Veterinary Research Forum

Abstract Testicular ischemia-reperfusion is accompanied by elevated production of reactive oxygen species. It has been reported that reactive oxygen species are highly reactive to cellular carbohydrates, DNA, lipids and proteins, and result in testicular ischemia-reperfusion injury. Gastrodin is the principal active ingredient isolated from the medicinal plant Gastrodia elata Blume and has anti-oxidative stress effect. In this study, we explored the potential protective activity of gastrodin in rat testicular ischemia-reperfusion injury model and underlying mechanism. Male rats were randomized into 3 groups (sham control, testicular ischemia-reperfusion injury, testicular ischemia-reperfusion injury along with gastrodin injection) (n = 20). Testicular ischemia-reperfusion injury group received 2-hour period of left testicular torsion (720°, counterclockwise) and 4-hour or 3-month period of testicular detorsion. At the onset of testicular detorsion, gastrodin-treated rats were given 100.00 mg kg-1 gastrodin by intraperitoneal route. Following testicular detorsion, testicular tissues were collected for enzymatic activity analysis, oxidative stress evaluation and histopathological examination. The ipsilateral testicular xanthine oxidase activity (source of reactive oxygen species production) and malondialdehyde level (a precise biomarker of reactive oxygen species) were significantly increased in testicular ischemia-reperfusion injury group versus sham control group, while testicular spermatogenic function was decreased. Furthermore, gastrodin administration reduced xanthine oxidase activity and malondialdehyde level in ipsilateral testicular tissue, while improving testicular spermatogenic function. Consequently, it is suggested that gastrodin plays a protective role in testicular torsion/detorsion-induced ischemia/reperfusion injury through inhibiting xanthine oxidase activity to decrease reactive oxygen species formation.

Abstract Bone tissue engineering offers a potential alternative to conventional grafting by combining biocompatible scaffolds with biological stimulants. Polycaprolactone (PCL) is a biodegradable polyester with good mechanical strength and biocompatibility. Hydroxyapatite (HA) is a calcium phosphate mineral that is a major component of bone and has excellent bioactivity and biocompatibility. This research investigated the histopathological effects of PCL-HA nanocomposite scaffolds along with their combination with platelet-rich fibrin (PRF), on the regeneration of bone in rabbit calvarial bone defects. Four circular full-thickness bone defects of 5.00 mm in diameter were created on the calvarial bone of 15 male New Zealand white rabbits. Three defects were filled with PRF, PCL-HA, and PCL-HA/PRF, and one defect was served as a control with no filler. Histopathological evaluations were conducted at 4-, 8-, and 12-weeks post-implantation. Data were evaluated using the Kruskal-Wallis and Mann-Whitney U tests. Significant differences were observed between the treatment and control groups regarding bone regeneration throughout all 12^th weeks studied. In the 4^th weeks, no significant differences in bone regeneration were noted among the treatment groups. In the 8^th weeks, most new bone formation was observed in the PCL-HA/PRF group. Both the PCL-HA and PCL-HA/PRF groups significantly improved bone regeneration compared to the control and PRF groups, with the PCL-HA/PRF group demonstrating the greatest bone formation, and vascularization and the lowest inflammation by the 12^th week. Thus, PCL-HA/PRF could be considered as a suitable alternative to bone grafts and could be increasingly utilized in orthopedic surgery and bone tissue engineering.

Abstract In recent years, liver transplantation has emerged as the standard therapy for several liver disorders. Throughout the procedure, the transplanted liver tissue is subjected to varying degrees of ischemia-reperfusion (IR) damage. Consequently, there has been a long-standing pursuit of substances that can alleviate the harm caused by IR. In our investigation, we employed dapagliflozin as a potential therapeutic agent. Eighteen Wistar rats were divided into three groups (n = 6), including treatment, IR, and control that did not undergo surgical intervention. Two days prior to surgery, the treatment group received dapagliflozin at a dosage of 10.00 mg kg^-1 orally. During surgery, liver ischemia was induced for 1 hr, followed by a 24-hr reperfusion period. The IR group exhibited elevated levels of alanine transaminase, aspartate transaminase, alkaline phosphatase, bilirubin, lactate dehydrogenase, and malondialdehyde compared to the control group. In contrast, the treatment group showed levels of these factors that were closer to those of the control group. While total protein, albumin, and total anti-oxidant capacity decreased in the IR group, this decline was less significant in the treatment group. Analysis of oxidative stress in liver tissue revealed that the treatment group had increased anti-oxidant capacity, and exhibited less oxidative stress compared to the IR group. Furthermore, dapagliflozin was found to reduce the degree of liver edema, necrosis, and vascular hyperemia following IR. Overall, dapagliflozin demonstrates the potential to lessen liver damage, enhance liver tissue regeneration, and mitigate the consequences associated with liver impairment.

