Repro-protective role of royal jelly in phenylhydrazine-induced hemolytic anemia in male mice: Histopathological, embryological, and biochemical evidence.

To estimate the repro-protective effect of royal jelly (RJ) on phenylhydrazine (PHZ)-induced anemia's detrimental effects, 24 mature mice were divided into control group (0.10 mL normal saline; intra-peritoneally), RJ group (100 mg/kg/day; orally), experimental anemia (EA) group that received only PHZ (6 mg/100 g/48 h; intra-peritoneally), and RJ + EA (according to the previous prescription) group. After 35 days, testicular histoarchitecture, RNA damage in germinal cells, sperm characteristics, testicular total anti-oxidant capacity and malondialdehyde as well as serum testosterone levels, pre-implantation embryo development and cyclin D1 and c-myc mRNA levels at two-cell, morula and blastocyst stages were analyzed. Spermatogenesis indices were ameliorated following RJ co-administration. Moreover, RJ co-treatment reduced germinal cells RNA damage, improved sperm characteristics, boosted pre-implantation embryo development and restored androgenesis, and oxidant/anti-oxidant status. Co-administration of RJ also decreased mRNA levels of cyclin D1 and up-regulated those of c-myc in two-cell embryos, morulas and blastocysts. The findings suggest that RJ can play a repro-protective role in PHZ-induced anemia in mice through anti-oxidant defense system reinforcement and androgenesis restoration as well as cyclin D1 and c-myc expressions regulation.

Angiogenic effects of cell therapy within a biomaterial scaffold in a rat hind limb ischemia model.

Critical limb ischemia (CLI) is a life- and limb-threatening condition affecting 1-10% of humans worldwide with peripheral arterial disease. Cellular therapies, such as bone marrow-derived mesenchymal stem cells (MSCs) have been used for the treatment of CLI. However, little information is available regarding the angiogenic potency of MSCs and mast cells (MC) in angiogenesis. The aim of this study was to evaluate the ability of MCs and MSCs to induce angiogenesis in a rat model of ischemic hind limb injury on a background of a tissue engineered hydrogel scaffold. Thirty rats were randomly divided into six control and experimental groups as follows: (a) Control healthy (b) Ischemic positive control with right femoral artery transection, (c) ischemia with hydrogel scaffold, (d) ischemia with hydrogel plus MSC, (e) ischemia with hydrogel plus MC and (f) ischemia with hydrogel plus MSC and MCs. 106 of each cell type, isolated from bone marrow stroma, was injected into the transected artery used to induce hind limb ischemia. The other hind limb served as a non-ischemic control. After 14 days, capillary density, vascular diameter, histomorphometry and immunohistochemistry at the transected location and in gastrocnemius muscles were evaluated. Capillary density and number of blood vessels in the region of the femoral artery transection in animals receiving MSCs and MCs was increased compared to control groups (P < 0.05). Generally the effect of MCs and MSCs was similar although the combined MC/MSC therapy resulted in a reduced, rather than enhanced, effect. In the gastrocnemius muscle, immunohistochemical and histomorphometric observation showed a great ratio of capillaries to muscle fibers in all the cell-receiving groups (P < 0.05). The data indicates that the combination of hydrogel and cell therapy generates a greater angiogenic potential at the ischemic site than cell therapy or hydrogels alone.

Possible protective effects of crocin on destructive side effects of cyclo-phosphamide in mice ovarian tissue: Evaluation of histomorphometrical and biochemical changes.

