Comparison of the prophylactic use of ibandronate and its use in early-stage osteonecrosis in rats with steroid-induced osteonecrosis of the femoral head.

OBJECTIVES: The aim of this study was to investigate the effects of ibandronate before and after the onset of osteonecrosis in rats with steroid-induced osteonecrosis of the femoral head. MATERIALS AND METHODS: A total of 24 female Sprague-Dawley rats were used in this study. Three groups were formed with eight rats in each group. The first group was the prophylactic group that received ibandronate treatment before and after the onset of osteonecrosis (Group PT). The second group received ibandronate treatment three weeks after the development of osteonecrosis (Group TAO). The third group was the control group in which osteonecrosis was created, but only normal saline (NS) was given. At the end of the study, all rats were sacrificed, and their right femoral heads were removed, fixed with formalin, and sent for micro-computed tomography. Hematoxylin-eosin (H&E) and immunohistochemical examinations of the right femoral head sections were performed. RESULTS: In the PT group, the trabecular thickness was significantly higher compared to those of the TAO and control groups (p<0.05). The trabecular thickness did not significantly differ between the TAO and control groups. The trabecular bone pattern factor was significantly higher in the PT group compared to the control and TAO groups (p<0.05); however, it showed no significant difference between the TAO and control groups. The incidence of type 2 osteonecrosis in the PT and TAO groups was significantly lower than that in the control group (p<0.05). The incidence of tissue-non-specific alkaline phosphatase (TNAPase) immunoreactivity of osteoblast positivity was significantly higher in the PT and TAO groups compared to the control group (p<0.05), whereas the incidence of TRAPase immunoreactivity of osteoclastic positivity was significantly lower in the PT and TAO groups compared to the control group (p<0.05). CONCLUSION: Intravenous administration of ibandronate before the onset of the disease was more effective in the treatment of osteonecrosis in rats with steroid-induced osteonecrosis of the femoral head.

Silhouette Lymphangioma: An Unknown Macular Form of Cutaneous Lymphangioma.

Unusual angiomatous or lymphangiomatous vascular malformations are rarely seen. One of them is lymphangioma (LA), which is a rare benign lymphovascular abnormality. LA is usually seen in the types of circumscriptum (or capillary), cavernous and cystic. Here, we report a unique case of LA with a patchy appearance. The patient presented due to unusual symptoms and eccentric clinical manifestation of the lesion. Here, we present a new lymphatic entity which was diagnosed as LA with its clinical, radiological and pathological findings. Written informed consent of the patient was obtained for this report. To the best of our knowledge this macular form of cutaneous LA has not been previously reported in literature. Macular LA should be kept in mind when faced with a colored long-term macular lesion on the skin.

Amelanotic nodular appearance within superficial spreading melanoma.

Usual clinical types of malignant melanoma (MM) are lentigo maligna, superficial spreading, nodular, and acral lentiginous MMs. Rarer variants of MM are amelanotic, neurotropic, desmoplastic, metastatic, invisible, pedunculatedmelanoma (PM), or nodal melanoma (NM), which constitute less than 5% of cases. Recognition and discrimination of these variants are important to avoid misdiagnosis of MM. We report a 42-year-old Caucasian woman presented with a superficial spreading malignant melanoma (SSM) with an amelanotic nodular center. The lesion simulated a NM or PM clinically, but its histopathology showed an SSM. This misleading appearance of SSM has not been reported previously.

The effects of valsartan on scar maturation in an experimental rabbit ear wound model.

INTRODUCTION: In our study, we aimed to search and compare the effects of valsartan and enalapril on the pathological scar formation on the basis of histomorphological parameters. MATERIALS AND METHODS: Nine New Zealand albino male rabbits, which were divided into three groups, were included in the study. A previously described rabbit ear wound model was used. Enalapril was administered 0.75 mg/kg/day on the first group and valsartan was administered 10 mg/kg/day on the second group for 40 days. The third group was the control group. Results were evaluated on the 40th day with scar elevation index calculation and histological studies. Histological studies were done by using Hematoxylin-eosin, Masson trichrome and Sirius Red stains. RESULTS: Enalapril and valsartan groups were both significantly effective on the prevention of pathological scar formation when compared to the control group in terms of fibroblast count, capillary count, type 1/3 collagen ratio, collagen organization, and epithelial thickness. There was no significant difference between the enalapril and control group on the scar elevation index. Valsartan group was more efficient than the enalapril group on the reduction of fibroblast count and epithelial thickness. CONCLUSION: Both Valsartan and Enalapril are found to be effective for the prevention of pathological scar formation.

