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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 Biology: Original Article
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Abstract

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.

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Abbreviations

COCs:

Cumulus-oocyte complex

ACNs:

Amphiphilic chitosan nanocarriers

B-ACN:

Blank nanocapsules

Mel:

Melatonin

TTN:

Tretinoin

Mel+TTN-ACN:

ACN entrapping Mel and TTN

EE%:

Encapsulation efficiency %

DL%:

Drug loading capacity

FSH:

Follicle-stimulating hormone

LH:

Luteinizing hormone

CS:

Chitosan

TPP:

Sodium tripolyphosphate solution

PMSG:

Pregnant mare serum gonadotrophin

BSA:

Bovine Serum Albumin

H2DCFDA:

2′,7′-Dichlorodihydrofluorescein diacetate

GV:

Germinal vesicle

GVBD:

Germinal vesicle breakdown

M I:

Metaphase I

M II:

Metaphase II

t RNA:

Complementary RNA

SD:

Standard deviation

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Authors and Affiliations

  1. Fertility and Infertility Melearch Center, Health Technology Institute, Kermanshah University of Medical Sciences, Kermanshah, Iran

    Faranak Aghaz, Asad Vaisi-Raygani & Mozafar Khazaei

  2. Nano Drug Delivery Melearch Center, Faculty of Pharmacy, Health Technology Institute, Kermanshah University of Medical Sciences, Kermanshah, Iran

    Faranak Aghaz, Elham Arkan & Soheila Kashanian

  3. Department of Clinical Biochemistry, Medical School, Kermanshah University of Medical Sciences, Kermanshah, Iran

    Asad Vaisi-Raygani

  4. Faculty of Chemistry, Sensor and Biosensor Research Center (SBRC) and Nanoscience and Nanotechnology Research Center (NNRC), Razi University, Kermanshah, Iran

    Soheila Kashanian

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  1. Faranak Aghaz
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Contributions

Processed data, evaluated data, and written the manuscript: F. A. and A. V.; performed experimental research and contributed to the study of the results: F. A. and M. K. H.; engaged in experimental work and analyzed data: A. V.; engaged in experimental work and updated manuscript: E. A. and S. K. H.; selected suitable animals, rated embryos, and gathered research data: F. A.; planned tests, analyzed data, and revised the manuscript: F. A., A. V., and M. K. H.

Corresponding authors

Correspondence to Asad Vaisi-Raygani or Mozafar Khazaei.

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This research was carried out in compliance with the rules laid down by the Faculty of Medicine Ethics Committee of Kermanshah University of Medical Science (IR.KUMS.REC.1396.665) and Iran National Science Foundation (INSF, 97008908).

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Aghaz, F., Vaisi-Raygani, A., Khazaei, M. et al. 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. Reprod. Sci. 28, 3361–3379 (2021). https://doi.org/10.1007/s43032-021-00670-8

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