One of the side effects of cyclophosphamide (CP) is low fertility. In this study, we investigated the protective role of crocin (Cr) against CP chemotherapy-induced changes in ovarian tissue. In the current study, we treated 15 female mice aged 6-8 weeks old for 21 days. The mice were distributed into three groups including control received normal saline (0.10 mL; IP), CP or sham-control group (CP once a week, 15.00 mg kg-1; IP) and experimental (CP + Cr) group received CP along with Cr (200 mg kg-1 daily; IP). After completing the procedure, levels of total anti-oxidant capacity (TAC), superoxide dismutase (SOD) and sex hormones in serum as well as malondialdehyde (MDA) in the left ovarian tissue were measured. The right ovaries were used for histological and morphological tests. The obtained data were statistically analyzed by SPSS software using ANOVA and Tukey follow-up studies. Results showed that in the CP group a significant decrease was observed in ovarian follicles, the number of corpus luteum, levels of TAC, SOD and sex hormones; while, there was a significant increase in the number of atretic follicles and mast cells and level of MDA compared to control group. Administration of Cr along with CP caused a significant ameliorative effect on the studied parameters. In conclusion, the Cr could significantly decrease the side effects caused by CP chemotherapy in mice ovarian tissue.

Spirulina platensis (Arthrospira platensis) attenuates Cyclophosphamide-induced reproductive toxicity in male Wistar rats: Evidence for sperm apoptosis and p53/Bcl-2 expression.

Cyclophosphamide is an antitumor agent that causes disorders in fertility. This study aimed to evaluate the protective effects of Spirulina platensis against Cyclophosphamide-induced testicular toxicity. 42 male Wistar rats were randomly divided into six groups. Experimental groups included three groups. The first experimental group received Cyclophosphamide at a dose of 5 mg/kg body weight (BW) orally. The second and third experimental groups received 5 mg/kg BW Cyclophosphamide and 500 and 1,000 mg/kg BW S. platensis orally, respectively. The control groups included a control group, and two S. platensis control groups. Following 28 days, two flow cytometry techniques were used to determine sperm apoptosis and testicular protein expression of tumor protein (p53) and B-cell lymphoma 2 (Bcl-2). p53 is a tumor suppressor protein that causes the cell to enter the apoptosis cycle after DNA damage and Bcl-2 is an anti-apoptotic protein that acts through the mitochondrial pathway of apoptosis. FITC-Annexin V assay was used for sperm apoptosis evaluation. For protein expression assay, primary and secondary antibodies staining were performed. The Cyclophosphamide group showed a significant increase in sperm apoptosis compared to the control group. Cyclophosphamide significantly increased p53 and decreased Bcl-2 expression compared to the control group. S. platensis co-treated groups exhibited a significant decrease in sperm apoptosis compared to the Cyclophosphamide group. Moreover, S. platensis co-treated groups displayed a significant decreasing in p53 and increasing in Bcl-2 expression compared to the Cyclophosphamide group. The results of this study indicated that S. platensis protected rats against Cyclophosphamide-induced reproductive toxicity. PRACTICAL APPLICATIONS: Cyclophosphamide is the chemotherapy agent used to treat different cancers. Cyclophosphamide has side effects on the male reproductive system. Spirulina plantesis has a protective effect because of its antioxidant and anti-apoptotic properties. Co-administration of Spirulina plantesis with Cyclophosphamide reduces sperm apoptosis also decreases P53 protein expression and increases Bcl-2 protein expression. This study validated the anti-apoptotic potential of Spirulina plantesis against Cyclophosphamide-induced male reproductive toxicity.

Histomorphometrical evaluation of extensor digitorum longus muscle in sciatic nerve regeneration using tissue engineering in rats.