Comparison on effects of platelet-rich plasma versus autologous conditioned serum on Achilles tendon healing in a rat model.

OBJECTIVE: The aim of this study was to compare the effects of local administrations of platelet-rich plasma (PRP) with autologous conditioned serum (ACS) on Achilles tendon healing in a rat model. METHODS: In this study, 40 male Sprague-Dawley rats, aged 12 months and weighing 350 to 400 g were used. The rats were divided into three groups (n=10 in each group): a control group and two treatment groups (PRP vs ACS). A standardized procedure was applied for the complete rupture and repair of the Achilles tendon in each group. The PRP group received one dose of PRP on the operative area, and ACS group received ACS at 24, 48, and 72 hours after the surgery. The control group received no injection. Animals were sacrificed 30 days after the operation, and tendon healing in each group was assessed histopathologically based on Bonar's semi-quantitative score and Movin's semi-quantitative grading scale. For the biomechanical analyses, unoperated Achilles tendons of all rats in the control and ACS groups were also harvested, and pulling tests were applied to the specimen to measure the longitudinal axis strength. The highest force value among the data obtained was defined as the maximum strength value (Fmax). RESULTS: The mean Bonar's score was significantly lower in the PRP group (3.8±0.8) than in the ACS (4.8±0.45) and control groups (5.2±0.837) (p=0.0028). The mean Movin's score was significantly lower in the PRP group (7.80±1.49) than in the ACS (9.8±1) and control groups (11.2±2.4) (p=0.029). The ratio of type I collagen was significantly higher in the PRP group (60±6) than in the ACS (52±4.5) and control groups (42±9) (p=0.005). Biomechanical results obtained from operated sites were comparable in terms of Fmax among groups (PRP, 33.93±2.61; ACS, 35.24±3.26; control, 35.69±3.62) (p=0.674). Similarly, the results obtained from unoperated sites were comparable among groups (PRP, 47.71±1.21; ACS, 48.14±2; control, 49.14.69±1.88) (p=0.395). CONCLUSION: In terms of histopathological results, PRP seems to be more effective than ACS for Achilles tendon healing in rats.

Histopathological, immunohistochemical, and biomechanical effects of splenectomy on Achilles tendon healing in rats.

PURPOSE: This study aimed to assess Achilles tendon repair in rats following splenectomy to simulate patients with musculoskeletal system injury who had splenectomy after spleen injury, a situation often seen in orthopedics and traumatology practice. MATERIALS AND METHODS: The study included 32 male Sprague-Dawley rats (10 months old; average weight, 394.5 ± 28.3 g). The rats were fed with standard rodent food ad libitum at 22°C in a dark environment for 12 h. They were divided into two groups, namely the splenectomy (total splenectomy and Achilles tendon repair) and control groups (only Achilles tendon repair; n = 16). Four weeks after the surgery, the rats were euthanized, and their Achilles tendons were examined histopathologically, immunohistochemically, and biomechanically. RESULTS: In the splenectomy group, proinflammatory cytokines, such as interleukin-1ß, tumor necrosis factor-α, and interferon-γ, showed significantly lower values than those in the control group (p ˂0.01); moreover, the levels of anti-inflammatory cytokines like vascular endothelial growth factor, transforming growth factor-ß1, interleukin-2, interleukin-10, and hepatocyte growth factor were significantly higher than in the control group (p ˂ 0.001). The average ultimate tensile strengths were 2.58 ± 0.5 in the splenectomy and 2.78 ± 0.3 in the control group (p = 0.043). The average εUTS values were 0.33 ± 0.5 in the splenectomy and 0.44 ± 0.1 in the control group (p = 0.021). CONCLUSION: Splenectomy may positively influence Achilles tendon healing through modification of the proinflammatory/anti-inflammatory ratio in favor of anti-inflammatory cytokines by causing a decrease in spleen-originated inflammatory cells.

Osteoprotegerin and 25-hydroxy vitamin D levels in patients with diabetic foot.

