Effects of maternal diazinon exposure on frontal cerebral cortical development in mouse embryo.

BACKGROUND: Diazinon including organophosphate (OP) that is widely used in agriculture and animal husbandry industry and the risk of human infection with the toxin and their toxicity. METHODS: Pregnant balb/c mice (30-35 g) were randomly divided into five groups of five: the control group (no intervention), two sham groups (emulsifier 0.52, and 5.2 liters/volume). From the seventh to the eighteenth day of pregnancy, two experimental groups received diazinon inhaled 1.3 (EXP1) and 13 liters/volume (EXP2) for 40 min every other day, respectively. On the 18th day of pregnancy, the animals were killed and their embryos were removed to appraisal the growth of fetus and development of the frontal cortex. A computer-assisted morphometric quantitative images analysis were performed on the frontal cerebral cortex (FCC) including cortical plate (CP), intermediate zone (IZ) and matrix (proliferative) zone (MZ) of the mouse embryos. FINDINGS: The average of crown-rump length and weight of the embryos in the experimental groups were increased without any significant difference. The mean fetal FCC thickness in the EXP2 group was significantly reduced compared to the control group, CP thickness was remarkably increased in fetuses exposed to diazinon. Comparing the mean thickness of MZ and IZ in EXP groups with the sham and control groups indicated a significant decrease. The positive K-67 cells in the FCC of the EXP2 group were significantly reduced. DISCUSSION: Exposing diazinon during pregnancy can reduce brain development and would be neurotoxic to the developing brain and can lead to behavioral changes in the offspring.

Curcumin and Biodegradable Membrane Promote Nerve Regeneration and Functional Recovery After Sciatic Nerve Transection in Adult Rats.

BACKGROUND: Curcumin has immunomodulatory anti-inflammatory, antioxidant, and neuroprotective properties. The goal of this study was to determine the effects of curcumin and biodegradable membrane on nerve healing in rat sciatic nerve transected injuries. METHODS: Rats were divided into groups: (1) control group (Ctrl), (2) curcumin group (Cur), (3) membrane group (Mem), and (4) membrane and curcumin group (Mem + Cur). Functional recovery was evaluated at 2, 4, 6, and 8 weeks after surgery. At the end of the eighth week after surgery, histological assessments were done. RESULTS: At the end of 8th week after surgery, functional assessments (sciatic nerve index, withdrawal reflex latency, and electromyography) in the Mem + Cur group improved compared with other groups (P < 0.05). Histological results (number of nerve fibers, diameter of nerve fibers, and myelin thickness) improved in the Mem + Cur group compared with the control, Cur, and Mem groups (P < 0.05). CONCLUSION: The present study showed the positive effects of Mem + Cur on nerve regeneration of transected sciatic nerve in rat model.

The role of apoptosis in the cellular response of liver and spleen of BALB/c mice in cutaneous leishmaniasis.

BACKGROUND: Cutaneous leishmaniasis is a common parasitic disease in Iran being mainly caused by Leishmania (L.) major. The aim of this study was to investigate the occurrence of apoptosis in the spleen and liver of female mice infected with L. major. METHODS: BALB/c mice were randomly assigned into the control and experimental groups (ten mice per group). The experimental groups were subcutaneously inoculated with promastigotes of L. major at stationary phase. The animals were sacrificed after 20, 40, 60, 90, and 120 days of injection. The liver and spleen were analyzed for various parameters of apoptosis. RESULTS: Activities of superoxide dismutase and caspase-3, levels of superoxide anion production and malondialdehyde, and the percent of DNA fragmentation were increased in the liver and spleen of the infected mice. Catalase activity in the liver was increased, while glutathione level in both tissues was decreased after 90 and 120 days of infection. The numbers of apoptotic nuclei in the spleen were higher than the liver at 90 and 120 days post-infection using the TUNEL method. CONCLUSION: L. major infection induces a time-dependent increase in apoptosis in the liver and spleen as evidenced by the production of ROS, increasing activation of caspase-3, elevated DNA fragmentation, and increasing lipid peroxidation. Induction of oxidative stress was observed in the liver and spleen after 90 and 120 days of initiation of the infection. However, the spleen tissue appears to be more sensitive to the infection to L. major on oxidative stress and apoptosis induction compared with the liver tissue.

