Investigation of the relationship between inflammation and microbiota in the intestinal tissue of female and male rats fed with fructose: Modulatory role of metformin.

BACKGROUND: It has been reported that High-Fructose (HF) consumption, considered one of the etiological factors of Metabolic Syndrome (MetS), causes changes in the gut microbiota and metabolic disorders. There is limited knowledge on the effects of metformin in HF-induced intestinal irregularities in male and female rats with MetS. OBJECTIVES: In this study, we investigated the sex-dependent effects of metformin treatment on the gut microbiota, intestinal Tight Junction (TJ) proteins, and inflammation parameters in HF-induced MetS. METHODS: Fructose was given to the male and female rats as a 20% solution in drinking water for 15 weeks. Metformin (200 mg/kg) was administered by gastric tube once a day during the final seven weeks. Biochemical, histopathological, immunohistochemical, and bioinformatics analyses were performed. Differences were considered statistically significant at p < 0.05. RESULTS: The metformin treatment in fructose-fed rats promoted glucose, insulin, Homeostasis Model Assessment of Insulin Resistance Index (HOMA-IR), and Triglyceride (TG) values in both sexes. The inflammation score was significantly decreased with metformin treatment in fructose-fed male and female rats (p < 0.05). Moreover, metformin treatment significantly decreased Interleukin-1 Beta (IL-1ß) and Tumor Necrosis Factor-Alpha (TNF-α) in ileum tissue from fructose-fed males (p < 0.05). Intestinal immunoreactivity of Occludin and Claudin-1 was increased with metformin treatment in fructose-fed female rats. HF and metformin treatment changed the gut microbial composition. Firmicutes/Bacteroidetes (F/B) ratio increased with HF in females. In the disease group, Bifidobacterium pseudolongum; in the treatment group, Lactobacillus helveticus and Lactobacillus reuteri are the prominent species in both sexes. When the male and female groups were compared, Akkermansia muciniphila was prominent in the male treatment group. CONCLUSION: In conclusion, metformin treatment promoted biochemical parameters in both sexes of fructose-fed rats. Metformin showed a sex-dependent effect on inflammation parameters, permeability factors, and gut microbiota. Metformin has partly modulatory effects on fructose-induced intestinal changes.

The combined use of carbon nanotubes with synthetic ceramics enhances posterolateral fusion: an experimental study in a rat spinal fusion model.

PURPOSE: The aim of the present study was to evaluate the effectiveness of carbon nanotubes (CNTs)/ HA-tricalcium phosphate (TCP) composite in a posterolateral spinal fusion model. METHODS: At first, CNTs and CNTs/HA-TCP composites were prepared. Twenty adult male Sprague Dawley rats were randomized into four groups with five rats in each group. Decortication was carried out in standard manner in all animals. Group 1 (only decortication), group 2 (CNTs), group 3 (HA-TCP) and group 4 (CNTs/HA-TCP) were formed. Eight weeks later, all animals were killed and obtained fusion segments were evaluated by manual palpation, histomorphometry and micro-computed tomography (mCT). RESULTS: In all evaluations, highest fusion values were obtained in Group 4. In mCT investigations, bone volume/ tissue volume (BV/TV) ratio was found to be significantly higher in composite group (group 4) only compared to ceramic group (group 3) (p < 0.001). Although in Group 2, in which only CNTs were used, the ratio was found to be statistically significantly higher than group 1(p < 0.001), the difference was not considered as significant in terms of fusion and in addition in group 2, CNTs were completely surrounded by fibrous tissue, i.e., no bone formation was observed. CONCLUSIONS: The CNTs/HA-TCP composite is a promising synthetic bone graft substitute for spinal fusion. Although CNTs are inadequate in producing spinal fusion when they are used alone, due to their high biocompatibility due to their high biocompatibility, and multiple effect on bone regeneration, they seem to increase fusion rates significantly when they are used in combination with ceramic-based synthetic grafts.

The therapeutic effects of coenzyme Q10 on surgically induced endometriosis in Sprague Dawley rats.