Abstract Sterilization in animals serves multiple purposes, such as behavior control, performance improvement, and population management. Chemical sterilization has emerged as a promising non-surgical alternative to traditional methods. This study aimed to investigate the effects of intra-testicular injection of zinc-doped carbon dots (Zn-CDs) nanoparticles as a chemical sterilant in mature rats. Twenty-five rats were randomly divided into five groups, including a control group without injection, a sham group receiving 0.50 mL distilled water, and three treatment groups administered respectively 0.50, 2.00, and 8.00 mg kg^-1 of Zn-CDs synthesized through a hydrothermal process. Following anesthesia with ketamine and xylazine, and aseptic preparation, intra-testicular injections were administered bilaterally. At 60th day post-injections, blood samples were collected to measure serum testosterone levels using chemiluminescence immunoassay. The rats were then surgically castrated to assess sperm parameters and testicular histopathology. Testicular oxidant/anti-oxidant status was also evaluated. The results revealed a dose-dependent reduction in sperm viability, membrane integrity, and motility, accompanied by increased sperm DNA damage. The highest Zn-CDs dose caused the most significant decrease in sperm concentration, as well as severe testicular tissue damage. In addition, anti-oxidant capacity, seminiferous tubules maturation, testosterone production, and spermatogenesis declined with increasing Zn-CDs concentrations in a dose-dependent manner. These findings indicate that intra-testicular injection of Zn-CDs effectively induces infertility in mature rats and holds potential as a chemical sterilization method. With further studies to evaluate safety and efficacy, this approach could be developed as a practical solution for large-scale in situ castration, offering a non-surgical alternative for over-population control programs.

Abstract Spinal cord injury (SCI) results in the demise of neural and glial cells, as well as extensive neuro- inflammation. Hydrogel formulation for prolonged release of melatonin (Mel) has demonstrated enhanced effectiveness and safety. In this study, SCI was induced in rats by contusion at the T₉ vertebrae. Chitosan (CH) /Mel hydrogel was fabricated and characterized using scanning electron microscopy (SEM) and Fourier transform infra-red to examine its specific effects on the apoptotic and histopathological markers of SCI. The scanning electron microscopy images revealed the presence of porosity in the CH/Mel hydrogel. Forty male Wistar rats were randomly divided into five groups (n = 8), including sham, control (SCI-induced treated locally with 100 µL CH hydrogel), and groups 3, 4, and 5 (treated locally immediately after SCI induction with 100 µL CH hydrogel containing 50.00, 100, and 200 mg kg^-1 Mel, respectively). The CH/Mel hydrogel at a dose of 25.00 mg mL^-1 significantly increased cell viability in the U87 cell line after 24 hr of exposure. However, at 48 and 72 hr after exposure, Bax and Bcl2 expressions were significantly increased and reduced in the SCI group, respectively, and CH/Mel hydrogel could alleviate their expressions, especially in higher doses. In addition, S100 protein expression was up-regulated in the SCI group. However, CH/Mel hydrogel down-regulated it in a dose-dependent manner. The histopathological findings demonstrated that CH/Mel hydrogel dramatically improved SCI outcomes, like vacuolar degeneration, necrosis, and severe cystic and axonal degenerations. In conclusion, CH/Mel hydrogel induced neuroprotection and it had the potential to be used as a therapeutic agent for the treatment of SCI.