Neuronal injury and cytogenesis after simple febrile seizures in the hippocampal dentate gyrus of juvenile rat.

PURPOSE: Although simple febrile seizures are frequently described as harmless, there is evidence which suggests that hippocampal damage may occur after simple febrile seizures. This study aimed to investigate possible neuronal damages as well as alterations in cytogenesis in the hippocampal dentate gyrus following simple febrile seizures. METHODS: Simple febrile seizure was modeled by hyperthermia-induced seizures in 22-day-old male rats. The brains were removed 2 or 15 days after hyperthermia in all rats with (n=20) and without (n=10) occurrence of seizures as well as in control animals (n=10). The sections were stained with hematoxylin and eosin to estimate the surface numerical density of dark neurons. Ki-67 immunohistochemistry was performed to evaluate changes of cytogenesis following simple febrile seizures. RESULTS: Hyperthermia induced behavioral seizure activities in 67 % of the rats. The numerical densities of dark neurons as well as the mean Ki-67 index (the fraction of Ki-67-positive cells) were significantly increased in dentate gyrus after induction of seizures by hyperthermia compared to both controls and rats without seizure after hyperthermia. Both the seizure duration and intensity were correlated significantly with numerical densities of dark neurons (but not with Ki-67 index). CONCLUSION: The data indicate that simple febrile seizures can cause neuronal damages and enhancement of cytogenesis in the hippocampal dentate gyrus, which were still visible for at least 2 weeks. These findings also suggest the correlation of febrile seizure intensity and duration with neuronal damage.

Minocycline Enhance the Restorative Ability of Olfactory Ensheathing Cells by the Upregulation of BDNF and GDNF Expression After Spinal Cord Injury.

Introduction: Spinal Cord Injury (SCI) is a global public health issue that results in extensive neuronal degeneration, axonal and myelin loss, and severe functional deficits. Neurotrophic factors are a potential treatment for reducing secondary damage, promoting axon growth; they are responsible for inducing myelination after injury. Olfactory Ensheathing Cells (OECs) and minocycline have promoted locomotor function after SCI. The present study investigated the neuroprotective effects of combined treatment with minocycline and OECs on spinal cord injury related to Brain-Derived Neurotrophic Factor (BDNF) and Glial Derived Neurotrophic Factor (GDNF) expressions after SCI. Methods: Adult female rats were used to experimental SCI by weight compression method. Rats received an intraperitoneal minocycline injection (90 mg/kg) immediately after SCI and 24 h after injury. OECs were transplanted one week after the injury. The hindlimb function was assessed using Basso Beattie Bresnahan (BBB) locomotor rating scale and Electromyography (EMG). After 5 weeks, the spinal cord segment centered at the injury site was removed for histopathological analysis. Immunohistological and western blot assays were performed to observe the expression of NeuN, BDNF, GDNF, and Myelin Basic Protein (MBP). Results: SCI induced the loss of locomotor function with decreased BDNF and GDNF expressions in the injury site. Minocycline+OECs increased the score of the BBB locomotor scale and increased spared tissue in the injury site. Immunohistochemical results suggested that NeuN expression significantly increased in the minocycline+OECs group than other groups. Moreover, electromyography amplitude in treated rats was increased compared to the control group. BDNF, GDNF, and MBP expressions and the number of ventral motor neurons increased further by minocycline+OECs in SCI rats. Conclusion: The present study provides evidence that minocycline may facilitate recovery of locomotor function by OECs by increasing BDNF and GDNF expressions following SCI. Highlights: Combined treatment with Minocycline and OECs increased the locomotor function.The results showed that BDNF and GDNF expression increased by combined treatment with minocycline and OECs. Plain Language Summary: This study examined the effect of combined treatment with minocycline and olfactory ensheathing cell on the BDNF and GDNF expression after spinal cord injury model in rat. The results showed that injection of minocycline before transplantation of OECs enhances expression of neurotrophic factors that lead to an appropriate environment for transplanted OECs and increases neuronal survival that promotes tissue sparing and functional recovery.

