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Impact of Vascular Endothelial Growth Factor on the Shape, Survival, and Osteogenic Transformation of Gingiva-Derived Stem Cell Spheroids

Background and Objectives: Vascular endothelial growth factor (VEGF) is a protein which stimulates the formation of new blood vessels, playing a crucial role in processes such as wound healing and tumor growth. Methods: This study investigated the effects of VEGF on cell viability and osteogenic differentiation in mesenchymal stem cell (MSC) spheroids. Stem cell spheroids were fabricated using concave microwells and cultured with VEGF at concentrations of 0, 0.01, 0.1, 1, and 10 ng/mL. Morphological assessments were conducted on days 1, 3, 5, and 7, while cell viability was evaluated using the LIVE/DEAD assay and Cell Counting Kit-8. Alkaline phosphatase activity (ALP) and calcium deposition were measured to assess osteogenic differentiation, and qPCR was used to analyze osteogenic marker expression. Results: The spheroids maintained their shape across all VEGF concentrations, with the largest diameter being at 0.01 ng/mL on day 1, which decreased over time. Cell viability was highest at 0.01 ng/mL VEGF, while calcium deposition peaked at 0.1 ng/mL. Osteogenic markers, including RUNX2, osteocalcin, and COL1A1, showed significant upregulation at 1 ng/mL VEGF. Conclusions: These results suggest that VEGF enhances early osteogenic differentiation in MSC spheroids, indicating its potential for bone repair and tissue regeneration. VEGF could be applied in clinical settings for bone healing, fracture repair, and regenerative dentistry treatments.

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Effects of Pelargonium Sidoides Extract on Apoptosis and Oxidative Stress in Human Neuroblastoma Cells

Background and Objectives: Neuroblastoma is the most common extracranial solid tumor in children, often presenting challenges in treatment due to its clinical and genetic heterogeneity. This study investigated the anticancer potential of Pelargonium sidoides root extract on the human neuroblastoma cell line (SH-SY5Y). Using XTT assays, ELISA-based oxidative stress markers, and RT-PCR analysis of apoptotic genes, the study explored the extract’s effects on cell proliferation, oxidative stress, and apoptosis. Materials and Methods: For the cell culture, SH-SY5Y human neuroblastoma cells were thawed, cultured, and maintained under appropriate conditions for experiments. The dose- and time-dependent activity of Pelorgonium sidoides extract on SH-SY5Y neuroblastoma cells was investigated by XTT assay. The change in the oxidative stress marker 8-Hydroxy-2′-deoxyguanosine (8-OhDG) level was determined by ELISA for the doses applied to the control group root extract at a concentration of 25 μg/mL. Total antioxidant status (TAS) and total oxidant status (TOS) were measured from the cells in the study group with the help of a commercial kit. The oxidative stress index (OSI) was calculated by dividing the TAS by the TOS and multiplying by 100. In order to evaluate the expression levels of apoptosis-related Bax, Bcl-2, Caspase-3, Caspase-8, and Caspase-9 genes at the mRNA level in control and dose group cells, RNA isolation was performed from the SH-SY5Y control and dose group cells (IC50 value). Results: It is observed that the P. sidoides substance inhibits proliferation in cells at 24 h (p < 0.05). As the dose increases, cell proliferation decreases (p < 0.05). The IC50 value was calculated to be 113.83 μg/mL at 24 h. The concentration of 8-OhDG increased in neuroblastoma cells as a result of P. sidoides extract treatment (p < 0.05). TOS levels increased in neuroblastoma cells treated with P. sidoides extract (p < 0.01). OSI levels increased in cells treated with P. sidoides extract (p < 0.001). BAX and Caspase-8 expression increased are statistically significant in the P. sidoides dose group (p < 0.05). Conclusions: P. sidoides extract induces apoptosis in neuroblastoma cells through oxidative stress and mitochondrial- and death receptor-mediated pathways. This study highlights the potential of P. sidoides as a complementary therapeutic agent for neuroblastoma, warranting further in vivo and clinical investigations to assess its safety and efficacy.

