Glycyrrhizic acid delivery system Chitosan-coated liposome as an adhesive anti-inflammation.

Nowadays, medicinal plants are used to overcome the side effects of prescription drugs in modern medicine. Glycyrrhizic acid (GA) derived from the root of the licorice plant is one of the plant compounds whose effectiveness has been confirmed in the treatment of inflammatory bowel disorders (IBD). Liposome thin film hydration method was used to synthesize chitosan-coated liposomes containing GA. In the present study, chitosan-coated liposome was characterized by dynamic light scattering (DLS), zeta potential, scanning electron microscope (SEM) and Fourier transforms infrared spectroscopy (FTIR). The FTIR spectrum confirmed the coating of liposomes by chitosan polymer.  Liposome coating leads to an increase in the size and values of zeta potential. The 3-(4, 5-dimethylthiazol-2-yl)-2, 5-diphenyltetrazolium bromide (MTT) assay of chitosan-coated liposomes containing GA confirmed that it has no cytotoxicity toward fibroblasts cell line, therefore confirming their cytocompatibility. Overall, drug loading, release and cytotoxicity were evaluated and it was found that chitosan decreased the release rate of GA. It seems; chitosan-coated liposomes may be a suitable system for delivering liposomal GA in the treatment of IBD.

The anti-cancer effect of resveratrol nano-encapsulated supplements against breast cancer via the regulation of oxidative stress.

AIM: Several shreds of evidence have supported the growth-inhibitory effect of resveratrol in breast cancer. Owing to the low efficiency, we aimed to fabricate an ACN nanoparticle containing resveratrol to target the proliferation of breast cancer cells. METHODS: Resveratrol encapsulation was characterized using spectrophotometry, FTIR, and SEM. The cytotoxicity and antioxidant activities of compounds were evaluated by MTT, NO, FRAP, and qRT-PCR assays on MCF7 and SKBr3 cells. RESULTS: Our result found that the encapsulation efficiency was 87%, the particle size was 200 ± 15 nm, and the zeta potential was 31 ± 0.4 mV. The prepared RES + ACN had controlled in vitro release. RES + ACN nanoparticle showed a significantly increased cytotoxicity in both cell lines. The reduced level of NO and enhance antioxidative content in both cells, especially the MCF7, were in line with increased expression of Nrf2 and SOD as well as more apoptotic effect. CONCLUSION: Decreased growth and increased expression of Nrf2 in MCF7 compared to SKBr3 cells proved that the upregulation of Nrf2 by nanoresveratrol is likely contributed to its relationship with ER/PR signaling factors, albeit its precise mechanism is needed to be more elucidated.

Beneficial effect of L-Proline supplementation on the quality of human spermatozoa.

L-Proline is a natural anti-oxidative and osmoprotectant agent, playing a versatile role in cell metabolism and physiology. The present study aimed to explore the antioxidant effects of L-Proline on human sperm function during incubation. Thirty healthy, normozoospermic men (27-40 years) were enrolled. Sperm samples were incubated in an unsupplemented sperm medium (control group), or supplemented with L-Proline (1, 2 and 4 mmol/L) to evaluate its effect during 0, 1, 4 and 24 h of incubation. Sperm were assessed in terms of motility, viability, morphology, chromatin and DNA integrity. Moreover, the levels of reactive oxygen species (ROS), malondialdehyde (MDA), and total antioxidant capacity (TAC) were determined in the sperm medium. The results indicated that 2 mmol/L of L-Proline significantly improved the maintenance of sperm motility, viability, normal morphology, chromatin and DNA integrity, and TAC levels compared to the control group during 24 h of incubation (p < 0.05). However, 1 and 4 mmol/L of L-Proline could not significantly preserve sperm parameters, chromatin quality, and antioxidant status during different incubation times compared to the control group (p > 0.05). Collectively, the inclusion of L-Proline (2 mmol/L) in the human sperm medium maintains sperm parameters and chromatin quality probably by modulating the oxidative status.

L-proline as a novel additive to cryopreservation media improved post-thaw quality of human spermatozoon via reducing oxidative stress.