Neuroprotective Effects of Combined Treatment with Minocycline and Olfactory Ensheathing Cells Transplantation against Inflammation and Oxidative Stress after Spinal Cord Injury.

OBJECTIVE: Traumatic spinal cord injury (SCI) is considered one of the most devastating injuries leading to neuronal disruption. Olfactory ensheathing cells (OECs) and minocycline have been shown to promote locomotor function after spinal cord injury. In this study, we have tested the efficacy of combined treatment with minocycline and OECs after contusive spinal cord injury. MATERIALS AND METHODS: In this experimental study, adult female Wistar rats were randomly divided into five groups. Rats received an intraperitoneal injection of minocycline immediately after SCI, and then 24 hours after the injury. Transplantations were performed 7 days after the injury. Functional recovery was evaluated using the Basso, Beattie and Bresnahan scale (BBB). After that, the animals were sacrificed, and T11 segment of the spinal cord was removed after 5 weeks, and then used for histopathological, immunohistochemical, and biochemical assessments. Western blot analysis was applied to determine the protein expression of tumor necrosis factor alpha (TNF-α), interleukin 1 beta (IL1ß) and caspase3. RESULTS: The results of this study showed that the combination of OECs graft and minocycline reduced the functional deficits and diminished cavitation and astrogliosis in spinal tissue. The analysis of protein expression by western blotting revealed that minocycline treatment along with OECs transplantation further decreased the level of IL-1ß, TNF-α, caspase-3, and the oxidative stress as compared with when minocycline or OECs transplantation was used alone. CONCLUSION: The combinatory treatment with OECs graft and minocycline induced a more effective response to the repair of spinal cord injury, and it is considered a therapeutic potential for the treatment of SCI.

Evaluation of nerve growth factor (NGF) treated mesenchymal stem cells for recovery in neurotmesis model of peripheral nerve injury.

BACKGROUND: Peripheral nerve damages are a relatively common type of the nervous system injuries. Although peripheral nerves show some capacity of regeneration after injury, the extent of regeneration is not remarkable. The present study aimed to evaluate the effect of NGF treated mesenchymal stem cells on regeneration of transected sciatic nerve. MATERIALS AND METHODS: In this experimental study, forty-two male Wistar.rats (180-200 g) were randomly divided into 6 groups (n = 7) including control, Membrane + Cell (Mem + Cell), NGF group, NGF + Cell group, NGF + Mem group and NGF + Mem + Cell group. Regeneration of sciatic nerve was evaluated using behavioral analysis, electrophysiological assessment and histological examination. RESULTS: The rats in the NGF + Mem + Cell group showed significant decrease in sciatic functional index (SFI) and hot water paw immersion test during the 2nd to 8th weeks after surgery. (p < 0.001). At 8 weeks after surgery, electrophysiological findings showed that amplitude increased and latency decreased significantly in NGF + Mem + Cell group (p < 0.001). Measured histological parameters showed that number of nerve fibers, number of vessels and percent of vessel area also increased significantly in NGF + Mem + Cell group (p < 0.05). CONCLUSION: The present study showed that NGF in accompany with mesenchymal stem cells improved electrophysiological and histological indices.

Evaluation of dexamethasone treated mesenchymal stem cells for recovery in neurotmesis model of peripheral nerve injury.

