scholarly journals Dynamic Characterization of the Biomechanical Behaviour of Bovine Ovarian Cortical Tissue and Its Short-Term Effect on Ovarian Tissue and Follicles

Materials ◽  
2020 ◽  
Vol 13 (17) ◽  
pp. 3759
Author(s):  
Giulia Pascoletti ◽  
Maddalena Di Nardo ◽  
Gionata Fragomeni ◽  
Vincenza Barbato ◽  
Teresa Capriglione ◽  
...  
Keyword(s):  
Mechanical Test ◽  
Cell Damage ◽  
Ovarian Tissue ◽  
Viscoelastic Behavior ◽  
Mechanical Tests ◽  
Cortical Tissue ◽  
Short Term ◽  
Tissue Properties

The ovary is a dynamic mechanoresponsive organ. In vitro, tissue biomechanics was reported to affect follicle activation mainly through the Hippo pathway. Only recently, ovary responsiveness to mechanical signals was exploited for reproductive purposes. Unfortunately, poor characterization of ovarian cortex biomechanics and of the mechanical challenge hampers reproducible and effective treatments, and prevention of tissue damages. In this study the biomechanical response of ovarian cortical tissue from abattoir bovines was characterized for the first time. Ovarian cortical tissue fragments were subjected to uniaxial dynamic testing at frequencies up to 30 Hz, and at increasing average stresses. Tissue structure prior to and after testing was characterized by histology, with established fixation and staining protocols, to assess follicle quality and stage. Tissue properties largely varied with the donor. Bovine ovarian cortical tissue consistently exhibited a nonlinear viscoelastic behavior, with dominant elastic characteristics, in the low range of other reproductive tissues, and significant creep. Strain rate was independent of the applied stress. Histological analysis prior to and after mechanical tests showed that the short-term dynamic mechanical test used for the study did not cause significant tissue tear, nor follicle expulsion or cell damage.

scholarly journals Physical and Biological Characterization of Ferromagnetic Fiber Networks: Effect of Fibrin Deposition on Short-Term In Vitro Responses of Human Osteoblasts

2015 ◽  
Vol 21 (3-4) ◽  
pp. 463-474 ◽  
Author(s):  
Rose L. Spear ◽  
Brajith Srigengan ◽  
Suresh Neelakantan ◽  
Wolfram Bosbach ◽  
Roger A. Brooks ◽  
...  
Keyword(s):  
Biological Characterization ◽  
Fibrin Deposition ◽  
Short Term ◽  
Human Osteoblasts ◽  
Fiber Networks

scholarly journals A novel long intergenic non-coding RNA, Nostrill, regulates iNOS gene transcription and neurotoxicity in microglia

2021 ◽  
Vol 18 (1) ◽  
Author(s):  
Nicholas W. Mathy ◽  
Olivia Burleigh ◽  
Andrew Kochvar ◽  
Erin R. Whiteford ◽  
Matthew Behrens ◽  
...  
Keyword(s):  
Nitric Oxide ◽  
No Production ◽  
Cortical Tissue ◽  
Non Coding Rna ◽  
Transcriptional Regulatory ◽  
Lncrna Locus ◽  
Rna Immunoprecipitation ◽  
Long Non Coding Rna

