Nuclear targeting peptide-modified,DOX-loaded,PHBV nanoparticles enhance drug efficacy by targeting to Saos-2 cell nuclear membranes

The aim of this study was to target nano sized (266 ± 25 nm diameter) poly(3-hydroxybutyrate-co-3-hydroxyvalerate) (PHBV) particles carrying Doxorubicin (DOX), an anticancer agent, to human osteosarcoma cells (Saos-2). A nuclear targeting molecule (Nuclear Localization Signal, NLS), a 17 a.a. peptide, was attached onto the doxorubicin loaded nanoparticles. NLS conjugated nanoparticles surrounded the cell nuclei, but did not penetrate them. Free doxorubicin and doxorubicin loaded nanoparticles entered the cytoplasm and were evenly distributed within the cytoplasm. The localization of the NLS-targeted particles around the nuclear membrane caused a significantly higher decrease in the cancer cell numbers due to apoptosis or necrosis than the untargeted and free doxorubicin formulations showing the importance of targeting the nanoparticles to the nuclear membrane in the treatment of cancer.     

Enhanced anticancer potency by thermo/pH-responsive PCL-based magnetic nanoparticles

Great efforts have been made to develop drug carriers with the aim of providing predictable therapeutic response. Moreover, combination therapies have become promising strategies for clinical cancer treatment with synergistic effects. The present study purposed to develop a new stimuli-responsive paramagnetic nanocarrier for the intracellular co-delivery of doxorubicin (DOX) and methotrexate (MTX) to the MCF7 cell line. A novel thermo/pH-sensitive amphiphilic paramagnetic nanocomposite comprised of hydrophobic and biodegradable PCL segments and a hydrophilic biocompatible P(NIPAAm-co-HEMA-co-MAA-co-TMSPMA) block was designed and synthesized by combining the ring opening and free radical polymerization methods. The structure and physic-chemical characterization of synthesized nanoparticles and intermediates were studied and revealed using FTIR, HNMR, CNMR, SEM, EDX, TGA, and VSM techniques. DOX and MTX on a nanocarrier achieved 95.04 and 97.29% encapsulation efficiency, respectively. The dual drug release profile revealed tumor niche-assisted release behavior (more drug release was observed at a temperature of 41 °C and pH ≤ 5.4). The antitumor ability of the DOX/MTX-loaded nanocomposite was significantly higher than that of free drugs, confirmed by MTT assay, DAPI staining, cell cycle, and real-time PCR analysis on MCF7 cell lines. Furthermore, the cytotoxicity assay of a nanocarrier to the MCF7 cell line revealed its suitability as an anticancer drug nanocarrier. The results indicated that this engineered dual anticancer drug delivery system ensures increased antitumor activity as well as decreased toxicity in comparison with the free drugs.     

Low-dose Methotrexate Inhibits Lung Metastasis and Lengthens Survival in Rat Osteosarcoma

Lung metastasis is the most crucial event affecting the treatment of osteosarcoma and is dependent on tumor angiogenesis. To improve the prognosis for patients with osteosarcoma, prevention of lung metastasis is essential. Low-dose methotrexate is a useful drug for treating a variety of diseases. Low-dose methotrexate reportedly plays a role in antiangiogenesis for the synovial blood vessels in rheumatoid arthritis. However, whether low-dose methotrexate is correlated with tumor angiogenesis and metastasis is unclear. We investigated the inhibitory effect of methotrexate on lung metastasis in a rat osteosarcoma cell line with high metastatic potential, S-SLM. Two weeks after inoculation of S-SLM cells into male Fischer 344 rats, low-dose methotrexate (1.2 mg/kg once or twice a week) or saline was intraperitonealy injected for 4 weeks and the antimetastatic effect was evaluated. Low-dose methotrexate significantly reduced the number of lung metastatic nodules and the wet weight of the lungs. Immunohistochemical staining showed a decrease in microvessel density in the metastatic nodules. We also evaluated the effect of methotrexate on the proliferation of endothelial cells and S-SLM osteosarcoma cells in vitro. Methotrexate significantly inhibited the proliferation of endothelial cells at a lower concentration than that of S-SLM osteosarcoma cells. These data suggest that low-dose methotrexate inhibited lung metastasis of osteosarcoma through its antiangiogenic activity. Our results indicate that low-dose methotrexate is a promising drug for tumor dormancy therapy in patients with osteosarcoma and lung metastasis.     

Suppression of inflammation in a mouse model of rheumatoid arthritis using targeted lipase-labile fumagillin prodrug nanoparticles

Nanoparticle-based therapeutics are emerging technologies that have the potential to greatly impact the treatment of many human diseases. However, drug instability and premature release from the nanoparticles during circulation currently preclude clinical translation. Herein, we use a lipase-labile (Sn 2) fumagillin prodrug platform coupled with a unique lipid surface-to-surface targeted delivery mechanism, termed contact-facilitated drug delivery, to counter the premature drug release and overcome the inherent photo-instability of fumagillin, an established anti-angiogenic agent. We show that α(v)β(3)-integrin targeted fumagillin prodrug nanoparticles, administered at 0.3?mg of fumagillin prodrug/kg of body weight suppress the clinical disease indices of KRN serum-mediated arthritis in a dose-dependent manner when compared to treatment with the control nanoparticles with no drug. This study demonstrates the effectiveness of this lipase-labile prodrug nanocarrier in a relevant preclinical model that approximates human rheumatoid arthritis. The lipase-labile prodrug paradigm offers a translatable approach that is broadly applicable to many targeted nanosystems and increases the translational potential of this platform for many diseases.     