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Prevalence and Risk Factors of Osteoporosis: A Cross-Sectional Study in a Tertiary Center

Background and Objectives: Osteoporosis is a common disease of the skeletal system that increases the risk of fracture. The prevalence of osteoporosis has been increasing as the aging population increases, affecting more than 200 million people worldwide. This study aimed to shed light on the clinical impact of osteoporosis on women’s health and quality of life by evaluating the prevalence and risk factors for this disease among postmenopausal women, using a 10-year dataset from a tertiary center. Materials and Methods: This retrospective cohort study was conducted at Dokuz Eylül University, Department of Obstetrics and Gynecology, between 2010 and 2022. A total of 3289 postmenopausal women aged 50–60 years who were undergoing routine gynecological checkups were included in the study. Patients with a prior diagnosis, a history of medical conditions, or who were taking medications affecting bone mineral density (BMD) were excluded. Data on demographics, smoking status, diabetes status, body mass index (BMI), parity, and fracture history were collected. BMD scores were classified as normal, osteopenia, or osteoporosis. Results: The prevalence of osteoporosis was 10.4%. The median age of the patients was 56.0 years. Smoking, diabetes, and a history of bone fractures were found to significantly increase the risk of osteoporosis. BMI was found to have a protective effect against osteoporosis. According to multivariate analysis, the risk of osteoporosis increased by 2.46 times in patients who smoke, 3.78 times in patients with diabetes, and 6.23 times in patients with a history of fractures. Conclusions: Awareness of modifiable risk factors such as smoking is crucial for preventing osteoporosis-related complications. Diabetes, even when it is not complicated, increases the risk of osteoporosis. Physical exercise, calcium, and vitamin D intake are important, especially during young adulthood, as they are the primary contributors to peak bone mass. Because neck fractures are more common in patients with osteopenia, early diagnosis and routine screening can mitigate future complications.

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Stem Cells Within Three-Dimensional-Printed Scaffolds Facilitate Airway Mucosa and Bone Regeneration and Reconstruction of Maxillary Defects in Rabbits

Background and Objectives: Current craniofacial reconstruction surgical methods have limitations because they involve facial deformation. The craniofacial region includes many areas where the mucosa, exposed to air, is closely adjacent to bone, with the maxilla being a prominent example of this structure. Therefore, this study explored whether human neural-crest-derived stem cells (hNTSCs) aid bone and airway mucosal regeneration during craniofacial reconstruction using a rabbit model. Materials and Methods: hNTSCs were induced to differentiate into either mucosal epithelial or osteogenic cells in vitro. hNTSCs were seeded into polycaprolactone scaffold (three-dimensionally printed) that were implanted into rabbits with maxillary defects. Four weeks later, tissue regeneration was analyzed via histological evaluation and immunofluorescence staining. Results: In vitro, hNTSCs differentiated into both mucosal epithelial and osteogenic cells. hNTSC differentiation into respiratory epithelial cells was confirmed by Alcian Blue staining, cilia in SEM, and increased expression levels of FOXJ1 and E-cadherin through quantitative RT-PCR. hNTSC differentiation into bone was confirmed by Alizarin Red staining, increased mRNA expression levels of BMP2 (6.1-fold) and RUNX2 (2.3-fold) in the hNTSC group compared to the control. Four weeks post-transplantation, the rabbit maxilla was harvested, and H&E, SEM, and immunohistofluorescence staining were performed. H&E staining and SEM showed that new tissue and cilia around the maxillary defect were more prominent in the hNTSC group. Also, the hNTSCs group showed positive immunohistofluorescence staining for acetylated α-tubulin and cytokerin-5 compared to the control group. Conclusions: hNTSCs combined with PCL scaffold enhanced the regeneration of mucosal tissue and bone in vitro and promoted mucosal tissue regeneration in the in vivo rabbit model.

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The Impact of Diet on Hidradenitis Suppurativa Severity: A Cross-Sectional Case–Control Study

Background and Objectives: Hidradenitis suppurativa (HS) is a chronic inflammatory skin disease primarily affecting hair follicles, characterized by painful nodules, abscesses, and sinus tract formation. Recent evidence suggests that weight management and nutritional factors may influence HS symptoms. This cross-sectional case–control study aimed to assess the impact of body composition and nutritional factors on HS severity. Materials and Methods: We included 50 patients with HS and 50 matched controls comparable in body mass index (BMI), sex, and age. The data collected included 3-day food records, a food frequency questionnaire, 24 h physical activity records, Mediterranean Diet Adherence Screener (MEDAS) scores, body composition, and anthropometric measurements. The macronutrient and micronutrient intake, as well as dietary glycemic index (GI), were analyzed. HS severity was assessed using the Hurley staging system and the International Hidradenitis Suppurativa Severity Score System (IHS4). Results: HS patients exhibited significantly lower adherence to the Mediterranean diet and a higher dietary GI compared to controls. Their micronutrient intake was also reduced, while the consumption of junk food, dairy products, and nightshade vegetables was more frequent. The MEDAS scores and physical activity levels were negatively correlated with the IHS4 scores, while higher anthropometric measurements, dietary energy, protein, total fat, and GI showed positive correlations. The Hurley stage was negatively correlated with the MEDAS scores and positively correlated with the GI and visceral fat. In multiple regression analysis, the MEDAS score emerged as the primary variable associated with disease severity. Conclusions: These findings suggest that an increased adherence to the Mediterranean diet, intake of food with a lower GI, and maintaining an ideal body weight may positively affect HS management. Long-term studies are warranted to corroborate our findings.

