Cryopreservation of ram semen: baicalein efficiency on oxidative stress, chromatin integrity, viability and motility post thaw.

Baicalein (B) has potential antioxidant properties, but it has not been tested as a ram semen extender. This study aimed to assess the impact of B on various sperm parameters and determine its potential influence on semen quality after the freeze-thawing process. During the breeding season, ejaculates were obtained from four rams with the aid of an artificial vagina. The collected mixed semen samples were divided into four groups: control (C; 0), B0.5 (0.5 mM), B1 (1 mM), and B2 (2 mM). After semen extension, the samples were loaded into 0.25 mL straws and stored for 2 h at 4°C prior to freezing in liquid nitrogen vapor and thawed in a water bath at 37°C. Among the groups, B0.5 demonstrated the highest progressive motility results, while B1 and B2 exhibited reduced motility (p < 0.05). In terms of high mitochondrial membrane potential, plasma membrane and acrosome integrity, and viability, B0.5 showed significantly superior outcomes to the other B groups (p < 0.05), although it was not significantly better than C. B1 displayed the highest plasma membrane integrity levels (p < 0.05). Notably, B2 displayed the lowest total antioxidant status levels among the groups (p < 0.05). The findings of this study suggested that the in vitro spermatological characteristics of ram spermatozoa such as progressive motility and chromatin integrity can be protected from the freeze-thawing process by using the 0.5 mM dose of baicalein as a semen extender. The treatment of sperm freezing might benefit from further in-depth research on the role of B in the improvement of cryoinjury and its underlying processes.

Investigation of the effects of syringic acid supplemented to Tris semen diluent on ram semen freezability.

To the best of our knowledge, no research has been conducted to test the effects of syringic acid (SA) on ram semen freezing within the scope of natural antioxidants added to semen extenders. Therefore, this study had two main objectives. First, to test whether adding SA to ram semen freezing extender has a protective effect and contributes positively to sperm kinetic, plasma and acrosome integrity, mitochondrial membrane potential, lipid peroxidation, oxidant and antioxidant and DNA damage parameters after thawing. Second, it was to determine at what concentration the SA supplemented to the extender could be applied by in vitro studies by preserving the fertilization ability of frozen semen at the highest level. In the study, six individuals of Sönmez rams were used. The semen was collected from the rams using an artificial vagina and pooled. The pooled semen was divided into five different groups and extended with 0, 0.5, 1, 2 and 4 mM SA (control C, SA0.5, SA1, SA2 and SA4, respectively). After dilution, the semen samples were kept at 4°C for 3 h, then loaded into 0.25 mL straws and frozen in liquid nitrogen vapour. The SA1 and SA2 groups were higher plasma membrane and acrosome integrity (PMAI), high mitochondrial membrane potential (HMMP), plasma membrane integrity and motility compared to other groups (p < .05). It was observed that SA supplemented to the Tris extender significantly reduced DNA damage, and the lowest values were obtained especially in the SA1 and SA2 treatments (p < .05). Also, lowest MDA level was determined at the SA1 and this was statistically significant compared to SA4 and C (p < .05). In conclusion, it was revealed that SA added to Tris semen extender at 1 and 2 mM treatment doses increased progressive and total motility and preserved PMAI, plasma membrane integrity, HMMP and DNA integrity.

Investigation of Changes in Spermatozoon Characteristics, Chromatin Structure, and Antioxidant/Oxidant Parameters after Freeze-Thawing of Hesperidin (Vitamin P) Doses Added to Ram Semen.

We conducted this study to determine the potential cryopreservative effects of different hesperidin (vitamin P; H) doses on ram semen after freeze-thawing. Semen samples were obtained from Sönmez rams using an artificial vagina. The samples were divided into six groups: control, 10, 50, 100, 250, and 500 µg/mL H (C, H10, H50, H100, H250, and H500, respectively). At the end of the study, sperm motility and kinetic parameters, acrosome integrity (AI), mitochondrial membrane potential (MMP), viability, lipid peroxidation levels (LPL), chromatin damage, oxidant parameters, and antioxidant parameters were assayed. None of the doses of H added to the semen extender showed any enhancing effects on progressive motility compared to C (p > 0.05). In fact, H500 had negative effects (p < 0.05). Moreover, AI was the highest at the H10 dose, while LPL values were the lowest at the same dose (p < 0.05). The doses of H10 and H50 added to the Tris extender medium showed positive effects on sperm cell chromatin damage. Consequently, we can say that H doses used in this study are not effective on semen progressive motility, but the H10 dose is effective on AI and chromatin damage by reducing LPL.