Abstract Buildup of reactive oxygen species during testicular torsion causes oxidative stress and ischemia-reperfusion (I/R) injury in testis. The purpose of this study was to investigate influence of β-cryptoxanthin (BCX) on I/R injury in testicular torsion/detorsion in mature rats. Thirty mature male Wistar rats were divided into five groups of six animals each, including sham group: In this group, midline incision of the scrotum was performed and the testicles were taken out for 2 hr with a 720-degree rotation, I/R group: In this group, midline incision of the scrotum was performed and the testicles were taken out and undergone ischemia for 2 hr with a 720-degree rotation, I/R/Oil group: In this group, a midline scrotum cut was performed, the testicles were taken out, ischemia was created for 2 hr with a 720-degree rotation, and at the end of ischemia 100 µL of corn oil (BCX solvent) was injected intraperitoneally, I/R/BCX10 group: The same as I/R/Oil group, as well as intraperitoneal administration of 100 µL of BCX (10.00 µg kg^-1) at the end of ischemia, and I/R/BCX40: The same as I/R/Oil group, as well as intraperitoneal administration of 100 µL of BCX (40.00 µg kg^-1) at the end of ischemia. Evaluations were based on histopathological and spermatological parameters and oxidative stress assessments. Histopathological spermatological and oxidative stress parameters values obtained from I/R/BCX40 were significantly different from those of other groups (p < 0.05). It could be concluded that BCX could ameliorate testicular injuries in acute testicular torsion/detorsion in mature rats.

Abstract Osteomyelitis is caused by the local spread of an infected source adjacent to the infection following trauma, bone surgery or joint replacement. The present study aimed to evaluate the effect of vancomycin (Van)/polycaprolactone (PCL) nanocomposite in comparison with curcumin (Cur)/PCL on the healing of experimental osteomyelitis in tibia in rabbits. After induction of osteomyelitis forty adult male New Zealand white rabbits were randomly divided into four groups. Control group: The animals were considered as controls and no scaffolds were used. In PCL/Van group, the created bone defects were filled with the combination of PCL and Van. In PCL/Cur, the created bone defects were filled with the combination of PCL and Cur. Polycaprolactone/Cur/Van group: The created bone defects were filled with the combination of PCL, Cur and Van. Bone samples were taken for histopathological evaluation on the 30^th and 60^th days. For radiological evaluations of the osteomyelitis, radiographs were prepared at time intervals zero (day of surgery), 15, 30, 45, and 60 days after surgery. In order to evaluate hematology, blood was taken on days 0 (day of surgery), 15, 30, 45, and 60. The results of the present study showed that Cur nanocomposite significantly improved the healing process of the rabbit tibia experimental osteomyelitis model compared to the control group. Also, the PCL/Cur/Van group showed the best healing results. In conclusion, PCL/Cur/Van nanocomposite scaffold showed positive effects on the healing process.

Abstract Significant advancements in imaging and surgical methodologies have led to more frequent performance of neurosurgical procedures such as laminectomy in both animal and human patients. Epidural fibrosis (EF) is defined as the excessive formation of scar tissue in the epidural space after lumbar laminectomy, often resulting in recurring postoperative pain. Given the association between postoperative hematoma accumulation at the laminectomy site and the development of EF, the present study aimed to evaluate the preventive impact of tranexamic acid (TXA), an antifibrinolytic agent with well-recognized hemostatic properties across various surgical fields. A rabbit laminectomy model was constructed to assess its effectiveness in reducing EF formation. A total number of 18 adult New Zealand White male rabbits were randomly divided into two groups: The control (saline) group and the treatment (topical TXA) group. Each rabbit underwent a two-level laminectomy at L3-L4. The treatment group received 5.00 mL of 100 mg mL^-1 TXA solution applied topically to the laminectomy site, while the control group received 5.00 mL of saline. Postoperative evaluations included magnetic resonance imaging at week six to assess EF, followed by histopathological examinations to evaluate fibroblast cell density in scar tissue, EF grading and thickness of the dura mater. The analysis of magnetic resonance imaging and histopathologic data revealed significant differences between the two groups indicating that topical administration of TXA might be a promising approach for preventing EF.

Abstract Surgery commonly causes post-operative pain that should be alleviated to prevent complications. In addition to the use of synthetic drugs, there has been a widespread desire to use medicinal plants for surgical pain management. Thymoquinone (TQ), a constituent of Nigella sativa black seeds, exhibits a potent anti-oxidant property. Celecoxib (CLX), a potent non-steroidal anti-inflammatory drug, is widely used in pain management. In the present study, the effects of TQ and CLX on pain caused by hind paw surgical incision were compared. Fifty-six rats were divided into four groups of 14 rats as intact, vehicle, TQ, and CLX groups. In each group, six rats were planned to record pain-related behaviors on days 1 - 10 and eight rats were designed for determination of serum biochemical alterations on days 1 (four rats) and 3 (four rats) after surgery. Oral administrations of TQ and CLX at a same dose of 10.00 mg kg^-1 alleviated paw lifting number (spontaneous pain) and paw withdrawal threshold evoked by von Frey filaments on metal mesh floor, improved the decreased contents of serum total anti-oxidant capacity and superoxide dismutase, and restored the increased levels of serum malondialdehyde and tumor necrosis factor-alpha. The results suggested that TQ by employing anti-oxidant and anti-inflammatory mechanisms, might relieve the pain induced by hind paw plantar incision, being comparable with CLX.