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Does Robotic Spine Surgery Add Value to Surgical Practice over Navigation-Based Systems? A Study on Operating Room Efficiency

Background and Objectives: Spine surgery has undergone significant advancements, particularly with regard to robotic systems that enhance surgical techniques and improve patient outcomes. As these technologies become increasingly integrated into surgical practice, it is essential to evaluate their added value and cost savings. Hence, this study compared robot-assisted and navigation-based spine surgery, focusing on surgical efficiency. Materials and Methods: We conducted a single-center, retrospective cohort study of patients undergoing single- and double-level transforaminal lumbar interbody fusion (TLIF) and oblique lumbar interbody fusion (OLIF) surgeries. Patients were divided into two groups: those who had robot-assisted and navigation-based surgeries, stratified by surgery type (TLIF or OLIF) and fusion levels (one or two). A comparative analysis of factors related to surgical efficiency, including operative duration, blood loss, and length of hospital stay, was conducted. Results: Our results showed a statistically significant reduction in operative duration for robot-assisted one- and two-level OLIF cases, with average time savings of 50 and 62 min, respectively, compared to navigation-based surgery. These time savings translated to an estimated cost reduction of SGD 1500 for the hospital for each patient for a two-level OLIF procedure and could be higher as the number of operated levels increase. Conclusions: These results indicated that robot-assisted spine surgery offers superior surgical efficiency and cost savings, particularly with increased numbers of surgical levels. As robotic technologies evolve, their integration into spine surgery is justified, promising improved patient outcomes and cost-effectiveness.

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Effects of Sodium-Glucose Cotransporter-2 Inhibitors on Body Composition and Fluid Status in Cardiovascular Rehabilitation Patients with Coronary Artery Disease and Heart Failure

Background and Objectives: Sodium glucose cotransporter-2 (SGLT-2) inhibitors have emerged as integral therapeutic tools in the management of patients with cardiovascular–kidney–metabolic (CKM) syndrome. In addition to their well-documented effects on lowering glucose levels and cardiovascular- and reno-protective actions, SGLT-2 inhibitors, through a reduction in body weight (BW), generate changes in the body composition and volume status that have not been clearly studied. Materials and Methods: This retrospective, observational longitudinal cohort, single-center study analyzed and compared body composition and fluid status measured by bioelectrical impedance analysis (BIA) from weeks 0 to 12 after the initiation of the cardiac rehabilitation (CR) program for coronary artery disease and heart failure in 59 patients who started treatment with SGLT-2 inhibitors (SGLT-2iG) and 112 patients without SGLT-2 inhibitors (non-SGLT-2iG). Results: Changes between the baseline and week 12 in the SGLT-2iG and non-SGLT-2iG were −0.3 L (p = 0.003) and −0.03 L (p = 0.82) in extracellular water (ECW) (p = 0.05), −0.39 L (p < 0.001) and −0.14 L (p = 0.33) in intracellular water (ICW) (p = 0.12), −0.69 (p < 0.001) and −0.16 (p = 0.52) in total body water (TBW) (p = 0.08), and −0.01 (p = 0.37) and −0.001 (p = 0.25) in the ECW/TBW ratio, respectively. After 3 months of exercise therapy in the CR program, patients in the SGLT-2iG showed a greater decrease than the non-SGLT-2iG in weight (−1.34 kg, p < 0.001 vs. −0.99, p = 0.02), body mass index (BMI) (−0.45 kg/m2, p < 0.001 vs. −0.38, p = 0.004), arm circumference (−0.57 cm, p = 0.008 vs. −0.12 cm, p = 0.21), waist circumference (−1.5 cm, p = 0.04 vs. −0.11 cm, p = 0.83), systolic blood pressure (SBP) (−8.9 mmHg, p = 0.049 vs. −4.19, p = 0.08), and diastolic blood pressure (DBP) (−5.15, p = 0.03 vs. −2.85, p = 0.01). The bioelectrical impedance analysis (BIA) revealed a significant decrease in body fat mass (BFM) and visceral fat area, without a loss of lean body mass (LBM) or skeletal muscle mass in the SGLT-2iG. Conclusions: SGLT-2 inhibitors exert beneficial effects on body compartments and volume status. Although they induce modest weight loss, this appears to be mainly directed at ECW, BFM, and visceral fat, without a loss of LBM nor skeletal muscle mass, which could contribute to the observed CKM benefits.