Thymoquinone improves motility, plasma membrane integrity and DNA integrity of frozen-thawed ram semen.

The aim of this study was to determine the effects of thymoquinone (TQ), which is the most essential active compound of Nigella sativa, on the spermatological parameters of ram semen during cryopreservation. Ejaculates were collected from five Sonmez rams using an artificial vagina and extended with Tris-based extender not containing TQ (control, 0 µg/ml TQ) and containing 10, 25, 50 and 100 µg/ml TQ. The extended semen samples were equilibrated in a + 4°C cold cabinet for 2 h. After 2 h, the samples were loaded into 0.25 ml French straws. The straws were frozen by liquid nitrogen vapour and stored in a liquid nitrogen container (-196°C). The frozen straws were thawed in a water bath (37°C for 30 s) and evaluated in terms of motility characteristics, plasma membrane and acrosome integrity, mitochondrial reactive oxygen species levels, lipid peroxidation levels, DNA damage and biochemical alterations (oxidative stress index, malondialdehyde and glutathione). TQ100 had higher total motility (53.59 ± 3.01) and progressive motility (19.84 ± 1.44; not significantly different from TQ25 and TQ50) compared to the control and TQ10 (p Ë‚ 0.05). According to the results of the analyses on motility characteristics, there were significant differences between the groups in terms of curvilinear velocity (VCL), amplitude of lateral head displacement (ALH) and linearity (LIN; p Ë‚ 0.05). The highest DNA damage was detected in the control group (p Ë‚ 0.05). TQ50 had higher plasma membrane and acrosome integrity (59.56 ± 5.92) compared to the control and TQ25 (p < 0.05) but not significantly different from TQ10 and TQ100. The lowest mitochondrial reactive oxygen species levels were detected in TQ50 and TQ100 (p Ë‚ 0.05). There were no significant differences among the groups in terms of their oxidative stress index, lipid peroxidation, malondialdehyde and glutathione levels (p > 0.05). According to the results, it could be concluded that supplementing 50 or 100 µg/ml TQ to Tris extenders that were used for ram semen cryopreservation showed a positive effect on motility, plasma membrane integrity and acrosome integrity, and it reduced DNA damage and mitochondrial reactive oxygen species levels.

Use of Infrared Thermography during Ejaculation Process and Its Link with Semen Quality and Freezability in Dogs.

This study aimed to describe the thermal variation of external reproductive tracts during ejaculation in relation to sperm quality in dogs. Forty-six adult fertile dogs were monitored using a thermal camera before, during and after the semen collection, taking into account penile and scrotal temperatures as reproductive thermal patterns while eye and perianal temperatures were recorded as complementary thermal patterns of behavioral response. The parameters were classified depending on age (≤4 years and >4 years), body weight (BW) (≤75 kg and >75 kg), sperm concentration (CON) (≤300 million and >300 million), total testicular volume (TTV) (≤600 cm3 and >600 cm3) and total ejaculation time (TET) (≤800 s and >800 s) of the animals from which semen was collected successfully. Heavier males (p < 0.05) that have more consistent testicles (p < 0.01) as well as quicker ejaculate responders (p < 0.001) and lower scrotal temperature had better semen (Δ motility) freezability. The lower eye temperature prior to the ejaculation (p < 0.01), lower scrotal temperature following ejaculation (p < 0.01), and conversely, higher penile temperature during the ejaculation (p < 0.001) had a higher sperm concentration. Furthermore, the sperm freezability was negatively correlated with total ejaculation time (r = -0.39, p < 0.05) and sperm abnormalities were lower in the ejaculate of dogs having a higher temperature of the scrotum, bulbus and penis. In conclusion, infrared monitoring throughout semen collection in dogs can provide information on behavioral reactions during human manipulation, as well as semen quality and testicular functionality.

Red pine (Pinus brutia Ten) bark tree extract preserves sperm quality by reducing oxidative stress and preventing chromatin damage.