Sperm cryopreservation as a routine technique in assisted reproductive technique (ART) laboratories has detrimental effects on spermatozoa. Various methods have been introduced to improve it. The aim of this research was to evaluate the effects of L-proline supplementation in cryopreservation medium on normozoospermic semen samples. A total of 30 semen samples were collected from normozoospermic men. Cryopreservation media were supplemented with different concentrations of L-proline (0, 1, 2 and 4 mmol/L). The semen samples were cryopreserved. After thawing, sperm parameters and chromatin integrity (aniline blue (AB), toluidine blue (TB), sperm chromatin dispersion test (SCD) and chromomycin A3 (CMA3)), reactive oxygen species (ROS), and total antioxidant capacity (TAC) and malondialdehyde (MDA) levels were evaluated. A total of 4 mmol/L L-proline significantly improved progressive motility and viability (p < 0.05). MDA and ROS levels significantly diminished in samples were cryopreserved by 4 mmol/L L-proline supplemented cryopreservation media (p < 0.001). Also, it significantly increased TAC level. Also, chromatin damages (AB, TB and CMA3) significantly improved in samples were cryopreserved by 4 mmol/L L-proline supplemented cryopreservation media (p < 0.05). The results support that the usage of L-proline supplemented cryopreservation media to improve sperm quality after cryopreservation.

The effect of air dust pollution on semen quality and sperm parameters among infertile men in west of Iran.

Background: Pollutants during haze and Asian dust storms are transported out of the Asian continent, affecting the regional climate and the hydrological and biogeochemical cycles. Nonetheless, no specific studies evaluated the dust particles influence on semen quality in a specific geographical area.Objective: In this article, we investigated the effect of dust particles on semen quality and sperm parameters among infertile men.Methods: A descriptive-analytic study was conducted among 850 infertile men between 2011 and 2015 years. Semen quality was assessed according to the WHO 2010 guidelines, including sperm concentration, progressive motility, and morphology. Four-year average dust particle concentrations were estimated at each participant's address using the Air Pollution Monitoring Station affiliated with the Department of Environment of Kermanshah city were gathered.Results: Dust particle levels were highest in the summer months, in Kermanshah province. Our results show that, dust pollution was found to be significantly negatively correlated with sperm morphology and sperm concentration before and after lab-processing, but sperm progressive motility is low sensitive to dust particles.Conclusions: Our findings showed that exposures to dust particle may influence sperm quantity in infertile men, consistent with the knowledge that sperm morphology and concentration are the most sensitive parameters of dust pollution.

Cryoprotective effect of sericin supplementation in freezing and thawing media on the outcome of cryopreservation in human sperm.

The destructive effects of sperm cryopreservation result in decreased sperm parameters and their fertilizing ability. Antioxidants supplementation can potentially improve cryopreservation outcomes. In this study, we tried to investigate the effects of sericin supplementation in freezing and thawing media on frozen-thawed human sperm motility, morphology, viability, and DNA fragmentation. In experiment 1, semen samples were collected from 30 healthy fertile men and were cryopreserved in the presence of freezing medium supplemented with different concentrations of sericin (0, 0.5, 1, 2.5, and 5%). The results showed that the addition of 2.5 and 5% sericin in freezing medium significantly increased sperm viability and total motility (A + B) and decreased DNA fragmentation (P < 0.05). In experiment 2, semen samples were collected from 21 fertile men and were cryopreserved in freezing medium without any supplementation for 48 h. Then, the samples were thawed in medium supplemented with different concentrations of sericin (0, 0.5, 1, 2.5, and 5%). The addition of 5% sericin to thawing medium increased the total motility, viability, and decreased DNA fragmentation compared with those in thaws without sericin. In nutshell, the results clearly indicate the feasibility of sericin as an cryoprotective supplement for freezing media in human spermatozoa.

Activities and polymorphisms of MMP-2 and MMP-9, smoking, diabetes and risk of prostate cancer.

