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PURPOSE: To investigate whether the DNA methylation profiles of GNAS(20q13.32), MEST(7q32.2), MESTIT1(7q32.2), IGF2(11p15.5), H19 (7q32.2), and CEP41(7q32.2) genes are related to the transcriptomic and epigenomic etiology of male infertility. METHODS: The DNA methylation levels of spermatozoa were obtained from fertile (n = 30), oligozoospermic (n = 30), and men with normal sperm count (n = 30). The methylation status of each CpG site was categorized as hypermethylated or hypomethylated. Expression levels of target gene transcripts were determined using real-time PCR. RESULTS: The oligozoospermia showed a higher frequency of hypermethylation at GNASAS 1st, 3rd, and 5th CpG dinucleotides (66.7%, 73.3%, 73.3%) compared to the fertile group (33.3%, 33.3%, 40%, respectively). The normal sperm count exhibited a higher frequency of hypermethylation at the 3rd CpG of CEP41 (46.7%) than the fertile group (16.7%). Normal sperm count was predicted by CEP41 hypermethylation (OR = 1.750, 95%CI 1.038-2.950) and hypermethylation of both CEP41 and GNASAS (OR = 2.389, 95%CI 1.137-5.021). Oligozoospermia was predicted solely by GNASAS hypermethylation (OR = 2.460, 95%CI 1.315-4.603). In sperms with decreased IGF2 expression in the fertile group, we observed hypomethylation in the 2nd CpG of IGF2 antisense (IFG2AS), and hypermethylation in the 1st, 2nd, and 4th CpGs of H19. No significant relationship was found between IGF2 expression and methylation status of IGF2AS and H19 in infertile groups. CONCLUSION: The disappearance of the relationship between IGF2 expression and IGF2AS and H19 methylations in the infertile group provides new information regarding the disruption of epigenetic programming during spermatogenesis. A better understanding of sperm GNASAS and CEP41 hypermethylation could advance innovative diagnostic markers for male infertility.
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BACKGROUND: Lower lateral cartilage reshaping is one of the basic steps in rhinoplasty. Hemitransdomal suture is frequently used for dome narrowing. Different suture materials can be used for hemitransdomal suture. In this study, we investigated the effectiveness of polypropylene and polydioxanone in hemitransdomal suture by designing a new experimental model in the rabbit ear cartilage. METHODS: Twelve young adult male New Zealand White rabbits were used. The bipedicled inverted-U-shaped cartilage was elevated in each ear of the rabbits. Two hemitransdomal sutures were applied using 5-0 polypropylene on one ear and 5-0 polydioxanone on the other ear randomly. A 5 mm high cartilage mound was created with two hemitransdomal sutures on each side. The sample size was twelve both in the polypropylene group and the polydioxanone group (n = 12). All animals were sacrificed after three months. Cartilage mound heights were measured. The samples were examined histologically regarding fibroadipose tissue, inflammation, foreign body granuloma, cartilage degeneration, and the presence of inclusion cyst. RESULTS: Cartilage mound height was significantly higher in the polypropylene group than in the polydioxanone group at the end of the third month [3.75 mm (± 0.68) vs. 3.03 mm (± 0.69); p < 0.05]. There was no significant difference between the two groups in histological analysis (p > 0.05). CONCLUSIONS: Polypropylene suture may be more effective in maintaining the shape given to the dome by hemitransdomal sutures in rhinoplasty. No Level Assigned This journal requires that authors assign a level of evidence to each submission to which Evidence-Based Medicine rankings are applicable. This excludes Review Articles, Book Reviews, and manuscripts that concern Basic Science, Animal Studies, Cadaver Studies, and Experimental Studies. For a full description of these Evidence-Based Medicine ratings, please refer to Table of Contents or the online Instructions to Authors www.springer.com/00266.