This study aimed to investigate the effectiveness of using red pine bark tree extract (P; Pinus brutia Ten) as a TRIS extender in an attempt to prevent oxidative stress in bull spermatozoa after freezing. Semen specimens were obtained from Simmental bulls via an artificial vagina and pooled. They were separated into five specimens and diluted with Tris extender consisting of P (200, 100, 50 and 25 µg/ml) and P free (control; C) up to a final concentration of 16 × 106 per straw. All specimens were equilibrated for a period of 4 hr at a temperature of 4°C, following which they were filled in 0.25-ml French straws and frozen. Addition of P resulted in favourable tail length in comparison with C (p < .05). The lowest malondialdehyde levels and the highest glutathione levels were detected in all P groups (p < .05). Supplementation with P did not show advanced results in terms of total, progressive sperm motility and total abnormality in comparison with C (p > .05). In conclusion, it has been shown that although P added to a Tris extender does not have a positive effect on sperm motility, it prevents chromatin damage by reducing oxidative stress, in addition to reducing head abnormalities when used at the amount of 50 µg/ml.

Genomewide association of male reproductive traits in Aksaray Malakli dogs.

Male reproductive parameters are often used for the functional examination and evaluation of predicted genetic values for future aspects. However, these traits are relatively reliable until the measurable effects are expressed on desired traits. Therefore, we aimed to associate the single nucleotide polymorphism (SNP) genotype of the investigated characteristics and reproductive loci. A total of 46 male dogs are divided into three age groups (I ≤ 3 years, n = 19; II 4-6 years; n = 19, and III ≥7 years, n = 8). The testis, scrotum and body weight, libido sexualis and ejaculation time for each fraction were monitored as functional traits, while the pH, fractional semen volume, motility, concentration, and abnormal and dead spermatozoa rate were recorded as spermatological traits. The Affymetrix Canine 127 K SNP genotyping array v2 (Affymetrix Inc., California, USA) was used for SNP genotyping. In the primary results, the scrotal circumference was found to be higher in group II compared to other groups (p < 0.05) and the lowest total abnormal spermatozoa rate was found in group I (p < 0.05). The normal spermatozoa rate was found to be significantly above the threshold in relation to the SNP in chromosome 17. In conclusion, this study represents an exciting first step towards SNP association with dog semen spermatological parameters. Future studies might be undertaken to evaluate this SNP region for gene-knockout and expression analysis and for fine mapping to validate and/or discover the exact position of the effect region.

Effect of cholesterol loaded cyclodextrin on semen cryopreservation of Aksaray Malakli shepherd dogs of different ages.

The objective of the study was to determine the effect of cholesterol-loaded cyclodextrin (CLC) on the quality parameters of semen from Aksaray Malakli Shepherd dogs of different age groups. Forty-eight male dogs were divided into 3 groupings according to their ages (young age (Y): ≤3 years, n: 20; middle age (M): 4-6 years, n: 20; old age (O): ≥7 years; n: 8). The sperm-rich portion of the ejaculate from each dog was divided into four aliquots and extended with either tris as a control (C) or tris loaded with 0.5, 1.0, and 1.5 mg/120 × 106 CLC as low (L), intermediate (I), and high (H) doses, respectively. Following equilibration for at least half an hour, the straws were frozen in nitrogen vapor and then stored in liquid nitrogen at least for 48 h. Later, the frozen straws were thawed in a water bath for spermatological evaluation. Significant differences were observed between different age groups in terms of the spermatological parameters (p < 0.05). The evidence suggests that increasing age is associated with poor in-vitro spermatological parameters and CLC was able to protect the acrosome integrity from cryo-damage during the freeze-thawing process. Better semen freezability characteristics were obtained at young ages, considering the overall parameters.

Cryopreservation of bull sperm: effects of extender supplemented with different cryoprotectants and antioxidants on sperm motility, antioxidant capacity and fertility results.