FA and cRGD dual modified lipid-polymer nanoparticles encapsulating polyaniline and cisplatin for highly effective chemo-photothermal combination therapy

A combination of chemotherapy and photothermal therapy as a promising strategy has exhibited noticeable therapeutic effect on cancer therapy. To ensure the exertion of synergistic effect on a tumor region, a multifunctional vehicle for selectively delivering therapeutic agent into tumor cells is highly desirable. Thus, folate-poly (ethylene glycol)-distearoylphosphatidylcholine (FA-PEG-DSPE), cRGD [cyclic (Arg-Gly-Asp-D-Phe-Lys)]-PEG-DSPE and lecithin were employed to develop dual modified nanoparticles (FA/cRGD-PNPs) encapsulating polyaniline and cisplatin by a film-ultrasonic dispersion method. The FA/cRGD-PNPs showed a uniform size of 102.7 nm, remarkable stability and monodispersity, and highly localized temperature respond. Compared to chemo or photothermal treatment alone, the combined treatment on cells in vitro significantly suppressed the survival rate of MDA-MB-231 cells (1.87%) and MGC-803 cells (2.37%) treated for 48 h. The results further indicated the induced cell apoptosis rate of MDA-MB-231 cells reached to 92.6% with treatment for 24 h. Hence, our research highlights the great potential in drug delivery and the combination of chemotherapy and photothermal therapy.     

Effects of vascular endothelial growth factor expression on pathological characteristics and prognosis of osteosarcoma

Vascular endothelial growth factor (VEGF) has been linked with tumor invasion and metastasis. However, the role of VEGF expression in osteosarcoma remains controversial. By searching the PubMed, Embase, and Google Scholar databases, we conducted a meta-analysis to evaluate the pathological and prognostic significance of VEGF in osteosarcoma. Studies were pooled, and the odds ratio (OR) and its corresponding 95 % confidence interval (CI) were calculated. Nine relevant articles were included in this meta-analysis study. We performed pooled analysis with available data on the association between VEGF expression and age, gender, tumor stages IIB–III versus I–IIA, tumor recurrence, response to chemotherapy, and tumor metastasis. Our results revealed that VEGF expression might be closely associated with metastasis of osteosarcoma (OR 4.74, 95 % CI 2.53–8.87, P < 0.001). Furthermore, our findings also demonstrated that patients with grade IIB–III osteosarcoma showed a higher frequency of VEGF expression than those with grade I–IIA osteosarcoma (OR 5.33, 95 % CI 2.03–13.98, P = 0.001). We failed to find the association between VEGF expression and age (OR 0.82, 95 % CI 0.44–1.53, P = 0.539), gender (OR 1.33, 95 % CI 0.52–3.42, P = 0.553), tumor recurrence (OR 1.47, 95 % CI 0.56–3.86, P = 0.429), and response to chemotherapy (OR 1.26, 95 % CI 0.14–11.72, P = 0.839). In conclusion, VEGF is related to the grade and metastasis of osteosarcoma. It may play a significant role in clinical guidelines for the treatment and prognostic evaluation.     

Pathological responses to preoperative high-dose methotrexate chemotherapy in osteosarcoma--experience in Korea cancer hospital.

During the last decade, many clinical investigators at various cancer centers have reported the efficacy of various chemotherapeutic agents in the treatment of osteosarcoma. The regimens using high-dose methotrexate (HDMTX) with citrovorum factor rescue are now considered to be one of the most effective treatments of choice. From December 1989 to May 1991, sixteen patients with Enneking''s stage (Enneking et al., 1980) IIB osteosarcoma of the extremities were treated with a high-dose methotrexate regimen. After two cycles of preoperative chemotherapy, an operation was performed; either limb salvage or amputation. The resected lesions were examined pathologically and classified according to Huvos'' criteria. On pathological examination, 8 (50%) cases showed Grade IV; 1 (6.25%) Grade III; 4 (25%) Grade II; and 3 (18.75%) Grade I. The types of surgery performed were tumor prosthesis replacement (11); wide resection with or without reconstruction (2); resection and arthrodesis (1); and amputation (2).     