Abstract Testicular ischemia-reperfusion (I/R) injury during testicular torsion is strongly influenced by oxidative stress caused by excessive accumulation of unscavenged reactive oxygen species. This study aimed to investigate the effects of intra-peritoneal administration of Mito-TEMPO (MT) on I/R injury in testicular torsion/detorsion (T/D) in mice. Forty-two male mice were divided into seven groups including 1 control and 6 treatment groups (360° T/D, 720° T/D, 360° T/D + 0.70 mg kg^-1 MT, 360° T/D + 1.00 mg kg^-1 MT, 720° T/D + 0.70 mg kg^-1 MT, and 720° T/D + 1.00 mg kg^-1 MT). After inducing 360° and 720° clockwise testicular torsions for 2 hr, sperm parameters, apoptosis-related genes expression, and in vivo fertility index were evaluated. The results showed that 720° T/D can lead to increased abnormal sperm morphology, sperm DNA damage, and Bax expression, while the Bcl-2 expression was reduced compared to the other groups. In addition, it also had negative effects on sperm total and progressive motilities as well as viability and plasma membrane functionality (PMF). The results also showed that administration of MT to T/D mice can result in a reduction in abnormal sperm morphology, DNA damage, and Bax expression. It could also increase sperm total and progressive motilities, viability and PMF, Bcl-2 expression, and in vivo fertility index. Based on our results, it is concluded that MT, when administered after spermatic cord torsion in mice, provides significant protection against acute testicular T/D injury.

Abstract Scaffolds play a crucial role in tendon healing by providing structural support, promoting cell infiltration, and guiding tissue regeneration. Polycaprolactone (PCL) has been used as a polymer in biological scaffolds for several tissue engineering studies. This study aimed to investigate the effects of curcumin-loaded PCL scaffold on Achilles tendon using a tenotomy model in rats. Twenty adult male Wistar rats were randomized into two groups. In control group, tenotomy and suture placement were performed. The identical intervention followed by the implantation of curcumin-loaded PCL scaffold around the tendon stumps was performed in the treatment group. The nanofibrous PCL scaffold containing 5.00% curcumin was fabricated by electrospinning. Walking track analysis was performed weekly. Then, after 6 weeks, histopathological examination and tendon mechanical tests were performed. The weekly walking track analysis revealed a significant improvement in Achilles functional index in scaffold-treated rats from week three to six. The rate of functional improvement was remarkably slower in the control group. Histopathological examination revealed aseptic inflammation and enhanced neovascularization in the treatment group. Also, collagen arrangement and density were significantly improved in this group compared to the control samples including less regular orientation and loose organization of collagen fibers. Significant increase in mechanical properties, except for strain, was observed in the treatment group. The present study demonstrated that implantation of curcumin-loaded PCL scaffold resulted in increased fibrillar architecture, as well as improved mechanical properties and Achilles functional index in rats. To reduce the biodegradation-induced inflammation, an anti-inflammatory treatment is recommended.

Abstract Cryopreservation and re-transplantation of ovarian tissue is a relatively new technique to preserve fertility. This study evaluated the preservation of ovarian follicles after the vitrification-thawing procedure by implanting ovarian fragments into the gluteus muscle of rabbit. Bilateral ovariectomy was performed on each rabbit. The cortices were separated from the medullae of ovaries and divided into four fragments. One fresh cortical fragment was used as a control, fixed in 10.00% formalin. Three fragments underwent vitrification-thawing for two weeks. Two of these vitrified-thawed fragments were auto-transplanted into separate locations within the opened gluteus muscle, while the final fragment was fixed in 10.00% formalin. Eight weeks after re-implantation, biopsies were collected from the ovarian fragments and fixed in 10.00% formalin. The numbers and areas (µm²) of morphologically normal follicles were evaluated on sections stained with Hematoxylin and Eosin through light microscopy. The follicular morphology in the ovarian fragments immediately after vitrification-thawing remained similar to that of the fresh ovary. The number of morphologically intact follicles was significantly lower in the ovarian fragments eight weeks after re-implantation than fresh ovary and ovarian tissue fragments after vitrification-thawing procedure. However, follicular development to the antral stage was observed in all samples eight weeks after re-implantation. There was no statistical difference in the areas (µm²) of primordial, primary, pre-antral, and antral follicles in the ovarian fragments before cryopreservation, immediately after vitrification-thawing, and following re-implantation procedures. Auto-transplantation of ovarian tissue into the gluteus muscle of rabbits could be a viable approach for preserving ovarian follicles after vitrification-thawing procedure.