OBJECTIVES: This study aims to compare the levels of osteoprotegerin (OPG) and 25-hydroxy vitamin D (25(OH)D) in patients with diabetic foot and patients with newly diagnosed type 2 diabetes mellitus (DM) and to investigate the prevalence and severity of 25(OH)D insufficiency in patients with diabetic foot. PATIENTS AND METHODS: This prospective study was conducted on 105 patients including 58 patients with diabetic foot (42 males, 16 females; mean age 63.6 years; range, 31 to 90 years), who applied to our hospital between June 2014 and May 2015, and 47 newly diagnosed type 2 DM patients (27 males, 20 females; mean age 51.4 years; range, 29 to 85 years) (control group). 25(OH)D and osteoprotegerin serum levels in both groups were measured and compared. RESULTS: Osteoprotegerin levels in diabetic foot group were significantly higher than the control group (p<0.05). The 25(OH)D levels in diabetic foot group were significantly lower than the control group (p<0.05). There were positive correlations between OPG levels and C-reactive protein (CRP) and creatinine levels in patients with diabetic foot. CONCLUSION: Elevated levels of OPG in patients with diabetic foot may display the severity of the clinical status due to its positive correlation with CRP and creatinine. We detected severe 25(OH)D deficiency in the majority of diabetic foot patients. Vitamin D supplementation may be required in diabetic foot patients to prevent unfavorable immunologic alterations.

Biomechanical, histological, and radiological effects of different phosphodiesterase inhibitors on femoral fracture healing in rats.

PURPOSE: To investigate the biomechanical, histological, and radiological effects of sildenafil and pentoxifylline on femoral fracture healing in rats. METHODS: Forty-eight Sprague-Dawley rats were divided into three groups equally according to the pharmacological agents to be investigated. Femoral shaft fractures were formed in the left side. Group 1 (control group), group 2, and group 3 were administered with saline, sildenafil, and pentoxifylline during the fracture healing process, respectively. Eight rats from each group were euthanized on days 15 and 30. X-ray images of the rats were taken after euthanasia for radiographical examination. Femur samples were subjected to histopathological and biomechanical (three-point bending) examinations. RESULTS: Radiologically, no difference between the Goldberg scores of the groups was found for day 15 ( p > 0.05), while higher Goldberg scores were obtained from group 2 than that of group 1 ( p > 0.05) and group 3 ( p < 0.05) for day 30. In the biomechanical analysis, higher mean breaking forces were found both for day 15 and day 30 from group 2 than those obtained from group 1 (for day 15 p > 0.05 and day 30 p > 0.05) and group 3 (for day 15 p < 0.05 and day 30 p < 0.01). Higher mean absorbed energy values were obtained from group 2 than those obtained from group 1 (for day 15 p > 0.05 and day 30 p < 0.05) and group 3 (for day 15 p < 0.01 and day 30 p < 0.01). A significant difference was not found between the histological scores of all groups ( p > 0.05) for day 15, while the histological score of group 1 on day 30 was found to be significantly lower than that of sildenafil and pentoxifylline groups ( p < 0.05). CONCLUSION: Sildenafil had a positive effect on fracture healing, while pentoxifylline did not provide consistent positive effect.

Blaschko-linear "Congenital Mixed Hemato-lymphangio-keratoma Serpiginosum" Naeviforme: A New Hybrid Entity or Various Morphological Reflections of the Two Different Vascular Abnormalities?

We present the case of a 15-year-old girl who presented with an unusual grouping of lesions on her upper left leg. The lesions had been present since birth. The patient had five different types of lesions: 1) transparent grouped or scattered yellowish vesicles; 2) keratotic-surfaced, grouped dark-yellowish papules; 3) bright-red grouped papules; 4) keratotic-surfaced grouped dark-red papules; and 5) patchy, punctate, and erythematous red macules. All of the lesions were intertwined along the lines of Blaschko and were in the form of irregular serpiginous plaques. Histopathological examinations of the lesions showed three main histological features, and diagnoses of the lesions were made as lymphangioma circumscriptum, lymphangiokeratoma, and verrucous hemangioma. To the best of our knowledge, such intertwined lesions have never been reported in the literature. Hence, we suggest that the name of this unique combination of lesions be "congenital nevoid mixed hemato-lymphangio-keratoma serpiginosum."

