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Functional evaluation of constructed pseudo-endogenous microRNA-targeted myocardial ultrasound nanobubble

BACKGROUND: Stem cell transplantation is one of the treatment methods for acute myocardial infarction (AMI). MicroRNA-1 contributes to the study of the essential mechanisms of stem cell transplantation for treating AMI by targeted regulating the myocardial microenvironment after stem cell transplant...

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Autores principales: Ainiwan, Ailifeire, Wei, Yuanyuan, Dou, Jing, Tang, Lingpeng, Mu, Yuming, Guan, Lina
Formato: Online Artículo Texto
Lenguaje:English
Publicado: Frontiers Media S.A. 2023
Materias:
Acceso en línea:https://www.ncbi.nlm.nih.gov/pmc/articles/PMC10556731/
https://www.ncbi.nlm.nih.gov/pubmed/37809333
http://dx.doi.org/10.3389/fmed.2023.1136304
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author Ainiwan, Ailifeire
Wei, Yuanyuan
Dou, Jing
Tang, Lingpeng
Mu, Yuming
Guan, Lina
author_facet Ainiwan, Ailifeire
Wei, Yuanyuan
Dou, Jing
Tang, Lingpeng
Mu, Yuming
Guan, Lina
author_sort Ainiwan, Ailifeire
collection PubMed
description BACKGROUND: Stem cell transplantation is one of the treatment methods for acute myocardial infarction (AMI). MicroRNA-1 contributes to the study of the essential mechanisms of stem cell transplantation for treating AMI by targeted regulating the myocardial microenvironment after stem cell transplantation at the post-transcriptional level. Thus, microRNA-1 participates in regulating the myocardial microenvironment after stem cell transplantation, a promising strategy for the Stem cell transplantation treatment of AMI. However, the naked microRNA-1 synthesized is extremely unstable and non-targeting, which can be rapidly degraded by circulating RNase. Herein, to safely and effectively targeted transport the naked microRNA-1 synthesized into myocardial tissue, we will construct pseudo-endogenous microRNA-targeted myocardial ultrasound nanobubble pAd-AAV-9/miRNA-1 NB and evaluate its characteristics, targeting, and function. METHODS: The pAd-AAV-9/miRNA-1 gene complex was linked to nanobubble NBs by the “avidin-biotin bridging” method to prepare cardiomyocyte-targeted nanobubble pAd-AAV-9/miRNA-1 NB. The shape, particle size, dispersion, and stability of nanobubbles and the connection of pAd-AAV-9/miRNA-1 gene complex to nanobubble NB were observed. The virus loading efficiency was determined, and the myocardium-targeting imaging ability was evaluated using contrast-enhanced ultrasound imaging in vivo. The miRNA-1 expression level in myocardial tissue and other vital organs ex vivo of SD rats was considered by Q-PCR. Also, the cytotoxic effects were assessed. RESULTS: The particle size of NBs was 504.02 ± 36.94 nm, and that of pAd-AAV-9/miRNA-1 NB was 568.00 ± 37.39 nm. The particle size and concentration of pAd-AAV-9/miRNA-1 NBs did not change significantly within 1 h at room temperature (p > 0.05). pAd-AAV-9/miRNA-1 NB had the highest viral load rate of 86.3 ± 2.2% (p < 0.05), and the optimum viral load was 5 μL (p < 0.05). pAd-AAV-9/miRNA-1 NB had good contrast-enhanced ultrasound imaging in vivo. Quantitative analysis of miRNA-1 expression levels in vital organs ex vivo of SD rats by Q-PCR showed that pAd-AAV-9/miRNA-1 NB targeted the myocardial tissue. Q-PCR indicated that the expression level of miRNA-1 in the myocardium of the pAd-AAV-9/miRNA-1 NB + UTMD group was significantly higher than that of the pAd-AAV-9/miRNA-1 NB group (p < 0.05). pAd-AAV-9/miRNA-1 NB had no cytotoxic effect on cardiomyocytes (p > 0.05). CONCLUSION: The pAd-AAV-9/miRNA-1 NB constructed in this study could carry naked miRNA-1 synthesized in vitro for targeted transport into myocardial tissue successfully and had sound contrast-enhanced imaging effects in vivo.
