Showing papers in "Tissue & Cell in 2018"
TL;DR: The results showed that administration of 100μM of H2O2 on HUVECs for 2, 6, 12 and 24’h induced a time-dependent increase in ICAM-1 and VCAM- 1 mRNA and protein expression levels with a significant increase observed from 6 h, suggesting that H2 O2 induced oxidative stress may be a reasonable for atherosclerosis.
Abstract: Alterations of Endothelial cells (ECs) play a critical role in different pathogenesis of many serious human diseases, and dysfunction of the vascular endothelium is an indicator for human disorders Endothelial dysfunction is considered to be an early indicator for atherosclerosis, which is characterised by overexpression of adhesion molecules, including intercellular adhesion molecule-1 (ICAM-1) and vascular cell adhesion molecule-1 (VCAM-1) Hydrogen peroxide (H2O2) released via neutrophils is an important mediator of endothelial cell function Ambient production of superoxide anion (O2−) and subsequently H2O2 at low levels is critical for regulating endothelial cell functions and proliferation In this study, we investigated the effects of H2O2 on the expression of adhesion molecules VCAM-1 and ICAM-1 in cultured human umbilical vein endothelial cells (HUVECs) Intracellular superoxide anion production was detected by using p-Nitro Blue Tetrazolium (NBT) assay Our results showed that administration of 100μM of H2O2 on HUVECs for 2, 6, 12 and 24 h induced a time-dependent increase in ICAM-1 and VCAM-1 mRNA and protein expression levels with a significant increase observed from 6 h HUVECs exposed to H2O2 exhibit increased O2−, suggesting that H2O2 induced oxidative stress may be a reasonable for atherosclerosis This increase can be reduced by the flavonoid, N-acetyl cysteine (NAC) The modulation of endothelial cell function through this mechanism may underlie the contribution of H2O2 to the development of vascular disease
66 citations
TL;DR: A two-stage approach is proposed for computing oral histology images, where 12-layered (7 × 7×3 channel patches) deep convolution neural network (CNN) are used for segmentation of constituent layers in the first stage and in the second stage the keratin pearls are detected from the segmented keratin regions using texture-based feature (Gabor filter) trained random forests.
Abstract: Identification of various constituent layers such as epithelial, subepithelial, and keratin of oral mucosa and characterization of keratin pearls within keratin region as well, are the important and mandatory tasks for clinicians during the diagnosis of different stages in oral cancer (such as precancerous and cancerous). The architectural variations of epithelial layers and the presence of keratin pearls, which can be observed in microscopic images, are the key visual features in oral cancer diagnosis. The computer aided tool doing the same identification task would certainly provide crucial aid to clinicians for evaluation of histological images during diagnosis. In this paper, a two-stage approach is proposed for computing oral histology images, where 12-layered (7 × 7×3 channel patches) deep convolution neural network (CNN) are used for segmentation of constituent layers in the first stage and in the second stage the keratin pearls are detected from the segmented keratin regions using texture-based feature (Gabor filter) trained random forests. The performance of the proposed computing algorithm is tested in our developed oral cancer microscopic image database. The proposed texture-based random forest classifier has achieved 96.88% detection accuracy for detection of keratin pearls.
60 citations
TL;DR: The 3D tissue engineered composite model closely simulated the native oral hard and soft tissues and has the potential to be used as a valuable in vitro model for the investigation of bone invasion of oral cancer and for the evaluation of novel diagnostic or therapeutic approaches to manage OSCC in the future.
Abstract: Recent advances in three-dimensional printing technology have led to a rapid expansion of its applications in tissue engineering. The present study was designed to develop and characterize an in vitro multi-layered human alveolar bone, based on a 3D printed scaffold, combined with tissue engineered oral mucosal model. The objective was to incorporate oral squamous cell carcinoma (OSCC) cell line spheroids to the 3D model at different anatomical levels to represent different stages of oral cancer. Histological evaluation of the 3D tissue model revealed a tri-layered structure consisting of distinct epithelial, connective tissue, and bone layers; replicating normal oral tissue architecture. The mucosal part showed a well-differentiated stratified oral squamous epithelium similar to that of the native tissue counterpart, as demonstrated by immunohistochemistry for cytokeratin 13 and 14. Histological assessment of the cancerous models demonstrated OSCC spheroids at three depths including supra-epithelial level, sub-epithelial level, and deep in the connective tissue-bone interface. The 3D tissue engineered composite model closely simulated the native oral hard and soft tissues and has the potential to be used as a valuable in vitro model for the investigation of bone invasion of oral cancer and for the evaluation of novel diagnostic or therapeutic approaches to manage OSCC in the future.
40 citations
TL;DR: The L. inermis loaded PLLA-Gelatin nanofibers showed a potential application as a wound dressing in order to control wound infections and biocompatibility of the nan ofibers was proven on 3T3 fibroblasts.