Skeletal muscle atrophy induced by denervation is one of the common disorders in traumatic nerve injuries. The aim of this study was the evaluation of histomorphometrical changes of extensor digitorum longus muscle after denervation and its regeneration by tissue engineering. Ninety adult male Wistar rats were randomly divided into six main groups (n = 15) in three time periods (2, 4 and 8 weeks; n = 5). Control group was treated without surgery, in transection (Tr) group left sciatic nerve was transected, in scaffold (S) group only collagen gel scaffold was used, in mast cell (MC) group mast cells were used, mesenchymal stem cell (MSC) group was treated with mesenchymal stem cells and in MC+MSC group, mast cells along with mesenchymal stem cells were used. In the cellular groups, the scaffold and cells were mixed and placed in the transected nerve gap. The average diameter of muscle fibers, ratio of the muscle fibers nuclei to the fibrocytes nuclei (mn/fn), ratio of the muscle fibers nuclei number to the muscle fibers number (mn/mf), the average ratio of blood vessels to muscle fibers number (v/mf) and muscles weight in Tr group were the lowest compared to the other groups; but, in cellular and S groups, amelioration was observed according to the time period. However, in MC+MSC group, there were the highest ameliorative results. This study revealed that simultaneous use of MCs and MSCs mixed with collagen gel scaffold can be considered as a suitable approach to improve denervated skeletal muscle atrophy associated with sciatic nerve injury.

Protective effect of ethyl pyruvate on testicular histology and fertilization potential in cyclophosphamide treated mice.

In the present study, we aimed to address the use of ethyl pyruvate (EP) against the harmful effects of cyclophosphamide (CP) treatment. Thirty-nine adult male mice were divided into three groups including control group received normal saline [0.20 mL per day; intraperitoneally (IP)], CP group received CP (15.00 mg kg-1 per week, IP) and CP+EP group received EP (40.00 mg kg-1 per day, IP) along with CP and treated for 35 days. After preparation of paraffin sections and Hematoxylin and Eosin staining, the histomorphometric studies were performed on the testicular tissue. Additionally, the serum superoxide dismutase (SOD) and testosterone level, testis malondialdehyde (MDA) and in vitro fertilization rate were assessed. The results showed an increase in the tubal differentiation index, repopulation index, spermiogenesis index, thickness of testicular capsule, mean distribution of active Sertoli cells, SOD and testosterone levels of the CP+EP group in comparison with the CP group. Moreover, the MDA levels in the CP+EP group were lower than the CP group. An increase occurred in the percentage of fertilization in the CP+EP group compared to the CP group. Results of this study revealed that the EP ameliorates deleterious side effects of CP on testicular histology and in vitro fertility.

Histological Evidence for Therapeutic Induction of Angiogenesis Using Mast Cells and Platelet-Rich Plasma within A Bioengineered Scaffold following Rat Hindlimb Ischemia.

OBJECTIVE: Peripheral arterial disease results from obstructed blood flow in arteries and increases the risk of amputation in acute cases. Therapeutic angiogenesis using bioengineered tissues composed of a chitosan scaffold that was enriched with mast cells (MCs) and/or platelet-rich plasma (PRP) was used to assess the formation of vascular networks and subsequently improved the functional recovery following hindlimb ischemia. This study aimed to find an optimal approach for restoring local vascularization. MATERIALS AND METHODS: In this experimental study, thirty rats were randomly divided into six experimental groups: a. Ischemic control group with right femoral artery transection, b. Ischemia with phosphate-buffered saline (PBS) control group, c. Ischemia with chitosan scaffold, d. Ischemia with chitosan and MCs, e. Ischemia with chitosan and PRP, and f. Ischemia with chitosan, PRP, and MCs. The left hind limbs served as non-ischemic controls. The analysis of capillary density, arterial diameter, histomorphometric analysis and immunohistochemistry at the transected locations and in gastrocnemius muscles was performed. RESULTS: The group treated with chitosan/MC significantly increased capillary density and the mean number of large blood vessels at the site of femoral artery transection compared with other experimental groups (P<0.05). The treatment with chitosan/MC also significantly increased the muscle fiber diameter and the capillary-to-muscle fiber ratio in gastrocnemius muscles compared with all other ischemic groups (P<0.05). CONCLUSION: These findings suggested that chitosan and MCs together could offer a new approach for the therapeutic induction of angiogenesis in cases of peripheral arterial diseases.

Ameliorative Effect of Crocin on Sperm Parameters and In Vitro Fertilization in Mice under Oxidative Stress Induced by Paraquat.

