Despite efforts in osteosarcoma (OS) research, the role of inductive moderate hyperthermia (IMH) in delivering and enhancing the antitumor effect of liposomal doxorubicin formulations (LDOX) remains unresolved. This study investigated the effect of a combination treatment with LDOX and IMH on Saos-2 human OS cells. We compared cell viability using a trypan blue assay, apoptosis and reactive oxygen species (ROS) measured by flow cytometry and pro-apoptotic Bax protein expression examined by immunocytochemistry in response to IMH (42 MHz frequency, 15 W power for 30 min), LDOX (0.4 μg/mL), and LDOX plus IMH. The lower IC₅₀ value of LDOX at 72 h indicated increased accumulation of the drug in the OS cells. LDOX plus IMH resulted in a 61% lower cell viability compared to no treatment. Moreover, IMH potentiated the LDOX action on the Saos-2 cells by promoting ROS production at temperatures of <42 °C. There was a 12% increase in cell populations undergoing early apoptosis with a less heterogeneous distribution of Bax after combination treatment compared to those treated with LDOX (p < 0.05). Therefore, we determined that IMH could enhance LDOX delivery and its antitumor effect via altered membrane permeabilization, ROS generation, and a lower level of visualized Bax heterogeneity in the Saos-2 cells, suggesting the potential translation of these findings into in vivo studies.

Magnetic nanocomposites (MNC) are promising theranostic platforms with tunable physicochemical properties allowing for remote drug delivery and multimodal imaging. Here, we developed doxorubicin-loaded Fe₃O₄–Au MNC (DOX-MNC) using electron beam physical vapor deposition (EB-PVD) in combination with magneto-mechanochemical synthesis to assess their antitumor effect on Walker-256 carcinosarcoma under the influence of a constant magnetic (CMF) and electromagnetic field (EMF) by comparing tumor growth kinetics, magnetic resonance imaging (MRI) scans and electron spin resonance (ESR) spectra. Transmission (TEM) and scanning electron microscopy (SEM) confirmed the formation of spherical magnetite nanoparticles with a discontinuous gold coating that did not significantly affect the ferromagnetic properties of MNC, as measured by vibrating-sample magnetometry (VSM). Tumor-bearing animals were divided into the control (no treatment), conventional doxorubicin (DOX), DOX-MNC and DOX-MNC + CMF + EMF groups. DOX-MNC + CMF + EMF resulted in 14% and 16% inhibition of tumor growth kinetics as compared with DOX and DOX-MNC, respectively. MRI visualization showed more substantial tumor necrotic changes after the combined treatment. Quantitative analysis of T₂-weighted (T₂W) images revealed the lowest value of skewness and a significant increase in tumor intensity in response to DOX-MNC + CMF + EMF as compared with the control (1.4 times), DOX (1.6 times) and DOX-MNC (1.8 times) groups. In addition, the lowest level of nitric oxide determined by ESR was found in DOX-MNC + CMF + EMF tumors, which was close to that of the muscle tissue in the contralateral limb. We propose that the reason for the relationship between the observed changes in MRI and ESR is the hyperfine interaction of nuclear and electron spins in mitochondria, as a source of free radical production. Therefore, these results point to the use of EB-PVD and magneto-mechanochemically synthesized Fe₃O₄–Au MNC loaded with DOX as a potential candidate for cancer magnetic nanotheranostic applications.

Evaluating intratumor heterogeneity with image texture analysis offers a more sophisticated understanding of sarcoma response to treatment. We examined the effects of inductive moderate hyperthermia (IMH) on sarcoma-45 growth and intratumor heterogeneity across tissue, cellular and molecular levels using magnetic resonance imaging (MRI), ultrasound and histology image analysis. IMH (42 MHz, 20 W) inhibited sarcoma-45 growth kinetics by 34% compared to the untreated control group. T₂-weighted MRI brightness was increased by 42%, reflecting more extensive tumor necrosis, while Young’s modulus increased by 37% due to more pronounced connective tissue replacement in response to IMH. Whereas calculations of Moran’s spatial autocorrelation index revealed distinctions in heterogeneity between tumor core, periphery and capsule regions of interest (ROIs) on MRI, ultrasound and histological examination in the untreated tumor-bearing animals, there was no significant difference between core and periphery after IMH. Exposure to IMH increased overall tumor ROI heterogeneity by 22% on MRI but reduced heterogeneity in the core and periphery on ultrasound and histology images. Ki-67 protein distribution was 25% less heterogeneous on the tumor periphery after IMH. Therefore, this study provides a quantitative characterization of IMH effects on different manifestations of intratumor sarcoma-45 heterogeneity using experimental imaging data.