Abstract: Since ancient times, some herbal medicines have been extensively used for burn and wound treatments, showing preference to the common synthetic medications by virtue of having less side effects and faster healing rate. In this study, hybrid nanofibrous scaffolds of poly- l -lactic-acid (PLLA) and gelatin incorporated L. inermis were fabricated via electrospinning technique. Morphology and characteristics of the scaffolds were studied by scanning electron microscopy (SEM) and Fourier transform infrared (FTIR), respectively. The release profile of the L. inermis from the nanofibers was also assessed in vitro. Moreover, the structural stability of the released L. inermis from the nanofibers was evaluated using high-performance liquid chromatography (HPLC). The nanofibers showed a gradual release of L. inermis up to two days while the intact structure was preserved. Furthermore, antibacterial assay demonstrated that L. inermis-loaded nanofibrous scaffolds could effectively kill E. coli and S. aureus within 2 h. Finally, biocompatibility of the nanofibers was proven on 3T3 fibroblasts. Therefore, the L. inermis loaded PLLA-Gelatin nanofibers showed a potential application as a wound dressing in order to control wound infections.
38 citations
TL;DR: It seems that LC could improve the aging-related features due to increasing the telomerase activity, decreasing aging, and changing the methylation status of hTERT promoter; it could potentially beneficial for enhancing the application of aged-MSCs in regenerative medicine.
Abstract: The identification of factors that reduce the senescent tendency of the mesenchymal stem cells (MSCs) upon expansion has great potential for cellular therapies in regenerative medicine. Previous studies have shown the aging protective effect of L-carnitine (LC). On the other hand, reduction in proliferation potential and age-dependent decline in number and functions of MSCs were accompanied by telomere shortening, reduction in telomerase activity and epigenetic changes. The aim of this study was to evaluate the effects of LC on aging of MSCs through telomerase activity assessment and the investigation of methylation status of the hTERT gene promoter. Telomerase activity and hTERT promoter methylation investigation was performed with PCR-ELISA TRAP assay and methylation specific PCR (MSP), respectively. Also, beta-galactosidase (SA-s-gal) staining was used to calculate the percentage of senescent cells. The results showed that the LC could efficiently promote the telomerase activity. In addition, the percentage of senescent cells had significantly decreased and changes in the methylation status of the CpG islands in the hTERT promoter region under treatment with LC were seen. In conclusion, it seems that LC could improve the aging-related features due to increasing the telomerase activity, decreasing aging, and changing the methylation status of hTERT promoter; it could potentially beneficial for enhancing the application of aged-MSCs in regenerative medicine.
32 citations
TL;DR: It is demonstrated that Rutin significantly ameliorates the side effects induced by restraint stress and expressional levels of cleaved PARP1 and cleaved Caspase-3 are significantly increased in testes after restraint stress.
Abstract: In order to investigate the effects of Rutin against restraint stress, 50 adult male mice were divided into five groups: control, restraint stress (RS), and RS with 2 doses of Rutin treatments. Mice were restrained in a conical tube for 4 h daily and Rutin was injected intraperitoneally for 15 consecutive days. Restraint stress significantly decreases body weights, testis and epididymis weights, thymus weights, visceral fats, serum concentrations of testosterone, sperm counts, sperm motility and sperm viability, while it increases serum epinephrine levels, adrenal gland weights and abnormal sperms. In addition, restraint stress severely damages the testicular histoarchitecture and spermatogenesis. Stressed groups also showed broken seminiferous tubules, few spermatozoa in lumen, less population of Leydig cells between the interstitial spaces, spermiation arrest in stage I–III and degenerated population of round spermatids in the lumen; as well as missing cells in stages IV–VI. Furthermore, lumen sizes increased in stages VII, VIII, IX and X. Residual bodies increased in stages IV–VI, VII–VIII and vacuoles found in stages XI–XII after restraint stress. PARP1 signaling is involved in apoptosis. In this study, expressional levels of cleaved PARP1 and cleaved Caspase-3 are significantly increased in testes after restraint stress. We demonstrate that Rutin significantly ameliorates the side effects induced by restraint stress.
28 citations
TL;DR: Non-mulberry silk fibroin fiber reinforced chitosan based scaffold might be suitable scaffold that can act as a potential artificial matrix for cartilage tissue engineering.
Abstract: Lack of potential regenerative medicine to reconstruct damaged cartilage tissue has accelerated investigation and development of potential biomaterial for cartilage tissue engineering. In this study, we fabricated micron-sized non-mulberry silk fibroin fiber (SFF) using N,N-Dimethylacetamide (DMAC)/10% LiBr solution and further used to develop SFF reinforced chitosan(CH) based porous scaffold with desired pore size, porosity, swelling and structural stability. The developed scaffold was characterized for its various physico-chemical, mechanical and biological properties. The developed CH/SFF composite scaffold facilitates human mesenchymal stem cell (hMSCs) attachment, colonization and extracellular matrix deposition. Furthermore, hMSCs shows significantly higher sulfated glycosaminoglycan deposition over CH/SFF in comparison to pure chitosan scaffold (control). Immunocytochemistry studies have shown enhanced expression of collagen type II and aggrecan by hMSCs over composite scaffold than chitosan scaffold. Thus, non-mulberry silk fibroin fiber reinforced chitosan based scaffold might be suitable scaffold that can act as a potential artificial matrix for cartilage tissue engineering.
24 citations
TL;DR: This study describes how the morphological, mechanical and biochemical properties of adenocarcinoma (A549) cells change in response to a 3D environment and varying substrates.