The aim of this study was to evaluate the effects of coenzyme Q10 in the treatment of endometriosis rat models. Twenty seven Sprague Dawley rats were divided into four groups; Control Group (n = 7; Endometriosis group), Reference Group (n = 6; Endometriosis + Buserelin acetate, 20 mg/kg), CoQ10 Group-I (n = 7; Endometriosis + CoQ10, 50 mg/kg) and CoQ10 Group-II (n = 7; Endometriosis + CoQ10, 100 mg/kg). At the end of the experiment, all the rats were sacrificed, and the volume and histoarchitecture of endometrial implants were evaluated. The mast cells were determined by Toluidine blue and collagen fiber density was analysed by Masson's Trichrome staining. Tumour necrosis factor and vascular endothelial growth factor (VEGF) levels were analysed by enzyme-linked immunosorbent assay in peritoneal fluid and VEGF and matrix metalloproteinase-9 (MMP-9) were evaluated by immunohistochemistry. Terminal deoxynucleotidil transferase-mediated dUTP Nick end labelling (TUNEL) was also used for the detection of apoptotic cells. The CoQ10 treatment significantly decreased the volume of endometriotic implants, VEGF, and MMP-9 immunoreactivity and increased TUNEL-positive cells. The findings of the study suggest that CoQ10 can be used in endometriosis treatment by suppressing the endometriotic implants.IMPACT STATEMENTWhat is already known on this subject? Endometriosis is a gynaecological disorder and previous studies have shown that different treatments with antioxidants cause significant regression in the endometriotic implants.What the results of this study add? In this study, CoQ10 reduced intra-abdominal adhesion scores and volume of the endometriotic implants. In addition, CoQ10 treatment affected mast cell, TNF-α, VEGF, and MMP-9.What of these findings for clinical practice and/or further research? CoQ10 treatments may be possible to apply, it can contribute to science in terms of a new therapeutic treatment for endometriosis. Further studies are required to evaluate the Coenzyme Q10's effects on pain and subfertility in endometriosis.

The effects of melatonin on possible damage that will occur on adipocytokines and liver tissue by coadministration of fructose and bisphenol a (BPA).

BPA, one of the environmental endocrine disruptors, and fructose, reason of liver steatosis which is frequently encountered in the daily diet, contribute to the formation of metabolic syndrome (MetS). This study examines the possible effects of concurrent fructose and BPA administration on MetS and determines the effects of melatonin on this process. In the seven identified groups, a total of forty-two adult male Sprague Dawley rats were treated by following fructose, BPA, and melatonin amounts, separately and together: group 1 (control), group 2 (10% aqueous fructose), group 3 (25 mg/kg BPA), group 4 (10% fructose + 25 mg/kg BPA), group 5 (10% fructose + 20 mg/kg melatonin), group 6 (25 mg/kg BPA + 20 mg/kg melatonin), and group 7 (10% fructose + 25 mg/kg BPA + 20 mg/kg melatonin). At the end of 60 days, histochemical, immunohistochemical, and biochemical procedures were performed on liver tissue. As a result, it was seen that BPA and fructose + BPA induced morphological alteration and inflammation and increased intracellular lipid quantity and amount of collagen and reticular fibers. The percentage of apoptotic liver cells stained by annexin V-FITC/PI was lower in group 7 compared to the group 4 (p < 0,001) and also in group 6 compared to the group 3 (p = 0.014). Both BPA and fructose application caused an increase in lipid peroxidation level due to the increase of oxidative stress. Application of melatonin induced antioxidant enzyme activity and reduced lipid peroxidation level. Our results indicate that fructose and BPA administration triggered the formation of MetS, whereas melatonin healed these variations, although not entirely.

The Effect of Hepcidin on Cardiac Ischemia-Reperfusion Injury.

Background/aim: Hepcidin is the main hormone in the regulation of iron metabolism which is also released from the heart. The aim of our study was to investigate the effects of hepcidin on the cardiac ischemia-reperfusion injury.Materials and methods: In this study, 12 Wistar albino rats were divided into two groups (n = 6 each): 1) The ischemia-reperfusion group (Group 1); 2) Hepcidin-treated group (Group 2). Rat hearts were perfused on Langendorff system with KH (Krebs-Henseleit) and subjected to 30 min stabilization, 30 min global ischemia, and 30 min reperfusion. Hepcidin (- M) was applied to group 2 at the onset of ischemia. Malondialdehyde (MDA), glutathione (GSH), and nitric oxide (NOx) levels were measured in heart tissue for NOx levels, viscosity, and ion content of perfusate were collected before ischemia and the 1st, 5th, 10th, 20th, and 30th minutes of reperfusion were determined. Apoptosis in heart was evaluated.Results: NOx and MDA levels significantly decreased in heart tissue in Hepcidin-treated group. NOx and viscosity of perfusate were not significantly different between the groups. Perfusate iron, calcium, magnesium, potassium, and sodium levels in group 2 were more homogeneous. Histologic structures of heart tissue were regularly in group 2. Apoptosis were increased in control group compared to hepcidin treated group.Conclusion: These results suggest that hepcidin may have a protective effect on the heart for the ischemia-reperfusion injury.