Matrix metalloproteinases (MMPs) are a group of zinc dependent enzymes that are involved in tumor cell invasion and metastasis. The role of MMP-2 and -9 genetic polymorphism in different malignancies has been the subject of numerous studies. The present research has attempted to discover any positive correlation between MMP-2 and MMP-9 SNPs and prostate cancer (PCa) in patients with a history of either diabetes or smoking habits. 112 PCa-patients and 150 unrelated healthy-controls that matched for age and sex were selected for present case-control study. MMP-2 -1575G/A and MMP-9 -1562 C/T polymorphisms detected by PCR-RFLP, serum tissue inhibitors of metalloproteinases (TIMP-1 and TIMP-2), testosterone, prostate-specific antigen (PSA), free-prostate-specific-antigen (fPSA), and fPSA/PSA levels were detected by ELISA and enzyme assay, respectively. MMP-2 and MMP-9 activities were measured by gelatin-zymography. Covariates were considered as age, status of cigarette smoking, and a possible history of diabetes mellitus (DM). The frequency of -1575 MMP-2 A/A + A/G and -1562 MMP-9 C/T + T/T genotypes were higher in PCa-patients with DM (74.3%,p = 0.003) and with smoking habits (72.5%,p = 0.005). These genotypes were associated with the increased risk of prostate cancer in smokers (3.52-folds) and in individuals with history of DM (4.34-folds). A significant positive association was found between level of TIMPs (TIMP -1 and TIMP-2) and BMI in PCa-patients and also between testosterone levels and MMP-9 activity in healthy control subjects. For the first time, this study demonstrated that activities of MMP-2 -1575G/A and MMP-9 -1562C/T variants in association with smoking and diabetes are considered significant risk factors for PCa.

Enhancing Cisplatin Efficacy with Low Toxicity in Solid Breast Cancer Cells Using pH-Charge-Reversal Sericin-Based Nanocarriers: Development, Characterization, and In Vitro Biological Assessment.

Platinum-based chemotherapeutic agents are widely employed in cancer treatment because of their effectiveness in targeting DNA. However, this indiscriminate action often affects both cancerous and normal cells, leading to severe side effects and highlighting the need for innovative approaches in achieving precise drug delivery. Nanotechnology presents a promising avenue for addressing these challenges. Protein-based nanocarriers exhibit promising capabilities in the realm of cancer drug delivery with silk sericin nanoparticles standing out as a leading contender. This investigation focuses on creating a sericin-based nanocarrier (SNC) featuring surface charge reversal designed to effectively transport cisplatin (Cispt-SNC) into MCF-7 breast cancer cells. Utilizing AutoDock4.2, our molecular docking analyses identified key amino acids and revealed distinctive conformational clusters, providing insights into the drug-protein interaction landscape and highlighting the potential of sericin as a carrier for controlled drug release. The careful optimization and fabrication of sericin as the carrier material were achieved through flash nanoprecipitation, a straightforward and reproducible method that is devoid of intricate equipment. The physicochemical properties of SNCs and Cispt-SNCs, particularly concerning size, surface charge, and morphology, were evaluated using dynamic light scattering (DLS) and scanning electron microscopy (SEM). Chemical and conformational analyses of the nanocarriers were conducted using Fourier-transform infrared spectroscopy (FTIR) and circular dichroism (CD), and elemental composition analysis was performed through energy-dispersive X-ray spectroscopy (EDX). This approach aimed to achieve the smallest nanoparticle size for Cispt-SNCs (180 nm) and high drug encapsulation efficiency (84%) at an optimal sericin concentration of 0.1% (w/v), maintaining a negative net charge at a physiological pH (7.4). Cellular uptake and cytotoxicity were investigated in MCF-7 breast cancer cells. SNCs demonstrated stability and exhibited a pH-dependent drug release behavior, aligning with the mildly acidic tumor microenvironment (pH 6.0-7.0). Efficient cellular uptake of Cispt-SNC, along with DNA fragmentation and chromatin condensation, was found at pH 6, leading to cell apoptosis. These results collectively indicate the potential of SNCs for achieving controlled drug release in a tumor-specific context. Our in vitro studies reveal the cytotoxicity of both cisplatin and Cispt-SNCs on MCF-7 cells. Cisplatin significantly reduced cell viability at 10 µM concentration (IC50), and the unique combination of sericin and cisplatin showcased enhanced cell viability compared to cisplatin alone, suggesting that controlled drug release is indicated by a gradient decrease in cell viability and highlighting SNCs as promising carriers. The study underscores the promise of protein-based nanocarriers in advancing targeted drug delivery for cancer therapy.