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Cartilagem da Orelha , Rinoplastia , Coelhos , Masculino , Animais , Cartilagem da Orelha/cirurgia , Polidioxanona , Polipropilenos , SuturasRESUMO
BACKGROUND: Studies have described various surgical maneuvers in rhinoplasty to limit thickening from excessive scarring in the supratip region. This study aimed to compare the effectiveness of three maneuvers-a simple suture, a U suture, and bolster dressing-used to avoid supratip deformity in a rabbit model. METHODS: Thirty-two male New Zealand white rabbits were included. The animals were divided into four groups, and dissection was performed in the supra-perichondrial plane up to the supratip region through an open rhinoplasty incision. After dissection, the following approaches were applied to the supratip region: Group 1, simple approximation suture; Group 2, U suture; Group 3, bolster dressing; and Group 4 (control group), no suture. All animals were sacrificed after 12 weeks. Histological analysis was performed. RESULTS: In Group 4, scar thickness was significantly greater than in the other groups (p < 0.05). Group 3 had greater scar thickness than Group 2 (p < 0.05). The ratio of scar thickness to skin thickness was higher in Group 4 compared with the other groups (p < 0.05). Finally, there was no difference in the ratio of scar thickness to skin thickness between Groups 1, 2, and 3 (p > 0.05). CONCLUSIONS: In this study, it was concluded that surgical methods using sutures in the supratip region reduced scar thickness in a rabbit model, and these surgical methods had similar levels of effectiveness. NO LEVEL ASSIGNED: This journal requires that authors assign a level of evidence to each submission to which Evidence-Based Medicine rankings are applicable. This excludes Review Articles, Book Reviews, and manuscripts that concern Basic Science, Animal Studies, Cadaver Studies, and Experimental Studies. For a full description of these Evidence-Based Medicine ratings, please refer to the Table of Contents or the online Instructions to Authors www.springer.com/00266 .
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BACKGROUND AND OBJECTIVES: Recently, the terms autophagy and apoptosis have been studied on implants, especially in cell culture and in vitro studies, but in vivo evaluations are limited. The aim of this study was to compare the differences in apoptosis and autophagy intensity at the molecular and cellular level in periodontal and peri-implant diseases. METHODS: Sixty-four biopsy samples were obtained from 52 patients, 36 female and 16 male, whose mean age was between 18 and 75, and were included in the study. The periodontitis group was defined as PG (n:30 sample) and the peri-implantitis group as IG (n:34 samples). Granulation tissues as biopsy materials were collected, and immunohistochemical analysis was performed with hematoxylin-eosin, Masson's trichrome, anti-MAP1LC3A, anti-beclin, and anti-active caspase-3 antibodies and terminal TdT-mediated dUTP-biotin nick end labeling (TUNEL) methods. The histological slide images were evaluated with the ImageJ software program. Inflammatory cell density in epithelial tissue, inflammatory cell density in connective tissue, the density of necrotic tissue debris, and collagen density in connective tissue were scored between 0 and 3 (0: none, 1: minimal, 2: moderate, 3: severe by hematoxylin-eosin and Masson's trichrome). The antibody binding reaction areas were evaluated per unit area (mm2 ) in connective tissue by immunohistochemical examination. RESULTS: As histochemical evaluations, there was no statistically significant differences the mean inflammatory cell density value in the epithelial tissue, inflammatory cell density value in the connective tissue, density value of necrotic tissue debris, collagen density value in the connective tissue between the groups. There was no statistically significant difference on immunohistochemical staining with LC3, caspase-3, Beclin-1 and TUNEL between the two groups (p > .05). CONCLUSIONS: A higher rate of inflammatory accumulation was shown on peri-implantitis, but no difference was found between periodontitis and peri-implantitis according to autophagy and apoptosis markers. Studies with high sample sizes with different markers are needed.
