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BACKGROUND: Bioscaffolds and cells are two main components in the regeneration of damaged tissues via cell therapy. Umbilical cord stem cells are among the most well-known cell types for this purpose. The main objective of the present study was to evaluate the effect of the pretreatment of the foreskin acellular matrix (FAM) by monophosphoryl lipid A (MPLA) and Lactobacillus casei supernatant (LCS) on the attraction of human umbilical cord mesenchymal stem cells (hucMSC). METHODS AND RESULTS: The expression of certain cell migration genes was studied using qRT-PCR. In addition to cell migration, transdifferentiation of these cells to the epidermal-like cells was evaluated via immunohistochemistry (IHC) and immunocytochemistry (ICC) of cytokeratin 19 (CK19). The hucMSC showed more tissue tropism in the presence of MPLA and LCS pretreated FAM compared to the untreated control group. We confirmed this result by scanning electron microscopy (SEM) analysis, glycosaminoglycan (GAG), collagen, and DNA content. Furthermore, IHC and ICC data demonstrated that both treatments increase the protein expression level of CK19. CONCLUSION: Pretreatment of acellular bioscaffolds by MPLA or LCS can increase the migration rate of cells and also transdifferentiation of hucMSC to epidermal-like cells without growth factors. This strategy suggests a new approach in regenerative medicine.
Subject(s)
Lacticaseibacillus casei , Lipid A , Mesenchymal Stem Cells , Humans , Mesenchymal Stem Cells/metabolism , Lacticaseibacillus casei/metabolism , Lipid A/metabolism , Lipid A/analogs & derivatives , Cell Movement/drug effects , Skin/metabolism , Tissue Scaffolds/chemistry , Male , Umbilical Cord/cytology , Umbilical Cord/metabolism , Foreskin/cytology , Cell Transdifferentiation/drug effects , Tissue Engineering/methods , Extracellular Matrix/metabolism , Keratin-19/metabolism , Keratin-19/geneticsABSTRACT
The importance of the tumor microenvironment in cancer progression has been well studied for many years. Immune checkpoint inhibitors (ICIs) are regarded as potential strategies in enhancing the immune responses in patients with cancer, particularly colorectal cancer (CRC). Notably, CRCs are extraordinarily heterogeneous and mostly are microsatellite-stable (MSS) or cold tumors, which means that the immune response is not usually as strong as that of foreign cells. T-cell immunoglobulin and ITIM domain (TIGIT) is a new immune checkpoint receptor overexpressed inside the CRC tumor-immune microenvironments. Moreover, several studies have shown that TIGIT in combination with other ICIs and/or conventional treatments, can lead to a robust anti-tumor response in CRC. This review looks deep inside TIGIT expression patterns, their various functions, and possible immunotherapy strategies to increase survival rates and decrease immune-related adverse events.
Subject(s)
Adenocarcinoma/therapy , Colorectal Neoplasms/therapy , Immune Checkpoint Inhibitors , Immune Checkpoint Proteins/immunology , Immunotherapy/methods , Receptors, Immunologic/antagonists & inhibitors , Adenocarcinoma/immunology , Adenocarcinoma/pathology , Amino Acid Motifs , Animals , Antigens, CD/immunology , CRISPR-Cas Systems , Colorectal Neoplasms/immunology , Colorectal Neoplasms/pathology , Colorectal Neoplasms/radiotherapy , Combined Modality Therapy , Gastrointestinal Microbiome , Gene Expression Regulation, Neoplastic , Humans , Mice , Prognosis , Protein Domains , Receptors, Immunologic/biosynthesis , Receptors, Immunologic/deficiency , Receptors, Immunologic/genetics , Receptors, Immunologic/immunology , Tumor MicroenvironmentABSTRACT
Osteosarcopenia is an increasingly recognized geriatric syndrome with a considerable prevalence which increases morbidity and mortality. Although osteosarcopenia is a result of age-related deterioration in muscle and bone, there are many risk factors that provoking osteosarcopenia. These risk factors should be considered by the clinicians to treat osteosarcopenia. We assessed the link between osteosarcopenia and conventional risk factors of cardiovascular diseases. This study was a cross-sectional study that has been conducted within the framework of Bushehr Elderly Health (BEH) program stage II in which participants aged ≥ 60 years were included. Osteopenia/osteoporosis was defined as a t-score ≤ - 1.0 standard deviation below the mean values of a young healthy adult. We defined sarcopenia as reduced skeletal muscle mass plus low muscle strength and/or low physical performance. Osteosarcopenia was considered as the presence of both osteopenia/osteoporosis and sarcopenia. We estimated the age-standardized prevalence of osteosarcopenia for men and women, separately. Using modified Poisson regression analysis, adjusted prevalence ratio (PR) with 95% CI was used to show the measure of associations in the final model. Among 2353 participants, 1205 (51.2%) were women. Age-standardized prevalence of osteosarcopenia was 33.8 (95% CI 31.0-36.5) in men and 33.9 (30.9-36.8) in women. In both sexes, the inverse association was detected with body mass index and having osteosarcopenia (PR 0.84, 95% CI 0.81-0.88 in men and 0.77, 95% CI 0.74-0.80 in women). In both sexes, high-fat mass was positively associated with osteosarcopenia [PR 1.46 (95% CI 1.11-1.92) in men, and 2.25 (95% CI 1.71-2.95) in women]. Physical activity had a significant inverse association in men (PR = 0.64, 95% CI 0.46, 0.88), but not in women. Diabetes was also showed a direct association with osteosarcopenia in men (PR 1.33, 95% CI 1.04-1.69). No associations were detected between the lipid profiles and osteosarcopenia. Results demonstrated a high prevalence of osteosarcopenia in both sexes suggesting a high disease burden in a rapidly aging country. Lifestyle and socioeconomic factors, as well as chronic diseases, were significantly associated with osteosarcopenia.