OBJECTIVE: Peripheral nerve injuries comprise significant portion of the nervous system injuries. Although peripheral nerves show some capacity of regeneration after injury, the extent of regeneration is not remarkable. The present study aimes to evaluate the regeneration of transected sciatic nerve by a therapeutic value of dexamethasone (DEX) associated with cell therapy (Cell) and biodegradable membrane (Mem) in rat. METHODS: Male Wistar rats (n = 42, 180-200g) were randomly divided into control (Ctrl), Membrane+ Cell, Mem, DEX, DEX+ Cell, DEX+ Mem and DEX+ Cell+ Mem groups. Functional recovery was evaluated at 2, 4, 6, 8 and 12 weeks after surgery using sciatic functional index (SFI), withdrawal reflex latency (WRL) test, electrophysiological and histological analyses. RESULTS: The rats in the DEX+ Cell+ Mem-treated group showed a significant improvement in SFI, WRL and electrophysiological findings during the 2nd to 12th weeks after surgery. In addition, histomorphological findings showed a significant improvement in the DEX+ Cell+ Memtreated group, at 12 weeks after surgery. DISCUSSION: Taken together, use of DEX associated with cell and biodegradable membrane could improve functional and histomorphological properties of the sciatic nerve after injury.

Chitosan-film associated with mesenchymal stem cells enhanced regeneration of peripheral nerves: A rat sciatic nerve model.

OBJECTIVE: Peripheral nerve injuries comprise significant portion of the nervous system injuries. Although peripheral nerves show some capacity of regeneration after injury, but the extent of regeneration is not remarkable. Regeneration might be through the activity of the mesenchymal stem cells (MSCs) which can release growth factors or extracellular matrix components or by the therapeutic effect of some material with the MSCs. The present study aimed to evaluate the regeneration of transected sciatic nerve by a therapeutic value of mesenchymal stem cells (MSCs) associated with chitosan-film (Cs) in rat. MATERIALS & METHODS: Male Wistar rats (n=42, 180-200g) were randomly divided into intact; control; sham; Cs; MSCs; MSCs + Cs groups. Functional recovery was evaluated at 2, 4, 6 and 8 weeks after surgery using sciatic functional index (SFI), hot water paw immersion test, electrophysiological, histological analyses. RESULTS: The rats in the MSCs+Cs group showed significant decrease in SFI and hot water paw immersion test during the 2nd to 8th weeks after surgery. Electrophysiological findings showed a significant decrease in latency time in the MSCs +Cs group. Amplitude of the nerve impulses also increased. Number of nerve fibers with more than 6 µm diameters increased significantly in MSCs+Cs. The number of nerve fibers with less than 4 µm diameters also increased significantly in MSCs+Cs group. CONCLUSION: Taken together, mesenchymal stem cells associated with Cs could improve functional and histomorphological properties of the sciatic nerve after injury which may have some clinical outcomes as well.

Bone Marrow Stromal Cells Associated with Poly L-Lactic-Co-Glycolic Acid (PLGA) Nanofiber Scaffold Improve Transected Sciatic Nerve Regeneration.

Background: Although peripheral nerves show capacity for regeneration after injury to a certain extent, the extent of regeneration is not remarkable. Previous studies have suggested that through the production of growth factors or extracellular matrix components, mesenchymal stem cells may enhance nerve regeneration. Objectives: In the present study, the therapeutic potency of the Bone Marrow Stromal Cells (BMSCs) associated with Poly L-lactic-co-glycolic acid (PLGA) nanofiber Scaffolds on rat sciatic nerve repair was evaluated. Material and Methods: Thirty adult male Wistar rats (220-250 g) were divided randomly into six groups, including control 1 (transected sciatic nerve), control 2 (transected sciatic nerve and stitched), Sham, PLGA, BMSCs, and PLGA+BMSCs. Functional recovery was evaluated at the end of 2nd, 4th, 6th, and 8th weeks after surgery using sciatic functional index (SFI) and hot water test. After killing all rats at the end of 8th week, their sciatic nerves were removed, fixed, and processed for the histological examination and analysis by the Motic software. Results: A significant recovery of the sciatic nerve function was observed in the PLGA+BMSCs transplanted group at the 8th week after surgery as demonstrated by SFI and hot water findings. Histological examinations also showed a significant improvement in the PLGA+BMSCs group compared to the control 1, 2, Sham, PLGA and BMSCs groups. Conclusion: BMSCs associated with PLGA nanofiber scaffold might be useful for improving the functional peripheral nerve repair having some clinical outcome.