How Shilajit-Based Nanocarriers Alter Classical Doxorubicin Delivery to Breast Cancer Cells (MCF-7 and ZR-75-1).

Chemotherapy has been ineffective in cancer treatment, and efficient delivery of chemotherapeutic agents remains a challenge. In this study, we developed a doxorubicin-loaded shilajit-based nanocarrier (SHN-Dox) using a nanoprecipitation method to enhance Dox uptake into breast cancer cells (MCF-7 and ZR-75-1). After confirmation of the physicochemical properties of the nanocarriers, the cytotoxic and pro-apoptotic effects of SHN-Dox and the production of reactive oxygen species (ROS) were evaluated on breast cancer cells. SHN-Dox showed a spherical shape with a size of 244 nm and a sustainable release profile of Dox. It exhibited high cytotoxicity against MCF-7 and ZR-75-1 cells, effectively inducing DNA fragmentation in these cells. After 24 h of treatment, SHN-Dox increased the apoptosis rate in MCF-7 cells and raised ROS levels. Therefore, SHN-Dox is a promising carrier that might reduce the side effects of Dox on healthy cells and provide a new strategy for clinical cancer treatment.

Effects of adding antioxidant nanoparticles on sperm parameters of non-human species after the freezing and thawing process: A systematic review and meta-analysis.

Cryopreservation is a widely used technique to store spermatozoa for a long time. Some Published articles have identified the cryoprotective effect of nanoparticles on sperm quality after the freeze-thaw process, but others have suggested the opposite results. PubMed, ISI Web of Science, and Scopus were systematically searched in animal studies by ("sperm" OR "spermatozoa") AND ("cryopreservation" OR "cooling storage" OR "freezing" OR "thawing") AND ("nanoparticle (lecithin nanoparticle, selenium nanoparticle, zinc nanoparticle, zinc oxide nanoparticle, nanoliposome, solid lipid nanoparticle (SLN), micelle, hydrogel, nanogel, silica nanoparticle, quantum dot, dendrimer, gold (Au) nanoparticle, silver nanoparticle, nanocomposite and mesoporous)"). Among 154 publications, data on sperm quality were extracted from 11 articles. The meta-analysis results demonstrated that nanoparticles had a positive impact on sperm progressive motility (WMD= 9.72, 95 % CI: 4.70, 14.75, p < 0.0001), total motility (WMD= 6.78, 95 % CI: 0.78, 12.78, p = 0.027), viability (WMD= 14.30, 95 % CI: 9.48, 19.13, p < 0.0001) and plasma membrane integrity (WMD = 13.74, 95 % CI: 8.20, 19.29, p < 0.0001). In conclusion, our results indicated the positive effects of nanoparticles as cryoprotectant agents on post-thawed sperm motility, viability, and membrane integrity.

Codelivery of resveratrol melatonin utilizing pH responsive sericin based nanocarriers inhibits the proliferation of breast cancer cell line at the different pH.