BACKGROUND: Paraquat (PQ) is an herbicide that is genotoxic and cytotoxic for male germ cells. In this study, we investigated the protective role of crocin (Cr) against the destructive effects of PQ on sperm quality and in vitro fertilization (IVF) outcomes. MATERIALS AND METHODS: In this experimental study, a total of 28 male mice (20-25 g) were divided into four groups: control, which received intraperitoneal (IP) injections of 0.1 ml normal saline per day; PQ group received IP injections of PQ (5 mg/kg/day); experimental (PQ+Cr group) received PQ along with IP injections of Cr (200 mg/kg/day); and positive control (Cr) received IP injections of Cr (200 mg/kg/day). In the last two weeks of the treatment period (35 days of treatment), 16 non-pregnant mice were stimulated to receive adult oocytes. At the end of the treatment period, after euthanizing the mice, the sperms were extracted from the epididymis of each mouse and prepared for evaluation of sperm parameters and IVF. RESULTS: In the PQ+Cr group, Cr caused a significant increase in the average number of sperms and the mean percentage of motile and viable sperms. There was a significant decrease in the mean number of immature and DNA-damaged sperms compared to the PQ group (P<0.001). IVF evaluation in the PQ+Cr group showed that the mean percentage of fertilization, two- and four-cell embryos, blastocysts, and hatched embryos significantly increased. Cr caused a significant decrease in the mean percentage of the arrested embryos compared to the PQ group (P<0.001). However, the Cr group did not have any toxic effects on sperm quality or IVF results. CONCLUSION: The findings of this study showed that Cr, due to its effective and potent antioxidant properties, could reduce or suppress the destructive effects on sperm parameters and IVF caused by PQ.

Anatomical and cytohistological study of the pituitary gland in adult turkey.

In order to conduct this study, eight adult turkey heads were obtained. Pituitary glands were harvested following cranial bones removal and examined morphologically and anatomically as well as topographically. Then, tissue sections were prepared and stained using Hematoxylin and Eosin, Alcian blue, orange G and periodic acid-Schiff staining techniques. The results showed that turkey pituitary gland as a pea-sized structure is located in the ventral part of the cerebrum and composed of adenohypophysis and neurohypophysis parts. Moreover, histological analyses revealed that sinusoids are well-developed at the distal part of the adenohypophysis and irregular masses of endocrine cells exist among them. Distributions of basophilic cells in the distal part of adenohypophysis were significantly higher than those of other endocrine cells, while the acidophilic cells had the lowest distribution. Lower and higher numbers of chromophobe cells were also found compared to those of basophilic and acidophilic cells, respectively. These findings were mostly similar to the other birds' pituitary gland anatomical and histological features, but there were also differences in cellular elements distributions along with infundibular cavity topography.

Histomorphometric and immunohistochemical evaluation of angiogenesis in ischemia by tissue engineering in rats: Role of mast cells.

The aim of this study was to find a proper method for improvement of ischemic condition in the rat hind limb and also to observe the efficacy of cell engraftment with alginate/gelatin three-dimensional scaffolds. Eighteen male Wistar rats weighing 200 to 250 g were randomly divided into three groups (n = 6) including a) ischemia group; in which femoral artery was removed after ligation at the distance of 5 mm, b) scaffold group; in which hydrogel scaffold was added to the site of transected femoral artery and c) test group; in which in addition to hydrogel scaffold, mast cells (MCs) were also added (1 × 106 cells). Analysis of capillary density, artery diameter, histomorphometric parameters and immunohistochemistry in transected location were done on day 14 after femoral artery transection. The average number of blood capillary was significantly higher in the test group than other groups. Also, the average number of medium and large blood vessels was significantly higher in the test group compared to ischemia and scaffold groups. Application of MCs through the use of hydrogel scaffolds (alginate/gelatin) can be considered as a new approach in the application of stem cells for therapeutic angiogenesis under ischemic conditions which can improve the angiogenesis process in patients with peripheral artery diseases.