Coadministration of Dexamethasone and Melissa officinalis Has Neuroprotective Effects in Rat Animal Model with Spinal Cord Injury.

OBJECTIVE: Spinal cord injury (SCI) causes inflammation, deformity and cell loss. It has been shown that Melissa officinalis (MO), as herbal medicine, and dexamethasone (DEX) are useful in the prevention of various neurological diseases. The present study evaluated combinational effects of DEX and MO on spinal cord injury. MATERIALS AND METHODS: Thirty six adult male Wistar rats were used in this experimental study. The weight-drop contusion method was employed to induce spinal cord injury in rats. DEX and MO were administrated alone and together in different treatment groups. Intra-muscular injection of DEX (1 mg/kg) was started three hours after injury and continued once a day for seven days after injury. Intra-peritoneal (I.P) injection of MO (150 mg/ kg) was started one day after injury and continued once a day for 14 days. RESULTS: Our results showed motor and sensory functions were improved significantly in the group received a combination of DEX and MO, compared to spinal cord injury group. Mean cavity area was decreased and loss of lower motor neurons and astrogliosis in the ventral horn of spinal cord was significantly prevented in the group received combination of DEX and Melissa officinalis, compared to spinal cord injury group. Furthermore, the findings showed a significant augmentation of electromyography (EMG) recruitment index, increase of myelin diameter, and up-regulation of myelin basic protein in the treated group with combination of DEX and MO. CONCLUSION: Results showed that combination of DEX and MO could be considered as a neuroprotective agent in spinal cord injury.

Study of the Effects of Diazinon on Fetal Liver in BALB/c Mice.

BACKGROUND: Diazinon is an organophosphate that is broadly used as a pesticide to control insects and environmental pollutions. This toxic material is absorbed via inhalation, contact, or digestion and affects different tissues. OBJECTIVES: This research was a histomorphometric and immunohistochemical study of the fetal liver of mice after exposure to Diazinon. MATERIALS AND METHODS: Twenty-five pregnant BALB/c mice (25 - 30 gr) were divided into five equal groups in the animal lab of Baqiyatallah University of Medical Sciences, Tehran, Iran. The normal group was without any intervention, and two sham groups received an emulsifier as 0.52 and 5.2 µL/volume (5000 cc in desiccator) and two experimental groups received Diazinon 1.3 and 13µL/volume from the seventh to eighteenth days of pregnancy every other day via forty minutes of inhalation. The pregnant mice were killed on the eighteenth day of gestation and their fetuses were removed and evaluated for fetal growth and liver development. Five fixed fetuses were dehydrated through a series of graded ethanol, embedded in paraffin wax and their whole bodies were sectioned sagittally and stained via the hematoxylin-eosin method. Quantitative computer-assisted morphometric studies were done on the fetal liver tissues occupied by hepatocytes, blood islands, liver sinusoids, and apoptosis. RESULTS: The mean crown-rump of the fetuses and their mean weight were increased in the experimental group as compared to the sham and normal groups, but the differences were not significant. The mean percentage of the hepatocyte area significantly increased in the experimental group as compared to the sham and control groups (P < 0.0001). However, the mean sinusoid area significantly decreased in the experimental group as compared to the sham and control groups. The mean percentage of the area occupied by apoptotic hepatocytes in the experimental group - 13 µL /volume (8.6143 ± 1.00945) and 1.3 µL /volume (6.1091 ± 0.93093) - significantly increased as compared to the normal and sham groups (P < 0.0001). CONCLUSIONS: Our data showed that inhalation of Diazinon during pregnancy increased the hepatocyte area and hepatocyte apoptosis while it decreased the sinusoid area of the fetal liver.

Lavandula angustifolia Extract Improves the Result of Human Umbilical Mesenchymal Wharton's Jelly Stem Cell Transplantation after Contusive Spinal Cord Injury in Wistar Rats.