Abstract: Alternative models such as three-dimensional (3D) cell cultures represent a distinct milestone towards capturing the realities of cancer biology in vitro and reduce animal experimentation in the preclinical stage of drug discovery. Significant work remains to be done to understand how substrates used in in vitro alternatives influence cancer cells phenotype and drug efficacy responses, so that to accurately link such models to specific in vivo disease scenarios. Our study describes how the morphological, mechanical and biochemical properties of adenocarcinoma (A549) cells change in response to a 3D environment and varying substrates. Confocal Laser Scanning (LSCM), He-Ion (HIM) and Atomic Force (AFM) microscopies, supported by ELISA and Western blotting, were used. These techniques enabled us to evaluate the shape, cytoskeletal organization, roughness, stiffness and biochemical signatures of cells grown within soft 3D matrices (PuraMatrix™ and Matrigel™), and to compare them to those of cells cultured on two-dimensional glass substrates. Cell cultures are also characterized for their biological response to docetaxel, a taxane-type drug used in Non-Small-Cell Lung Cancer (NSCLC) treatment. Our results offer an advanced biophysical insight into the properties and potential application of 3D cultures of A549 cells as in vitro alternatives in lung cancer research.
24 citations
TL;DR: Results revealed, ERRs indirectly regulate Leydig cell proliferation while ERRα and β affect cell monolayer formation, and ERRs should be considered important factors in developing of innovating approaches that target pathological processes of testicular Leydigs cells.
Abstract: In this study, we demonstrate, for the first time, estrogen-related receptor (ERR) regulation of the physiological and biochemical status of testicular tumor Leydig cells. In a mouse tumor Leydig cells, ERRs (α, β, and γ) were silenced via siRNA. Cell morphology and cell physiology (proliferation and observation of monolayer formation) were performed by inverted phase-contrast microscope. Leydig cell functional markers (steroid receptors and signaling molecules) were examined by immunofluorescence and Western blotting. Additionally, progesterone secretion was assessed. Mitochondrial mass and membrane potential were analyzed by flow-cytometry while cGMP and Ca2+ concentrations were analyzed using immunoenzymatic and colorimetric assays, respectively. These results revealed, ERRs indirectly regulate Leydig cell proliferation while ERRα and β affect cell monolayer formation. ERRs interact with canonical and membrane estrogen receptors (ERα, ERβ, and GPER), androgen receptor, metalloproteinase (MMP 9), protein kinase A (PKA), extracellular-regulated kinase (ERK), and neurogenic locus notch homolog protein 2 (Notch2). Depending on the type of ERR knocked down, coupled with estradiol treatment, changes in progesterone concentration and cGMP and Ca2+ concentrations constitute a microenvironment that may effect tumor Leydig cell characteristics. ERRs should be considered important factors in developing of innovating approaches that target pathological processes of testicular Leydig cells.
22 citations
TL;DR: Investigation of the possible role of zinc oxide nanoparticles as a new strategy to protect against CP induced testicular injury revealed that CP induced deleterious testicular histopathological, biochemical and genetic alterations that were effectively prevented by ZnO NPs.
Abstract: Despite its wide range of application, cyclophosphamide (CP) exhibits a wide range of adverse effects including reproductive toxicity. The emerging field of zinc oxide nanoparticles (ZnO NPs) therapy may provide a new hope for prevention of CP induced gonadal toxicity. Herein, we aim to investigate the possible role of ZnO NPs as a new strategy to protect against CP induced testicular injury. Sixty adult male albino rats were divided into 3 groups; control, CP treated and CP + ZnO NPs treated groups. CP group was injected with CP (5 mg/kg/day), whereas CP + ZnO NPs group was concomitantly injected with CP and ZnO NPs (5 mg/kg/day). Testicular specimens were processed for histological, ultrastructural and c-kit immunohistochemical study. Biochemical analysis for tissue malondialdehyde and serum testosterone was done in addition to sperm morphology assay and cytogenetic study. Our results revealed that CP induced deleterious testicular histopathological, biochemical and genetic alterations that were effectively prevented by ZnO NPs.
21 citations
TL;DR: It has been demonstrated in this study that depletion of Treg ameliorates bleomycin-induced lung fibrosis by immune modulating Th17 and other important T cell subsets response in lung, and suggested that Treg depletion led to considerable upregulation of IFN-γ after bleomyzin administration.
Abstract: Idiopathic pulmonary fibrosis is an irreversible, progressive and lethal lung disease. Regulatory T cells (Tregs) and Th17 cells both are involved in lung fibrosis. But there are only few reports regarding the effect of Treg on other T cell subsets in experimental lung fibrosis. The aim of this study was to investigate the impact of Treg on Th17, CD4+CD28-T, CD4+CD28+T and CD8 + T cell subsets that could drive lung fibrosis. To reach the goal of our study, first we depleted Tregs by anti-CD25 mAb injection in experimental C57BL/6 mice model. It has been demonstrated in our study that depletion of Treg ameliorates bleomycin-induced lung fibrosis by immune modulating Th17 and other important T cell subsets response in lung. Our flow cytometry data revealed that the percentages of Th17, CD4+CD28-T, CD4+CD28+T and CD8 + T cell subsets were decreased in experimental lung fibrosis after Treg depletion. We also observed significant downregulation of IL-17 A in Treg-depleted mice after bleomycin delivery. In addition, the study also suggested that Treg depletion led to considerable upregulation of IFN-γ after bleomycin administration. Therefore, Th17 cells, CD8 + T cells, CD4+CD28- and CD4+CD28+ T cell subsets all are controlled by regulatory T cell, help in progression of fibrosis in experimental lung fibrosis.
TL;DR: In this paper, a total of 74 Chinese Alligators (30 males and 44 females) were examined to establish reference range values of hematology and serum biochemistry parameters during the active and hibernating periods.