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Implantes Dentários , Peri-Implantite , Periodontite , Humanos , Masculino , Feminino , Peri-Implantite/metabolismo , Caspase 3 , Amarelo de Eosina-(YS) , Hematoxilina , Periodontite/metabolismo , Colágeno , Apoptose , AutofagiaRESUMO
BACKGROUND: Ketamine (KET) is a commonly used anesthetic agent. However, several previous studies reported that KET leads to neuronal damage in neurodevelopmental stages and has neuroprotective effects. The present experimental study aimed to determine the undesirable histopathological effects of KET in the cerebral cortex, striatum, and hippocampus after recurrent KET administration in juvenile rats. METHODS: After ethical approval was obtained, 32 juvenile male Wistar Albino rats were randomized into four groups: 1 mg/kg serum saline intraperitoneally (i.p.), 5 mg/kg KET i.p., 20 mg/kg KET i.p., and 50 mg/kg KET i.p. KET was administered for three consecutive days at three-h intervals in three doses. Ten days after the last KET dose, the rats were sacrificed. Cerebral hemispheres were fixed. Hematoxylin and eosin stain was used for morphometric analysis. Hippocampi were evaluated by immunohistochemistry with anticleaved caspase-3 antibodies. Statistical analysis was conducted with SPSS 21 software using the ANOVA test and Bonferroni post hoc analysis method. RESULTS: The experimental study findings revealed no difference between the groups' cell counts or sizes in cortical morphometry. No degenerative changes were observed in pyramidal and granular cells in the striatum. Mild gliosis was observed in the 20 mg/kg and 50 mg/kg KET administration groups. Immuno-histo-chemical analysis was conducted to determine apoptosis in the CA1 region of the hippocampus and revealed that caspase-3 positivity increased with the KET dose. However, there was no statistical difference between the groups. While it was lower than the control group in the 5 mg/kg KET group, it was similar to the control group in the 20 mg/kg KET group and higher in the 50 mg/kg KET group (p > 0.05). DISCUSSION: : Repetitive KET exposure did not significantly affect juvenile cerebral morphology and apoptosis in hippocampal cells.
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Ketamina , Animais , Ratos , Masculino , Ketamina/farmacologia , Caspase 3 , Ratos Wistar , Hipocampo , EncéfaloRESUMO
BACKGROUND: The use of fat obtained from ultrasound-assisted liposuction is popular. However, no study has considered the effect of different energy levels on fat grafts. OBJECTIVES: We hypothesized that different ultrasonic energy levels could change the fat graft viability. METHODS: Both flanks of 15 CD1 nude mice (30 experimental areas) were used, with experimental areas randomly distributed into five groups. Using different energy settings, fat grafts were obtained from a patient's abdominoplasty material and applied to the mouse flank regions. Device settings were intermittent mode with 50% vibration amplitude in group 1, continuous mode with 50% vibration amplitude in group 2, intermittent mode with 90% vibration amplitude in group 3, and continuous mode with 90% vibration amplitude in group 4. The control group was grafted with fat obtained via the conventional method. After 6 weeks, all mice were sacrificed, and fat grafts were excised. Sections were stained with hematoxylin-eosin, Masson's trichrome, and anti-perilipin A antibody. RESULTS: The perilipin A immunostaining result was lowest in group 4, indicating the lowest viable cell count (p < 0.01). There was no significant difference between groups for the other parameters (p > 0.05). CONCLUSION: High ultrasonic energy may affect fat graft survival. If fat injection is planned, avoiding high energy settings (our recommendation is not to exceed 16 Watts.) should be considered. We also recommend increasing the vibration amplitude rather than switching from intermittent to continuous mode in body parts that are relatively resistant to liposuction. NO LEVEL ASSIGNED: This journal requires that authors assign a level of evidence to each submission to which Evidence-Based Medicine rankings are applicable. This excludes Review Articles, Book Reviews, and manuscripts that concern Basic Science, Animal Studies, Cadaver Studies, and Experimental Studies. For a full description of these Evidence-Based Medicine ratings, please refer to the Table of Contents or the online Instructions to Authors www.springer.com/00266 .
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Lipectomia , Animais , Camundongos , Amarelo de Eosina-(YS) , Hematoxilina , Lipectomia/métodos , Camundongos Nus , UltrassomRESUMO
Male infertility affects approximately 30% of infertile couples. As spermatozoa mature in the epididymal lumen, their potential for mobility increases, and their protein, lipid and small RNA (sRNA) content changes, whereas capacitation and fertilisation take place in the female reproductive tract. Both of the latter processes are affected by maturation, because impaired maturation causes premature capacitation and fertilization. The epididymis produces a suitable environment for sperm maturation via ion transport, vesicle secretion and protein matrix formation. The microenvironment for sperm maturation varies in three broad segments: the caput, the corpus and the cauda epididymis. Epididymosomes transfer proteins, lipids and sRNAs from the epididymal epithelium to spermatozoa and genetic alterations of epididymal genes can lead to decreased sperm motility, morphological abnormalities of spermatozoa and subfertility. Genetic factors are involved in all aetiological categories in male infertility. However, studies conducted on the genes involved in epididymal functions are limited. The sRNA content of spermatozoa changes during epididymal migration, and these sRNAs play a role in embryo development and epigenetic inheritance. This review aims to clarify the role of the epididymal epithelium in the maturation of spermatozoa in light of the current molecular genomic knowledge.