Introduction. The primary trauma of spinal cord injury (SCI) results in severe damage to nervous functions. At the cellular level, SCI causes astrogliosis. Human umbilical mesenchymal stem cells (HUMSCs), isolated from Wharton's jelly of the umbilical cord, can be easily obtained. Previously, we showed that the neuroprotective effects of Lavandula angustifolia can lead to improvement in a contusive SCI model in rats. Objective. The aim of this study was to investigate the effect of L. angustifolia (Lav) on HUMSC transplantation after acute SCI. Materials and Methods. Sixty adult female rats were randomly divided into eight groups. Every week after SCI onset, all animals were evaluated for behavior outcomes. H&E staining was performed to examine the lesions after injury. GFAP expression was assessed for astrogliosis. Somatosensory evoked potential (SEP) testing was performed to detect the recovery of neural conduction. Results. Behavioral tests showed that the HUMSC group improved in comparison with the SCI group, but HUMSC + Lav 400 was very effective, resulting in a significant increase in locomotion activity. Sensory tests and histomorphological and immunohistochemistry analyses verified the potentiation effects of Lav extract on HUMSC treatment. Conclusion. Transplantation of HUMSCs is beneficial for SCI in rats, and Lav extract can potentiate the functional and cellular recovery with HUMSC treatment in rats after SCI.

Study of Transected Sciatic Nerve Repair by Amniotic Membrane with Betamethasone in Adult Albino Wistar Rats.

BACKGROUND: The aim of this study was to determine the effects of amniotic membrane impregnated with betamethasone on regeneration of transected sciatic nerve injury in adult albino Wister rats. METHODS: In this research, 42 male adult rats were divided into six equal groups. 1) Normal (intact) group: healthy rats without any injury; 2) CONTROL GROUP: sciatic nerve was cut and sutured; 3) Sham group: 0.2 mL culture medium was injected on the epineurium in the injury; 4) Amniotic membrane group (AM): Acellular amniotic membrane was used around the damaged sciatic nerve; 5) Betamethasone group (B): 0.2 mL Betamethasone (4 mg/mL) was injected in the site of damaged nerve and 6) Amniotic membrane group and Betamethasone (AM/B) group: Acellular amniotic membrane impregnated with 0.2 mL betamethasone was used around the damaged sciatic nerve. The rate of recovery was studied by Sciatic Functional Index (SFI), withdrawal reflex latency (WRL) test and electroctrophysiological assessments at 2, 4, 6 and 8 weeks after surgery. Histological assessment was done 8 weeks after surgery. RESULTS: At 8 weeks after surgery, SFI, WRL test and electrophysiological values in AM/B group were significantly improved compared to control and sham groups (P < 0.05). Histological results showed improvement in therapeutic groups, especially AM/B group compared to control and sham groups and other therapeutic groups (P < 0.05). CONCLUSION: The present study showed the positive effects of Amniotic membrane and Betamethasone on nerve regeneration of transected sciatic nerve in a rat model.

Assessment of Neuroprotective Properties of Melissa officinalis in Combination With Human Umbilical Cord Blood Stem Cells After Spinal Cord Injury.

INTRODUCTION: The pathophysiology of spinal cord injury (SCI) has a classically bad prognosis. It has been demonstrated that human umbilical cord blood stem cells (hUCBSCs) and Melissa officinalis (MO) are useful for the prevention of neurological disease. METHODS: Thirty-six adult male rats were randomly divided into intact, sham, control (SCI), MO, hUCBSC, and MO-hUCBSC groups. Intraperitoneal injection of MO (150 mg/kg) was commenced 24 hr post-SCI and continued once a day for 14 days. Intraspinal grafting of hUCBSCs was commenced immediately in the next day. The motor and sensory functions of all animals were evaluated once a week after the commencement of SCI. Electromyography (EMG) was performed in the last day in order to measure the recruitment index. Immunohistochemistry, reverse transcription-polymerase chain reaction, and transmission electron microscopy evaluations were performed to determine the level of astrogliosis and myelination. RESULTS: The results revealed that motor function (MO-hUCBSC: 15 ± 0.3, SCI: 8.2 ± 0.37, p < .001), sensory function (MO-hUCBSC: 3.57 ± 0.19, SCI: 6.38 ± 0.23, p < .001), and EMG recruitment index (MO-hUCBSC: 3.71 ± 0.18, SCI: 1.6 ± 0.1, p < .001) were significantly improved in the MO-hUCBSC group compared with SCI group. Mean cavity area (MO-hUCBSC: 0.03 ± 0.03, SCI: 0.07 ± 0.004, p < .001) was reduced and loss of lower motor neurons (MO-hUCBSC: 7.6 ± 0.43, SCI: 3 ± 0.12, p < .001) and astrogliosis density (MO-hUCBSC: 3.1 ± 0.15, SCI: 6.25 ± 1.42, p < 0.001) in the ventral horn of spinal cord were prevented in MO-hUCBSC group compared with SCI group. CONCLUSION: The results revealed that the combination of MO and hUCBSCs in comparison with the control group has neuroprotective effects in SCI.