Abstract Background Microglia are resident immunocompetent and phagocytic cells in the CNS. Pro-inflammatory microglia, stimulated by microbial signals such as bacterial lipopolysaccharide (LPS), viral RNAs, or inflammatory cytokines, are neurotoxic and associated with pathogenesis of several neurodegenerative diseases. Long non-coding RNAs (lncRNA) are emerging as important tissue-specific regulatory molecules directing cell differentiation and functional states and may help direct proinflammatory responses of microglia. Characterization of lncRNAs upregulated in proinflammatory microglia, such as NR_126553 or 2500002B13Rik, now termed Nostrill (iNOS Transcriptional Regulatory Intergenic LncRNA Locus) increases our understanding of molecular mechanisms in CNS innate immunity. Methods Microglial gene expression array analyses and qRT-PCR were used to identify a novel long intergenic non-coding RNA, Nostrill, upregulated in LPS-stimulated microglial cell lines, LPS-stimulated primary microglia, and LPS-injected mouse cortical tissue. Silencing and overexpression studies, RNA immunoprecipitation, chromatin immunoprecipitation, chromatin isolation by RNA purification assays, and qRT-PCR were used to study the function of this long non-coding RNA in microglia. In vitro assays were used to examine the effects of silencing the novel long non-coding RNA in LPS-stimulated microglia on neurotoxicity. Results We report here characterization of intergenic lncRNA, NR_126553, or 2500002B13Rik now termed Nostrill (iNOS Transcriptional Regulatory Intergenic LncRNA Locus). Nostrill is induced by LPS stimulation in BV2 cells, primary murine microglia, and in cortical tissue of LPS-injected mice. Induction of Nostrill is NF-κB dependent and silencing of Nostrill decreased inducible nitric oxide synthase (iNOS) expression and nitric oxide (NO) production in BV2 and primary microglial cells. Overexpression of Nostrill increased iNOS expression and NO production. RNA immunoprecipitation assays demonstrated that Nostrill is physically associated with NF-κB subunit p65 following LPS stimulation. Silencing of Nostrill significantly reduced NF-κB p65 and RNA polymerase II recruitment to the iNOS promoter and decreased H3K4me3 activating histone modifications at iNOS gene loci. In vitro studies demonstrated that silencing of Nostrill in microglia reduced LPS-stimulated microglial neurotoxicity. Conclusions Our data indicate a new regulatory role of the NF-κB-induced Nostrill and suggest that Nostrill acts as a co-activator of transcription of iNOS resulting in the production of nitric oxide by microglia through modulation of epigenetic chromatin remodeling. Nostrill may be a target for reducing the neurotoxicity associated with iNOS-mediated inflammatory processes in microglia during neurodegeneration.

scholarly journals The activin-follistatin system and in vitro early follicle development in goats

2006 ◽  
Vol 189 (1) ◽  
pp. 113-125 ◽  
Author(s):  
J R V Silva ◽  
T Tharasanit ◽  
M A M Taverne ◽  
G C van der Weijden ◽  
R R Santos ◽  
...  
Keyword(s):  
Ovarian Tissue ◽  
Primordial Follicle ◽  
Activin A ◽  
Terminal Deoxynucleotidyl Transferase ◽  
Follicle Development ◽  
Cortical Tissue ◽  
Primary Follicle ◽  
Growth And Survival ◽  
Primordial Follicles