Effects of Pyrroloquinoline Quinone (PQQ) on Ischemia-Reperfusion Injury in Rat Ovaries: Histological and Biochemical Assessments.

OBJECTIVE: To evaluate effects of pyrroloquinoline quinone (PQQ) on ischemia-reperfusion injury using a rat ovary model. METHOD: Thirty healthy female Wistar rats with 250g were randomized into five experimental groups (n = 6): Group SHAM: The rats underwent only laparotomy. Group Ischemia: A 3- hour ischemia only. Group I/R: A 3-hour ischemia and a 3-hour reperfusion. 30 min before termination of reperfusion 20 µL soybeen oil (Solvent of PQQ) was administered. Group I/PQQ: A 3-hour ischemia only and 20 µL (10 mg/kg) intraperitoneal administration (IP) of PQQ 2.5 hours after induction of ischemia. Group I/R/PQQ: A 3-hour ischemia, a 3-hour reperfusion and 20 µL (10 mg/kg) IP of PQQ 2.5 hours after induction of ischemia. RESULTS: Animals treated with PQQ showed significantly ameliorated development of ischemia and reperfusion tissue injury compared to those of other groups (p=0.001). The significant higher values of SOD, GPO and GST were observed in I/R/PQQ animals compared to those of other groups (p=0.001). Damage indicator (MDA) was significantly lower in I/R/PQQ animal compared to those of other groups (p=0.001). CONCLUSION: Intraperitoneal administration of PQQ could be helpful in minimizing ischemia-reperfusion injury in ovarian tissue exposed to ischemia.

Ameliorative effects of crocin on paraquat-induced oxidative stress in testis of adult mice: An experimental study.

BACKGROUND: Paraquat (PQ), as a pyridine compound, is widely used worldwide to control annual weeds. The oxidative stress caused by PQ can cause deleterious changes in the testicular tissue. OBJECTIVE: An investigation on the protective effects of Crocin (CCN) against PQ-induced oxidative damages and apoptotic indices in testicular tissue. MATERIALS AND METHODS: Twenty-eight adult male albino mice (20-25 gr) were divided into four groups (n = 7/each). The control group received 0.1 ml/day of normal saline by intraperitoneal injection (IP); sham-control group received PQ 5 mg/kg/day, IP, and the experimental groups received PQ (CCN+PQ) and CCN-sole (200 mg/kg/day, IP), respectively, for 35 continuous days. At the end of the treatment period, the testes were dissected out and used for biochemical, molecular, and histological analyses. The expressions of tumor suppressor p53, B-cell lymphoma 2 (bcl-2), and caspase-3 were considered as hallmark factors of mitochondria-dependent apoptosis. Moreover, the testicular superoxide dismutase (SOD) and malondialdehyde (MDA) were evaluated as key biomarkers for oxidative stress. RESULTS: The PQ significantly (p < 0.02, p < 0.01) diminished the spermatogenesis indices and SOD, increased MDA levels, and enhanced the apoptosis-related gene expression. However, the co-administration of CCN and PQ significantly (p < 0.01, p < 0.01, p < 0.02) ameliorated the spermatogenesis ratio, upregulated the SOD level as well as bcl-2 expression, and reduced the MDA content and apoptosis vs the PQ-sole group. CONCLUSION: This study showed that the antioxidant properties of CCN enable to ameliorate the PQ-induced destructive effects by upregulating the testicular structure, antioxidant and apoptotic status.

Protective effects of Cornus mas fruit extract on methotrexate-induced alterations in mice testicular tissue: Evidences for histochemical and histomorphometrical changes in an animal model study.