Abstract: The Chinese alligator Alligator sinensis is an endangered freshwater crocodilian species endemic to China. Hematology and serum biochemistry reference range are useful in the assessment and management of animal health condition. In this study, a total of 74 Chinese Alligators (30 males and 44 females) were examined to establish reference range values of hematology and serum biochemistry parameters during the active and hibernating periods. We measured and analyzed 9 hematology and 21 serum biochemistry parameters including 4 serum electrolyte parameters, and described the morphology of different types of blood cells. No statistical differences between the sexes were found for hematology parameter, while significant differences were noted for some serum biochemistry parameters, with males having greater alkaline phosphatase activity level and lower globulin concentration value than females. There were some significant differences between the two different periods with alligators during the active period possessing lower values for mean corpuscular volume, mean corpuscular hemoglobin, total bilirubin and creatine kinase, but higher values for red blood cell and white blood cell counts, monocyte percentage, aspartate aminotransferase, a-amylase, blood urea nitrogen, creatinine, triglycerides, and cholesterol. These baseline data were essential for health condition evaluation and disease diagnosis of this endangered species.
TL;DR: Evidence is provided of the structure-modifying abilities of ASU via its anti-oxidative and anti-inflammatory properties, in addition to its ability to modulate MMP-13 activity, which suggests that ASU can be used as a potential disease- modifying treatment for OA.
Abstract: Osteoarthritis (OA), the most common type of arthritis, is a disabling progressive disease mainly affecting the elderly and is becoming a major public health problem. Current therapies for OA provide only palliative pain relief and therapeutic candidates that are able to slow the progression of structural deterioration is a major unmet need for this disorder. Avocado soybean unsaponifiables (ASU) has proven its safety and effectiveness in clinical studies of knee osteoarthritis (OA); however, whether ASU exerts structure-modifying effects is still to be elucidated. There are limited studies that have explored the underlying mechanisms of ASU’s beneficial effects in animal models of OA. To this end, this study is the first to evaluate the effects of ASU in a rat model of mono-iodoacetate (MIA)-induced knee OA. OA was induced in rats by knee intra-patellar injection of MIA. Oral administration of ASU (27.5 mg/kg per day for 3 weeks) was initiated 3 weeks after MIA injection. We analysed the knee samples using light and electron microscopy. In addition, we used immunohistochemistry to investigate the expression of inducible nitric oxide synthase (iNOS), tumour necrosis factor-α (TNF-α), and matrix metalloproteinase-13 (MMP-13) in OA cartilage and subchondral bone. ASU significantly attenuated the synovium, cartilage, and subchondral degeneration. In addition, it reduced the expression of TNF-α and MMP-13 in OA cartilage and the expression of iNOS in both OA cartilage and subchondral bone. These results provide evidence of the structure-modifying abilities of ASU via its anti-oxidative and anti-inflammatory properties, in addition to its ability to modulate MMP-13 activity. This work suggests that ASU can be used as a potential disease-modifying treatment for OA.
TL;DR: The natural characteristic of the present bone scaffold, its cell adhesion features and capability to induce bone mineralization, even in the standard culture medium, provides a potentially optimal bone scaffolds for bone tissue engineering.
Abstract: Extracellular matrix (ECM) controls cellular behaviors such as proliferation, migration, and differentiation. The decellularized matrix of mammalian tissues has been used as a scaffold for the repair and reconstruction of tissue defects. In this study, for the first time demineralized and decellularized human epiphyseal bone matrix was used as a scaffold for a bone generation. Human epiphyseal bone was demineralized by hydrochloric acid and then decellularized by three methods of physical (slow freezing and snap freeze-thaw), enzymatic (trypsin 0.25%, 18 h) and chemical sodium dodecyl sulfate (SDS),)2.5%, 26 h). The scaffolds were cultured with rat adherent bone marrow cells (RABMC). Then, the histological studies were performed on days 7, 14, 21, and 28 of the culture to observe the distribution and morphology of cells. Bone formation was also investigated using Alizarin red staining. The results of this study indicated that RABMC migrated, proliferated and separated by forming lacuna in this three-dimensional bone scaffold. In addition, the Alizarin red staining indicated the calcium deposition on the scaffold in both bone differentiation and standard culture medium. The natural characteristic of the present bone scaffold, its cell adhesion features and capability to induce bone mineralization, even in the standard culture medium, provides a potentially optimal bone scaffold for bone tissue engineering.
TL;DR: The data obtained suggest that interferon signaling pathway activation by PRP treatment in combination with BCG immunotherapy may provide novel therapeutic approaches for non-muscle invasive bladder cancer.
Abstract: This study describes the effects of a promising therapeutic alternative for non-muscle invasive bladder cancer (NMIBC) based on Bacillus Calmette-Guerin (BCG) intravesical immunotherapy combined with Platelet-rich plasma (PRP) in an animal model. Furthermore, this study describes the possible mechanisms of this therapeutic combination involving Toll-like Receptors (TLRs) 2 and 4 signaling pathways. NMIBC was induced by treating female Fischer 344 rats with N-methyl-N-nitrosourea (MNU). After treatment with MNU, the animals were distributed into four experimental groups: Control (without MNU) group, MNU (cancer) group, MNU + PRP group, MNU + BCG group and MNU + PRP + BCG group. Our results demonstrated that PRP treatment alone or associated with BCG triggered significant cytotoxicity in bladder carcinoma cells (HTB-9). Animals treated with PRP associated to BCG clearly showed better histopathological recovery from the cancer state and decrease of urothelial neoplastic lesions progression in 70% of animals when compared to groups that received the same therapies administered singly. In addition, this therapeutic association led to distinct activation of immune system TLRs 2 and 4-mediated, resulting in increased MyD88, TRIF, IRF3, IFN-γ immunoreactivities. Taken together, the data obtained suggest that interferon signaling pathway activation by PRP treatment in combination with BCG immunotherapy may provide novel therapeutic approaches for non-muscle invasive bladder cancer.
