Assessment of apoptosis and oxidative stress in cryopreserved ovary after grafting in fibrin-alginate scaffold with endothelial cells and melatonin in wistar rats.

OBJECTIVE: Infertility is a significant public health concern affecting 10-15% of couples. Young women undergoing gonadotoxic treatment are at higher risk of ovarian dysfunction and infertility. To mitigate this risk, ovarian tissue freezing and transplantation have been developed as a novel strategy. However, challenges such as follicular loss and dysfunction during the freezing process, and ovarian damage during transplantation, persist. This study aimed to investigate the potential of using appropriate antifreeze, antioxidant, wound healing, and biological hydrogels to reduce these injuries. Specifically, the effect of fibrin scaffold with endothelial cells and melatonin on apoptotic gene expression and antioxidants in cryopreserved ovaries after transplantation was examined. METHODS: A total of 36 adult female wistar rats) 6-8-week-old and weighing from 200 to 220 g) were divided equally into six groups (n = 6): 1) control group (C), 2) transplanted ovarian tissue after vitrification and thawing process (Group 1), 3) transplanted vitrified/thawed ovarian tissue while encapsulated in Fib/Alg hydrogel (Group 2), 4) transplanted vitrified/thawed ovarian tissue while encapsulated in Fib/Alg hydrogel in addition with melatonin (Group 3), 5) transplanted vitrified/thawed ovarian tissue while encapsulated in Fib/Alg hydrogel in addition with endothelial cells (Group 4) and 6) transplanted vitrified/thawed ovarian tissue while encapsulated in Fib/Alg hydrogel in addition with melatonin endothelial cells (Group 5). The ovaries were auto-transplanted in the rats' lumbar region. After 14 days, the ovaries were removed. Antioxidant levels (SOD, GPx, MDA, and TAC) were evaluated using ELISA, and apoptotic gene expressions (Bax/Bcl2 and caspase 3) were analyzed by real-time RT-PCR to determine apoptosis. RESULTS: In the transplanted frozen ovary group, Bax/Bcl2 and caspase 3 gene expression increased significantly (P<0.05), while antioxidant levels (SOD, GPx, MDA, and TAC) decreased. The encapsulated frozen ovary group showed decreased gene expression and increased antioxidant levels. The ovary group encapsulated with fibrin scaffold, endothelial cells, and melatonin had the most significant decrease in gene expression and increase in antioxidant levels (P<0.05). CONCLUSION: Coordinated action of Fibrin-based scaffold with endothelial cells and melatonin could decrease apoptosis gene expression and increase antioxidant levels in cryopreserved ovaries after transplantation, providing valuable insights into preserving fertility in young women undergoing gonadotoxic treatment.

Melatonin and endothelial cell-loaded alginate-fibrin hydrogel promoted angiogenesis in rat cryopreserved/thawed ovaries transplanted to the heterotopic sites.

BACKGROUND: Ischemic niche can promote follicular atresia following the transplantation of cryopreserved/thawed ovaries to the heterotopic sites. Thus, the promotion of blood supply is an effective strategy to inhibit/reduce the ischemic damage to ovarian follicles. Here, the angiogenic potential of alginate (Alg) + fibrin (Fib) hydrogel enriched with melatonin (Mel) and CD144+ endothelial cells (ECs) was assessed on encapsulated cryopreserved/thawed ovaries following transplantation to heterotopic sites in rats. METHODS: Alg + Fib hydrogel was fabricated by combining 2% (w/v) sodium Alg, 1% (w/v) Fib, and 5 IU thrombin at a ratio of 4: 2: 1, respectively. The mixture was solidified using 1% CaCl2. Using FTIR, SEM, swelling rate, and biodegradation assay, the physicochemical properties of Alg + Fib hydrogel were evaluated. The EC viability was examined using an MTT assay. Thirty-six adult female rats (aged between 6 and 8 weeks) with a normal estrus cycle were ovariectomized and enrolled in this study. Cryopreserved/thawed ovaries were encapsulated in Alg + Fib hydrogel containing 100 µM Mel + CD144+ ECs (2 × 104 cells/ml) and transplanted into the subcutaneous region. Ovaries were removed after 14 days and the expression of Ang-1, and Ang-2 was monitored using real-time PCR assay. The number of vWF+ and α-SMA+ vessels was assessed using IHC staining. Using Masson's trichrome staining, fibrotic changes were evaluated. RESULTS: FTIR data indicated successful interaction of Alg with Fib in the presence of ionic cross-linker (1% CaCl2). Data confirmed higher biodegradation and swelling rates in Alg + Fib hydrogel compared to the Alg group (p < 0.05). Increased viability was achieved in encapsulated CD144+ ECs compared to the control group (p < 0.05). IF analysis showed the biodistribution of Dil+ ECs within hydrogel two weeks after transplantation. The ratio of Ang-2/Ang-1 was statistically up-regulated in the rats that received Alg + Fib + Mel hydrogel compared to the control-matched groups (p < 0.05). Based on the data, the addition of Mel and CD144+ ECs to Alg + Fib hydrogel reduced fibrotic changes. Along with these changes, the number of vWF+ and α-SMA+ vessels was increased in the presence of Mel and CD144+ ECs. CONCLUSIONS: Co-administration of Alg + Fib with Mel and CD144+ ECs induced angiogenesis toward encapsulated cryopreserved/thawed ovarian transplants, resulting in reduced fibrotic changes.