The aim of the present study was to investigate the effects of activin-A and follistatin on in vitro primordial and primary follicle development in goats. To study primordial follicle development (experiment 1), pieces of ovarian cortex were cultured in vitro for 5 days in minimal essential medium (MEM) supplemented with activin-A (0, 10 or 100 ng/ml), follistatin (0, 10 or 100 ng/ml) or combinations of the two. After culture, the numbers of primordial follicles and more advanced follicle stages were calculated and compared with those in non-cultured tissue. Protein and mRNA expression of activin-A, follistatin, Kit ligand (KL), growth differentiation factor-9 (GDF-9) and bone morphogenetic protein-15 (BMP-15) in non-cultured and cultured follicles were studied by immunohistochemistry and PCR. To evaluate primary follicle growth (experiment 2), freshly isolated follicles were cultured for 6 days in MEM plus 100 ng/ml activin-A, 100 ng/ml follistatin or 100 ng/ml activin-A plus 200 ng/ml follistatin. Morphology, follicle and oocyte diameters in cultured tissue and isolated follicles before and after culture were assessed. Terminal deoxynucleotidyl transferase-mediated dUTP nick-end labelling (TUNEL) reactions were performed to study DNA fragmentation in follicles. In experiment 1, it was found that goat primordial follicles were activated to develop into more advanced stages, i.e. intermediate and primary follicles, during in vitro culture, but neither activin-A nor follistatin affected the number of primordial follicles that entered the growth phase. Activin-A treatment enhanced the number of morphologically normal follicles and stimulated their growth during cortical tissue culture. The effects were, however, not counteracted by follistatin. The follicles in cultured goat tissue maintained their expression of proteins and mRNA for activin-A, follistatin, KL, GDF-9 and BMP-15. Fewer than 30% of the atretic follicles in cultured cortical tissue had TUNEL-positive (oocyte or granulosa) cells. Activin-A did not affect the occurrence of TUNEL-positive cells in follicles within cortical tissue. In experiment 2, addition of activin-A to cultured isolated primary follicles significantly stimulated their growth, the effect being counteracted by follistatin. Absence of such a neutralizing effect of follistatin in the cultures with ovarian cortical tissue can be due to lower dose of follistatin used and incomplete blockage of activin in these experiments. In contrast to cortical enclosed atretic follicles, all atretic follicles that had arisen in cultures with isolated primary follicles had TUNEL-positive cells, which points to differences between isolated and ovarian tissue-enclosed follicles with regard to the followed pathways leading to their degeneration. In summary, this in vitro study has demonstrated that cultured goat primordial follicles are activated to grow and develop into intermediate and primary follicles. During in vitro culture, the follicles maintain their ability to express activin-A, follistatin, KL, GDF-9 and BMP-15. The in vitro growth and survival of activated follicles enclosed in cortical tissue and the in vitro growth of isolated primary follicles are stimulated by activin-A.

scholarly journals Steroids and miRNAs in assessment of ovarian tissue damage following cryopreservation

2019 ◽  
Vol 62 (4) ◽  
pp. 207-216 ◽  
Author(s):  
Nadia Islam ◽  
Ugwoke Sunday Paul ◽  
Rana Alhamdan ◽  
Juan Hernandez-Medrano ◽  
Bruce K Campbell ◽  
...  
Keyword(s):  
Tissue Damage ◽  
Culture Media ◽  
Cell Damage ◽  
Ovarian Tissue ◽  
Expression Patterns ◽  
Cortical Tissue ◽  
Steroid Production ◽  
Primordial Follicles ◽  
Non Invasive ◽  
Novel Approach

Ovarian cortical tissue cryopreservation is a relatively novel approach to preserving fertility in women diagnosed with cancer. However, the effects of freezing-thawing are not fully understood, mainly due to the lack of suitable methods to assess tissue’s survival after thawing. Disparities in steroid production have been associated with ovarian failure by disrupting folliculogenesis, ovulation and oocyte apoptosis. Moreover, specific miRNAs, identified in human ovarian follicles, are thought to play a fundamental role in folliculogenesis. In this study, we investigated the possible interplay between the ovarian steroidal production and miRNA expression patterns in spent culture media, as potential non-invasive markers for ovarian tissue damage after cryopreservation. Cryopreservation of ovarian cortical tissue decreased (P < 0.05) both steroid production (oestradiol and progesterone) and expression of miRNA-193b and 320A in spent culture media over 5 days; however, expression of miRNA-24 increased (P < 0.05). The number of primordial follicles was also reduced (P < 0.05) in fresh-cultured and cryopreserved-cultured cortical tissues when compared with fresh tissues. Downregulation of miRNA-193b and miRNA-320A together with upregulation of miRNA-24 could have a synergistic role in cell apoptosis, and consequently leading to reduced oestradiol and progesterone production. Thus, there appears to be an interplay between these miRNAs, ovarian steroid production and cell damage, which can be further explored as novel non-invasive markers of cell damage following cryopreservation.

