Enhanced Synergistic-Antioxidant Activity of Melatonin and Tretinoin by Co-encapsulation into Amphiphilic Chitosan Nanocarriers: During Mice In Vitro Matured Oocyte/Morula-Compact Stage Embryo Culture Model,Reproductive Sciences

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Enhanced Synergistic-Antioxidant Activity of Melatonin and Tretinoin by Co-encapsulation into Amphiphilic Chitosan Nanocarriers: During Mice In Vitro Matured Oocyte/Morula-Compact Stage Embryo Culture Model
Reproductive Sciences ( IF 2.6 ) Pub Date : 2021-07-06 , DOI: 10.1007/s43032-021-00670-8
Faranak Aghaz _{1,

2} , Asad Vaisi-Raygani _{1,

3} , Mozafar Khazaei ₁ , Elham Arkan ₂ , Soheila Kashanian _{2,

4}

Affiliation

The use of exogenous antioxidants or the combination of them during in vitro oocyte/embryo culture media is reasonable. Co-delivery by nanocarrier has been designed to overcome the limitations of combining them traditionally. In this work, amphiphilic chitosan nanocarrier (ACN) was applied to co-encapsulate melatonin (Mel) and tretinoin (TTN) by the self-assembled method and evaluate their synergistic antioxidant efficacy in mice oocytes/embryos. The formation of single/dual-ACN was confirmed by Fourier-transformed infrared spectroscopy (FT-IR). The average particle diameter, size distribution, polydispersity index (PDI), and zeta potential of them were measured by dynamic light scattering (DLS), and the morphology was evaluated by TEM and SEM technologies. Also, the encapsulation efficiency (EE%) and drug loading content (DL%) of the nanocapsules were determined by UV-vis spectrophotometry. Studies of the in vitro release showed a continued drug release without any bursting effect of Mel+TTN-ACNs compared with single Mel/TTN-ACNs. Then, in both experiments, nuclear staining (Aceto-orcein and Hoechst 33342), fluorescent staining of H2DCFDA, chemiluminescence test, and qRT-PCR technique were performed as in vitro toxicity studies. The results of all these evaluations demonstrated that the dual delivery of Mel and TTN could accumulate a safety (without high-dose toxicity) synergistic anti-oxidative effect in oocyte/embryo by passive controlled, and inhibit intra/extracellular ROS levels by an enhanced intracellular penetration.

Graphical abstract

中文翻译：

通过共同封装到两亲壳聚糖纳米载体中增强褪黑激素和维甲酸的协同抗氧化活性：在小鼠体外成熟卵母细胞/桑葚紧凑阶段胚胎培养模型中

在体外卵母细胞/胚胎培养基中使用外源性抗氧化剂或它们的组合是合理的。纳米载体的共同递送旨在克服传统上将它们结合的局限性。在这项工作中，将两亲性壳聚糖纳米载体（ACN）通过自组装法共包裹褪黑激素（Mel）和维甲酸（TTN），并评估它们在小鼠卵母细胞/胚胎中的协同抗氧化功效。傅里叶变换红外光谱 (FT-IR) 证实了单/双 ACN 的形成。采用动态光散射法(DLS)测定了它们的平均粒径、粒度分布、多分散指数(PDI)和zeta电位，采用TEM和SEM技术评价了其形貌。还，采用紫外-可见分光光度法测定纳米胶囊的包封率（EE%）和载药量（DL%）。体外释放研究表明，与单一的 Mel/TTN-ACNs 相比，Mel+TTN-ACNs 的持续药物释放没有任何爆发效应。然后，在这两个实验中，进行了核染色（乙酰紫红和 Hoechst 33342）、H2DCFDA 的荧光染色、化学发光测试和 qRT-PCR 技术作为体外毒性研究。所有这些评估的结果表明，Mel和TTN的双重递送可以通过被动控制在卵母细胞/胚胎中积累安全（无大剂量毒性）协同抗氧化作用，并通过增强细胞内抑制细胞内/细胞外ROS水平。渗透。体外释放研究表明，与单一的 Mel/TTN-ACNs 相比，Mel+TTN-ACNs 的持续药物释放没有任何爆发效应。然后，在这两个实验中，进行了核染色（乙酰紫红和 Hoechst 33342）、H2DCFDA 的荧光染色、化学发光测试和 qRT-PCR 技术作为体外毒性研究。所有这些评估的结果表明，Mel和TTN的双重递送可以通过被动控制在卵母细胞/胚胎中积累安全（无大剂量毒性）协同抗氧化作用，并通过增强细胞内抑制细胞内/细胞外ROS水平。渗透。体外释放研究表明，与单一的 Mel/TTN-ACNs 相比，Mel+TTN-ACNs 的持续药物释放没有任何爆发效应。然后，在这两个实验中，进行了核染色（乙酰紫红和 Hoechst 33342）、H2DCFDA 的荧光染色、化学发光测试和 qRT-PCR 技术作为体外毒性研究。所有这些评估的结果表明，Mel和TTN的双重递送可以通过被动控制在卵母细胞/胚胎中积累安全（无大剂量毒性）协同抗氧化作用，并通过增强细胞内抑制细胞内/细胞外ROS水平。渗透。H2DCFDA的荧光染色、化学发光测试和qRT-PCR技术作为体外毒性研究进行。所有这些评估的结果表明，Mel和TTN的双重递送可以通过被动控制在卵母细胞/胚胎中积累安全（无大剂量毒性）协同抗氧化作用，并通过增强细胞内抑制细胞内/细胞外ROS水平。渗透。H2DCFDA的荧光染色、化学发光测试和qRT-PCR技术作为体外毒性研究进行。所有这些评估的结果表明，Mel和TTN的双重递送可以通过被动控制在卵母细胞/胚胎中积累安全（无大剂量毒性）协同抗氧化作用，并通过增强细胞内抑制细胞内/细胞外ROS水平。渗透。

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更新日期：2021-07-07

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