Epidermal growth factor enhances spinal fusion: Posterolateral lumbar fusion model on rats.

OBJECTIVE: The aim of this study was to investigate the effects of human recombinant epidermal growth factor (EGF) on posterolateral lumbar fusion in a rat model. METHODS: 36 male Sprague Dawley rats underwent posterolateral fusion at L4-5 level. They were randomly assigned to 3 groups: 1- Sham control group where no local augmentation was made, 2- Local Hydoxyapatite ß-tricalcium phosphate (HA/ß-TCP) augmentation group and 3- Local HA/ß-TCP + EGF augmentation group. Rats were euthanized at 8 weeks post-surgery. 6 rats from each group were selected for manual palpation examination, micro-computed tomography analysis and histologic analysis; and the rest was used for biomechanical analysis. RESULTS: Based on manual palpation, there was no fusion in the sham control group. Fusion rate was 33.3% in the HA/ß-TCP group and 66.7% in the HA/ß-TCP + EGF group (p = 0.085). Micro-CT results revealed that new bone formation was higher in the HA/ß-TCP + EGF group (BV/TV: 40% vs. 65%) (p = 0.004). Histologically newly formed bone tissue was more pronounced in the EGF group and compacted and bridging bone spicules were observed. The median maximum bending moment values were 0.51 Nmm (0.42-0.59), 0.73 Nmm (0.49-0.88) and 0.91 Nmm (0.66-1.03) in the sham control, HA/ß-TCP and HA/ß-TCP + EGF groups, respectively (p = 0.013). The median stiffness values were 1.69 N/mm (1.12-2.18), 1.68 N/mm (1.13-2.74) and 3.10 N/mm (1.66-4.40) as in the previous order (p = 0.087). CONCLUSION: This study demonstrates that EGF enhances posterolateral lumbar fusion in the rat model. EGF in combination with ceramic grafts increased the fusion rates. Our findings may provide insights to further studies, investigating EGF's clinical usage as an alternative fusion enhancer.

The protective role of melatonin and curcumin in the testis of young and aged rats.

We aimed to investigate the effect of melatonin and curcumin treatment on oxidative stress, apoptosis, and histology of testicular tissue in our study. Four groups were formed using young (4 months old, n = 6) and aged (20-22 months old, n = 18) male Wistar albino rats: (a) Young control (1% ethanol:phosphate-buffered saline [PBS], subcutaneously [s.c.]); (b) Aged control (CTL; n = 6, 1% ethanol:PBS, s.c.); (c) Aged Melatonin (MLT; n = 6, 10 mg/kg, s.c.); (d) Aged Curcumin (CUR; n = 6, 30 mg/kg, i.p.). At the end of 21 days, the rats were sacrificed, and testicular tissues were removed. Malondialdehyde (MDA) in the testicular tissue was determined with thiobarbituric acid reactive substances formation, and glutathione (GSH) was determined with modified Ellman method; testosterone level was determined with chemiluminescence method and histologic changes were determined with Haematoxylin-Eosin and Johnsen's scoring; Apoptotic cell counts were made with TUNEL staining of seminiferous tubule in testis. With ageing, MDA level increased in testicular tissue, but GSH and blood testosterone levels decreased. Melatonin treatment for aged rats significantly decreased Paired total testicular/body weight ratio compared to aged control group (p < 0.05). Curcumin treatment for aged rats significantly increased GSH level compared to the aged control group (p < 0.05). Besides, melatonin and curcumin treatment significantly decreased the number of apoptotic cells and significantly increased Johnsen's score (p < 0.05).

Protective effects of Nigella sativa oil against carboplatin-induced liver damage in rats.

