the peak expression rate of Ang-1 in PRPCryogel-MSCs and MSCs groups on day 14. The peak expression rate of VEGF was also seen for PRPCryogel-MSCs and PRPCryogel groups on day 28. Conclusion: The treatment of burn wounds with PRPCryogel-MSCs promoted the wound closure and re-epithelialization rates. It also increased the expression rate of Ang-1 and VEGF genes which is consistent with pathological assessments. In conclusion, combination of MSCs with PRP can be considered as a promising approach for burn healing. Lay Summary and Future Work: Application of different natural substances for wound healing returns to a long period of time ago. Nature has been always generous and among these, mesenchymal stem cells (MSCs) and platelet-rich plasma (PRP) are such prominent gifts. MSCs have been well-identified for tissue regeneration which can be derived from bone marrow, umbilical cord, adipose tissue, etc. By differentiation into key cells such as fibroblasts and keratinocytes, contributing in vascularization, re-epithelialization, and collagen synthesis, MSCs accelerate the regeneration of damaged tissue. PRP is another spectacular substance derived from blood which is a rich suspension of platelets, growth factors with pain relieving and antimicrobial effects. We assessed the effects of MSCs which were placed within the PRP gel (PRPCryogel) on healing of grade-II burn wounds. Results showed that PRPCryogel-MSCs promote burn healing by increasing the rate of wound closure, angiogenesis, and re-epithelialization compared to other groups. This verifies the impressive effects of MSCs with PRP when combined together for burn treatment. Finally, this method can be also examined on regeneration of grade-III burn and/or chronic wounds as a future study in this field