Disease-associated regulation of gene expression by resveratrol: Special focus on the PI3K/AKT signaling pathway.

Resveratrol (3,5,4'-trihydroxy-trans-stilbene) is a natural phenol that is present in the skin of the grape, blueberry, raspberry, mulberry, and peanut. This substance is synthesized in these plants following injury or exposure to pathogens. Resveratrol is used as a dietary supplement for a long time and its effects have been assessed in animal models of human disorders. It has potential beneficial effects in diverse pathological conditions such as diabetes mellitus, obesity, hypertension, neoplastic conditions, Alzheimer's disease, and cardiovascular disorders. Notably, resveratrol has been found to affect the expression of several genes including cytokine coding genes, caspases, matrix metalloproteinases, adhesion molecules, and growth factors. Moreover, it can modulate the activity of several signaling pathways such as PI3K/AKT, Wnt, NF-κB, and Notch pathways. In the current review, we summarize the results of studies that reported modulatory effects of resveratrol on the expression of genes and the activity of signaling pathways. We explain these results in two distinct sections of non-neoplastic and neoplastic conditions.

Nrf2-Related Therapeutic Effects of Curcumin in Different Disorders.

Curcumin is a natural polyphenol with antioxidant, antibacterial, anti-cancer, and anti-inflammation effects. This substance has been shown to affect the activity of Nrf2 signaling, a pathway that is activated in response to stress and decreases levels of reactive oxygen species and electrophilic substances. Nrf2-related effects of curcumin have been investigated in different contexts, including gastrointestinal disorders, ischemia-reperfusion injury, diabetes mellitus, nervous system diseases, renal diseases, pulmonary diseases, cardiovascular diseases as well as cancers. In the current review, we discuss the Nrf2-mediated therapeutic effects of curcumin in these conditions. The data reviewed in the current manuscript indicates curcumin as a potential activator of Nrf2 and a therapeutic substance for the protection of cells in several pathological conditions.

Review of ovarian tissue cryopreservation techniques for fertility preservation.

Ovarian failure and ovarian malfunction are among major fertility problems in women of reproductive age (18-35 years). It is known that various diseases, such as ovarian cancer and premature ovarian failure, besides certain treatments, such as radiotherapy and chemotherapy of other organs, can affect the normal process of folliculogenesis and cause infertility. In recent years, various procedures have been proposed for the treatment of infertility. One of the newest methods is the use of cryopreservation ovarian fragments after cancer treatment. According to some studies, this method yields very satisfactory results. Although ovarian tissue cryopreservation (OTC) is an accepted technique of fertility preservation, the relative efficacy of cryopreservation protocols remains controversial. Considering the controversies about these methods and their results, in this study, we aimed to compare different techniques of ovarian cryopreservation and investigate their advantages and disadvantages. Reviewing the published articles may be possible to identify appropriate strategies and improve infertility treatment in these patients.

The impact of non-coding RNAs on normal stem cells.

Self-renewal and differentiation into diverse cells are two main characteristics of stem cells. These cells have important roles in development and homeostasis of different tissues and are supposed to facilitate tissue regeneration. Function of stem cells is regulated by dynamic interactions between external signaling, epigenetic factors, and molecules that regulate expression of genes. Among the highly appreciated regulators of function of stem cells are long non-coding RNAs (lncRNAs) and microRNAs (miRNAs). Impact of miR-342-5p, miR-145, miR-1297, miR-204-5p, miR-132, miR-128-3p, hsa-miR-302, miR-26b-5p and miR-10a are among miRNAs that regulate function of stem cells. Among lncRNAs, AK141205, ANCR, MEG3, Pnky, H19, TINCR, HULC, EPB41L4A-AS1 and SNHG7 have important roles in the regulation of stem cells. In the current paper, we aimed at reviewing the importance of miRNAs and lncRNAs in differentiation of stem cells both in normal and diseased conditions. For this purpose, we searched PubMed/Medline and google scholar databases using "stem cell" AND "lncRNA", or "long non-coding RNA", or "microRNA" or "miRNA".