Methotrexate (MTX) as a chemotherapeutic agent, has adverse effects on reproductive organs by enhancing oxidative stress. In this study, the protective effects of Cornus mas fruit extract (CMFE) against MTX side effects were evaluated. Forty-eight mature male NMRI mice were divided into six groups: group 1 (control) received 0.10 mL per day of normal saline intraperitoneally (IP), group 2 received MTX (20.00 mg kg-1 per week, IP), group 3 received MTX along with CMFE 250 mg kg-1 per day by oral gavage, group 4 received MTX along with CMFE 500 mg kg-1 per day by oral gavage, group 5 received MTX plus 1000 mg kg-1 per day of CMFE by oral gavage, and group 6 received 1000 mg kg-1 per day of CMFE extract, orally. All animals were treated for 35 consecutive days. Thickness of testicular capsule and germinal epithelium and diameter of seminiferous tubules were measured. Intra-cytoplasmic levels of carbohydrate, unsaturated fatty acid (UFA) and alkaline phosphatase were assessed. Serum level of testosterone and testicular total antioxidant capacity (TAC) were also evaluated. The results demonstrated that MTX administration caused morphometrical parameters except the thickness of testicular capsule were significantly different in comparison to control group and decreased cytoplasmic concentration of carbohydrate in the first three layers of germinal epithelium and increased the UFA levels. Contrarily, CMFE ameliorates the condition. Moreover, CMFE increased testosterone level and increased the MTX-reduced TAC level. In conclusion, it was revealed that CMFE decreased the cellular atrophy by controlling the energy substrate utilization based on lipids and carbohydrates via provoking the testicular antioxidant status.

Developing Rat Bone Marrow Derived Mast Cells by the Splenic Cells Culture Supernatant of Rat and Mouse.

BACKGROUND: Mast cells play a critical role in the pathogenesis of various immunological and non-immunological diseases. It is now accepted that culturing primary mast cells considered as a tool for investigation role of mast cells in diseases. Development of various animal primary mast cells and their function could be used for the translational studies in the pathogenesis of human diseases. The aim of the study was to develop simple and cost-efficient method for differentiation and culture of rat mast cells from bone marrow by using rat and mouse spleen supernatant. MATERIALS AND METHODS: Bone marrow cells from 10 to15-weeks-old male rats was obtained and cultured for three weeks on cell culture medium. After that, purity of cells was approved by FCɛRI and CD117 antibodies, toluidine blue and Immunohistochemistry (IHC). RESULTS: After 3 weeks continuous culturing, high purity of cells was found. CD117, CD34 expression and tryptase were 80.1, 76.89 and 87.9%, respectively by rat splenic supernatant, whereas 85.4, 83.07 and 82.1%, respectively with mouse splenic supernatants. Besides, rat spleen supernatant developed 91.4% and mouse splenocyte supernatant developed 89.7% mast cells based on surface markers. CONCLUSION: The data presented in this study indicated equal maturation and differentiation of bone marrow derived rat mast cells by using both spleen supernatants.

The effect of vitamin C on mice hemolytic anemia induced by phenylhydrazine: an animal model study using histological changes in testis, pre-implantation embryo development, and biochemical changes.

OBJECTIVES: The aim of the present study was to assess the effects of vitamin C (Vit C) on hemolytic anemia induced by phenylhydrazine (PHZ). MATERIALS AND METHODS: Twenty-four healthy male mice were divided into four groups, randomly: Control group (0.1 ml/day, normal slaine, IP), PHZ group that received only PHZ 8 mg/100 g/48 hr, IP, PHZ+Vit C group that received PHZ 8 mg/100 g/48 hr, IP and Vit C 100 mg/kg BW-1/day by gavage and Vit C group that received 100 mg/kg BW-1/day Vit C by gavage. After 35 days, germinal cells, RNA damage, sperm parameters, testis malondialdehyde (MDA) content, serum total antioxidant capacity (TAC), pre-implantation embryo development and mRNA levels of cyclin D1 and c-myc in two-cell, and morula and blastocyst stages were assessed. RESULTS: Vit C reduced the RNA damage, enhanced sperm quality, promoted pre-implantation embryo development and improved testicular antioxidant and endocrine status (P<0.05). Vit C reduced cyclin D1 expression and up-regulated c-myc mRNA level in two-cell, morula, and blastocyst embryonic cells. CONCLUSION: Vit C enhanced the fertilizing potential by ameliorating the endocrine status, antioxidant capacity, and sperm quality. Finally, the cyclin D1 and c-myc gene expressions were regulated in PHZ+Vit C treated group that promoted the embryo development.