Abstract This study investigated the effect of captopril (Cap) on spinal cord ischemia-reperfusion injury (SCII) in rats. Twenty-four adults male Wistar rats were randomly divided into four groups of six animals each: spinal cord ischemia-reperfusion (SCI-R) with Cap (SCI-R + Cap), SCI-R, sham-operated with Cap (SHAM + Cap), and SHAM. The 24 hr and 90 min before ischemia induction, Cap was administered intragastrically (100 mg kg^-1) to the SHAM + Cap and SCI-R + Cap groups. Abdominal aortic clamping was performed in the SCI-R and SCI-R + Cap groups for 40 min. Hindlimb motor function was evaluated using the Tarlov Scale at 4, 6, 12, 24, 48, and 60 hr after SCII. The malondialdehyde (MDA), the ferric-reducing ability of plasma (FRAP) and prooxidant-antioxidant balance (PAB) values were also measured. Throughout the study period, the SCI-R group had significantly lower motor function scores compared to the other groups. The MDA and PAB levels were higher and the FRAP value was lower in the SCI-R group compared to in the SHAM group. The SCI-R + Cap had higher motor function scores compared to the SCI-R group at all time points. There were no significant differences in MDA concentration, FRAP and PAB values between the SCI-R + Cap and SCI-R groups. Captopril may act as a protective agent against SCII in rats based on hind limb motor function assessment.

Abstract Spinal cord injury (SCI) presents challenging and unpredictable neurological recovery. During inflammatory conditions, the amount of serum albumin and nutrition consumption decreases. Currently, it is proposed to measure serum albumin and glucose content in human or animal subjects to predict the recovery rate and the efficiency of treatments following SCI. In this study, the effect of extra-cellular vesicles (EVs) from immortalized human adipose tissue-derived mesenchymal stem cells (hTERT-MSCs) equipped with the ectopic expression of the human indoleamine 2,3-dioxygenase-1 (IDO1) gene on serum albumin and glucose levels was investigated. After pre-clearing steps of 72-hr conditioned media, small EVs (sEVs) were isolated based on the ultra-filtration method. They were encapsulated with a chitosan-based hydrogel. Five experimental groups (female rats, N = 30, ~ 230 g) were considered, including SCI, sham, hydrogel, control green fluorescent protein (GFP)-EVs and IDO1-EVs. The 60.00 µL of hydrogel or hydrogels containing 100 µg sEVs from GFP or IDO1-EVs were locally injected immediately after SCI (laminectomy of the T10 vertebra and clip compression). After 8 weeks, non-fasting serum glucose and albumin levels were measured. The results indicated that the level of serum albumin in the animals received IDO1-EVs (3.52 ± 0.04) was increased in comparison with the SCI group (3.00 ± 0.94). Also, these animals indicated higher glucose levels in their serum (250.17 ± 69.61) in comparison with SCI ones (214 ± 45.34). Although these changes were not statistically significant, they could be considered as evidence for the beneficial effects of IDO1-EVs administration in the context of SCI to reduce hypoalbuminemia and improve energy consumption. More detailed experiments are required to confirm these results.