The Impact of lncRNAs and miRNAs in Regulation of Function of Cancer Stem Cells and Progression of Cancer.

Stem cells have two important features, namely the ability for self-renewal and the capacity to differentiate into some cell kinds with specialized functions. These two features are also present in cancer stem cells (CSCs). These cells have been detected in almost all kinds of cancers facilitating their tumorigenicity. Molecular cascades that control self-renewal of stem cells, namely the Wnt, Notch, and Hedgehog pathways have been suggested to influence CSCs functions as well. Moreover, non-coding RNAs can regulate function of CSCs. Function of miRNAs in the regulation of CSCs has been mostly assessed in breast cancer and hepatocellular carcinoma. miR-130a-3p, miR-600, miR-590-5p, miR-142-3p, miR-221, miR-222, miR-638, miR-375, miR-31, and miR-210 are among those regulating this feature in breast cancer. Moreover, miR-206, miR-192-5p, miR-500a-3p, miR-125, miR-125b, miR-613, miR-217, miR-194, and miR-494 regulate function of CSCs in hepatocellular carcinoma. DILC, lncTCF7, MUF, HAND2-AS1, MALAT1, DLX6-AS1, HOTAIR, and XIST are among lncRNAs that regulate function of CSCs. In the present paper, we explain the effects of these two classes of non-coding RNAs in the regulation of activity of CSCs.

Functional roles of non-coding RNAs in atrophy.

Atrophy is defined as a reduction in cell, organ, or tissue size after reaching their normal mature sizes because of loss of organelles, cytoplasmic compartments, and proteins. This process is also involved in the pathogenesis of human disorders. Inadequate nourishment, poor circulation, inadequate hormonal support, defects in nerve supply of the tissue, disproportionate induction of apoptosis in the tissue, and absence of exercise are some underlying causes of atrophy. Recently, several non-coding RNAs (ncRNAs) have been identified that regulate atrophy, thus participating in the pathobiology of related disorders such as neurodegenerative/ neuromuscular diseases, age-related muscle atrophy, and cardiac tissue atrophy. In the current review, we have focused on two classes of ncRNAs namely long ncRNAs (lncRNAs) and microRNAs (miRNAs) to unravel their participation in atrophy-associated disorders.

An update on the role of miR-379 in human disorders.

miR-379 is a miRNA transcribed from the MIR379 locus on 14q32.31. This miRNA is located in an evolutionarily conserved miRNA cluster in an imprinted region that contains DLK1 and DIO3 genes. The mouse homolog of this miRNA has been shown to be under-expressed in response to glucocorticoid receptor deficiency. Moreover, miR-379 has a tumor-suppressive role in a wide variety of tissues including the brain, breast, lung, and liver. In addition to restraining cell proliferation and migration, miR-379 can suppress the epithelial-mesenchymal transition process. Abnormal expression of this miRNA implies the pathogenesis of Duchene muscular dystrophy, spinal cord injury, diabetic nephropathy, acute myocardial infarction, and premature ovarian failure. This review aims to the summarization of the role of miR-379 in neoplastic and non-neoplastic conditions.

The interaction between miRNAs/lncRNAs and nuclear factor-κB (NF-κB) in human disorders.

Nuclear factor-κB (NF-κB) represents a group of inducible transcription factors (TFs) regulating the expression of a great variety of genes implicated in diverse processes, particularly modulation of immune system responses. This TF has functional interactions with non-coding RNAs, constructing a complicated network through which NF-κB, miRNAs, and lncRNAs coordinately regulate gene expression at different facets. This type of interaction is involved in the pathophysiology of several human disorders including both neoplastic disorders and non-neoplastic conditions. MALAT1 and NKILA are among lncRNAs whose interactions with NF-κB have been vastly assessed in different conditions including cancer and inflammatory conditions. In addition, miR-146a/b has functional interactions with this TF in different contexts. Although miRNAs have mutual interactions with NF-κB, the regulatory role of miRNAs on this TF has been more clarified. The aim of the current review is to explore the function of NF-κB-related miRNAs and lncRNAs in these two types of human disorders.

The role of H19 lncRNA in conferring chemoresistance in cancer cells.