The objectives of this study were to compare glycerol (G) and ethylene glycol (EG) at different concentrations and trehalose (T) or cysteine (C; with/without) in Tris extender for cryopreservation of bull semen. Twenty-four ejaculates obtained from three bulls were included in the study. Each ejaculate was divided into four equal aliquots and diluted using both of the Tris extenders with G (5% or 7%) or EG (3% or 5%). After that, each extenders were divided into three equal aliquots and diluted using both of the 5 mM C or 25 mM T, and control (without additives) was cooled to 4 ° C and frozen in 0.25 ml French straws. The addition of 3% and 5% EG without antioxidants resulted in the least Computer-Assisted Sperm motility Analysis (CASA) motility as compared with the other groups. Treatment with 25 mM T in 3% EG beneficially effected acrosome morphology as compared with the other groups. Also, treatment with 3% EG with 25 mM T and 5% EG resulted in a greater rate of total abnormalities. Treatment with 3% G yielded a slightly greater percentage of membrane integrity by Hypo-Osmotic Swelling Test (HOST) assessment than that of the other groups. Treatment with 3% EG with 5 mM C resulted in the greatest concentration of malondialdehyde (MDA). The glutathione peroxidase (GPx) antioxidant activity was increased in the C-treatment groups when compared to the other groups. Treatment with 5% EG and 5 mM C resulted in less chromatin damage and detrimental impacts on tail moment. Treatment with 5% EG led to greater non-return rates of inseminated cows. However, this result was not considered to be statistically important.

Comparing ethylene glycol with glycerol and with or without dithiothreitol and sucrose for cryopreservation of bull semen in egg-yolk containing extenders.

There are few studies performed for investigating the roles of different ratio and cryoprotectants with dithiothreitol or sucrose on sperm motility characteristics and antioxidant capacities of post-thawed bull spermatozoa. The objectives of this study were to compare glycerol (G) and ethylene glycol (EG) at different concentrations as cryoprotectants and dithiothreitol (D) or sucrose (S) (with/without) as antioxidants in Tris extender for cryopreservation of bull semen. Twenty-four ejaculates obtained from three bulls were included in the study. Each ejaculate was split into four equal aliquots and diluted using both of the Tris extenders with glycerol (5% or 7%) or ethylene glycol (3% or 5%). After that, each extenders were split into three equal aliquots and diluted using both of the dithiothreitol 5mM or sucrose 25 mM, and control (without additives) was cooled to 4 °C and frozen in 0.25-ml French straws. when compared to control, different doses cryoprotectants and antioxidants addition no significantly increased the percentages of post-thaw sperm progressive and motitilities, acrosome abnormality and plasma membrane integrity (P>0.05). However, EG3+S yielded the greatest percentages of the total abnormality (P<0.05). As regard to antioxidant activities G7 and EG5 led to lowest MDA activity with or without D or S but, these results were not supported to the GPx activity (P<0.01). The sperm motion characteristics such as VAP, VCL, ALH and BCF gave significantly different results (P<0.05). When compared the DNA integrity, different doses cryoprotectants without antioxidants addition significantly increased the percentages of the tail intensity and tail moment (P<0.05). There were no significant differences observed in non-return rates among all treatment groups (P>0.05).

Can linoleic acid improve the quality of frozen thawed bull sperm?

BACKGROUND: Cryopreservation is known to have a detrimental effect on the motility, viability and membrane integrity of sperm cells. OBJECTIVE: The aim of this study was to investigate the effect of various amount of linoleic acid supplementation to the Tris extender, on bull sperm parameters, DNA integrity and oxidative stress after freeze-thawing. METHODS: Ejaculates were split into five aliquots and extended to a final concentration of 18x10(6) spermatozoa per ml with the base extender containing different doses of linoleic acid 0.125 ml, (L125); 0.250 ml (L250); 0.5 ml (L500), 1 ml (L1000) and no additive (control; L0). The extended samples were equilibrated slowly to 4 degree C for 4 h and then froze using a digital freezing machine. Frozen straws were thawed individually in water bath at 37 degree C for 30 s to analyse progressive motility and sperm motion characteristics as well as membrane integrity. Biochemical assays were performed in a spectrophotometer using commercial kits. DNA damage was evaluated by Comet Assay. RESULT: The addition of various linoleic acid did not improve the sperm subjective, CASA and progressive motilities, sperm motility characteristics and DNA integrity (P>0.05). L500 exhibited the greatest values for membrane integrity than that of the other groups (P<0.001). All supplementation groups led to lower percentages of tail abnormalities in comparison to the control (P<0.001). L500 and L1000 significantly decreased total abnormalities. In conclusion, our findings showed that L500 linoleic acid supplementation in semen extender was of great beneficial effect on frozen-thawed bull semen in terms of morphology and plasma membrane integrity.

Effects of various cryoprotectants on bull sperm quality, DNA integrity and oxidative stress parameters.