Abstract The torsion model of testis in a rat was adopted for evaluation of possible effects of propolis (Prop) on ischemia-reperfusion (IS/REP) injury. The healthy male Wistar rats (totally 24 animals) were randomized into four groups (n = 6) and animals experienced bilateral testicular torsions as follows: In sham group just, laparotomy was performed and in IS group, animals experienced a 3 hr period testicular IS. In IS/REP group, a 3 hr period of IS followed by a 3 hr period of testicular REP for left testis and a one-week testicular REP for right testis were done. In this group animals were gavaged by 1.00 mL normal saline 1 hr before the onset of IS. In IS/REP/ Prop group, the same procedures for IS/REP animals were followed as well as gavage of 1.00 mL Prop extract solution 1 hr before the onset of IS. Analyses of biochemistry, histology, inflammatory biomarkers and sperm parameters were carried out. In IS/REP/Prop group, nitric oxide synthase malondialdehyde, myeloperoxidase and 8-hydroxy-2 deoxyguanine in IS/REP/Prop group were significantly decreased and, superoxide dismutase, total glutathione, glutathione peroxidase, glutathione reductase and glutathione S-transferase were significantly increased compared to the other animals. In IS/REP/Prop group, seminiferous tubules (with normal spermatogenesis) showed all stages of spermatogenic cells with plentiful spermatozoa. Tubular deterioration and atrophy and spermatogenic cell loss in were seen in a limited extent. The mean concentrations of Interleukin-1 beta and tumor necrosis factor alpha in IS/REP/Prop were significantly decreased. Sperm quality was significantly improved by Prop in IS/REP/Prop group. It was concluded that Prop could be supportive in diminishing IS/REP injury in testicular tissue exposed to ischemia.

Abstract The present study was designed to evaluate the effects of platelet growth factors and periosteal mesenchymal stem cells on bone healing process, radiographically. Forty male White New Zealand rabbits in five equal groups were used in this study. A 2 mm full thickness bone defect was made in left radial bone of each animal. In group A (control) the defect was left with no medical intervention. In group B the defect was covered by a fibrin membrane. In group C the defect was covered by a fibrin membrane plus platelet growth factors. In group D the defect was covered by a fibrin membrane plus periosteal mesenchymalstem cells, and in group E the defect was covered by a fibrin membrane enriched with platelet growth factors and periosteal mesenchymalstem cells. Radiological evaluation was done immediately after surgery (week 0) and then at the 1^st, 2^nd, 4^th, 6^th and 8^th weeks after operation. At the end of the eighth week, bone samples were taken to evaluate the histopathology. The radiological and histopathological observations showed a superior bone healing in the groups D and E, after eight weeks in comparison with the groups A, B and C. According to this study, it could be concluded that the platelet growth factors and periosteal mesenchymalstem cells could promote bone regeneration in long bone defects in a rabbit model.

Abstract Chitosan bears numerous properties, such as biocompatibility, biodegradability and non-toxicity making it suitable for use in different biomedical fields. Zinc (Zn) is required for fibroblasts proliferation and collagen synthesis as essential elements of wound healing. Its nanoparticles are well known for their capability to enhance wound healing by cell adhesion and migration improvement through growth factors-mediated mechanisms. Poor blood supply and unique histological characteristics of tendon make its regeneration always slow. Also, adhesion formation between tendon and its surrounding tissues is another problem for neotendon to return to its normal structure and functional activities. In this study, a novel tubular scaffold of zinc oxide (ZnO) nanoparticles loaded chitosan has been fabricated for tendon repair. Experimental complete tenotomy of deep digital flexor tendon in a rabbit model was done and scaffolds were placed in the transected area after two ends suturing. After four and eight weeks, adhesion formation around the tendons and tissue reaction to the scaffolds were evaluated macroscopically. Inflammation, angiogenesis and collagen fibers arrangement were also analyzed in histopathological evaluations. After eight weeks, the scaffolds were absorbed completely, adhesions around the tendon were decreased and there was no sign of significant tissue reaction and/or infection in histopathological analyses. The reduced adhesion formation, improved gliding function and better histopathological characteristics suggest this scaffold application as a potential therapy in treatment of tendon acute injuries.

Abstract Operation on the thoraco-abdominal aorta may lead to paraplegia or paraparesis is after spinal ischemia/reperfusion (I/R) injury. In this study, we investigated the protective effect of the spinach extract on spinal cord I/R injury. Thirty-five male Sprague-Dawley rats were divided into five groups: Intact, sham surgery, normal saline (NS), low dose spinach extract (20 mg kg^-1), high dose spinach extract (50 mg kg^-1). Neurological function, biochemical and histological evaluations were performed in 72 hr after ischemia. The mean motor deficit index scores of the spinach extract groups were significantly lower than in the NS group at 72hr after spinal cord ischemia. In addition, Spinach extract groups significantly increased plasma level of total antioxidative capacity and decreased the plasma level of malondialdehyde than the NS group. The spinach extract groups displayed a significantly large number of normal motor neurons compared with the NS group. In conclusion, the present study showed that the spinach extract may preserve more neurons in a rat model of spinal cord I/R injury.