OBJECTIVE: This study aimed to investigate the protective effects of nigella sativa oil (NSO) against liver damage due to intraperitoneal (i.p.) usage of carboplatin which is commonly used as a chemotherapeutic agent. MATERIAL AND METHOD: Twenty four female Wistar-albino rats (about 200-350 grams each) were divided into 4 groups. Group 1 (n = 6) was administered 4 ml/kg intraperitoneal (i.p.) saline 48 and 24 h before. Group 2 (n = 6) was i.p. administered 4 ml/kg NSO 48 h before and 4 ml/kg saline 24 h before. Group 3 (n = 6) was i.p. administered 4 ml/kg saline 48 h before and 80 mg/kg carboplatin 24 h before. Group 4 (n = 6) was i.p. administered 4 ml/kg NSO 48 h before and 80 mg/kg carboplatin 24 h before. At the end of 48 h, all rats were sacrificed, and liver tissues were put into 10% neutral formalin. After the routine tissue follow-up, histopathological changes and collagen fiber density were evaluated with Hematoxylin-Eosin and Masson's Trichrome staining. Apoptotic index was determined with TUNEL staining. RESULTS: The degeneration in hepatocytes, fiber distribution and density around central vein and portal space was observed in the carboplatin group compared to the control and NSO groups, hepatocyte cords preserved integrity, partial degeneration in hepatocytes and decreased collagen fiber distribution around central vein was noted in the NSO-carboplatin group compared to the carboplatin group. The apoptosis was lower in the NSO-carboplatin group compare with the carboplatin group, but no statistically significant difference was found between the two groups (p = 0.449). CONCLUSION: When used NSO before carboplatin exposure, it may protect against liver damage.

Comparative evaluation of various low-level laser therapies on bone healing following tooth extraction: An experimental animal study.

PURPOSE: The aim of this study was to evaluate the effects of helium-neon and gallium-aluminum-arsenide lasers with various doses on bone healing following tooth extraction. MATERIALS AND METHODS: Maxillary right incisor teeth of 30 female albino Wistar rats were extracted. Five groups were established: four groups treated with helium-neon or gallium-aluminum-arsenide lasers and a control group. Both laser groups' rats received energy doses of 6 J/cm2 and 10 J/cm2 for 7 days. At the end of 30 days, all subjects were sacrificed for histological and morphological evaluations. RESULTS: Laser groups showed faster bone healing and gallium-aluminum-arsenide lasers increased vascular immunoreactivity. The most widespread organized bone formation in the extraction socket was observed in the gallium-aluminum-arsenide laser group with the energy dose of 10 J/cm2 (p < 0.05). CONCLUSION: This study demonstrated that low-level laser therapies were effective on alveolar bone healing and that an energy dose of 10 J/cm2 did not have an inhibition effect on bone regeneration.

Comparison of Resorbable Mesh (Poly L-Lactide/Glycolic Acid) and Porous Polyethylene in Orbital Floor Fractures in an Experimental Model.

BACKGROUND: Resorbable mesh and porous polyethylene are frequently used alloplastic materials for the treatment of the orbital blowout fractures. The literature lacks reports comparing their long-term effects on experimental models. OBJECTIVE: Our aim was to radiologically and histologically evaluate the effectiveness and safety of porous polyethylene and resorbable mesh in a rabbit orbital blowout fracture model. METHODS: Twelve New Zealand white rabbits (24 orbits) were randomized to 4 groups. In group 1, only orbital floor dissection was done. In group 2, following orbital floor dissection, a 10-mm defect was created without any extra procedure. In group 3, following a 10-mm defect creation, a 12-mm-round cut porous polyethylene was placed on the defect. In group 4, following a 10-mm defect creation, a 12-mm-round cut resorbable mesh was placed on the defect. Computed tomographic analysis was performed during follow-up period. Orbital floors were evaluated histologically at month 6. RESULTS: No clinical complications were observed during follow-up period. In radiological evaluation, there was no statistically significant difference between groups regarding bone formation. In histological evaluation, the connective tissue was denser, and organized and better bone formation was observed in group 3 and 4 when compared with other groups. CONCLUSION: Although no significant radiological changes were present, porous polyethylene and resorbable mesh performed better histologically. They were effective and well tolerated for reconstruction of the isolated orbital floor defects.