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spelling pubmed-105567312023-10-07 Functional evaluation of constructed pseudo-endogenous microRNA-targeted myocardial ultrasound nanobubble Ainiwan, Ailifeire Wei, Yuanyuan Dou, Jing Tang, Lingpeng Mu, Yuming Guan, Lina Front Med (Lausanne) Medicine BACKGROUND: Stem cell transplantation is one of the treatment methods for acute myocardial infarction (AMI). MicroRNA-1 contributes to the study of the essential mechanisms of stem cell transplantation for treating AMI by targeted regulating the myocardial microenvironment after stem cell transplantation at the post-transcriptional level. Thus, microRNA-1 participates in regulating the myocardial microenvironment after stem cell transplantation, a promising strategy for the Stem cell transplantation treatment of AMI. However, the naked microRNA-1 synthesized is extremely unstable and non-targeting, which can be rapidly degraded by circulating RNase. Herein, to safely and effectively targeted transport the naked microRNA-1 synthesized into myocardial tissue, we will construct pseudo-endogenous microRNA-targeted myocardial ultrasound nanobubble pAd-AAV-9/miRNA-1 NB and evaluate its characteristics, targeting, and function. METHODS: The pAd-AAV-9/miRNA-1 gene complex was linked to nanobubble NBs by the “avidin-biotin bridging” method to prepare cardiomyocyte-targeted nanobubble pAd-AAV-9/miRNA-1 NB. The shape, particle size, dispersion, and stability of nanobubbles and the connection of pAd-AAV-9/miRNA-1 gene complex to nanobubble NB were observed. The virus loading efficiency was determined, and the myocardium-targeting imaging ability was evaluated using contrast-enhanced ultrasound imaging in vivo. The miRNA-1 expression level in myocardial tissue and other vital organs ex vivo of SD rats was considered by Q-PCR. Also, the cytotoxic effects were assessed. RESULTS: The particle size of NBs was 504.02 ± 36.94 nm, and that of pAd-AAV-9/miRNA-1 NB was 568.00 ± 37.39 nm. The particle size and concentration of pAd-AAV-9/miRNA-1 NBs did not change significantly within 1 h at room temperature (p > 0.05). pAd-AAV-9/miRNA-1 NB had the highest viral load rate of 86.3 ± 2.2% (p < 0.05), and the optimum viral load was 5 μL (p < 0.05). pAd-AAV-9/miRNA-1 NB had good contrast-enhanced ultrasound imaging in vivo. Quantitative analysis of miRNA-1 expression levels in vital organs ex vivo of SD rats by Q-PCR showed that pAd-AAV-9/miRNA-1 NB targeted the myocardial tissue. Q-PCR indicated that the expression level of miRNA-1 in the myocardium of the pAd-AAV-9/miRNA-1 NB + UTMD group was significantly higher than that of the pAd-AAV-9/miRNA-1 NB group (p < 0.05). pAd-AAV-9/miRNA-1 NB had no cytotoxic effect on cardiomyocytes (p > 0.05). CONCLUSION: The pAd-AAV-9/miRNA-1 NB constructed in this study could carry naked miRNA-1 synthesized in vitro for targeted transport into myocardial tissue successfully and had sound contrast-enhanced imaging effects in vivo. Frontiers Media S.A. 2023-09-22 /pmc/articles/PMC10556731/ /pubmed/37809333 http://dx.doi.org/10.3389/fmed.2023.1136304 Text en Copyright © 2023 Ainiwan, Wei, Dou, Tang, Mu and Guan. https://creativecommons.org/licenses/by/4.0/This is an open-access article distributed under the terms of the Creative Commons Attribution License (CC BY). The use, distribution or reproduction in other forums is permitted, provided the original author(s) and the copyright owner(s) are credited and that the original publication in this journal is cited, in accordance with accepted academic practice. No use, distribution or reproduction is permitted which does not comply with these terms.
spellingShingle Medicine
Ainiwan, Ailifeire
Wei, Yuanyuan
Dou, Jing
Tang, Lingpeng
Mu, Yuming
Guan, Lina
Functional evaluation of constructed pseudo-endogenous microRNA-targeted myocardial ultrasound nanobubble
title Functional evaluation of constructed pseudo-endogenous microRNA-targeted myocardial ultrasound nanobubble
title_full Functional evaluation of constructed pseudo-endogenous microRNA-targeted myocardial ultrasound nanobubble
title_fullStr Functional evaluation of constructed pseudo-endogenous microRNA-targeted myocardial ultrasound nanobubble
title_full_unstemmed Functional evaluation of constructed pseudo-endogenous microRNA-targeted myocardial ultrasound nanobubble
title_short Functional evaluation of constructed pseudo-endogenous microRNA-targeted myocardial ultrasound nanobubble
title_sort functional evaluation of constructed pseudo-endogenous microrna-targeted myocardial ultrasound nanobubble
topic Medicine
url https://www.ncbi.nlm.nih.gov/pmc/articles/PMC10556731/
https://www.ncbi.nlm.nih.gov/pubmed/37809333
http://dx.doi.org/10.3389/fmed.2023.1136304
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