Subject(s)
Aging , Osteoporosis/complications , Osteoporosis/epidemiology , Sarcopenia/epidemiology , Aged , Bone Density/physiology , Cross-Sectional Studies , Exercise/physiology , Female , Hand Strength/physiology , Heart Disease Risk Factors , Humans , Male , Prevalence , Risk FactorsABSTRACT
PURPOSE: Hyperthermia can modulate inflammation and the immune response. Based on the recruitment of mesenchymal stem cells (MSCs) to inflamed tissues and the immunomodulatory properties of these cells, the aim of this study was to examine the effects of hyperthermia on the immunomodulatory properties of MSCs in a mixed lymphocyte reaction (MLR). MATERIALS AND METHODS: Passages 4-6 of human umbilical cord vein mesenchymal stem cells were co-cultured in a two-way MLR. Cells in the hyperthermia groups were incubated at 41 °C for 45 min. A colorimetric assay was employed to examine the effects of MSCs on cell proliferation. The levels of IL-4 and TNF-α proteins in the cell culture supernatant were measured, and non-adherent cells were used for RNA extraction, which was then used for cDNA synthesis. RT-PCR was utilised to assess levels of IL-10, IL-17A, IL-4, TNF-α, TGF-ß1, FOX P3, IFN-γ, CXCL12 and ß-actin mRNA expression. RESULTS: UCV-MSCs co-cultured in an MLR reduced lymphocyte proliferation at 37 °C, whereas hyperthermia attenuated this effect. Hyperthermia increased expression of IL-10, TGF-ß1 and FOXP3 mRNAs in co-culture; however, no effects on IL-17A and IFN-γ were observed, and it reduced CXCL12 expression. In co-culture, IL-4 mRNA and protein increased at 37 °C, an effect that was reduced by hyperthermia. No considerable change in TNF-α mRNA expression was found in hyperthermia-treated cells. CONCLUSION: Hyperthermia increases cell proliferation of the peripheral blood mononuclear cells and modifies the cytokine profile in the presence of UCV-MSCs.
Subject(s)
Cytokines/immunology , Hyperthermia, Induced , Leukocytes, Mononuclear/immunology , Mesenchymal Stem Cells/immunology , Umbilical Cord/cytology , Cell Differentiation , Cell Proliferation , Cells, Cultured , Coculture Techniques , Cytokines/genetics , Humans , Immunomodulation , Lymphocyte Culture Test, MixedABSTRACT
INTRODUCTION: Chronic pancreatitis (CP) is an intractable and multi-factorial disorder. Developing appropriate animal models is an essential step in pancreatitis research, and the best ones are those which mimic the human disorder both aetiologically and pathophysiologically. The current study presents an optimised protocol for creating a murine model of CP, which mimics the initial steps of chronic pancreatitis in alcohol chronic pancreatitis and compares it with two other mouse models treated with cerulein or ethanol alone. MATERIAL AND METHODS: Thirty-two male C57BL/6 mice were randomly selected, divided into four groups, and treated intraperitoneally with saline (10 ml/kg, control group), ethanol (3 g/kg; 30% v/v), cerulein (50 µg/kg), or ethanol + cerulein, for six weeks. Histopathological and immunohistochemical assays for chronic pancreatitis index along with real-time PCR assessments for mRNA levels of inflammatory cytokines and fibrogenic markers were conducted to verify the CP induction. RESULTS: The results indicated that CP index (CPI) was significantly increased in ethanol-cerulein mice compared to the saline, ethanol, and cerulein groups (p < 0.001). Interleukin 1ß (IL-1ß), tumor necrosis factor α (TNF-α), transforming growth factor ß (TGF-ß), α-smooth muscle actin (α-SMA), and myeloperoxidase activity were also significantly greater in both cerulein and ethanol-cerulein groups than in the saline treated animals (p < 0.001). Immunohistochemical analysis revealed enhanced expression of TGF-ß and α-SMA in ethanol-cerulein mice compared to the saline group. CONCLUSIONS: Intraperitoneal (IP) injections of ethanol and cerulein could successfully induce CP in mice. IP injections of ethanol provide higher reproducibility compared to ethanol feeding. The model is simple, non-invasive, reproducible, and time-saving. Since the protocol mimics the initial phases of CP development in alcoholics, it can be used for investigating basic mechanisms and testing new therapies.