HISTORIQUE: Le treillis résorbable et le polyéthylène poreux sont des matériaux alloplastiques souvent utilisés pour traiter les fractures isolées du plancher de l'orbite. Les publications ne contiennent pas de rapports sur les effets à long terme de ces matériaux dans des modèles expérimentaux. OBJECTIF: Les chercheurs visaient à évaluer l'efficacité et la sécurité du polyéthylène poreux et du treillis résorbable sur le plan radiologique et histologique dans un modèle de fracture isolée du plancher de l'orbite chez un lapin. MÉTHODOLOGIE: Douze lapins blancs néo-zélandais (24 orbites) ont été répartis au hasard en quatre groupes. Le groupe 1 a seulement subi la dissection du plancher de l'orbite. Dans le groupe 2, après cette dissection, une anomalie de 10 mm a été créée sans intervention supplémentaire. Dans le groupe 3, après la création d'une anomalie de 10 mm, une coupe ronde de polyéthylène poreux de 12 mm a été placée sur l'anomalie. Dans le groupe 4, après la création d'une anomalie de 10 mm, une coupe ronde de treillis résorbable de 12 mm a été placée sur l'anomalie. Les chercheurs ont effectué une analyse tomodensitométrique pendant la période de suivi. Au sixième mois, ils ont évalué les planchers orbitaux à l'histologie. RÉSULTATS: Les chercheurs n'ont observé aucune complication clinique pendant la période de suivi. À l'évaluation radiologique, la formation osseuse ne présentait aucune différence statistiquement significative entre les groupes. À l'évaluation histologique, les tissus conjonctifs étaient plus denses et la formation osseuse était organisée et de meilleure qualité dans les groupes 3 et 4 que dans les autres groupes. CONCLUSION: Malgré l'absence de modification significative à la radiologie, le polyéthylène poreux et le treillis résorbable donnaient de meilleurs résultats sur le plan histologique. Ces matériaux étaient efficaces et bien tolérés pour la reconstruction des anomalies isolées du plancher orbital.

Possible effects of melatonin against rat uterus exposure to bisphenol A during neonatal period.

The aim of this study was to investigate the possible effects of melatonin on rat uterine tissue against exposure with bisphenol A (BPA) in the neonatal period. Twenty-four female rats were divided into four groups, (n=6) per group. Group I was used as a control (sesame oil + ethanol), group II was injected daily with (100 mg/kg) BPA by subcutaneously (sc) daily postnatal days (PND 0-10), group III was injected daily with (10 mg/kg) melatonin by sc for 10 days (PND 20-30), and group IV was injected daily with (100 mg/kg) BPA (PND 0-10) and (10 mg/kg) melatonin (PND 20-30). All rats were sacrificed in the same day of metestrus cycle, approximately PND 70. Histological analyses, immunostaining of B cell lymphoma 2 (Bcl-2), and cytochrome c and TUNEL assays were performed. According to our results, neonatal exposure to BPA accelerates onset of puberty, causes degenerative and morphometric changes on rat uterus, and increases apoptotic reaction rates. The immunoreactivity of Bcl-2 was decreased after BPA administration. In addition, immunoreactivity of Bcl-2 showed an increase after melatonin treatment. However, cytochrome c immunoreactivity was decreased after melatonin administration. Our results suggest that melatonin may have positive effects against BPA-induced degenerative changes on rat uterus.

Effects of Atorvastatin on Experimental Spinal Cord Ischemia-Reperfusion Injury in Rabbits.

AIM: Extent of secondary injury is the determinant of tissue destruction and functional worsening after primary spinal cord injury (SCI). Data have accumulated on alleviation of secondary injury in SCI from many studies on the subject. Besides its cholesterol lowering effects, statins are known to have anti-inflammatory and anti-oxidant effects which are the main targets of spinal cord research. This study aims to evaluate the effects of atorvastatin on experimental spinal cord ischemia-reperfusion injury. MATERIAL AND METHODS: Thirty adult male New Zealand rabbits were allocated into control, ischemia-reperfusion (I/R) and treatment groups. Treatment group received 5 mg/kg of atorvastatin via lavage for the preceding 14 days. Other groups received placebo during the same time period. After two weeks, animals in the I/R and treatment groups underwent abdominal temporary aorta occlusion for 30 minutes. Neurological condition of the animals was recorded during the 48 hours of observation. Afterwards, animals were sacrificed and levels of malondialdehyde, glutathione and nitric oxide in spinal cord tissue and plasma and the histopathological tissue changes were determined. RESULTS: Animals in the treatment groups demonstrated significantly better results than the I/R group regarding biochemical markers. Neurological evaluation using the Tarlov scale demonstrated significantly better results at the 48th hour in treatment group. Histopathological results were also better in the treatment groups. CONCLUSION: Results of this study demonstrate the neuroprotective effects of atorvastatin. Atorvastatin has favorable effects on biochemical markers of oxidative stress in SCI. Further studies with larger cohorts and different time periods are also needed.

