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.

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.

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.

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.

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.

In vitro culture medium (IVC) supplementation with sericin improves developmental competence of ovine zygotes.

This study was carried out to investigate the effects of supplementation of potassium simplex optimized medium (KSOM-aa) with various sericin concentrations (0, 0.1, 0.5, 1 and 2.5%) on ovine zygotes. The results indicate that the supplementation of oocyte in vitro culture medium with optimal concentration of sericin (0.1 and 0.5%) may have beneficial effects on developmental competence of in vitro-derived ovine embryos.