Background: Osteosarcoma (OS) is the most common primary malignant bone tumor. Doxorubicin (DOX) is extensively used in OS chemotherapy, yet improving patient outcomes remains challenging. This study investigated the effect of free DOX combined with inductive moderate hyperthermia (IMH) on Saos-2 human OS cells. Methods: Cell viability was assessed by trypan blue exclusion. Flow cytometry analyzed apoptosis, necrosis, and reactive oxygen species (ROS) in cells exposed to control (no treatment), IMH (42 MHz frequency, 500 μT magnetic field induction, 564 V/m electric field strength, 15 W output power, and 30 min duration) alone, DOX (0.06 μg/mL) alone, or DOX combined with IMH. The expression of p14^ARF tumor suppressor and epidermal growth factor receptor (EGFR) was evaluated by immunocytochemistry. Spatial autocorrelation analysis quantified the heterogeneity of p14^ARF and EGFR distributions in acquired images. Results: The half maximal inhibitory concentration (IC₅₀) of DOX in Saos-2 cells had minimal variation between 48 h (0.060 ± 0.01 μg/mL) and 72 h (0.055 ± 0.003 μg/mL). DOX + IMH resulted in a 15% increase in early apoptosis and a 20% elevation in ROS levels compared with DOX alone. Immunocytochemical analysis revealed a 37% increase in p14^ARF and a 32% reduction in EGFR expression following combined treatment in comparison to DOX alone. Image analysis showed that DOX + IMH treatment caused the highest Moran’s index values for p14^ARF and EGFR, reflecting less heterogeneous spatial distributions (p < 0.05). Conclusions: IMH enhanced DOX-induced cytotoxicity in Saos-2 cells by initiating ROS-mediated apoptosis and reducing heterogeneity of cellular responses.

The objective of this study was to determine the sensitivity of tartrate-resistant acid phosphatase (Bone TRAP-5b) for early detection of bone metastases (BM) and to investigate the efficacy of bisphosphonates (BF) (zolendronic acid-ZA) in prevention of bone metastases in patients with Renal Cell Carcinoma (RCC). The 60 patients with RCC with proven BM were investigated to assess the sensitivity and specificity of Bone TRAP-5b. 95 patients with RCC with high level of tartrate-resistant acid phosphatase (Bone TRAP-5b) (8,5±0,2 IU/L) after radical surgical treatment were divided into two groups: 1-st group: (n=44) received zolendronic acid (ZA) (BF +), and 2-nd group (n=51) patients didn’t receive ZA (BF-). Patients of both subgroups were similar by age, sex, stage of disease. The levels of Bone TRAP-5b, Ca++, alkaline phosphatase, LDG were accessed every 3 months, and MRI imaging, bone scan with 99mTc every 6 month in both groups. We determined the high correlation between bone TRAP-5b and the presence of bone metastases (r=0,9; p <0,05), but its level wasn’t dependent with the number of BM. The results showed the high sensitivity and specificity of Bone TRAP-5b at the critical value of 5.2 IU/L (98,3 % and 90,0 %), (χ2=64,6; p<0.01). Using BF for the prevention of bone metastases in high risk group patients with RCC provides a significant difference in the incidence of bone metastases in patients.

BACKGROUND: Skin grafts may be used to reconstruct large skin defects after the excision of malignant skin tumors. Vacuum-assisted closure (VAC) leads closer approximation of the free flap. AIM: The objective of this study was to estimate the engraftment of grafts using VAC and explore factors influencing this process. MATERIALS AND METHODS: This study included 31 patients with skin cancers after a wide tumor excision. The wound defects were closed using a split-thickness skin graft and the VAC dressing. RESULTS: Complete and partial engraftment was observed in 17 (54.8 %) and 14 (45.2%) cases, respectively. The mean area of the wound defects among patients with complete engraftment was 24.3 ± 20.6 cm2, while that of the group with partial engraftments was 56.5 ± 58.8 cm2 (p = 0.0017). In the complete engraftment group, the average number of days with the VAC dressing was 7.2 ± 0.44 days, versus 6.9 ± 0.27 days in the partial engraftment group (p = 0.032). CONCLUSION: VAC dressing after skin grafting in the surgical management of malignant skin tumors provided good results. The area of the wound defect and the average number of days with VAC dressing is significant factors influencing the outcomes of skin grafting.