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Human keratinocytes could be used in the repair of damaged skin, in tissue engineering applications, gene therapy and recently, the generation of iPS cells. We isolated human keratinocytes from foreskin and subsequently cultured them on fibronectin, collagen type I, gelatin and laminin-coated dishes that contained three different types of serum-free medium (epilife, KSM or CnT). We developed improved conditions for efficient transfection of these human keratinocytes by testing three common transfection methods and a GFP plasmid vector. The isolated cells showed typical keratinocyte morphology and expressed the epithelial cell specific antigen, cytokeratin 14. Collagen type 1, epilife medium and lipofectamin 2000 gave the best results for isolation and transfection of human keratinocytes. Our protocol can be used as a reproducible, simple and efficient method for isolation, cultivation and genetic manipulation of human keratinocytes, which may be useful in cell and gene therapy applications.
Subject(s)
Keratinocytes/cytology , Antigens, Surface/metabolism , Cell Proliferation/drug effects , Cell Separation , Cells, Cultured , Collagen Type I/chemistry , Collagen Type I/pharmacology , Culture Media/chemistry , Culture Media/pharmacology , Fibronectins/chemistry , Foreskin/cytology , Green Fluorescent Proteins/genetics , Green Fluorescent Proteins/metabolism , Humans , Keratin-14/metabolism , Keratinocytes/metabolism , Keratinocytes/pathology , Lipids/chemistry , Male , TransfectionABSTRACT
BACKGROUNDS AND AIMS: Prostate cancer is the most common malignant cancer among men and is the second deadliest cancer in men after lung cancer. Understanding the molecular mechanisms involved in development and progression of prostate cancer is essential to improve both diagnostic and therapeutic strategies in this regard. In addition, using novel gene therapy-based methods for treatment of cancers has gotten increasing attention during the recent years. Accordingly, this study was aimed to evaluate the inhibitory effect of MAGE-A11 gene, as an important oncogene involved in the pathophysiology of prostate cancer invitro model. The study was also aimed to evaluate the downstream genes related to MAGE-A11. MATERIALS AND METHODS: First, MAGE-A11 gene was knocked out in PC-3 cell line using "Clustered regularly interspaced short palindromic repeats" (CRISPR)/ "CRISPR-associated genes 9" (CRISPR/Cas9) method. Next, the expression levels of MAGE-A11, survivin and Ribonucleotide Reductase Small Subunit M2 (RRM2) genes were determined by quantitative polymerase chain reaction (qPCR) technique. The levels of proliferation and apoptosis were also analyzed in PC-3 cells using CCK-8 and Annexin V-PE/7-AAD assays. RESULTS: The results showed that the disruption of MAGE-A11 by CRISPR/Cas9 method significantly decreased proliferation (P< 0.0001) and enhanced apoptosis (P< 0.05) in PC-3 cells compared to control group. Moreover, the disruption of MAGE-A11 significantly down regulated the expression levels of survivin and RRM2 genes (P< 0.05). CONCLUSION: Our results demonstrated that knocking out MAGE-11 gene by CRISPR/CAS9 technique could efficiently inhibit cell proliferation and induce apoptosis in PC3 cells. Survivin and RRM2 genes might also participated in these processes.