Protein-based nanocarriers have demonstrated good potential for cancer drug delivery. Silk sericin nano-particle is arguably one of the best in this field. In this study, we developed a surface charge reversal sericin-based nanocarrier to co-deliver resveratrol and melatonin (MR-SNC) to MCF-7 breast cancer cells as combination therapy. MR-SNC was fabricated with various sericin concentrations via flash-nanoprecipitation as a simple and reproducible method without complicated equipment. The nanoparticles were subsequently characterized for their size, charge, morphology and shape by dynamic light scattering (DLS) and scanning electron microscope (SEM). Nanocarriers chemical and conformational analysis were done by fourier transform infrared spectroscopy (FT-IR) and circular dichroism (CD) respectively. In vitro drug release was determined at different pH values (7.45, 6.5 and 6). The cellular uptake and cytotoxicity were studies using breast cancer MCF-7 cells. MR-SNC fabricated with the lowest sericin concentration (0.1%), showed a desirable 127 nm size, with a net negative charge at physiological pH. Sericin structure was preserved entirely in the form of nano-particles. Among the three pH values we applied, the maximum in vitro drug release was at pH 6, 6.5, and 7.4, respectively. This pH dependency showed the charge reversal property of our smart nanocarrier via changing the surface charge from negative to positive in mildly acidic pH, destructing the electrostatic interactions between sericin surface amino acids. Cell viability studies demonstrated the significant toxicity of MR-SNC in MCF-7 cells at all pH values after 48 h, suggesting a synergistic effect of combination therapy with the two antioxidants. The efficient cellular uptake of MR-SNC, DNA fragmentation and chromatin condensation was found at pH 6. Nutshell, our result indicated proficient release of the entrapped drug combination from MR-SNC in an acidic environment leading to cell apoptosis. This work introduces a smart pH-responsive nano-platform for anti-breast cancer drug delivery.

Composites based on alginate containing formylphosphazene-crosslinked chitosan and its Cu(II) complex as an antibiotic-free antibacterial hydrogel dressing with enhanced cytocompatibility.

Hydrogels based on chitosan or alginate biopolymers are believed to be desirable for covering skin lesions. In this research, we explored the potential of a new composite hydrogels series of sodium alginate (Alg) filled with cross-linked chitosan to use as hydrogel wound dressings. Cross-linked chitosan (CSPN) was synthesized by Schiff-base reaction with aldehydated cyclophosphazene, and its Cu(II) complex was manufactured and identified. Then, their powder suspension and Alg were transformed into hydrogel via ion-crosslinking with Ca2+. The hydrogel constituents were investigated by using FTIR, XRD, rheological techniques, and thermal analysis including TGA (DTG) and DSC. Moreover, structure optimization calculations were performed with the Material Studio 2017 program based on DFT-D per Dmol3 module. Examination of Alg's interactions with CSPN and CSPN-Cu using this module demonstrated that Alg molecules can be well adsorbed to the particle's surface. By changing the dosage of CSPN and CSPN-Cu, the number and size of pores, swelling rate, degradation behavior, protein absorption rate, cytotoxicity and blood compatibility were changed significantly. Subsequently, we employed erythromycin as a model drug to assess the entrapment efficiency, loading capacity, and drug release rate. FITC staining was selected to verify the hydrogels' intracellular uptake. Assuring the cytocompatibility of Alg-based hydrogels was approved by assessing the survival rate of fibroblast cells using MTT assay. However, the presence of Cu(II) in the developed hydrogels caused a significant antibacterial effect, which was comparable to the antibiotic-containing hydrogels. Our findings predict these porous, biodegradable, and mechanically stable hydrogels potentially have a promising future in the wound healing as antibiotic-free antibacterial dressings.

Comment on "Effect of Loaded Glycyrrhizic Acid on PLGA Nano-particle on Treatment of Allergic Asthma".

No Abstract.

A Novel Biocompatible and Biodegradable Electrospun Nanofibers Containing M. Neglectum: Antifungal Properties and In Vitro Investigation.