Placental vascularization and apoptosis in Type-1 and gestational DM.

OBJECTIVE: The objective of this study is to analyze the alteration in vascularization and apoptosis in the placentas of patients with Type 1 or gestational diabetes mellitus. METHODS: Placental samples drawn from normal (n = 6), GDM (n = 6), and Type 1 DM (n = 6) pregnancies were rinsed in PBS and fixed in 4% paraformaldehyde. The obtained sections were examined by both light and electron microscopy. Subsequently, immunohistochemical staining was performed to evaluate apoptosis and vascularization with caspase-9 and VEGF antibodies. RESULTS: Capillary structures in various sizes, both in free and in stem villi, were observed to be denser in the GDM group than in the control and Type-1 DM groups, utilizing electron microscopy. Similarly, when compared with Type-1 DM and controls, a decreased amount of microvilli with more irregularity and blunting on the villus surface was detected. GDM group showed increased immunoreactivity in capillaries of stem villi, free villi, and endothelial cells when compared with Type-1 DM and control groups. Regarding the immunohistochemical staining with VEGF, Type-1 DM, and GDM groups showed stronger immunoreactivity than the control group, especially in syncytiotrophoblastic cell nuclei and stromal cell nuclei. However, there was no significant difference between Type-1 DM and GDM groups. CONCLUSION: Type-1 DM and GDM placentas showed increased villous stromal capillarization, increased immunoreactivity with VEGF and caspase-9, and increased syncytial nodes, which may develop secondary to placental hypoxia-ischemia. However, more participants are needed to confirm these conclusions.

Effects of kinesiologic taping on epidermal-dermal distance, pain, edema and inflammation after experimentally induced soft tissue trauma.

PURPOSE: In sports medicine, the use of kinesiologic tape has recently gained popularity. Although widely used, there is no study examining the effects of kinesiologic tape on soft tissue after a contusion injury. The aim of this study was to examine the effects of kinesiologic taping on epidermal-dermal distance, edema, pain and inflammation after experimentally induced contusion injury. METHODS: Twelve adult female Wistar albino rats were divided into two groups: (1) 30 min group: n = 6, weight range: 182.0-199.4 g; and (2) 6 h group: n = 6, weight range: 186.9-200.8 g. After soft-tissue trauma, tape was applied to the right sides of each rat. In one group, tape was applied for 30 min while 6 h in the other. To assess the epidermal-dermal distance and edematous area, tissue sections were stained with hematoxylin and eosin and examined. Tissue sections were stained with nerve growth factor (NGF) and B-cell lymphoma 2 (Bcl-2) immunohistochemically to evaluate the effect of taping on pain and inflammation respectively. RESULTS: Epidermal-dermal distances were found to be significantly higher than controls' in both groups (p < 0.05). Notable decreases were seen in edematous areas in both groups (p < 0.05). NGF and Bcl-2 immune reactivity were decreased in all tape applied sides. CONCLUSIONS: After soft-tissue trauma, it was histologically shown that kinesiologic taping increases epidermal-dermal distance, and may reduce the sensation of pain, edema and inflammation. For better, faster and comfortable tissue healing with protection of soft-tissue integrity, kinesiologic taping may be a valuable treatment after contusion injury. However, these results should be supported by clinical studies.

Improving fat graft survival through preconditioning of the recipient site with microneedling.

Although fat grafts are considered the ideal soft-tissue fillers, the main concern dealing with this technique is not being able to predict long-term graft survival due to high absorption rates. The purpose of this study was to investigate the angiogenic effects of preconditioning the recipient area with micro-needling and to determine its overall impact on fat graft survival. The study consisted of a sham, control and study group. The source of fat was the Wistar albino rat inguinal fat pad while the recipient area was a dorsal subcutaneous pouch. The dorsal area was preconditioned with standard technique micro-needling 1-week prior to fat graft transfer in the study group while the control group did not undergo micro-needling. At the end of 15 weeks, morphological, biochemical, histological and immunohistochemical evaluation was carried out. Fat grafts in the study group had better integrity and a higher level of vascularity compared to the control group. Volume analysis demonstrated higher graft survival in the study group in comparison to the control group. Histomorphometric and immunohistochemical evaluation showed better graft integrity and uniform adipocytes, less fibrosis, less vacuolisation and inflammation and better vascularisation in the study group. Although higher triglyceride concentrations were measured for the study group, the difference between the two groups was statistically insignificant. In conclusion, fat grafting performed in an area preconditioned with micro-needling results in higher graft volume, better integrity and vascularisation and an overall higher graft survival rate.