Subject(s)
Antigens, Neoplasm , CRISPR-Cas Systems , Prostatic Neoplasms , Humans , Male , Apoptosis/genetics , Prostatic Neoplasms/genetics , Prostatic Neoplasms/pathology , Survivin/genetics , Antigens, Neoplasm/geneticsABSTRACT
OBJECTIVE: Acellular matrices of different allogeneic or xenogeneic origins are widely used as structural scaffolds in regenerative medicine. The main goal of this research was to optimize a method for decellularization of foreskin for skin regeneration in small wounds. MATERIALS AND METHODS: In this experimental study, the dermal layers of foreskin were divided into two sections and subjected to two different decellularization methods: the sodium dodecyl sulfate method (SDS-M), and our optimized foreskin decellularization method (OFD-M). A combination of non-ionic detergents and SDS were used to decellularize the foreskin in OFD-M. The histological, morphological, and biomechanical properties of both methods were compared. In addition, human umbilical cord mesenchymal stem cells (hucMSCs) were isolated, and the biocompatibility and recellularization of both scaffolds by hucMSC were subsequently determined. RESULTS: We observed that OFD-M is an appropriate approach for successful removal of cellular components from the foreskin tissue, without physical disturbance to the acellular matrix. In comparison to SDS-M, this new bioscaffold possesses a fine network containing a high amount of collagen fibers and glycosaminoglycans (GAG) (P≤0.03), is biocompatible and harmless for hucMSC (viability 91.7%), and exhibits a relatively high tensile strength. CONCLUSION: We found that the extracellular matrix (ECM) structural integrity, the main ECM components, and the mechanical properties of the foreskin are well maintained after applying the OFD-M decellularization technique, indicating that the resulting scaffold would be a suitable platform for culturing MSC for skin grafting in small wounds.
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OBJECTIVE: Membrane type-matrix metalloproteinases (MT-MMPs) are known as key regulators of cancer progression/metastasis. However, their roles in the growth and progression of multiple myeloma (MM) have not been yet elucidated. METHODS AND MATERIALS: The expression of 6 MT-MMPs in MM, B cell lines, and normal peripheral blood (PB) cells were measured by RT-PCR, qRT-PCR, flow cytometry, western blotting, and immunocytochemistry. B lymphocytes, CD19-/CD138-, and CD19-/CD138+ cells, known as malignant plasma cells (MPC), were sorted from bone marrow (BM) aspirations of 10 MM patients, and MT2-MMP expression was examined in these cells using qRT-PCR, flow cytometry and immunohistochemistry, and western blotting. Moreover, the expression of MT2-MMP in BM biopsies from 13 normal individuals and 14 MM patients was analyzed by immunohistochemistry. MT2-MMP was also knocked down in U266 cells using siRNA technology and the adhesion, invasion, migration abilities, and cell proliferation were determined and compared with scrambled ones in both in vitro and in vivo studies. RESULTS: Our results showed that MT2-MMP expression is significantly higher in MM cell lines and MPC cells than B cell lines and other PB- or BM-derived cells. MT2-MMP is expressed in BM biopsies from all 14 patients with MM, and 67.85% ± 32.38 of BM cells were positive for MT2-MMP. In contrast, only 0.38 ± 0.76 of BM biopsies from normal individuals were positive for MT2-MMP. Importantly, MT2-MMP was expressed in all the patients' BM biopsies at the diagnosis, but not in the remission phase. MT2-MMP siRNA significantly decreased adhesion, invasion, migration, and 3D cell proliferation of U266 cells. Moreover, in the xenographic model, MT2-MMP siRNA prevented the growth and development of plasmacytoma. Taken together, these data demonstrate that MT2-MMP is strongly expressed in MM cells and plays important role in the growth and progression of these cells, suggesting that MT2-MMP is an appropriate biomarker in diagnosis and therapeutic interventions of MM.
Subject(s)
Matrix Metalloproteinase 15/metabolism , Multiple Myeloma , Cell Movement , Humans , Immunohistochemistry , Matrix Metalloproteinase 15/genetics , Matrix Metalloproteinases, Membrane-Associated , Multiple Myeloma/genetics , Multiple Myeloma/metabolismABSTRACT
Purpose: Acute pancreatitis (AP) which is distinguished by local pancreatic necrosis, followingby systemic organ failure is known as an inflammatory disease. Up to now, there are only a fewtreatment options accessible for patients suffering from AP. In this study, we aimed to examinethe anti-inflammatory capacities of human bone marrow-derived mesenchymal stromal cells(hBM-MSCs) in a detailed AP model experiment. Methods: AP was induced in C57BL/6 mice by intraperitoneal administration of cerulein (100µg/kg/h × 7 doses) at intervals of 1 hour. Then, 2×105 MSCs were infused in the AP mice bytail vein 6 hours after the last cerulein injection. Mice were sacrificed 12 hours following theinjection of hBM-MSC, and blood samples and pancreas tissues were obtained. Results: We first determined the presence of transplanted hBM-MSC in the pancreas of micewith AP, but not the control mice. Our data indicate that administration of hBM-MSCs to micewith AP lead to (i) decreased serum levels of amylase, lipase and myeloperoxidase activities, (ii)downregulation of proinflammatory cytokine, macrophage inflammatory protein 2 (MIP-2), and(iii) upregulation of the anti-inflammatory cytokine, interleukin 10 (IL-10). Moreover, hBM-MSCadministration results in notably attenuated cerulein-induced histopathological alternationsand edema. Conclusion: we demonstrate that hBM-MSC attenuates AP signs and indicating that hMB-MSCtherapy could be a suitable approach for the treatment of inflammatory disease such as AP.