Effects of high-dose vitamin c and hyaluronic acid on tendon healing / Efeitos de altas doses de vitamina c e ácido hialurônico sobre a cicatrização do tendão

ABSTRACT Objective: To assess the histopathologic and biomechanical effects of hyaluronic acid (HA) and high-dose vitamin C (VC) on rat Achilles tendon healing. Methods: Forty-eight Sprague-Dawley rats were randomized to HA and VC and control groups with equal numbers. Each group was further divided into two subgroups to be sacrificed on Day 15 (n=8) and Day 30 (n=8). The Achilles tendons were cut and repaired. While the control rats remained untreated, HA and VC were administered after repair. The repaired tendons were removed for biomechanical and histopathologic analyses. In the biomechanical tests, the tendons were stretched to failure and maximum forces were measured. For histopathologic examination, the specimens were interpreted semiquantitatively using Movin's grading scale and Bonar scores. Results: The highest mean forces were obtained in the HA group on Day 15 and in the VC group on Day 30, with a significant difference between HA and VC on Day 15 between control and VC on Day 30 (p<0.05). Histological examination showed both Movin and Bonar scores decreased in all groups on Day 30, with significant improvements in the HA and VC groups (p<0.05). Conclusion: Our results demonstrated that both VC and HA had therapeutic effects on tendon healing, especially in the late phase. Level of Evidence I; High quality randomized trial with statistically significant difference.

RESUMO Objetivo: Avaliar os efeitos histopatológicos e biomecânicos do ácido hialurônico (AH) e altas doses de vitamina C (VC) na cicatrização do tendão do calcâneo em ratos. Métodos: Quarenta e oito ratos Sprague-Dawley foram randomizados em grupo AH, grupo VC e grupo controle, iguais em número. Cada grupo foi dividido em dois subgrupos a serem sacrificados no Dia 15 (n = 8) e no Dia 30 (n = 8). Os tendões do calcâneo foram cortados e reparados. Enquanto os ratos do grupo controle permaneceram não tratados, os do grupo AH/VC receberam AH e VC após o reparo. Os tendões reparados foram removidos para análises biomecânicas e histopatológicas. Nos testes biomecânicos, os tendões foram estirados até a falha e as forças máximas foram medidas. No exame histopatológico, as amostras foram interpretadas semiquantitativamente usando os escores de Movin e de Bonar. Resultados: As forças médias mais altas foram obtidas no grupo AH no Dia 15 e no grupo VC, no Dia 30, com diferença significativa entre os grupos HA e VC no Dia 15 e entre os grupos controle e VC (p < 0,05). No exame histológico, os escores de Movin e Bonar diminuíram em todos os grupos no dia 30, com melhora significativas nos grupos AH e VC (p < 0,05). Conclusões: Nossos resultados demonstraram que tanto a VC quanto o AH tiveram efeitos terapêuticos sobre cicatrização do tendão, especialmente na fase tardia. Nível de Evidência I; Estudo clínico randomizado de alta qualidade com ou sem diferença estatisticamente significante.

Effects of low molecular weight heparin and rivaroxaban on rat Achilles tendon healing.

OBJECTIVES: This study aims to compare the histopathological and biomechanical effects of low molecular weight heparin (LMWH) and rivaroxaban, which are frequently used in orthopedic surgery for thromboembolic prophylaxis, on rat Achilles tendon healing. MATERIALS AND METHODS: In the study, 36 adult, male Sprague Dawley rats weighing between 300 g and 400 g were used. Three groups were formed including 12 rats in each. Achilles tendons of all rats were cut and repaired. Nadroparin calcium was administered subcutaneously for 21 days at a dose of 170 IU AXa to the first group (LMWH group). Rivaroxaban was administered daily at a dose of 3 mg/kg for 21 days as gastric lavage to the second group (rivaroxaban group). The third group was identified as the control group and no medication was administered in this group. At the end of three weeks, tendons extracted from the groups were examined histopathologically and biomechanically. RESULTS: Bonar's and Movin's scores obtained as a result of histopathological examination were statistically significantly higher in the control group (p=0.003 and p=0.004, respectively) (high scores indicate that tendon healing is not sufficient). When type I and type III collagen ratios were examined, type I collagen ratio, which should be found at a higher ratio in mature tendon, was statistically significantly higher in rivaroxaban and LMWH groups compared with the control group (p=0.002). As a result of biomechanical examination, higher mean maximum force values were obtained from the rivaroxaban group compared with the LMWH group (p=0.31). Mean maximum force values obtained from the control group were higher than those obtained from the LMWH group (p=0.03) and the rivaroxaban group (p=0.18). CONCLUSION: Histopathological examination revealed that both LMWH and rivaroxaban have positive effects on tendon healing. However, the same positive effects were not detected in biomechanical examination.