Characterization of [11C]vorozole binding in ovarian tissue in rats throughout estrous cycle in association with conversion of androgens to estrogens in vivo and in vitro

Steroids ◽  
2003 ◽  
Vol 68 (14) ◽  
pp. 1139-1146 ◽  
Author(s):  
Dmitrijus Kirilovas ◽  
Tord Naessen ◽  
Mats Bergström ◽  
Elizabeth Bergström-Petterman ◽  
Kjell Carlström ◽  
...  
Keyword(s):  
Estrous Cycle ◽  
Ovarian Tissue

scholarly journals Development of an in vitro tissue culture system for hammer coral (Fimbriaphyllia ancora) ovaries

2021 ◽  
Vol 11 (1) ◽  
Author(s):  
Yi-Ling Chiu ◽  
Ching-Fong Chang ◽  
Shinya Shikina
Keyword(s):  
Structural Integrity ◽  
Culture Method ◽  
Culture Conditions ◽  
Short Term ◽  
Wide Range ◽  
Fetal Bovine ◽  
Short Term Culture ◽  
Mesenterial Filaments

AbstractIn vitro gonad culture systems have proven useful to investigate intrinsic mechanisms of sexual reproduction in animals. Here we describe development of an in vitro culture method for coral ovaries. Mesenterial tissues containing both ovaries and mesenterial filaments were microscopically isolated from the scleractinian coral, Fimbriaphyllia ancora, and culture conditions were optimized. M199 diluted 10× (10% M199, pH 8.1) and supplemented with 25 mM HEPES and the antibiotics, ampicillin, penicillin and streptomycin, supported oocyte survival and maintained the structural integrity of ovaries during short-term culture (~ 6 days). Addition of a commercial antibiotic–antimycotic solution (Anti–Anti) and fetal bovine serum adversely affected ovary maintenance and caused tissue disintegration. Characterization of cultured ovaries showed that there is no difference in cell proliferation of ovarian somatic cells between culture Days 1 and 6. Moreover, the presence of oogonia and expression of a major yolk protein, vitellogenin, were confirmed in ovaries cultured for 6 days. This system will be useful for studying effects of a wide range of substances on coral oogenesis.

47 THE AVIAN CHORIO-ALLANTOIC MEMBRANE: A SUITABLE SHORT-TERM CULTURE SYSTEM FOR BOVINE OVARIAN TISSUE

2015 ◽  
Vol 27 (1) ◽  
pp. 116
Author(s):  
K. L. Beck ◽  
J. Singh ◽  
M. Anzar
Keyword(s):  
In Vitro Culture ◽  
Blood Vessels ◽  
Culture System ◽  
Ovarian Tissue ◽  
Short Term ◽  
In Ovo ◽  
Ovarian Cortex ◽  
In Vitro Culture System ◽  
Short Term Culture