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CXCR7 binds chemokines CXCL11 (I-TAC) and CXCL12 (SDF-1) but does not act as a classical chemoattractant receptor. Using CCX771, a novel small molecule with high affinity and selectivity for CXCR7, we found that, although CXCR7 is dispensable for "bare filter" in vitro chemotaxis, CXCR7 plays an essential role in the CXCL12/CXCR4-mediated transendothelial migration (TEM) of CXCR4(+)CXCR7(+) human tumor cells. Importantly, although CXCL11 is unable to stimulate directly the migration of these cells, it acts as a potent antagonist of their CXCL12-induced TEM. Furthermore, even though this TEM is driven by CXCR4, the CXCR7 ligand CCX771 is substantially more potent at inhibiting it than the CXCR4 antagonist AMD3100, which is more than 100 times weaker at inhibiting TEM when compared with its ability to block bare filter chemotaxis. Far from being a "silent" receptor, we show that CXCR7 displays early hallmark events associated with intracellular signaling. Upon cognate chemokine binding, CXCR7 associates with beta-arrestin2, an interaction that can be blocked by CXCR7-specific mAbs. Remarkably, the synthetic CXCR7 ligand CCX771 also potently stimulates beta-arrestin2 recruitment to CXCR7, with greater potency and efficacy than the endogenous chemokine ligands. These results indicate that CXCR7 can regulate CXCL12-mediated migratory cues, and thus may play a critical role in driving CXCR4(+)CXCR7(+) tumor cell metastasis and tissue invasion. CXCR7 ligands, such as the chemokine CXCL11 and the newly described synthetic molecule CCX771, may represent novel therapeutic opportunities for the control of such cells.
Subject(s)
Cell Migration Inhibition/immunology , Endothelium, Vascular/immunology , Endothelium, Vascular/pathology , Receptors, CXCR4/antagonists & inhibitors , Receptors, CXCR4/physiology , Receptors, CXCR/antagonists & inhibitors , Receptors, CXCR/physiology , Signal Transduction/immunology , Animals , CHO Cells , Cell Line , Cell Line, Tumor , Chemokine CXCL12/antagonists & inhibitors , Chemokine CXCL12/metabolism , Chemotaxis, Leukocyte/immunology , Cricetinae , Cricetulus , Endothelium, Vascular/metabolism , Humans , Ligands , Receptors, CXCR/biosynthesis , Receptors, CXCR/metabolism , Receptors, CXCR4/biosynthesis , U937 CellsABSTRACT
AIM: Osteoarthritis (OA) is the most common chronic joint disorder, resulting from the breakdown of joint cartilage. It occurs in the knees, hands, and hips, leading to pain, stiffness, inflammation, and swelling. METHODS: In this study, 100 hand and knee OA patients, meeting the American College of Rheumatology criteria were included in the case group, and 100 healthy individuals were allocated to the control group. Blood samples were collected from the participants. After DNA extraction, genotyping was carried out for GDF5 rs143383 C/T polymorphism by allele-specific polymerase chain reaction (ASPCR) and for D-repeat alleles of asporin (ASPN) by conventional PCR assay. RESULTS: The results showed that the frequency of the D14 allele of ASPN was significantly higher than other alleles in the case group (P = .0001). Also, the frequency of the D14 allele among women was significantly higher than in men (P = .004). Moreover, the frequency of the TT allele in GDF5 rs143383 C/T polymorphism was significantly higher than the CC and CT alleles in the case group, compared with the control group (P = .001). A significant difference was found between the TT allele and other alleles in female and male patients compared with the control group (P = .02). CONCLUSIONS: The D14 allele of the ASPN gene and TT allele of the GDF5 gene (rs143383 + 104T/C) are associated with hand and knee OA in the Kurdish population, indicating that these alleles could be risk factors for OA, at least in our populations.