Investigation of the biomechanical and histopathological effects of autologous conditioned serum on healing of Achilles tendon.

OBJECTIVE: The aim of this study to evaluate the effects of autologous conditioned serum (ACS) on the healing of transected rat Achilles tendons via the assessment of biomechanical and histological parameters. METHODS: The study was conducted on 45 male Sprague-Dawley rats. Five rats were used as donors for ACS preparation. Animals were randomly assigned to the experimental or control group. In both groups, the Achilles tendon was cut transversally and then sutured. In the placebo control and ACS-treated groups, saline or ACS, respectively, was injected into the repair zone three times after surgery. Ten rats from each group (ACS group, n = 20; control group, n = 20) were euthanized at days 15 and 30 after surgery for histopathological (n = 5) and biomechanical (n = 5) testing. The histopathological findings were interpreted using the Bonar and Movin scales. Tendon remodelling was evaluated via the immunohistochemical staining of collagen type 3. Biomechanical effects were assessed by tensile testing. RESULTS: The Bonar and Movin scale scores were significantly better in the ACS-treated group on both day 15 (p = 0.003 and p = 0.003, respectively) and day 30 (p = 0.005 and p = 0.004, respectively). The immunohistochemical density of collagen type 3 was significantly lower in the ACS-treated group on day 30 (p = 0.018). The type 1/3 collagen ratios of the groups were similar on days 15 and 30, as determined by Sirius Red staining (p = 0.910 and p = 0.133, respectively). In the biomechanical assessment results, the ACS-treated group's maximum load to failure values were significantly higher on day 15 (p = 0.049). CONCLUSION: Injection of ACS had a positive effect on the histopathological healing of rat Achilles tendons on days 15 and 30 and on biomechanical healing on day 15. ACS treatment contributed to lowering the collagen type 3 density by day 30. According to our study, ACS may be favourable for the treatment of human Achilles tendon injuries and tendinopathies.

Comparison of the early period effects of bone marrow-derived mesenchymal stem cells and platelet-rich plasma on the Achilles tendon ruptures in rats.

INTRODUCTION: This study aims to histopathologically, biomechanically, and immunohistochemically compare the fourth-week efficiencies of local platelet-rich plasma (PRP) and bone marrow-derived mesenchymal stem cell (rBM-MSC) treatments of the Achilles tendon ruptures created surgically in rats. MATERIALS AND METHODS: The study included 35 12-month-old male Sprague Dawley rats, with an average weight of 400-500 g. Five rats were used as donors for MSC and PRP, and 30 rats were separated into MSC, PRP, and control groups (n = 10). The Achilles tendons of the rats were cut transversely, the MSC from bone marrow was administered to the MSC group, the PRP group received PRP, and the control group received physiological saline to create the same surgical effect. In previous studies, it was shown that this physiological saline does not have any effect on tendon recovery. Thirty days after the treatment, the rats were sacrificed and their Achilles tendons were examined histopathologically, immunohistochemically, and biomechanically. RESULTS: The use of rBM-MSC and PRP in the Achilles tendon ruptures when the tendon is in its weakest phase positively affected the recovery of the tendon in histopathologic, immunohistochemical, and biomechanical manners compared to the control group (p < 0.05). While the levels of pro-inflammatory cytokines TNF-α, IFNγ, and IL 1ß were significantly low, the levels of anti-inflammatory cytokines and growth factors playing key roles in tendon recovery, such as IL2, VEGF, transforming growth factor-beta, and HGF, were significantly higher in the MSC group than those of the PRP and control groups (p < 0.05). In the MSC group, the [Formula: see text] (mm) value was significantly higher (p ˂ 0.05) than that in the PRP and control groups. CONCLUSION: rBM-MSC and PRP promote the recovery of the tendon and increase its structural strength. The use of PRP and MSC provides hope for the treatment of the Achilles tendon ruptures that limit human beings' functionalities and quality of life, particularly for athletes. It is thought that the use of MSC can be more effective for tendon healing; hence, more extensive and advanced studies are needed on this topic.