The objectives of this study was to compare the effects of type and concentration of cryoprotectants glycerol (G), ethylene glycol (EG) and dimethyl sulfoxide (DMSO) on the plasma membrane and DNA integrity as well as antioxidant activity of cryopreserved Eastern Anatolian red bull sperm. Ejaculates were collected from the three bulls using an artificial vagina twice a week. The ejaculates were pooled to increase the semen volume for replication and to eliminate variability among the evaluated samples. The pooled ejaculates were also split into seven equal experimental groups and diluted with the modified base extender to a final spermatozoa concentration of 15×10(6)/ml. The extended samples were cooled slowly to 4°C and equilibrated for 4h. They were then loaded into 0.25ml French straws and frozen using a digital freezing machine at 3 programmed rates: -3°C/min from +4°C to -10°C, -40°C/min from -10°C to -100°C, and -20°C/min from -100°C to -140°C. Thereafter, the straws were plunged into liquid nitrogen at -196°C. Frozen straws were thawed individually at 37°C for 30s in a water bath to analyse progressive motility and sperm motion characteristics as well as membrane integrity using hypo-osmotic swelling test. Biochemical assays were performed in a spectrophotometer using commercial kits. DNA damage was evaluated by Comet Assay using Image Analysis System. 6% G exhibited the greatest percentages of CASA (43.7±2.92%) and progressive (26.4±2.64%) motilities when compared to the other groups (P<0.001). 6% G and 6% EG showed the greatest values of preserved membrane integrity (P<0.001). 6% DMSO and 3% EG + 3% DMSO resulted in greater chromatin damage than the other groups (P<0.001). The antioxidant activities of GPx, GSH, and CAT as well as the total antioxidant activity were affected by the type of cryoprotectant; notably, 2% G+2% EG+2% DMSO yielded the lowest activities when compared to the other groups (P<0.001). In conclusion, no advantages were found in using EG or DMSO to replace G in bull sperm cryopreservation. Freezing with cryoprotectant 6% G yielded the best post-thaw sperm characteristics for Eastern Anatolian Red bull spermatozoa.

Formulation of zolmitriptan sublingual tablets prepared by direct compression with different polymers: in vitro and in vivo evaluation.

First-pass metabolism can be overcome by sublingual drug delivery, and quick drug entry into the systemic circulation can be obtained. In certain diseases such as migraine therapy, taking fast pharmacological response is an important criteria. In this study, zolmitriptan sublingual tablets were prepared by direct compression method using different mucoadhesive polymers such as hydroxypropyl methyl cellulose, chitosan and sodium carboxy methyl cellulose at a concentration range of 0.5-5% to reduce flushing action of saliva and provide enough time for drug to be absorbed. Tablets were evaluated for the physical properties, and optimum formulations were chosen for in vivo studies to carry on sheep model. The tablets disintegrated rapidly, and dissolution tests revealed that zolmitriptan was dissolved from the formulation within the compendial limits. This especially showed us that the concentration range of polymers is in acceptable limit. It was also concluded that microcrystalline cellulose, spray-dried lactose and sodium starch glycolate are the appropriate excipient and formulated in good proportions. In vivo studies indicated that formulation containing 5% chitosan has the maximum C(max) and AUC and minimum t(max) values (p<0.05). As a result, sublingual tablet administration of zolmitriptan formulated with appropriate excipients and especially with chitosan seems promising alternative to traditional routes.

Ovarian response and embryo yield of Angora and Kilis goats given the day 0 protocol for superovulation in the non-breeding season.

The aim of this study was to compare ovarian response and embryo yield of Day 0 protocol in Angora goats (AG) and indigenous Kilis goats (KG) in the non-breeding season. A total of 16 Angora goats (AG group) and 11 Kilis goats (KG group) were used in this study. In the synchronization process, after controlled internal drug release withdrawal, when estrus signs were observed, natural mating was performed. Ovarian response was determined by synchronized laparotomy 6 days after natural mating, and number of corpora lutea (CL) was recorded. Embryos were collected and morphologically evaluated by stereomicroscope. Synchronization rates did not differ between AG (88%, 14/16) and KG group (91%, 10/11). In AG and KG groups, the proportion of CL on the right (44% and 53%, respectively) and left (56% and 47%, respectively) ovaries were similar. The CL number per animal did not differ significantly between the two breeds and was determined as 4.4 ± 0.90 in AG group and 6.4 ± 1.44 in KG group. Transferable embryo yields were significantly higher in AG group (31/42, 74%) compared to KG group (16/46, 35%) in the non-breeding season (P < 0.01). In conclusion, it is suggested that the day 0 protocol can be used for goat superovulation in the non-breeding season; however, transferable embryo yields are affected by the breed.