Subject(s)
Cartilage, Articular/physiopathology , Extracellular Matrix Proteins/genetics , Genetic Predisposition to Disease/genetics , Growth Differentiation Factor 5/genetics , Hand/physiopathology , Osteoarthritis, Knee/genetics , Polymorphism, Genetic/genetics , Aged , Alleles , Case-Control Studies , Extracellular Matrix Proteins/metabolism , Female , Genetic Predisposition to Disease/ethnology , Growth Differentiation Factor 5/metabolism , Humans , Iran/epidemiology , Male , Middle Aged , Osteoarthritis, Knee/epidemiology , Polymorphism, Single NucleotideABSTRACT
BACKGROUND: Inflammation and inflammatory mediators have been proposed to be key players in the pathobiology of Irritable bowel syndrome (IBS. The chemokine CCL28 plays a role in the trafficking of inflammatory cells into mucosal tissues. However, its levels in patients with IBS has not been yet elucidated. METHOD: In this study, the levels of CCL28 were measured in the serum of 41 patients with IBS and 41 age- and gender-matched normal individuals using Elisa. Then, the receiver operating characteristic (ROC) curve was conducted to assess the diagnostic value of CCL28. RESULTS: Our data showed that the levels of CCL28 are significantly elevated in patients with IBS compared to the control donors. Moreover, we observed that the level of CCL28 is associated with many clinical symptoms such as constipation, diarrhea, and abdominal pain. The area under the ROC curve was 0.71 (95% confidential interval, 0.598-0.823), the sensitivity and specificity of CCL28 for the diagnosis of IBS patients were 68.3% and 70.7%, respectively with a cut off of 278.9 ng/mL. CONCLUSIONS: We demonstrated that CCL28 is elevated in patients with IBS and correlates with clinical findings, indicating that CCL28 might be an appropriate biomarker for the diagnosis of IBS; however, further studies are necessary.
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BACKGROUND: Mesenchymal stem cells have restorative effects on premature ovarian failure (POF). The aim of this study was to investigate the effects of human umbilical cord vein MSCs (hUCV-MSCs) on follicular quantitative parameters and histological changes of ovaries in the cyclophosphamide (CTX)-induced POF in mice. MATERIALS AND METHODS: C57BL/6 mice were divided into three groups (10 mice in each group). In the control group, phosphate-buffered saline (PBS) was injected via tail vein following 15 days injection of PBS intraperitoneally (IP). In the CTX group, CTX was administered IP for 15 days and then PBS was injected via tail vein. In the CTX + hUCV-MSCs group, following CTX administration, single dose of the 1 × 106 of hUCV-MSCs were injected into tail vein. H&E, trichrome and PAS staining as well as TUNEL assay were performed on the ovaries tissue sections. The number of follicles, follicular quantitative parameters and apoptotic index were obtained. The serum levels of estradiol and FSH were measured in the mice. RESULTS: In the CTX + hUCV-MSCs group, degenerative changes were decreased and follicular quantitative parameters increased in the ovarian follicles compared to the CTX group. In this group number of follicles was increased, apoptotic index was decreased, estradiol and FSH levels were decreased and increased, respectively, all of them improved compared to the CTX group. The mean percentage areas of collagen fibers content were decreased compared to the CTX group. CONCLUSION: Results showed that, hUCV-MSCs administration increases follicular quantitative parameters and improve degenerative changes in the follicles following CTX injury.
Subject(s)
Cyclophosphamide/adverse effects , Mesenchymal Stem Cell Transplantation , Mesenchymal Stem Cells/metabolism , Ovarian Follicle/metabolism , Primary Ovarian Insufficiency , Umbilical Cord/metabolism , Animals , Cyclophosphamide/pharmacology , Disease Models, Animal , Female , Heterografts , Humans , Mesenchymal Stem Cells/pathology , Mice , Ovarian Follicle/pathology , Primary Ovarian Insufficiency/chemically induced , Primary Ovarian Insufficiency/metabolism , Primary Ovarian Insufficiency/pathology , Primary Ovarian Insufficiency/therapy , Umbilical Cord/pathologyABSTRACT
PURPOSE: T-cell immunoglobulin and ITIM domain (TIGIT) is an immune checkpoint that is overexpressed on both immune cells and some cancer cells. TIGIT can alter the anti-tumor responses inside the tumor microenvironment. Hypoxia-inducible factor 1-alpha (HIF-1α) plays a significant role in the TME and involves suppressing the anti-tumor responses. Under hypoxic conditions, HIF-1α can enhance the expression of different immune checkpoints. Accordingly, hypoxic TME and TIGIT overexpression cause cancer development. Thus, we decided to inhibit tumor cell expansion by inhibiting TIGIT and HIF-1α molecules and discovering the relationship between TIGIT and HIF-1α. METHODS: In this research, we utilized superparamagnetic iron oxide-based NPs (SPIONs) combined with chitosan lactate (CL) and folic acid (FA) nanoparticles (NPs) loaded with TIGIT-siRNA and HIF-1α- siRNA for suppressing TIGIT and HIF-1α in tumor cells and evaluated the consequences of this treatment strategy on tumor growth, apoptosis, and metastasis. RESULTS: The results showed that cancer cells treated with TIGIT and HIF-1α siRNA-loaded SPIONs-CL-FA NPs, strongly suppressed the TIGIT and HIF-1α expression, colony formation ability, angiogenesis, and the growth rate of cancer cells. CONCLUSIONS: Present data suggest the combination treatment of TIGIT and HIF-1α as a novel treatment strategy against colorectal and breast cancer, but more researches are required to realize the complete role of TIGIT and HIF-1α inside the TME.