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Hepatitis C Virus Infection in Hemodialysis Patients in the Era of Direct-Acting Antiviral Treatment: Observational Study and Narrative Review

Background and Objectives: Hepatitis C virus (HCV) infection is a major global public health concern, particularly in hemodialysis (HD) patients. This study aims to evaluate the demographic, clinical, and laboratory characteristics of HCV-positive patients undergoing HD and assess the long-term impact of direct-acting antivirals (DAAs) on patient outcomes. Moreover, a narrative review aims to summarize the current knowledge regarding HCV treatment in HD patients. The search in the PubMed, Google Scholar, and Scopus databases identified 48 studies relevant to our topic, 18 regarding clinical history and 29 related to HCV treatment. Methods: A retrospective analysis was performed on 165 HD patients from Bihor County HD centers, Romania, between 2014 and 2024. The cohort was divided into two groups: 54 patients who tested positive for HCV and 111 controls who were HCV-negative. Data collected from GPs included demographic information, comorbidities, laboratory parameters, and psychological assessments. Outcomes were evaluated at over 5 years after DAA treatment. A literature review was conducted using PubMed and Google Scholar to identify relevant studies on HCV in HD patients from 1989 to 2024. Results: Laboratory results showed similar parameters across groups, except for lower serum cholesterol levels in the HCV-positive DAA-treated group vs. HCV-positive non-treated ones (155.607 mg% vs. 170.174 mg%, p = 0.040) and increased ALT levels when comparing the same groups (29.107 vs. 22.261, p = 0.027), whereas comorbidities did not differ significantly. The incidence of malignancies was significantly higher among HCV-positive compared to HCV-negative patients (20.3% vs. 8.1%, p = 0.023), mainly among those treated with DAAs, highlighted by the multivariate analysis. Cardiovascular disease remains the leading cause of mortality regardless of HCV status or the use of antiviral therapy. Psychological assessments revealed more severe depression in HCV-positive patients compared to their HCV-negative counterparts. Conclusions: HCV infection in the hemodialysis population typically follows a subclinical course. At over five years after DAA therapy, the results indicate a stabilization of the liver function and the absence of major complications. However, the incidence of malignancies remains high in HCV-positive patients.

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Metabolic Dysfunction-Associated Steatotic Liver Disease in Type 2 Diabetes Patients—The Relationship with Platelets Indicators

Background and Objectives: Metabolic dysfunction-associated steatotic liver disease (MASLD) is an important chronic liver disease with major health risks, especially in the presence of T2DM, but the pathophysiology of this condition is not fully understood. This study aimed to investigate the platelet hematometric indices in patients with T2DM and MASLD. Materials and Methods: Demographic and medical (including anthropometric) data were collected from 271 participants, from whom blood samples were also drawn in fasting conditions for complete blood count, liver and metabolic panel, ferritin, haptoglobin, creatinine, and fibrosis markers. The correlations of main platelet parameters with clinical and laboratory data were investigated by bivariate and multiple regression analyses. Results: The median platelets number was 235·103/μL, and thus, the study population was divided into two subgroups: with higher and lower numbers (group 1 (mean): 286.38 ± 43.29·103/μL and group 2 (mean): 188.12 ± 39.77·103/μL). Despite similar BMIs, group 2 had higher fatty liver index (FLI) (84.44 ± 18.04 vs. 79.85 ± 17.98; p = 0.0088) and insulin resistance (HOMA-IR: 3.16 ± 1.50 vs. 2.63 ± 1.31; 0.0008), higher direct bilirubin, transaminases, uric acid, and ferritin concentrations. Higher percentages of males and subjects with HOMA-IR values >2.5 were accounted for in this group. In the multiple regression analyses, the platelet count and plateletcrit (PTC) correlated independently with sex, leucocyte count, HOMA-IR, and bilirubin concentrations (p < 0.0001). The platelet distribution width (PDW) was positively correlated with insulin resistance in two separate analyses (β = 0.060; p = 0.0004, and β = 0.052; p = 0.0025), and with GGT, while the mean platelet volume presented a weak but significant positive association with FLI. Patients with higher HOMA-IR had higher PDW and a lower platelet count and PTC. Conclusions: Male patients with T2DM and MASLD had lower platelet count and PTC and larger PDW. Higher insulin resistance was associated with lower platelet count and PTC and higher PDW.

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