Investigation of the nerve distribution pattern of leg muscles in rat.

Information about the distribution of intramuscular nerve fibres within the skeletal muscles will enhance the understanding of their morphological structure and functions. This study was designed to examine the entire intramuscular nerve distribution pattern in rat leg muscles. The tibialis cranialis, tibialis caudalis, extensor digitorum longus, flexor digitorum longus, gastrocnemius, peroneus longus and brevis muscles were dissected from origo and insertion points under a surgical microscope in seven rats. These skeletal muscles from right hindlimbs were stained with Sihler's stain. The diameter of extramuscular and major nerve branches, number of major and minor nerve branches and anastomoses were measured and photographed under a stereomicroscope. In addition, serial sections were obtained from the left hindlimb muscles with S100 immunohistochemical staining and transferred to the computer to reconstruct images. A significant difference was found between the gastrocnemius and tibialis caudalis (p < 0.001), flexor digitorum longus and tibialis caudalis (p < 0.003), and peroneus longus and tibialis caudalis (p < 0.049) with regard to the diameter of major branches. The gastrocnemius was significantly different from the flexor digitorum longus, peroneus longus, extensor digitorum longus, tibialis caudalis and tibialis cranialis with regard to the number of minor nerve branches (p < 0.001). Knowledge of the branching pattern and some key landmarks, such as the number and diameter of major and minor nerve branches and the number of anastomoses between the nerve branches of skeletal muscles, is helpful in surgical or therapeutic interventions and botulinum toxin injections in areas of high extramuscular and intramuscular nerve density.

Reconstruction of cranial bone defects using Struthio camelus eggshell.

BACKGROUND: There are many synthetic materials for the treatment of bone defects, which have their own advantages and disadvantages. We aimed to compare the efficacy of ostrich eggshell, which is cheap and easily available, and demineralized bone matrix in healing of cranial bone defects. METHODS: A full-thickness circular bone defect was created in the frontal bone of 40 Wistar rats. Group 1 was the operative control group. In group 2, demineralized bone matrix applied into the defects; in group 3, Struthio camelus (ostrich) eggshell implants (OSIs) were applied into the defects; and in group 4, ostrich eggshell powders were applied into the defects. Computed tomographic analysis was performed to evaluate the healing of bone defects, the bone density, the OSI area measurements, and the OSI volume and density. At the end of the 24th week, all rats were killed. New bone formation, infection, resorption, and tissue reactions were evaluated. RESULTS: Ostrich eggshell implants were slightly resorbed, integrated with bone, stable, and supplied good cranial completeness. Ostrich eggshell powders were totally resorbed at the sixth month. There were no significant differences between control and ostrich eggshell groups in new bone formation. CONCLUSIONS: Ostrich eggshell did not seem to be an osteoproductive material, but it has some important advantages as an implant. Ostrich eggshell has a strong structure, is cheap, is shaped easily, and does not cause tissue reaction or infection. Ostrich eggshell could be a good alternative graft material for craniomaxillofacial procedures. Further studies are required to find out the potential use of the ostrich eggshell in craniomaxillofacial reconstructions.

Feasibility of demineralized bone matrix for craniomaxillofacial contour restoration.