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Epididimo , Maturação do Esperma , Epigênese Genética , Feminino , Humanos , Masculino , Motilidade dos Espermatozoides , EspermatozoidesRESUMO
PURPOSE: To compare the biomechanical and histological properties of Achilles tendons repaired at different time points during the acute injury period. METHODS: Thirty-six skeletally mature Sprague-Dawley rats underwent bilateral mid-substance Achilles tenotomy. The Achilles tendons were repaired either in the first 24 h (group 1), 24-48 h (group 2), 48-72 h (group 3), or > 72 h (mean: 120 ± 5.2 h) (group 4) after tenotomy. Six weeks after repair, nine tendons per group were assessed biomechanically and histologically. The Stoll histological scoring system was used for histological examination. The groups were compared with each other and native tendons (control group). The correlations between biomechanical and histological results were analysed. RESULTS: There were no significant differences between groups 1, 2 and 3 regarding the mean load to failure; it was significantly lower in group 4. Healed tendons in groups 1, 2 and 3 had significantly greater stiffness than native tendons and group 4 tendons. All healed tendons had a larger cross-sectional area than native tendons. There was no significant difference in tendon length between the groups. There was no significant difference in Young's modulus between the groups; Young's modulus was lower in all the groups than in the control group. Group 1 had significantly higher extracellular matrix organization, cell alignment, cell distribution and nucleus morphology scores and total scores than group 4. Group 1 had significantly higher extracellular matrix organization, cell distribution, vascularization and inflammation scores and total scores than group 3. A significant positive correlation was detected between the maximum load to failure and total histological score. CONCLUSION: Repair of acute Achilles tendon rupture within 48 h, and especially in the first 24 h, provides better biomechanical and histological outcomes. In the clinical practice, the data could be used to decrease re-rupture rates, to achieve more anatomical tendon healing and to implement more effective post-operative rehabilitation programme.
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Tendão do Calcâneo/cirurgia , Traumatismos dos Tendões/cirurgia , Cicatrização , Tendão do Calcâneo/lesões , Tendão do Calcâneo/fisiologia , Animais , Fenômenos Biomecânicos , Masculino , Ratos , Ratos Sprague-Dawley , Ruptura/cirurgia , Tenotomia , Fatores de TempoRESUMO
BACKGROUND: Microneedling promotes percutaneous collagen induction; cupping therapy creates negative pressure and leads to increased blood flow in the applied area. The addition of cupping therapy to microneedling is thought to contribute positively to microneedling's effects. This study was carried out to investigate the histologic effects of adding cupping therapy to microneedling. METHODS: Thirty Wistar rats were divided into five groups, with six rats in each group. One control group and four experimental groups were formed, which are defined as follows: the control group; the single-session microneedling applied to the dorsal trunk group; the 15-minute cupping therapy added to the single-session microneedling group; the microneedling applied over a total of three sessions at 3-week intervals group; and the microneedling with cupping therapy applied over a total of three sessions at 3-week intervals group. Each animal was euthanized at the end of the fourth week following the last treatment, and skin samples were evaluated histologically with hematoxylin and eosin stain and type I and III collagen antibody immunostaining. RESULTS: The addition of cupping therapy to microneedling increased the thickness of the epidermis and dermis. A significant increase in type I collagen immunostaining and the type-I-to-type III collagen ratio was seen only in the single-session microneedling applied to the dorsal trunk group. Cupping therapy did not generate a significant difference in type I collagen immunostaining. No treatment was found to produce a significant increase in type III collagen immunostaining. CONCLUSION: Cupping therapy can be added to microneedling therapy and used to increase certain desired effects on skin. CLINICAL RELEVANCE STATEMENT: Microneedling is an easy and effective method to improve skin quality in clinical practice.