Subject(s)
Antineoplastic Combined Chemotherapy Protocols/pharmacology , Hypoxia-Inducible Factor 1, alpha Subunit/antagonists & inhibitors , Nanoparticle Drug Delivery System/chemistry , Neoplasms/drug therapy , Receptors, Immunologic/antagonists & inhibitors , Animals , Antineoplastic Combined Chemotherapy Protocols/therapeutic use , Cell Line, Tumor/transplantation , Cell Proliferation/drug effects , Disease Models, Animal , Drug Synergism , Female , Humans , Hypoxia-Inducible Factor 1, alpha Subunit/metabolism , Immune Checkpoint Inhibitors/pharmacology , Immune Checkpoint Inhibitors/therapeutic use , Magnetic Iron Oxide Nanoparticles/chemistry , Mice , Neoplasm Invasiveness/prevention & control , Neoplasms/immunology , Neoplasms/pathology , Receptors, Immunologic/metabolism , Tumor Microenvironment/drug effects , Tumor Microenvironment/immunologyABSTRACT
The emergence of three-dimensional (3D) printing promises a disruption in the design and on-demand fabrication of smart structures in applications ranging from functional devices to human organs. However, the scale at which 3D printing excels is within macro- and microlevels and principally lacks the spatial ordering of building blocks at nanolevels, which is vital for most multifunctional devices. Herein, we employ liquid crystal (LC) inks to bridge the gap between the nano- and microscales in a single-step 3D printing. The LC ink is prepared from mixtures of LCs of nanocellulose whiskers and large sheets of graphene oxide, which offers a highly ordered laminar organization not inherently present in the source materials. LC-mediated 3D printing imparts the fine-tuning required for the design freedom of architecturally layered systems at the nanoscale with intricate patterns within the 3D-printed constructs. This approach empowered the development of a high-performance humidity sensor composed of self-assembled lamellar organization of NC whiskers. We observed that the NC whiskers that are flat and parallel to each other in the laminar organization allow facile mass transport through the structure, demonstrating a significant improvement in the sensor performance. This work exemplifies how LC ink, implemented in a 3D printing process, can unlock the potential of individual constituents to allow macroscopic printing architectures with nanoscopic arrangements.
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BACKGROUND: Previously, we reported that the complement cleavage fragments C3a and C5a are important modulators of trafficking of hematopoietic stem/progenitor cells (HSPCs). The aim of this study was to examine a possible role for complement component 1, subcomponent q (C1q) in HSPC migration. STUDY DESIGN AND METHODS: CD34+ HSPCs isolated from cord blood (CB), bone marrow (BM), and granulocyte-colony-stimulating factor (G-CSF)-mobilized peripheral blood (mPB) were evaluated for the expression of C1q and its receptor for phagocytosis (C1qRp) using reverse transcription-polymerase chain reaction, Western blotting, and fluorescence-activated cell sorting. Chemotactic responses and chemoinvasiveness toward stromal cell-derived factor (SDF)-1 and expression of matrix metalloproteinase (MMP)-9 were also examined after C1q stimulation. Moreover, G-CSF- and zymosan-induced mobilization was evaluated in C1q-deficient mice. RESULTS: C1q was expressed in CD34+ cells from mPB, but not from CB or steady-state BM; however, stimulation of the latter with G-CSF induced C1q expression. C1qRp receptor was found on BM, CB, and mPB CD34+ cells and more mature ex vivo expanded myeloid and megakaryocytic precursors. Although C1q itself was not a chemoattractant for HSPCs, it primed/enhanced the chemotactic response of CD34+ cells to a low SDF-1 gradient and their chemoinvasion across the reconstituted basement membrane Matrigel and increased secretion of MMP-9 by these cells. Moreover, in in vivo studies C1q-deficient mice were found to be easy G-CSF mobilizers compared to wild-type mice and normal zymosan mobilizers. CONCLUSION: We demonstrated that C1q primes the responses of CD34+ HSPCs to an SDF-1 gradient, which may enhance their ability to stay within BM niches, suggesting that the C1q/C1qRp axis contributes to HSPC homing/retention in BM.