Nasal administration of metoclopramide from different dosage forms: in vitro, ex vivo, and in vivo evaluation.

Nasal drug delivery is an interesting route of administration for metoclopramide hydrochloride (MTC) in preventing different kind of emesis. Currently, the routes of administration of antiemetics are oral or intravenous, although patient compliance is often impaired by the difficulties associated with acute emesis or invasiveness of parenteral administration. In this perspective, nasal dosage forms (solution, gel, and lyophilized powder) of MTC were prepared by using a mucoadhesive polymer sodium carboxymethylcellulose (NaCMC). In vitro and ex vivo drug release studies were performed in a modified horizontal diffusion chamber with cellulose membrane and excised cattle nasal mucosa as diffusion barriers. The tolerance of nasal mucosa to the formulation and its components were investigated using light microscopy. In vivo studies were carried out for the optimized formulations in sheep and the pharmacokinetics parameters were compared with oral solution and IV dosage form. The release of MTC from solution and powder formulations was found to be higher than gel formulation (p < 0.05). Histopathological examination did not detect any severe damage. Hydroxypropyl-beta-cyclodextrin (HPbetaCD) used in powder formulations was found to be effective for enhancing the release and absorption of MTC. In contrast to in vitro and ex vivo experiments nasal bioavailability of gel is higher than those of solution and powder (p < 0.05). In conclusion, the NaCMC gel formulation of MTC with mucoadhesive properties with increased permeation rate is promising for prolonging nasal residence time and thereby nasal absorption.

Nasal absorption of metoclopramide from different Carbopol 981 based formulations: In vitro, ex vivo and in vivo evaluation.

There is a need for nasal drug delivery of metoclopramide HCI (MTC) in specific patient populations where the use of commercially available intravenous and oral dosage forms may be inconvenient and/or unfeasible. In this perspective, nasal dosage forms (solution, gel and lyophilized powder) of MTC were prepared by using a mucoadhesive polymer Carbopol 981 (CRB 981). The drug release studies of formulations were performed by using a modified horizontal diffusion chamber with cellulose membrane and excised cattle nasal mucosa as diffusion barriers. After the ex vivo experiments, the morphological appearances of the nasal mucosa were analyzed with the light microscopic studies. In vivo experiments were carried on sheep model. The release of MTC from solution and powder formulations was found higher than gel formulation (p < 0.05) and no severe damage was found on the integrity of nasal mucosa after ex vivo experiments. The penetration enhancing effect of dimethyl-beta-cyclodextrin (DM-beta-CD) used in powder formulations was observed in ex vivo and in vivo experiments. In contrast to in vitro and ex vivo experiments the nasal bioavailability of gel formulation was found higher than those of the solution and powder (p < 0.05) and might represent a promising novel tool for the systemic delivery of MTC.

Comparison of hyaluronate/carboxymethylcellulose membrane and melatonin for prevention of adhesion formation in a rat model.

BACKGROUND: To investigate the effectiveness of melatonin in preventing post-operative adhesion formation and to compare it with the efficacy of hyaluronate/carboxymethylcellulose membrane in a rat model. MATERIALS AND METHODS: Following pilot studies, 35 rats were operated on in the full study. In 15 animals (group one), 10 standard lesions were inflicted in each uterine horn (total 30 horns) and melatonin was applied before closure of the abdomen. In the second group, 20 animals were operated on and one of the uterine horns (total 20 horns) with standard lesions was treated with hyaluronate/carboxymethylcellulose membrane and the other uterine horn served as a control. Second-look operations were performed 1 week later and adhesion scores were compared. RESULTS: The adhesion scores of uterine horns treated with melatonin and of uterine horns treated with hyaluronate/carboxymethylcellulose membrane were significantly lower than the scores of the controls (P < 0.001). There was no statistically significant difference between the adhesion scores of uterine horns treated with melatonin and of uterine horns treated with hyaluronate/carboxymethylcellulose membrane (P > 0.05). CONCLUSIONS: Both melatonin and hyaluronate/carboxymethylcellulose membrane were effective in prevention of adhesion formation in a rat uterine horn model.