Demineralized bone matrix (DBM) could be a good alternative for craniomaxillofacial contour restoration, especially in perialar, malar, temporal, and frontal regions. In this study, the histologic behavior of DBM was investigated in different tissue planes to determine its proper application plane for restoration of craniomaxillofacial contour deformities and defects.Forty Wistar rats were divided into 6 groups: (1) 0.3 mL of 0.9% saline was injected into the subperiosteal plane of the cranium, (2) 0.3 mL of DBM was implanted into the subperiosteal plane of the cranium, (3) 0.3 mL of 0.9% saline was injected into the subdermal plane on the left inguinal region, (4) 0.3 mL of DBM was implanted into the subdermal plane on the right inguinal region, (5) 0.3 mL of 0.9% saline was injected between the left external and internal oblique muscles, and (6) 0.3 mL of DBM was implanted between the right external and internal oblique muscles. At the 8th week half of the rats and at 16th week the remaining rats were killed in each group, and tissue samples were harvested. Histological and immunohistochemical evaluation revealed new bone tissue and bone marrow formation in all planes that DBM was given.Demineralized bone matrix can provide satisfactory results in craniomaxillofacial contour deformities including forehead, temporal, and malar augmentations, as well as mental and perialar augmentations and saddle nose corrections, with supraperiosteal or deep subcutaneous applications. However, superficial applications must be avoided because of the possibility of palpation, because it induces hard bone tissue formation in all tissue planes.

Mutagenic and morphologic impacts of 1.8GHz radiofrequency radiation on human peripheral blood lymphocytes (hPBLs) and possible protective role of pre-treatment with Ginkgo biloba (EGb 761).

The mutagenic and morphologic effects of 1.8GHz Global System for Mobile Communications (GSM) modulated RF (radiofrequency) radiation alone and in combination with Ginkgo biloba (EGb 761) pre-treatment in human peripheral blood lymphocytes (hPBLs) were investigated in this study using Sister Chromatid Exchange (SCE) and electron microscopy. Cell viability was assessed with 3-(4, 5-dimethylthiazol-2-yl)-2, 5-diphenyltetrazolium bromide (MTT) reduction assay. The lymphocyte cultures were exposed to GSM modulated RF radiation at 1.8GHz for 6, 8, 24 and 48h with and without EGb 761. We observed morphological changes in pulse-modulated RF radiated lymphocytes. Longer exposure periods led to destruction of organelle and nucleus structures. Chromatin change and the loss of mitochondrial crista occurred in cells exposed to RF for 8h and 24h and were more pronounced in cells exposed for 48h. Cytoplasmic lysis and destruction of membrane integrity of cells and nuclei were also seen in 48h RF exposed cells. There was a significant increase (p<0.05) in SCE frequency in RF exposed lymphocytes compared to sham controls. EGb 761 pre-treatment significantly decreased SCE from RF radiation. RF radiation also inhibited cell viability in a time dependent manner. The inhibitory effects of RF radiation on the growth of lymphoctes were marked in longer exposure periods. EGb 761 pre-treatment significantly increased cell viability in RF+EGb 761 treated groups at 8 and 24h when compared to RF exposed groups alone. The results of our study showed that RF radiation affects cell morphology, increases SCE and inhibits cell proliferation. However, EGb 761 has a protective role against RF induced mutagenity. We concluded that RF radiation induces chromosomal damage in hPBLs but this damage may be reduced by EGb 761 pre-treatment.

Protective effect of beta-glucan on contrast induced-nephropathy and a comparison of beta-glucan with nebivolol and N-acetylcysteine in rats.

BACKGROUND: It has been shown that beta-glucan (BG), which has antioxidant and immunomodulatory effects, attenuats renal ischemia-reperfusion injury. We aimed to investigate whether BG might have a preventive role against the development of contrast-induced nephropathy and to compare its effect with nebivolol (Nb) and N-acetylcysteine (NAC). METHODS: Thirty-six Wistar albino female rats were randomly divided into six groups (n = 6 each): control, contrast media (CM), BG, BG + CM, Nb + CM, and NAC + CM. With the exception of control and CM groups, the others were given drugs orally once a day for 5 days. Kidney function parameters, inflammatory parameters, and serum and renal tissue oxidative stress markers were measured. RESULTS: Increases of serum creatinine and blood urea nitrogen levels were significantly higher (p < 0.05) in the CM group only. Absolute changes of serum creatinine levels in BG, BG + CM and Nb + CM groups were significantly lower than those in the CM group (p < 0.05). Serum levels of advanced oxidation protein products and malondialdehyde were significantly less (p < 0.05) in the BG group compared to the CM group. Histopathological lesions in the CM group were more advanced (p < 0.05). No significant differences between the BG + CM, Nb + CM and NAC + CM groups were found with regard to histopathological findings. CONCLUSION: This study suggests that BG protects or ameliorates against contrast-induced nephropathy. Its beneficial effects may be similar to or greater than those of Nb or NAC.