Presentation of a novel model of chitosan- polyethylene oxide-nanohydroxyapatite nanofibers together with bone marrow stromal cells to repair and improve minor bone defects.

OBJECTIVES: Various methods for repairing bone defects are presented. Cell therapy is one of these methods. Bone marrow stromal cells (BMSCs) seem to be suitable for this purpose. On the other hand, lots of biomaterials are used to improve and repair the defect in the body, so in this study we tried to produce a similar structure to the bone by the chitosan and hydroxyapatite. MATERIALS AND METHODS: In this study, the solution of chitosan-nanohydroxyapatite-polyethylene oxide (PEO) Nanofibers was produced by electrospinning method, and then the BMSCs were cultured on this solution. A piece of chitosan-nanohydroxyapatite Nanofibers with BMSCs was placed in a hole with the diameter of 1 mm at the distal epiphysis of the rat femur. Then the biomechanical and radiographic studies were performed. RESULTS: Biomechanical testing results showed that bone strength was significantly higher in the Nanofiber/BMSCs group in comparison with control group. Also the bone strength in nanofiber/BMSCs group was significant, but in nanofiber group was nearly significant. Radiographic studies also showed that the average amount of callus formation (radio opacity) in nanofiber and control group was not significantly different. The callus formation in nanofiber/BMSCs group was increased compared to the control group, and it was not significant in the nanofiber group. CONCLUSION: Since chitosan-nanohydroxyapatite nanofibers with BMSCs increases the rate of bone repair, the obtained cell-nanoscaffold shell can be used in tissue engineering and cell therapy, especially for bone defects.

Improved viability of random pattern skin flaps with the use of bone marrow mesenchymal-derived stem cells and chicken embryo extract.

OBJECTIVES: Covering tissue defects using skin flaps is a basic surgical strategy for plastic and reconstructive surgery. The aim of this study was to evaluate the effects of chicken embryo extract (CEE) and bone marrow derived mesenchymal stem cells (BM-MSCs) on random skin flap survival (RSF) in rats. Using chicken embryo extract can be an ideal environment for the growth and proliferation of transplanted cells. MATERIALS AND METHODS: Forty albino male Wistar rats were divided into 4 groups; each group consisted of 10 rats. BM-MSCs and CEE were transplanted into subcutaneous tissue in the area, where the flap would be examined. On the 7(th) postoperative day, the survival areas of the flaps were measured by using digital imaging with software assistance, and tissue was collected for evaluation. RESULTS: Survival area was 19.54±2 in the CEE group and 17.90±2 in the CEE/BM-MSC group when compared to the rates of the total skin flaps, which were significantly higher than the control group (13.47±2) (P<0.05). The biomechanical assessment showed a slight difference, although there was no statistically significant difference between the experimental groups and the control group (P>0.05). CONCLUSION: The findings from this study demonstrated that in operative treatment with BM-MSCs and CEE transplantation could promote flap survival, but the biomechanical parameters were not contrasted with a saline injection.

Evaluation of chitosan-gelatin films for use as postoperative adhesion barrier in rat cecum model.