Subject(s)
Complement C1q/metabolism , Complement C1q/pharmacology , Hematopoietic Stem Cells/cytology , Hematopoietic Stem Cells/metabolism , Animals , Antigens, CD34/metabolism , Blotting, Western , Cells, Cultured , Flow Cytometry , Granulocyte Colony-Stimulating Factor/pharmacology , Hematopoietic Stem Cells/drug effects , Humans , Mice , Mice, Inbred C57BL , Reverse Transcriptase Polymerase Chain ReactionABSTRACT
INTRODUCTION: Electroconvulsive therapy (ECT) is a non-pharmacological method for the treatment of psychiatric disorders. The precise biochemical mechanism of the effects of ECT is not clear, and since the two factors including matrix metalloproteinase-9 (MMP-9) and stromal cell-derived factor-1 alpha (CXCL12) play an important role in improving nerve damage, the effects of ECT and its relation with serum levels of MMP-9 and CXCL12 in patients with mania were investigated in this study. METHODS: In this before and after intervention study, the patients with mania, referring to the Qods Hospital in Sanandaj, were selected by the census method during the years 2015-2018. Young's test was performed 24 hrs before and after the first, third, and sixth sessions of ECT. For biochemical analysis, 3 mL of peripheral blood were taken prior to any anesthesia and 6 hrs after the first, third, and sixth sessions. Data were analyzed by two-way ANOVA and Pearson correlation coefficient by using the SPSS16 software. RESULTS: The results showed a significant decrease in Young's test scores during the first to the sixth session of ECT (P≤0.05). Although the levels of CXCL12 were slightly increased after the sixth course of ECT, they were not significant. Moreover, there were no significant relationship between the Young's test score and the serum levels of both MMP-9 and CXCL12 (P≥0.05). CONCLUSION: ECT improved patients clinically, but this effect was independent of serum levels of MMP-9 and CXCL12, and possibly other biochemical factors are involved in this pathway.
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BACKGROUND: The correlation of Helicobacter pylori infection with gastritis, peptic ulcer, and gastric cancer has been proven. The aim of this study was to determine the effects of cagA + and cagA - genotypes of H. pylori on genes expression of interleukin (IL) -10 and tumor growth factor (TGF) ß1 in gastric epithelial cells of patients with gastritis and H. pylori infection. METHODS: In all, 45 gastric biopsy samples were collected from patients with gastritis and H. pylori infection admitted to Tohid Hospital in Sanandaj city. Status of urease and cagA genes of H. pylori were directly determined from the biopsy samples using polymerase chain reaction (PCR) method. Expression of IL-10 and TGF-ß1 genes in gastric epithelial cells of patients with gastritis and cagA + and cagA- genotypes of H. pylori infection was serveyed using real-time PCR method. RESULTS: Overall, 25 samples had infection with H. pylori cagA + and 20 with cagA - genotypes. This study showed that there is a positive correlation between cagA - genotypes of H. pylori and increasing of IL-10 gene expression in gastric epithelial cells of patients with gastritis (P = 0.001). CONCLUSIONS: Level of gene expression of IL-10 as an anti-inflammatory cytokine in gastric epithelial cells of patients with H. pylori infection is connected to cagA- genotypes.
ABSTRACT
Matrix metalloproteinases (MMPs) play important roles in cancer progression and, despite their inhibitors have failed in the clinical trials, they have always been considered as suitable targets for the treatment of tumor. We have recently shown that membrane type (MT) 2-MMPs, is selectively expressed in multiple myeloma (MM) cells and mediates the metastatic characteristics of these cells. In this study, we designed efficient inhibitors against MT2-MMP using state-of-art molecular modeling methods. First, the 3D structure of MT2-MMP was predicted. Then, the proposed potent inhibitors against two regions of the catalytic domain of MT2-MMP (active site and MT-LOOP) were identified through molecular docking, QM-MM and molecular dynamics simulations from a set of compounds in Analyticon library, IBS library, Maybridge screening fragment library and drugbank library. Moreover, ADME estimation showed that pharmacokinetic properties of inhibitors are in the acceptable range for humans. Finally, our data suggested that compounds 'structures.722' (dobutamine) and 'M2' are suitable candidates to inhibit MT2-MMP for further examination in the laboratory.Communicated by Ramaswamy H. Sarma.