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Colágeno Tipo I , Ventosaterapia , Ratos , Animais , Colágeno Tipo III , Ratos Wistar , Colágeno/uso terapêutico , AgulhasRESUMO
BACKGROUND: Erythropoietin has neuroregenerative effects. Fibrin glue may be used for nerve repair and controlled release of substances. In this study, the authors investigated the effects of erythropoietin-containing fibrin glue on nerve repair, based on the hypothesis that erythropoietin-containing fibrin glue would positively affect nerve regeneration. METHODS: Thirty-six Long-Evans rats were used. The animals were divided into six groups. Their left sciatic nerves were isolated, transected, and repaired with saline-containing fibrin glue in group 1, with erythropoietin-containing fibrin glue in group 2, with saline-containing fibrin glue and two sutures in group 3, with erythropoietin-containing fibrin glue and two sutures in group 4, with two sutures in group 5, and with four sutures in group 6. Sciatic Functional Index calculation, pin-prick test, and toe-spread test were performed on days 21, 42, and 63. All animals were killed on day 63. The nerve sections were analyzed histologically. RESULTS: The Sciatic Functional Index, pin-prick test, and toe-spread test results were the best in group 4 and the worst in group 5. Group 4 showed superior Schwann cell proliferation (p < 0.05). Groups with epineural suture use (groups 3, 4, 5, and 6) had higher endoneurial collagen synthesis scores than the groups without suture use (groups 1 and 2) (p < 0.05). The myelin protein zero immunostaining results were significantly higher in the erythropoietin-treated groups (groups 2 and 4) (p < 0.05). CONCLUSION: The combined use of erythropoietin-containing fibrin glue and two epineural sutures (group 4) showed a statistically significant improvement in many parameters. CLINICAL RELEVANCE STATEMENT: Fibrin glue is already used in nerve repair. Adding erythropoietin to fibrin glue could be a safe and easy option to improve nerve regeneration.
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Eritropoetina/administração & dosagem , Adesivo Tecidual de Fibrina/administração & dosagem , Regeneração Nervosa/efeitos dos fármacos , Traumatismos dos Nervos Periféricos/terapia , Animais , Modelos Animais de Doenças , Ratos , Ratos Long-Evans , Nervo Isquiático/lesõesRESUMO
BACKGROUND: In the Ponseti technique, the residual equinus deformity is corrected with percutaneous tenotomy. This experimental study aimed to compare the safety and effectiveness of a large-gauge needle, a corneal knife, and a No. 11 blade in percutaneous achillotomy performed in rats. METHODS: Ninety Achilles tendons of 45 Sprague-Dawley rats were analyzed, following division into three study groups. In the study, group I (needle), group II (corneal knife), and group III (No. 11 blade) were compared on the basis of bleeding, incision length, requirement for primary suture, range of motion, and resulting neurovascular injury at day 0. Moreover, the groups were compared in terms of range of motion, macroscopic and microscopic adhesions, and tenocyte morphology at days 21 and 42 postoperatively. RESULTS: On day 0, one suture was required in group III, whereas in groups I and II, no sutures were required. Postoperative bleeding was greater in group III and similar in groups I and II. Neurovascular injury was not observed in any of the groups. Three incomplete tenotomies were observed in group III and one incomplete tenotomy was observed in group II. Importantly, all tenotomies were complete in group I. In all groups, the range of motion was similar. The macroscopic adhesion score revealed high adhesion in group III (P = .009). According to Tang's criteria, microscopic adhesion was significantly higher on day 21 in group III compared with the other groups (P <0.001). No significant differences were observed in tenocyte morphology based on the Bonar criteria (P = .850). CONCLUSIONS: In the results obtained from this animal study, we observed less bleeding, less adhesion, and less incomplete tenotomy in the large-gauge needle and corneal knife groups compared with the No. 11 blade group during the percutaneous Achilles tenotomy performed in rats.