Successful cryopreservation of bovine ovarian tissue holds enormous potential for long-term maintenance of female gametes to preserve genetic diversity by tissue banking. Traditionally, in vitro culture followed by histopathological examination has been used to assess the post-thaw viability of cryopreserved tissues. Recently, in ovo transplantation of mammalian tissues on the chorio-allantoic membrane (CAM) of a growing chicken embryo has emerged as an alternative method for short-term culture. The purpose of this experiment was to compare CAM culture of bovine ovarian tissue over a 5-day period with the in vitro culture system. Fertilized White Leghorn eggs were incubated at 37°C and 62% relative humidity. A window (1 × 2 cm) was cut into the eggshell on Day 3 of incubation. Ovaries were retrieved from a local abattoir and brought to the laboratory within 6 h. Ovarian cortex pieces (1–2 mm3) were randomly assigned to control, CAM-culture, or in vitro-culture groups. Control-group tissues were fixed immediately in 4% paraformaldehyde. The CAM was traumatized on Day 10 of incubation to expose the underlying blood vessels, and tissue pieces were grafted at the site (one graft per egg). For in vitro culture, the ovarian cortex pieces were placed on tissue culture inserts within 6-well plates containing TCM199 with 1% insulin-transferrin-selenium, 100 mIU mL–1 of FSH, 100 IU mL–1 of penicillin, and 50 μg mL–1 of streptomycin and incubated at 38°C in 5% CO2. Ovarian tissues from the CAM and in vitro culture group were removed on Day 1, 3, and 5 of grafting/culture, fixed, embedded in paraffin, sectioned at 5 μm, stained with hematoxylin-eosin, and analysed under a light microscope. The numbers of normal and degenerated follicles (indicating follicle survival) and number of blood vessels containing bovine and avian red blood cells (indicating angiogenesis) were counted using standard stereological procedures. All ovarian cortex grafts from surviving chick embryos showed adhesion with the CAM and a marked neo-vascularization in the graft areas. Gross and histological examination revealed the circulation of avian blood cells in ovarian stromal vessels with a concomitant decrease in the number of bovine blood vessels over the incubation period. Total follicle densities (mean ± s.e.m.) on Day 1, 3, and 5 were 13.3 ± 5.9, 27.9 ± 6.7, and 36.9 ± 7.3 in the in vitro-cultured group and 36.7 ± 13.0, 73.6 ± 24.0, and 44.02 ± 12.67 per millimeter cubed in the CAM-cultured group, respectively. Overall, total follicle density was higher in the CAM-cultured group (P < 0.05). Likewise, the normal follicle densities on Day 1, 3, and 5 were 10.4 ± 4.9, 15.5 ± 3.6, and 20.7 ± 6.3 in the in vitro-cultured group and 30.5 ± 8.5, 45.7 ± 18.4, and 22.7 ± 7.3 per millimeter cubed in the CAM-cultured group (P > 0.05). In conclusion, in ovo CAM grafting system was as successful as the in vitro-culture system and may be considered an acceptable alternative to the traditional in vitro-culture system for bovine ovarian tissue.

A sunscreen nanoparticles polymer based on prolonged period of protection

2021 ◽  
pp. 088391152110617
Author(s):  
Ebtesam A Mohamad ◽  
Monira M Rageh ◽  
Mirhan Mostafa Darwish
Keyword(s):  
Electron Microscopy ◽  
Cell Damage ◽  
Aloe Vera ◽  
Mouse Skin ◽  
Blocking Agent ◽  
Transmission Electron ◽  
Skin Epidermis ◽  
Uv Rays

UV rays are one of the most dangerous factors that harm the skin. There is continuous improvement in getting an effective sunscreen that protects the skin from excessive exposure to UV rays. Typically, phenylbenzimidazole-5-sulfonic acid (PBSA) is used as a sun blocking agent, but its disadvantage is that it can photodegrade and cause cell damage. In our work, PBSA was encapsulated in niosomes nanoparticles then coated with chitosan-aloe vera (CS-nio-aloe/PBSA) to form a carrier polymer with novel and potent properties. This polymer controls PBSA release and epidermal penetration. Characterization of CS-nio-aloe/PBSA polymer nanoparticles through transmission electron microscopy (TEM), scanning electron microscopy (SEM), Fourier transform infrared (FTIR) spectroscopy, and dynamic light scattering (DLS). The carrier polymer release rate was studied in vitro and epidermal permeability to coated PBSA was assessed using mouse skin. The nanoparticle polymer containing sunscreen was effectively prepared with an encapsulation efficiency of 80%. The formulation (CS-nio-aloe/PBSA) was completely deposited on the surface of the skin. This supports its use to protect the skin, and its nanostructures stimulate the release of PBSA for a longer period. Encapsulation of PBSA in CS-nio-aloe nanoparticles could allow for further cellular preservation, UV protection, control of free PBSA, and limited penetration through the mouse skin epidermis.

Morphologic characterization of isolated bovine early preantral follicles during short-term individual in vitro culture

2015 ◽  
Vol 84 (2) ◽  
pp. 301-311 ◽  
Author(s):  
E.P.A. Jorssen ◽  
A. Langbeen ◽  
W.F.A. Marei ◽  
E. Fransen ◽  
H.F.M. De porte ◽  
...  
Keyword(s):  
In Vitro Culture ◽  
Short Term ◽  
Preantral Follicles
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