TL;DR: A novel triple negative canine mammary cancer cell line, CMT-7364, was successfully established, which could be used as a promising model for the research of immunotherapy and Epithelial-Mesenchymal Transition (EMT) mechanism of the triple-negative breast cancer both in canine and human.
Abstract: Canine mammary tumor (CMT) has always been an ideal animal model for human breast cancer (HBC) research, however, there is a lack of various established CMT cell lines corresponding to HBC cell lines. This study was designed to establish a new type of CMT cell line. The primary tumor, CMT-7364, was identified as the intraductal papillary carcinoma, and showed negative immunoreactivity to estrogen receptors (ER), progesterone receptors (PR), and human epidermal growth factor receptor-2 (HER-2) by immunohistochemistry (IHC) analysis. The CMT-7364 cell line from this primary tumor also shows a negative immunoreactivity to ER, PR, and HER-2, and was negative to epithelial cell markers and positive to mesenchymal cell markers by immunocytochemistry (ICC) analysis. This cell line, which has been stably cultured for more than 115 passages, and was characterized by epithelial origin with the expression of the epithelial antigen by ICC analysis and invasion ability by transwell analysis. In vivo, tumor mass and metastases in the lung were found after inoculating the CMT-7364 cells in the nude mice model, and the immune-complete mice model respectively. The tissues from the xenograft tumor were also negative to ER, PR, and HER-2 by IHC analysis. Thus, a novel triple negative canine mammary cancer cell line, CMT-7364, was successfully established, which could be used as a promising model for the research of immunotherapy and Epithelial-Mesenchymal Transition (EMT) mechanism of the triple-negative breast cancer both in canine and human.
TL;DR: Gemcitabine (GEM) in combination with TRA was more effective than TRA alone in inhibition of pancreatic cancer patient-derived orthotopic xenograft tumor growth, and the clinical potential of the combination of TRA and GEM is suggested.
Abstract: Pancreatic cancer is resistant to treatment and needs precision individualized therapy to improve the outcome of this disease. Previously, we demonstrated that trametinib (TRA), a MEK inhibitor, could inhibit a pancreatic cancer patient-derived orthotopic xenograft (PDOX). In the present study, we show that gemcitabine (GEM) in combination with TRA was more effective than TRA alone. We implanted a patient pancreatic cancer orthotopically in the pancreatic tail of nude mice to establish the PDOX model. After seven weeks of tumor growth, we divided 32 pancreatic-cancer PDOX nude mice into 4 groups of eight: untreated control; GEM (once a week for 2 weeks); TRA (14 consecutive days); GEM + TRA (GEM: once a week for 2 weeks, TRA:14 consecutive days). We found that treated mice on day 14 had significantly reduced tumor volume in comparison to untreated control. TRA and the combination of GEM + TRA therapy significantly inhibited tumor development in comparison to GEM alone. However, GEM + TRA inhibited the PDOX tumor growth significantly greater than TRA alone. These results suggest the clinical potential of the combination of TRA and GEM for pancreatic cancer.
TL;DR: It is confirmed that allicin improves the cognitive function of APP/PS1 double transgenic mice by reducing the expression levels of Aβ, oxidative stress, and improving mitochondrial dysfunction.
Abstract: In middle and old age, Alzheimer's disease (AD) is a progressive neurodegenerative disorder of brain. As an increasingly aging population, AD represents a huge burden for the patients' family and the country. However, current therapeutical strategies have shown limited effectiveness. Allicin, which is the main composition of garlic, was reported to prevent the learning and memory impairment of AD mouse model. As the mechanism is not clear, in this study, we used the APP (amyloid precursor protein)/PS1 (presenilin 1) double transgenic mice, which express human mutant APP and PS1, to determine the protective effect of allicin on neurons. AD involves a broad range of clinical, cellular, and biochemical manifestations. This has led to many views of AD, e.g. the amyloid, presenilin, oxidative stress, and mitochondrial dysfunction. We confirmed that allicin improves the cognitive function of APP/PS1 double transgenic mice by reducing the expression levels of Aβ, oxidative stress, and improving mitochondrial dysfunction. Application of behavioral, morphological and molecular biology, and other means were conducted to investigate the effect of allicin, which provide an experimental basis for the reliable application of allicin in the treatment of neurodegenerative diseases.
TL;DR: The aberrant crypt foci were attenuated by approximately 43% and 20% in the kefir group compared to AOM group, suggesting that kefIR treatment may contribute to prevent and control the growth of intestinal neoplastic cells.