In the present study, biocompatible nanofibers containing aqueous extracts from Muscari neglectum (M. neglectum) plants (produced nanofiber) were prepared and their antifungal and cytotoxicity effects were investigated. For this purpose, the extracts obtained from flowers, stem leaves, and fresh onion from M. neglectum were lyophilized at various concentrations. Produced nanofibers were prepared using electrospinning techniques. During the electrospinning process, two auxiliary natural polymers including gelatin and chitosan were used. After synthesis, the physicochemical properties of the nanofibers were confirmed by Scanning Electron Microscopy (SEM), Fourier-transform infrared spectroscopy (FTIR), and X-ray energy diffraction spectroscopy (EDS or EDX), and Differential Scanning Calorimetry (DSC). The electrospun produced nanofibers have continuous and uniform structures. The cytotoxicity assay of these electrospun nanofibers were done on Human dermal fibroblast cell (HDF) and HUVEC cell (Human Umbilical Endothelial Cells) lines and results showed that nanofiber doesn't have any toxicity to normal cell lines. For anti-fungal activity tests, the appropriate amounts of nanofibers containing M. neglectum were placed in media with five different fungal species utilizing two methods: disc diffusion and well diffusion. In vitro results showed that all electrospun nanofibers containing M. neglectum had strong antifungal activity against Candida albicans, Glabrata, Parapacillus, Guillermoides, Crocus fungi species. Our findings also showed that nanofibers containing 86.88% polyvinyl alcohol/ gelatin/ chitosan/ M. neglectum root extract (produced nanofibers) were had better swelling and physicochemical properties and stronger antifungal activity than others (fiber formed with plant root). In a nutshell, natural nanofibers can be used as a beneficial drug delivery system.

Wound Healing Activities of Gundelia tournefortii L Extract and Milk-Cream Ointment on Second-Degree Burns of Rat Skin.

Skin burn is a major health problem in the community and seeking new and suitable treatment is suggested. In this regard, traditional remedies were consider in many countries. Regarding clinical application of herbal medicine in the healing of burn wounds, the present study was designed to evaluate the effect of Gundelia (Gundelia tournefortii L) extract with milk-cream on the healing of second-degree burn in a rat model. Thirty-six male Wistar rats (220 ± 30 g) were divided into 3 groups (n = 12), after establishment of second-degree burn: group1, were left without any intervention; group 2, were treated topically with silver sulfadiazine; and group 3, were treated with Gundelia tournefortii L extract composite with milk-cream once a day for 21 days. Macroscopically and histological examinations were conducted on 7, 14, and 21 days of therapy. Data analyses were done using 1-way analysis of variance and post hoc Tukey tests. Macroscopically, evaluation of wounds' sizes on the 14th and 21st days indicated that the wound surface was reduced significantly (P < .001) in group 3 compared with groups 1 and 2. Histological findings also showed that burn healing was significantly improved in group 3 compared with the other groups. Gundelia tournefortii L extract composite with milk-cream has an effective role on healing of second-degree burn in rat skin and it could a complementary and/or alternative medicine in wound healing.

Enhanced Cryoprotective Effect of Melatonin and Resveratrol by Coencapsulation: Improved In Vitro Development of Vitrified-Warmed Mouse Germinal Vesicle Oocytes.

Oocyte vitrification, as a vital step in reproductive medicine, is strongly associated with lower development caused by cryodamaging factors, such as oxidative stress. In this study, we evaluated the antioxidative synergistic effects of Melatonin (Mel) and Resveratrol (RES) coencapsulated by solid lipid nanocarriers (SLNs) against the pure antioxidant combination (Mel+RES). In this research, the formation of Mel+RES-SLN was confirmed by Fourier-transformed infrared spectroscopy. The average mean diameter, size distribution, polydispersity index, and zeta potential of particles were measured by Zetasizer, and the morphology was evaluated by scanning electron microscopy. In addition, the encapsulation efficiency (EE%) or drug loading capacity (DL%) of the nanocapsule was determined by spectrophotometric methods. Germinal vesicle (GV)-stage oocytes harvested from 6- to 12-week-old female NMRI mice were randomly divided into seven groups for in vitro studies. In these groups, (0, 10-12 M + 0.5 µM, 10-9 M + 2 µM, or 10-6 M + 10 µM) of Mel+RES/Mel+RES-SLN were added into vitrification media. After thawing, oocytes were matured, fertilized, and cultured for 3 days. Extra/intracellular reactive oxygen species (ROS) levels were measured in in vitro maturation medium after 24 hours. Our results revealed a significant improvement in the normal morphology of warmed GV-stage oocytes, GV breakdown (GVBD) rate, Metaphase II (MII)-stage oocyte formation, fertilization rate, early embryo development, and a significant reduction in intra/extracellular ROS level when vitrification media was supplemented with the lowest Mel+RES-SLN concentration. In vitro studies also demonstrated that the highest concentration of Mel+RES-SLN was safe, without a detrimental effect on embryonic development upon treatment. In conclusion, the lowest concentration of Mel+RES-SLN supplementation in GV-stage oocyte vitrification media improved maturation, fertilization, and embryo development rate and decreased extra/intracellular ROS level through an enhanced/controlled intracellular penetration compared to the pure Mel+RES.