Abstract This study was carried out to assess the different ovarian transplantation sites after short-time autografting. Female rats were randomized into five groups, with six rats in each group, including control (intact), cervical subcutaneous transplanted (CST), back subcutaneous transplanted (BST), subfascial transplanted (SFT) and intramuscular transplanted (IMT) groups. In all experimental groups, the right ovary was removed and transplanted into different sites. After three weeks, ovaries were removed for morphology assessment, follicular counting and the rates of corpus luteum (CL) and cyst formation. Transplanted ovaries in BST and SFT groups were full of cysts and did not have sufficient numbers of intact follicles and were excluded from experiments. In IMT and CST groups, re-anastomosis, follicular development and good homogeneity of the stromal tissue were seen. However, the difference in intact antral follicles between CST (7.92 ± 0.02%) and CST-Op (opposite ovary of CST group) (30.99 ± 0.03%) was significant as well as the difference between CST (7.92 ± 0.02%) and control (10.08 ± 0.01%) groups. In addition, the number of intact primordial follicles in the CST-Op (16.58 ± 0.02%) group was significantly less than that of the control (40.40 ± 0.03%) group. Interestingly, the number of CL was significantly increased in the CST-Op (11.71 ± 0.01%) and IMT-Op (9.16 ± 0.02%) groups compared to the control and experimental groups. Although both intramuscular and subcutaneous sites effectively preserved ovarian follicles after three weeks, cervical subcutaneous site was better suited for auto-transplantation in rat.

Abstract Application of smoke condensate derived from an indirect heating of jennet feces (Sargin) had been recommended by Iranian ancient scientists as a therapeutic agent. The present study was done to evaluate the healing effects of Guajol^® ointment on burn wound in rat. The Guajol^® ointment was prepared from the smoke condensate of Sargin samples. Wistar Rats (n = 50) were randomized into six groups including normal saline, silver sulfadiazine and 1.25%, 2.50%, 5.00% and 10.00% concentrations of Guajol^® ointment. Under general anesthesia, dorsum of the rats were shaved and burn wounds were created using hot plate. Area of wounds and percent of healing were measured. Normal saline had the highest area of wound, followed by 1.25% Guajol^® and silver-sulfadiazine groups. The group treated with 5.00% Guajol^® showed the highest percent of healing. Percent of healing in NS, SSD and 5.00% Guajol^® ointment groups on day 21 were 38.47%, 75.00% and 98.51%, respectively. Microscopic examination of wounds sections of rats treated with 5.00% Guajol^® showed more collagen fibers and fibroblasts cells on day 7. Wounds of 5.00% Guajol^® treated group was covered with healthy epithelial and epidermis tissues and hair follicles on day 21. This was the first report of using Sargin to heal the burn wound of rat. Further studies are recommended for investigation of the other effects of Guajol^® ointment and its possible application in medicine.

Abstract The microorganisms have been noted as the main cause of delayed wound healing.The most common pathogen causing the wound infections is Staphylococcus aureus. Silver nanoparticles (AgNPs) show ample antibacterial activities. In the present study, the effect of AgNPs on mouse wounds inoculated with S. aureus was investigated. Sixty male mice (20 to 30 g) were anesthetized, full-thickness skin wounds were made on their back and then the bacterial suspension was added to each wound bed. Treatments were administered on wound bed topically including gentamicin (8 mg kg^-1), AgNPs (0.08 mg kg^-1, 0.04 mg kg^-1 and 0.02 mg kg^-1) and normal saline in the control group. Wound healing was monitored macroscopically by taking digital photographs on days 0, 7, 14 and 21 of the experiment. Topical application of gentamicin and AgNPs (0.08 and 0.04 mg kg^-1) significantly increased the rate of wound healing more than treatment with AgNPs at a dose of 0.02 mg kg^-1and normal saline. The presence of silver nanoparticles in AgNPs groups (especially 0.08 mg kg^-1) improved wound appearance better than other groups without silver nanoparticles (gentamicin and control groups) and led to lesser wound scars. According to data analysis, healing rate of treated mice with gentamicin and AgNPs (0.08 mg kg^-1) was significantly (p < 0.001) faster than treated mice with other AgNPs doses and normal saline. The results of current study introduced an in vivo nanosilver accelerating effects on the treatment of on S. aureus infected skin wounds.