Abstract: Kefir is a probiotic fermented milk product produced from grains with a complex composition of bacteria and yeasts that live in a symbiotic association. Anti-proliferative, anti-inflammatory, and anti-mutagenic effects are some of the health beneficial properties of kefir grains. The present study was conducted to evaluate whether regular consumption of kefir milk would be capable of preventing the development of pre-neoplastic lesions induced by azoxymethane (AOM). Aberrant crypt foci were induced in BALB-c mice via 2 subcutaneous injections of azoxymethane (15 mg/kg) and kefir was administered by daily gavage for 8 weeks (5 ml/kg). Additionally, bacterial growth was monitored in pasteurized and ultra-high temperature (UHT) treated milk to compare different fermentation conditions. Our results showed that UHT milk presented better growth of Lactobacillus acidophilus colonies. The aberrant crypt foci were attenuated by approximately 43% (height) and 20% (width) in the kefir group compared to AOM group, suggesting that kefir treatment may contribute to prevent and control the growth of intestinal neoplastic cells.
TL;DR: The results suggest that direct culture of whole menstrual blood cells is a high throughput, straightforward and cost-effective method for MenSCs isolation using no special growth factors and these cells are a good producer of exosome which can offer a cell-free therapy approach.
Abstract: Background Mesenchymal stem cells (MSCs) are currently the most promising candidates in regenerative medicine. Nonetheless, there are several limitations associated with the MSC tissue source such as infrequent and invasive bone marrow sampling methods. To overcome these limitations, we have procured MSCs from the menstrual blood (MenSCs) as a non-invasive source using a straightforward and cost-effective method. Moreover, we isolated MenSCs-derived exosomes as a safe and highly effective approach to exert the paracrine effects of MSCs. Methods MSCs were isolated from menstrual blood through two different culture methods: ficoll-isolated mononuclear cells (MNCs) and whole blood culture. These cells were characterized by their plastic adherence, flow cytometry analysis of the surface markers and the differentiation potential. The exosomes were isolated from conditioned media using ultracentrifugation and characterized by different microscopy techniques, western blotting, and ELISA. Results Both Methods resulted in the rapid isolation of cells with MSC properties. However, the cellular yield of the whole blood culture method was remarkably more than MNCs culture. MenSCs also produced a substantial amount of extracellular vesicles (EVs) possessed the minimum criteria for exosome definition. Conclusion The results suggest that direct culture of whole menstrual blood cells is a high throughput, straightforward and cost-effective method for MenSCs isolation using no special growth factors. Moreover, these cells are a good producer of exosome which can offer a cell-free therapy approach.
TL;DR: It is suggested that EC from different tooth tissue have unique characteristics related to their target tissue and function as well as ex vivo sprouting angiogenesis from various tissues.
Abstract: Dental pulp is a highly vascularized tissue with a high regenerative capacity. This is attributed to its unique blood supply and the presence of progenitor or postnatal dental pulp stem cells. Here we aimed to isolate and compare the angiogenic properties of endothelial cells (EC) prepared from mouse dental pulp and periodontal ligament (PDL). EC were isolated from 4-week-old wild type immorto mice. Mice were sacrificed and after mandible isolation, the molar and incisor teeth and the PDL from molar teeth were dissected. EC were prepared by collagenase digestion of tissues and affinity purification using magnetic beads coated with platelet/endothelial cell adhesion molecule-1 (PECAM-1/CD31) antibody. EC prepared from incisor and molar pulps and PDL were examined for expression of appropriate markers by fluorescence-activated cell sorting (FACS) analysis. The proliferation, migration, and capillary morphogenesis of EC were evaluated. Ex vivo sprouting angiogenesis from various tissues was also compared. Data were analyzed at the level of significance of P<0.05. Pulp EC prepared from incisors proliferated and migrated significantly faster than molar and PDL EC (P<0.05). In addition, molar and PDL EC formed a more extensive capillary network when plated on Matrigel. This is consistent with the lower proliferative and migratory characteristics of these cells compared with incisor EC (P<0.05). However, PDL tissue showed significantly more sprouting area than molar and incisor pulp tissues (P<0.05). Thus, pulp EC from molar and incisor and PDL EC present different proangiogenic properties. Collectively our results suggest that EC from different tooth tissue have unique characteristics related to their target tissue and function.
TL;DR: The results show that new cells in the midgut arise from the anterior and posterior ends of this organ and either migrate or divide toward its middle, and these cells divide at a constant rate, suggesting that the cell turnover of these organs may be extremely slow or dependent on changing environmental conditions.
Abstract: The taxon Tardigrada, commonly called "water bears", consists of microscopic, eight-legged invertebrates that are well known for their ability to tolerate extreme environmental conditions. Their miniscule body size means that tardigrades possess a small total number of cells, the number and arrangement of which may be highly conserved in some organs. Although mitoses have been observed in several organs, the rate and pattern of cell divisions in adult tardigrades has never been characterized. In this study, we incubated live tardigrades over a period of several days with a thymidine analog in order to visualize all cells that had divided during this time. We focus on the midgut, the largest part of the digestive system. Our results show that new cells in the midgut arise from the anterior and posterior ends of this organ and either migrate or divide toward its middle. These cells divide at a constant rate and all cells of the midgut epithelium are replaced in approximately one week. On the other hand, we found no cell divisions in the nervous system or any other major organs, suggesting that the cell turnover of these organs may be extremely slow or dependent on changing environmental conditions.
TL;DR: It is hypothesize that eEF2 may be activated in a positive feedback cycle through inactivation of e EF2K via the PI3K/Akt/mTOR pathway through the PI2K-Akt-MTOR pathway.