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.

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.

Cryoprotective Effect of Tretinoin-Loaded Solid Lipid-Core Nanocapsules During Fresh and Freeze/Thaw Media on NMRI Mouse Sperm Parameters, DNA Damage, and Reactive Oxygen Species Production.

Due to the induced oxidative stress that exists in sperm freezing/thawing procedures and handling media, the use of exogenous antioxidant agents seems necessary. Drug delivery by nanocarriers has been designed to overcome the limitations of antioxidants, such as high-dose toxicity and short biological half-life. In this study, we tried to investigate the effects of tretinoin-loaded solid lipid core nanocapsules (TTN-SLN) added to freezing/thawing and handling media (in three experimental groups) on sperm motility (total/progressive), viability, DNA fragmentation, and extracellular reactive oxygen species (ROS) levels. Sperm samples from at least 30 adult male NMRI mice were evaluated in this study. The results of experiments 1 and 2 showed that the addition of 0.5 µM TTN-SLN in freezing and thawing medium significantly increased sperm viability and total/progressive motility and decreased DNA fragmentation and extracellular ROS levels (p < 0.05). Adding 0.25 and 0.5 µM of TTN-SLN to the handling medium (experiment 3), increased sperm parameters and decreased DNA fragmentation and extracellular ROS levels significantly (p < 0.05) compared with the control group. Briefly, our results indicate that SLN can deliver the lowest concentrations of tretinoin in a controlled release mechanism into the intracellular space of sperm. Also, high-dose TTN-SLN is safe during freezing/thawing and handling processes of mouse sperm.

The Anti-oxidative Effects of Encapsulated Cysteamine During Mice In Vitro Matured Oocyte/Morula-Compact Stage Embryo Culture Model: a Comparison of High-Efficiency Nanocarriers for Hydrophilic Drug Delivery-a Pilot Study.

Although it is well-recognized that antioxidant nano-encapsulation has many benefits such as minimizing side effects (e.g., high-dose toxicity), the most attention was paid to the hydrophobic antioxidant not hydrophilic. In this regard, we sought to compare two hydrophilic model nanocarriers to deliver the optimal dose of cystamine (Cys) into the in vitro matured oocyte and the first cleavage stages until morula-compact stage embryonic cells. The formation of Cys-loaded solid self-emulsifying lipid (Cys + SLN) and Cys-loaded chitosan shell (Cys-CS-NC) were confirmed by FT-IR and UV-Vis spectrophotometry, dynamic light scattering (DLS), transmission electron microscopy (TEM), and scanning electron microscopy (SEM) technologies. In two experiments, the oocytes/presumptive zygotes were cultured under various concentrations of Cys-SLN and Cys-CS-NC. The results of nuclear staining (aceto-orcein and Hoechst 33342), H2DCFDA fluorescent staining, chemiluminescence test, and quantitative reverse transcription-PCR (qRT-PCR) technique as in vitro toxicity studies demonstrated that adding the lowest dose of Cys-encapsulated in both nanocarriers [Cys-SLN (5 µM) and Cys-CS-NC (10 µM)] to maturation or culture medium could accumulate a strong anti-oxidative effect in oocyte/embryo by controlled release and enhanced intracellular penetration of Cys. In comparison, Cys-SLN (5 µM) is more effective than Cys-CS-NC (10 µM) groups to improve the expression of antioxidant genes (SOD, CAT, GPx) or anti-apoptotic (BCL-2) gene and decreased apoptosis (BAX and caspase-3) or intra-/extracellular ROS levels. In a nutshell, both nanocarriers (CS-NC or SLN) can deliver the lowest dose of Cys into the oocyte/embryo, thus encouraging a better expansion of antioxidant genes and enhancing the development of in vitro oocyte/embryo.