H19 is an oncofetal transcript with crucial roles in the development and progression of several neoplastic cells. With anti-apoptotic, pro-proliferative, and pro-migratory functions, H19 affects the carcinogenic process from different functional points. In addition, H19 has central roles in the induction of chemoresistance in breast cancer, lung cancer, glioma, liver cancer, and other types of cancers. Induction of EMT, activation of oncogenic signaling pathways, and changes in the tumor microenvironment are among mechanisms of participation of H19 in chemoresistance. Paclitaxel, doxorubicin, tamoxifen, erlotinib, gefitinib, temozolomide, and methotrexate are among therapeutic agents whose efficacy is influenced by the expression of H19. In the present paper, we discuss the impact of H19 in conferring resistance to chemotherapeutic agents in different cancers.

microRNA-140: A miRNA with diverse roles in human diseases.

MicroRNA-140 (miR-140) has been shown to be associated with the pathogenesis of a wide range of pathologies including osteoarthritis, osteoporosis, renal fibrosis, ischemic conditions, and most importantly neoplasia. This miRNA has been shown to be down-regulated in a diversity of cancers namely breast cancer, gastrointestinal cancers, lung cancer, and prostate cancer. miR-140 has a lot of immune-related targets. Moreover, several miR-140 targets regulate cell proliferation, cell cycle transition, and apoptosis. This miRNA has been shown to be sponged by a number of lncRNAs and circ-RNAs. miR-140 has essential roles in the determination of the sensitivity of neoplastic cells to chemotherapeutic agents such as temozolomide, doxorubicin, and cisplatin. Besides, expression quantities of miR-140 in cancer tissues can be used for the prediction of clinical outcomes of patients with neoplasia. In the present paper, we describe the impact of miR-140 in neoplastic and non-neoplastic disorders.

The interplay between non-coding RNAs and Twist1 signaling contribute to human disorders.

Twist-related protein 1 (Twist1) is a basic helix-loop-helix (bHLH) transcription factor (TF) being coded by the TWIST1 gene. This TF has a fundamental effect on the normal development and in the pathogenesis of various diseases especially cancer. Twist1 has interactions with some long non-coding RNAs and miRNAs. The interactions between this TF and various miRNAs such as miR-16, miR-26b-5p, miR-1271, miR-539, miR-214, miR-200b/c, miR-335, miR-10b, and miR-381 are implicated in the carcinogenic processes. TP73-AS1, LINC01638, ATB, NONHSAT101069, CASC15, H19, PVT1, LINC00339, LINC01385, TANAR, SNHG5, DANCR, CHRF, and TUG1 are among long non-coding RNAs which interact with Twist1 and participate in the carcinogenesis. This review aims at depicting the interaction between these non-coding transcripts and Twist1 and the consequence of these interactions in human neoplasms.

An update on the role of miR-124 in the pathogenesis of human disorders.

MicroRNA-124 (miR-124) is a copious miRNA in the brain, but it is expressed in a wide range of human/animal tissues participating in the pathogenesis of several disorders. Based on its important function in the development of the nervous system, abnormal expression of miR-124 has been detected in nervous system diseases including Alzheimer's disease, Parkinson's disease, Hypoxic-Ischemic Encephalopathy, Huntington's disease, and ischemic stroke. In addition to these conditions, miR-124 contributes to the pathogenesis of cardiovascular disorders, hypertension, and atherosclerosis. Besides, it has been shown to be down-regulated in a wide range of human cancers such as colorectal cancer, breast cancer, gastric cancer, glioma, pancreatic cancer, and other types of cancer. Yet, few studies have reported upregulation of miR-124 in some cancer types. In the current study, we describe the role of miR-124 in these malignant and non-malignant conditions.

miR-1: A comprehensive review of its role in normal development and diverse disorders.

MicroRNA-1 (miR-1) is a conserved miRNA with high expression in the muscle tissues. In humans, two discrete genes, MIRN1-1 and MIRN1-2 residing on a genomic region on 18q11.2 produce a single mature miRNA which has 21 nucleotides. miR-1 has a regulatory role on a number of genes including heat shock protein 60 (HSP60), Kruppel-like factor 4 (KLF4) and Heart And Neural Crest Derivatives Expressed 2 (HAND2). miR-1 has critical roles in the physiological processes in the smooth and skeletal muscles as well as other tissues, thus being involved in the pathogenesis of a wide range of disorders. Moreover, dysregulation of miR-1 has been noted in diverse types of cancers including gastric, colorectal, breast, prostate and lung cancer. In the current review, we provide the summary of the data regarding the role of this miRNA in the normal development and the pathogenic processes.