BACKGROUND: Postoperative adhesions remain a significant complication of abdominal surgery and can result in pain, infertility and potentially lethal bowel obstruction. Pharmacotherapy and barrier devices have reduced adhesion formation to varying degrees in preclinical studies or clinical trials. MATERIALS AND METHODS: In this study, we produced blends between chitosan (Ch) and gelatin (G) with various compositions (Ch/G 100/0, 75/25, 50/50, 25/75 w/w) as candidate materials for prevention of postoperative abdominal adhesion. For in vivo analysis, 30 female rats weighing 200-250 g were divided into 5 groups (One control and 4 treatment groups). Under general anesthesia, the anterior surface of serous membrane in rat was scraped slightly with sterile gauze until obvious congestion and small bleeding drops appeared, then sample films set on the cecum in treatment groups and the intestine was put back into the abdominal cavity, which were then closed. After 4 weeks, the abdominal cavity was reopened and the grades of peritoneal adhesion were studied by macroscopic and pathologic assessments. RESULTS: Our results showed Ch1/G3 films had an insignificant reduction effect on postoperative adhesion, but surprisingly, the sample with more than 25% by weight of chitosan did not have any effect on reducing adhesion formation but also increased inflammation near the cecum. CONCLUSION: Administration of chitosan-gelatin films with higher than 25% weight of chitosan had no effect on reduction of adhesion formation in the rat cecum model.

Macroscopic and pathological assessment of methylene blue and normal saline on postoperative adhesion formation in a rat cecum model.

BACKGROUND: Adhesion formation after abdominal surgery is a major cause of postoperative bowel obstruction, infertility, and chronic abdominal pain. In this study, we evaluated the effect of normal saline and methylene blue (MB) on postoperative adhesion formation in a rat cecum model. METHODS: A total of 30 Wistar female rats in 2 treatment and 1 control groups underwent midline laparotomy and standardized abrasion of the visceral peritoneum. Normal saline and methylene blue were administrated intraperitoneally at the end of the surgical procedure in 2 treatment groups. Fourteen days after surgery, a re-laparotomy was performed for macroscopic and pathological assessment. RESULTS: The adhesion grade and extent of the normal saline group was lower than control and MB groups in macroscopic assessment (P<0.05 for both). A comparison of adhesion stages in pathological assessment showed increment in abdominal adhesion by usage methylene blue 1% and demonstrated significant difference between MB and 2 other groups (P<0.05). CONCLUSIONS: Administrated normal saline individually reduce the adhesion grade near cecum. Conversely, usage of methylene blue 1% may unpredictably increase risk of adhesion formation.

Effects of maternal oral administration of morphine sulfate on developing rat fetal cerebrum: a morphometrical evaluation.

Intrauterine morphine exposure is a risk factor for neurological and behavioral deficit in children, although the precise underlying biological correlate for this is unclear. Female pregnant rats were orally treated with 0.1 mg/ml of morphine solution on the 21st day of gestation. Pregnant rats were killed on the 21st day of gestation and their fetuses were taken out and evaluated for growth and cerebral development. The fetuses were fixed and followed by dehydration through graded ethanol solutions and were then embedded and their heads were coronally sectioned through the frontal cerebral cortex. Quantitative computer-assisted morphometric study was done on the frontal cerebral cortex (FCC) which consists of cortical plate (CP), intermediate (migratory) zone (IZ) and matrix (proliferative) zone (MZ) in the rat embryos. The results showed that morphine exposure caused a significant reduction of fetal weight and crown-to-rump length in morphine exposure group. The present study showed that animals with intrauterine morphine exposure, induced by a period of reduced placental blood flow during the second week of pregnancy, demonstrate reduced both cortical thickness and the numbers of neurons in the developing fetal frontal cerebral cortex (FCC). Histomorphometric evaluation revealed that the thickness of the CP was significantly decreased in the morphine-exposed embryos. In addition, neuronal counting showed that cell proliferation in the CP was suppressed after morphine administration and that the migration of neurons from the matrix zone (MZ) to the cortex was decelerated. In conclusion, these results showed that morphine exposure during the second week of pregnancy could affect brain development in a way, which could lead to neurological and behavioral deficits in the postnatal animal.