Antioxidant effects of Spirulina platensis (Arthrospira platensis) on cyclophosphamide-induced testicular injury in rats.

Cyclophosphamide (CP) is known to reduce fertility. The protective effects of Spirulina plantesis (SP) against CP-induced testicular toxicity were investigated. Male Wistar rats were categorized into eight groups (n = 7). Four groups of rats were administered CP at a dose of 5 mg in 5 mL distilled water kg-1 per day orally. Two of these groups were received SP (500 and 1000 mg kg-1 per day) orally after CP administration. One of these groups was also received vitamin E (100 mg kg-1 per day) intraperitoneally. A vehicle treated control group, two SP control groups (500 and 1000 mg kg-1 per day) and a vitamin E control group were also included. Body and testes weights, sperm count, serum levels of glutathione peroxidase (GPx), malondialdehyde (MDA), histological and histomorphometric alternations in testes were investigated after four weeks. The CP-treated group exhibited significant decreases in the body and testes weights and spermatogenic activities. Several histological alterations were observed in this group. The CP treatment caused a significant reduction in sperm count, in serum level of GPx, as well increased serum concentration of MDA. The SP co-administration caused an increase in GPx serum level, a decrease in MDA serum level and improvements in histological and histomorphometric alternations. Vitamin E co-treatment showed partial recovery in above-mentioned parameters. These results suggest that SP due to a reduction in oxidative stress has more effective protection against CP-induced reproductive damages in rat than vitamin E.

Local Xenotransplantation of Bone Marrow Derived Mast Cells (BMMCs) Improves Functional Recovery of Transected Sciatic Nerve in Cat: A Novel Approach in Cell Therapy.

OBJECTIVE: To determine the effects of bone marrow derived mast cells (BMMCs) on functional recovery of transected sciatic nerve in animal model of cat. METHOD: A 20-mm sciatic nerve defect was bridged using a silicone nerve guide filled with BMMCs in BMMC group. In Sham-surgery group (SHAM), the sciatic nerve was only exposed and manipulated. In control group (SILOCONE) the gap was repaired with a silicone nerve guide and both ends were sealed using sterile Vaseline to avoid leakage and the nerve guide was filled with 100 µL of phosphate-buffered saline alone. In cell treated group ([SILOCONE/BMMC) the nerve guide was filled with 100 µL BMMCs (2× 106 cells/100 µL). The regenerated nerve fibers were studied, biomechanically, histologically and immunohiscochemically 6 months later. RESULTS: Biomechanical studies confirmed faster recovery of regenerated axons in BMMCs transplanted animals compared to control group (p<0.05). Morphometric indices of the regenerated fibers showed that the number and diameter of the myelinated fibers were significantly higher in BMMCs transplanted animals than in control group (p<0.05). In immunohistochemistry, location of reactions to S-100 in BMMCs transplanted animals was clearly more positive than that in control group. CONCLUSION: BMMCs xenotransplantation could be considered as a readily accessible source of cells that could improve recovery of transected sciatic nerve.

Mouse Bone Marrow-Derived Mast Cells Induce Angiogenesis by Tissue Engineering in Rats: Histological Evidence.