Abstract: Eukaryotic elongation factors 2 (eEF2) plays an essential role in the GTP-dependent translocation of the ribosome along mRNA. Previous studies have shown that eEF2 is overexpressed in various tumors. However, little is known about the role of eEF2 in ovarian cancer. The aim of the present study is to examine the effect of eEF2 on ovarian cancer proliferation. We first measured eEF2 protein expression by western blot using six fresh ovarian cancer tissues from G1 to G3. The results showed that eEF2 expression gradually increased from G1 to G3. Additionally, eEF2 expression correlated significantly with grade (P = 0.045), FIGO stage (P = 0.035) and Ki67 (P
TL;DR: It is implied that Foxc1 might regulates fibroblast-like synoviocytes proliferation by reducing PI3K/AKT signaling pathway and all above findings provide novel therapeutic effects in the treatment for RA patients.
Abstract: Forkhead box c1 (Foxc1) is a vital member of the Fox family of transcription factors which play important roles in numerous biological processes including metabolism, differentiation, proliferation, apoptosis, migration, invasion and longevity. However, up to date, the role of Foxc1 in the development of Rheumatoid Arthritis (RA) has not been fully elucidated. In the present study, the markedly higher expression of Foxc1 was observed in fibroblast-like synoviocytes (FLSs) of RA compared to control. Besides, we found that Foxc1 had a co-localization with THY1 (a marker for fibroblast-like synoviocytes). Moreover, during the process of TNF-α-induced inflammatory response model, Foxc1 was progressively accumulated in FLSs which was in parallel with MMP-1, MMP-13. Consistently, cell inflammatory response was distinctly hindered by small interfering RNA. Even more importantly, we discovered that Foxc1 promoted cell proliferation by upregulation PI3K/AKT signaling, which was inflammation-dependent. In summary, these data implied that Foxc1 might regulates fibroblast-like synoviocytes proliferation by reducing PI3K/AKT signaling pathway and all above findings provide novel therapeutic effects in the treatment for RA patients.
TL;DR: There is a great need for a comprehensive summary of these systems for researchers seeking proper in vitro models and several key points for the culture procedure are clarified and its utility in retinal research is summarized.
Abstract: Reliable disease models are essential for investigations on mechanisms and therapies. The Retinal organotypic culture can maintain the architecture and cellular connections within the tissue in vitro. The system is a refined retinal experiment platform. It narrows the gap between cell line studies and in vivo models and is flexible enough for sophisticated experimental procedures. It greatly reduces the consumption of time or resources. Retinas of many species in various development stages have been used for diverse explorations based on their morphologies and physical characteristics. But the culture time course and the viability of the cultured tissue restrict the utility of the system. Recently, researchers have made increasing attempts to improve the culture conditions and applications of this systems for retina experiments in vitro. Accordingly, there is a great need for a comprehensive summary of these systems for researchers seeking proper in vitro models. In this review, we clarify several key points for the culture procedure and summarize its utility in retinal research.
TL;DR: Combining 3D-cultured MSC spheroid technology with immune evasion stem cell composite sheet improved the outcome and strengthened the protection against allogenic immune rejection.
Abstract: Mesenchymal stem cells (MSCs) are widely used in transplantation therapy due to their multilineage differentiation potential, abundance, and immuno-modulating ability. However, the risk of allograft rejection limits their application. Here, we proposed a novel method to facilitate MSC transplantation with enhanced applicability and efficacy. We cultured human adipose-derived MSCs in a 3D culture under in vitro expansion conditions and under conventional 2D adherent culture conditions. MSC spheroids promoted extracellular matrix molecules that stimulate MSC proliferation, and produced more angiogenic cytokines such as vascular endothelial growth factor, hepatocyte growth factor, and fibroblast growth factor than 2D-cultured MSCs. Further, MSC spheroids showed increased IDO expression, increased proportion of M2 macrophages, and decreased macrophage proliferation, compared to 2D-cultured MSCs. Next, we proposed the wrapping of autologous cell sheets from the recipient around in-vitro-grown MSC spheroids to prevent allogenic immune rejection during transplantation. Myoblasts from C57BL/6 mice were used to prepare a stem cell composite sheet containing human-derived MSC spheres. The transplantation of MSC spheroids increased the survival rate and decreased the inflammatory response of the immunocompetent C57BL/6 ischemic mice. Thus, combining 3D-cultured MSC spheroid technology with immune evasion stem cell composite sheet improved the outcome and strengthened the protection against allogenic immune rejection.
TL;DR: It was showed that PDGFRA amplification could be effectively targeted by PAZ and the PDOX model can identify effective targeted therapy as well as standard therapy and at the same time, identify ineffective drugs, even if they are first-line.
