Modulating the lipid profile of blastocyst cell membrane with DPPC multilamellar vesicles

Rossi, Hugo De; Costa, Ciniro; Rodrigues-Rossi, Luana Teixeira; Nunes, Giovana Barros; Chéles, Dóris Spinosa; Pereira, Isabella Maran; Rocha, Daniele F. O.; Feitosa, Eloi; Simionato, Ana Valéria Colnaghi; Mingoti, Gisele Zoccal; Aoki, Pedro H.B.; Nogueira, Marcelo Fábio Gouveia

doi:10.1080/21691401.2022.2088545

Cited by 2 publications

(2 citation statements)

References 59 publications

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Section: Methodsmentioning

confidence: 99%

“…This protocol was developed in the Laboratory of Reproductive Physiology, School of Veterinary Medicine, Universidade Estadual Paulista (UNESP), Araçatuba, SP, Brazil, based on the experience and studies published by our group in cellular oxidative stress. [34][35][36][37] In addition, we used validated protocols for the in vitro evaluation of oxidative stress in oral cells. 28,30,33,38,39 For cell collection, eight volunteers were subdivided into two groups so that the collections were performed on two different days, with four patients per collection day (Table 1).…”

Section: Second Series Of the Experiment: Evaluation Of Oxidative Str...mentioning

confidence: 99%

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Development and validation of a minimally invasive protocol for assessing oxidative stress markers in exfoliated oral cells

de Abreu,

da Silva,

Bortoleto

et al. 2023

Cytopathology

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ObjectivesThis study aimed to develop and validate a minimally invasive protocol for characterizing oxidative stress markers in exfoliated oral cells.Materials and MethodsExfoliated oral cells were collected from healthy volunteers. The protocol included the utilization of specific fluorescent probes to measure intracellular reactive oxygen species (ROS), mitochondrial membrane potential (ΔΨm) and reduced glutathione (GSH). Cells from each volunteer were divided into the positive and negative control groups, which were, respectively, exposed or not to hydrogen peroxide (H2O2) aiming to induce the oxidative stress. Measurements of cell fluorescence were performed using a microscope equipped with epifluorescence.ResultsThe results showed that cells exposed to H2O2 exhibited significantly higher intracellular expression of ROS compared to unexposed cells (positive control: 3851.25 ± 1227.0 vs, negative control: 1106.07 ± 249.6; p = 0.0338). On the contrary, cells exposed to H2O2 displayed decreased expression of ΔΨm (p = 0.0226) and GSH (p = 0.0289) when compared to the negative control group (ΔΨm positive control: 14634.39 ± 1529.0 vs, negative control: 18897.60 ± 2338.0; and GSH positive control: 9011.08 ± 1900.0 vs, negative control: 15901.79 ± 2745.0).ConclusionsThe developed protocol proved to be effective in detecting and quantifying oxidative stress biomarkers, such as ROS, ΔΨm and GSH, in exfoliated oral cells. This minimally invasive approach offers a promising method to assess oxidative stress expression and may be clinically relevant in the evaluation of oral diseases associated with oxidative stress.

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Section: Methodsmentioning

confidence: 99%

Section: Second Series Of the Experiment: Evaluation Of Oxidative Str...mentioning

confidence: 99%

Development and validation of a minimally invasive protocol for assessing oxidative stress markers in exfoliated oral cells

de Abreu,

da Silva,

Bortoleto

et al. 2023

Cytopathology

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Developmental and Molecular Effects of C-Type Natriuretic Peptide Supplementation in In Vitro Culture of Bovine Embryos

Costa,

Silva,

Silva

et al. 2024

IJMS

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The use of C-type natriuretic peptide (CNP) in the interaction with the oocyte and in the temporary postponement of spontaneous meiosis resumption has already been well described. However, its action in pre-implantation developmental-stage embryos is yet to be understood. Thus, our study aimed to detect the presence of the canonical CNP receptor (natriuretic peptide receptor, NPR2) in germinal vesicle (GV)-, metaphase II (MII)-, presumptive zygote (PZ)-, morula (MO)-, and blastocyst (BL)-stage embryos and, later, to observe possible modulations on the embryos when co-cultured with CNP. In Experiment I, we detected and quantified NPR2 on the abovementioned embryo stages. Further, in Experiment II, we intended to test different concentrations (100, 200, or 400 nM of CNP) at different times of inclusion in the in vitro culture (IVC; inclusion from the beginning, i.e., day 1, or from day 5). In Experiment III, 400 nM of CNP was used on day 1 (D1) in the IVC, which was not demonstrated to be embryotoxic, and it showed potentially promising results in the blastocyst production rate when compared to the control. Thus, we analyzed the embryonic development rates of bovine embryos (D7) and hatching kinetics (D7, D8, and D9). Subsequently, morula and blastocyst were collected and evaluated for transcript abundance of their competence and quality (apoptosis, oxidative stress, proliferation, and differentiation) and lipid metabolism. Differences with probabilities less than p < 0.05, and/or fold change (FC) > 1.5, were considered significant. We demonstrate the presence of NPR2 until the blastocyst development stage, when there was a significant decrease in membrane receptors. There was no statistical difference in the production rate after co-culture with 400 nM CNP. However, when we evaluated the abundance of morula transcripts, there was an upregulated transcription in ADCY6 (p = 0.057) and downregulated transcripts in BMP15 (p = 0.013), ACAT1 (p = 0.040), and CASP3 (p = 0.082). In addition, there was a total of 12 transcriptions in morula that presented variation FC > 1.5. In blastocysts, the treatment with CNP induced upregulation in BID, CASP3, SOX2, and HSPA5 transcripts and downregulation in BDNF, NLRP5, ELOVL1, ELOVL4, IGFBP4, and FDX1 transcripts (FC > 1.5). Thus, our study identified and quantified the presence of NPR2 in bovine pre-implantation embryos. Furthermore, 400 nM of CNP in IVC, a concentration not previously described in the literature, modulated some transcripts related to embryonic metabolism, and this was not embryotoxic morphologically.

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Modulating the lipid profile of blastocyst cell membrane with DPPC multilamellar vesicles

Cited by 2 publications

References 59 publications

Development and validation of a minimally invasive protocol for assessing oxidative stress markers in exfoliated oral cells

Development and validation of a minimally invasive protocol for assessing oxidative stress markers in exfoliated oral cells

Developmental and Molecular Effects of C-Type Natriuretic Peptide Supplementation in In Vitro Culture of Bovine Embryos

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