OBJECTIVES: Therapeutic angiogenesis is employed to induce vascular network formation and improve functional recovery in ischemia. The aim of this study is to find an appropriate method to recover local ischemic conditions. MATERIALS AND METHODS: In this experimental survey, 20 male Wistar rats weighing approximately 200-250 g were randomly divided into four experimental groups respectively: ischemia group in which the femoral artery was transected; phosphate buffer solution group (PBS) in which the femoral artery transected location was immersed with PBS; chitosan (CHIT) group in which the transected location was immersed in a 50 µL CHIT solution; and mast cell transplanted group in which the transected location was immersed with a mixture of 50 µL CHIT and 50 µL PBS that contained 1×106 mast cells. RESULTS: On day 14 after surgery, mean numbers of blood vessels of different sizes in the CHIT/mast cell group significantly increased compared to the other experimental groups (P<0.05). CONCLUSIONS: Our data suggest that mast cell reconstitution could offer a new approach for therapeutic angiogenesis in cases of peripheral arterial diseases.

Protective effect of ethyl pyruvate on mice sperm parameters in phenylhydrazine induced hemolytic anemia.

The aim of the present study was to assess the protective effect of ethyl pyruvate (EP) on sperm quality parameters, testosterone level and malondialdehyde (MDA) in phenylhydrazine (PHZ) treated mice. For this purpose, 32 NMRI mice with the age range of 8 to 10 weeks, weight average 26.0 ± 2.0 g, were randomly divided into four equal groups. The control group (1) received normal saline (0. 1 mL per day) by intraperitoneal injection (IP). Group 2 (PHZ group) was treated with initial dose of PHZ (8 mg 100 g(-1), IP) followed by 6 mg 100 g(-1) , IP every 48 hr. Group 3, (Group PHZ+EP) received PHZ (according to the previous prescription) with EP (40 mg kg(-1), daily, IP). Ethyl pyruvate group (4) received only EP (40 mg kg(-1), daily, IP). Treatment period was 35 days. After euthanasia, sperms from caudal region of epididymis were collected and the total mean sperm count, sperm viability, motility and morphology were determined. Testis tissue MDA and serum testosterone levels of all experimental groups were also evaluated. A considerable reduction in mean percentage of number, natural morphology of sperm, sperm motility and viability and serum testosterone concentration besides DNA injury increment among mice treating with PHZ in comparison with control group were observed. However, in PHZ+EP group the above mentioned parameters were improved. This study showed that PHZ caused induction of toxicity on sperm parameters and reduction of testosterone as well as the increment of MDA level and EP as an antioxidant could reduce destructive effects of PHZ on sperm parameters, testosterone level and lipid peroxidation.

Ethyl Pyruvate Ameliorates The Damage Induced by Cyclophosphamide on Adult Mice Testes.

BACKGROUND: Cyclophosphamide (CP) is a chemotherapy drug which causes deleterious effects on testicular tissue and increases free radicals in the body. The aim of this study is to investigate the protective effects of ethyl pyruvate (EP) on testicular improvement in CP treated animals. MATERIALS AND METHODS: In this experimental study, 15 male mice (6-8 weeks) were divided into 3 groups. The control group received normal saline (0.1 ml/day), intraperitoneal (IP), CP group received CP (15 mg/kg/week, IP), and the CP+EP group received EP (40 mg/kg/day, IP) plus CP. After 35 days, we assessed serum total antioxidant capacity (TAC) along with histomorphometric and histochemical analyses of the testicles. RESULTS: The mean thickness of the germinal epithelium, diameter of seminiferous tubules, and the number of Leydig cells in the CP+EP group were higher than those of the CP group (P<0.05). The number of the mast cells in the CP+EP group significantly reduced compared with the CP group (P<0.05). Alkaline phosphatase (ALP), periodic acid-schiff (PAS) positive reactions and lipid granules in cytoplasm of the Leydig cells in the CP group increased compared with the other groups (P<0.05). TAC in the CP group significantly reduced compared with the other groups (P<0.05). CONCLUSION: This study showed the ability of EP to reduce the destructive side effects of CP in the adult mice reproductive system.