Abstract: Pleomorphic liposarcoma (PLPS) is a heterogeneous resistant group of tumors. Complete surgical resection is the only known way to treat PLPS. PLPS is reristant to both radiation and chemotherapy. Therefore, precise individualized therapy is needed to improve outcome of advanced PLPS patients. In this study, a patient-derived orthotopic xenograft (PDOX) model of a PDGFRA-amplified PLPS was established in the biceps femoris of nude mice by surgical orthotopic implantation (SOI) in order to match the patient. The PLPS PDOX was treated with pazopanib (PAZ) which targets PDGFRA, as well as with temozolomide (TEM) and first-line therapy doxorubicin (DOX). The PLPS PDOX was resistant to DOX and responded very well to PAZ as well as TEM. The tumor volume on treatment day-14 relative to day-1 was as follows: DOX (4.50 ± 2.6, p = 0.8087); PAZ (1.29 ± 0.9, p = 0.0008 compared to the control, p = 0.0167 compared to DOX); TEM (1.07 ± 0.8, p = 0.0079 compared to the control, p = 0.0079 compared to DOX). There was no significant difference in body weight between any treated group or control. The PAZ- and TEM-treated tumors showed extensive necrosis compared to the DOX-treated and untreated PDOX tumors. The present study showed that PDGFRA amplification could be effectively targeted by PAZ. The PLPS PDOX model also identified the efficacy of TEM which does not target PDGFRA, indicating that the PDOX model can identify effective targeted therapy as well as standard therapy and at the same time, identify ineffective drugs, even if they are first-line.
TL;DR: Recruitment of the cell types of the neurovascular unit is a prerequisite for proper retinal vascularization and BRB formation in neonatal mouse retina is suggested.
Abstract: The inner blood-retinal barrier (BRB) is made up by the neurovascular unit, consisting of endothelial cells, pericytes and glial cells. The BRB maintains homeostasis of the neural retina, but in pathological eye conditions the neurovascular unit is often disrupted, causing BRB loss. Here, we investigated in detail temporal and spatial recruitment of the neurovascular unit in the neonatal mouse retina from postnatal day (P)3 to P25 employing immunohistochemical staining of vascular endothelium (isolectin B4), pericytes (α-SMA and NG2) and astrocytes (GFAP). In addition, we investigated gene expression of polarized astrocytic end-feet markers aquaporin-4 and laminin α2 chain with qPCR. We observed GFAP-positive cells migrating ahead of the retinal vasculature during the first postnatal week, suggesting that the retinal vasculature follows an astrocytic meshwork. From P9 onwards, astrocytes acquired a mature phenotype, with a more stellate shape and increased expression of aquaporin-4. NG2-positive cells and tip cells co-localized at P5 and invaded the retina together as a vascular sprouting front. In summary, these data suggest that recruitment of the cell types of the neurovascular unit is a prerequisite for proper retinal vascularization and BRB formation.
TL;DR: Both treatments were able to accelerate injury repair, and the association of both presented better results than the isolated applications, but the FD method showed no sensitivity to differentiate the treatments, either in the histological aspects or the injury in collagen.
Abstract: Objectives To evaluate alterations from different therapies in muscular injury using the Fractal Dimension (FD) method Methods 35 animals were allocated in Control Group (C), Injury Control Group (IC), Injury Low Level Laser Therapy Group (ILT), Injury Platelet Rich Plasma Group (IP), and Injury LLLT and PRP Group (ILP) The animals suffered a stretch injury in gastrocnemius muscle and after that IP and ILP groups received PRP application The ILT and ILP groups received daily LLLT applications for seven days After seven days the animals were euthanized and the gastrocnemius muscle removed and frozen The muscles were stained with Hematoxylin and Eosin (HE) and Picrosirius Red, for observation of the morphology of the injury and semi-quantitative and quantitative analysis through the Fractal Dimension (FD) method Results In the qualitative and semi-quantitative analysis, in relation to IC group, the ILT presented a reduction in rounded fibers and the IP in angular fibers The ILP group demonstrated a reduction in both polymorphic fibers and inflammatory infiltrate The FD of the muscles stained with HE was higher in the groups that suffered the injury when compared to the C group (p Conclusion Both treatments were able to accelerate injury repair, and the association of both presented better results than the isolated applications However, the FD method showed no sensitivity to differentiate the treatments, either in the histological aspects or the injury in collagen
TL;DR: Results indicated that Dyrk1A might promote FLSs proliferation, migration and invasion by suppressing Spry2 expression and activating the ERK MAPK signaling pathway in RA.
Abstract: Fibroblast-like synoviocytes (FLSs) play an essential role in rheumatoid arthritis (RA) by promoting synovitis, pannus growth and cartilage/bone destruction. Increased proliferation, migration and invasion of FLSs greatly contribute to RA initiation and progression. Dual-specificity tyrosine-regulated kinase 1A (Dyrk1A), a serine/threonine kinase, regulates MAPK pathway activation, and governs the proliferation and differentiation of neuronal progenitor cells and cancer cells. Till now, the expression and possible function of Dyrk1A in RA FLSs have not been explored. In this study, we detected an increased expression of Dyrk1A both in the synovial tissues of RA patients and in a TNF-α-induced FLSs activation model. CCK-8 and Edu assays revealed that Dyrk1A knockdown inhibited TNF-α-induced FLSs proliferation. Moreover, inhibiting Dyrk1A expression apparently prevented the migration and invasion capability of FLSs accompanied by a decreased MMP-3 and -9 expression. To investigate the molecular mechanism through which Dyrk1A modulates FLSs activities, we evaluated the effects of Dyrk1A on Spry2, a negativity modulator of ERK MAPK pathway. Western blot assay demonstrated that Dyrk1A silencing significantly increased Spry2 expression and suppressed the phosphorylation of ERK in TNF-α-treated FLSs. Taken together, our results indicated that Dyrk1A might promote FLSs proliferation, migration and invasion by suppressing Spry2 expression and activating the ERK MAPK signaling pathway in RA.