Aven‐mediated checkpoint kinase control regulates proliferation and resistance to chemotherapy in conventional osteosarcoma

Conventional high‐grade osteosarcoma is the most common primary bone sarcoma, with relatively high incidence in young people. In this study we found that expression of Aven correlates inversely with metastasis‐free survival in osteosarcoma patients and is increased in metastases compared to primary tumours. Aven is an adaptor protein that has been implicated in anti‐apoptotic signalling and serves as an oncoprotein in acute lymphoblastic leukaemia. In osteosarcoma cells, silencing Aven triggered G₂ cell‐cycle arrest; Chk1 protein levels were attenuated and ATR–Chk1 DNA damage response signalling in response to chemotherapy was abolished in Aven‐depleted osteosarcoma cells, while ATM, Chk2 and p53 activation remained intact. Osteosarcoma is notoriously difficult to treat with standard chemotherapy, and we examined whether pharmacological inhibition of the Aven‐controlled ATR–Chk1 response could sensitize osteosarcoma cells to genotoxic compounds. Indeed, pharmacological inhibitors targeting Chk1/Chk2 or those selective for Chk1 synergized with standard chemotherapy in 2D cultures. Likewise, in 3D extracellular matrix‐embedded cultures, Chk1 inhibition led to effective sensitization to chemotherapy. Together, these findings implicate Aven in ATR–Chk1 signalling and point towards Chk1 inhibition as a strategy to sensitize human osteosarcomas to chemotherapy. Copyright © 2015 Pathological Society of Great Britain and Ireland. Published by John Wiley & Sons, Ltd.     

Effects of blood purification therapy on a patient with ifosfamide-induced neurotoxicity and acute kidney injury

Ifosfamide combined with other antineoplastic agents has been effective in the treatment of osteosarcoma, although adverse effects are reported in the increasing use of ifosfamide. The most serious complications among the ifosfamide intoxications are neurotoxicity and nephrotoxicity. We report on a patient who suffered from ifosfamide-induced neurotoxicity and nephrotoxicity and rhabdomyolysis after chemotherapy, and was successfully treated with blood purification therapy. The patient had osteosarcoma with multiple lung metastases, wherein the chemotherapy included ifosfamide (3 g/m²) and VP-16 (60 mg/m²) per day for 3 days. The first day after chemotherapy, the patient experienced impaired consciousness and renal function. Based on the clinical course and laboratory data, the diagnosis was ifosfamide-induced neurotoxicity and the acute kidney injury caused by ifosfamide-induced nephrotoxicity and rhabdomyolysis. As a detoxification treatment, blood purification procedures were performed daily for 3 days. Thirty-six hours after the first hemodialysis session, the symptoms of neurotoxicity disappeared. In the lead-up to the 10th day following intoxication, the serum creatinine recovered to the baseline level. Serum ifosfamide concentration decreased from 41.9 to 12.1 ng/ml by the second session of blood purification. Despite the absence of an established detoxification method when complications present simultaneously, blood purification therapy should be considered for treating severe concurrent neurotoxicity and nephrotoxicity and rhabdomyolysis.     

高分子纳米药物治疗骨肉瘤

背景:近年来研究者开发了各种高分子纳米粒子作为抗肿瘤药物载体,并利用纳米粒子的优势,例如血液循环时间延长、肿瘤内选择性聚集等,提高对骨肉瘤的疗效。目的:基于最新的相关研究,对高分子纳米药物在骨肉瘤治疗方面的应用及其发展前景作以综述。方法:作者应用计算机检索Web of Science、NCBI和PubMed生物医学数据库,检索时间为1900年至2019年6月,以“osteosarcoma;polymer;nanoparticle;controlled drug delivery;tumor therapy”为检索关键词,初检文章265篇,筛选后将107篇文章纳入高分子纳米药物治疗骨肉瘤的相关研究报道。结果与结论:骨肉瘤是最常见的恶性骨肿瘤,主要影响儿童和青少年,早期远处肺转移和局部高侵袭性使骨肉瘤患者长期生存率降低。虽然化疗提高了骨肉瘤患者的生存率,但其应用潜力因严重不良反应和耐药性受到限制。与传统化疗相比,高分子纳米药物不仅降低了对正常组织的毒副作用,而且还能够延长体内循环时间,使化疗药物在肿瘤部位持续缓慢释放,从而提高了治疗效果。因此高分子纳米药物对于骨肉瘤的治疗具有巨大的应用前景。     

The clinical importance of recognizing capecitabine-induced hypertriglyceridemia: A case report and review of the literature

This case report describes a patient who developed a mixed hyperlipidemia and severe hypertriglyceridemia (>20 mmol/L or 1772 mg/dL) while receiving capecitabine chemotherapy. After lipid-lowering treatment (statin and omega-3 acid ethyl esters) and completion of chemotherapy, her lipid levels had been significantly reduced. Other secondary causes of hyperlipidemia were also investigated. In view of the abnormal lipid results worsened by capecitabine, we suggest that careful lipid monitoring and thorough lipid management are important in such patients to avoid acute pancreatitis.     

Can Molecular Imaging Enable Personalized Diagnostics? An Example Using Magnetomotive Photoacoustic Imaging

The advantages of photoacoustic (PA) imaging, including low cost, non-ionizing operation, and sub-mm spatial resolution at centimeters depth, make it a promising modality to probe nanoparticle-targeted abnormalities in real time at cellular and molecular levels. However, detecting rare cell types in a heterogeneous background with strong optical scattering and absorption remains a big challenge. For example, differentiating circulating tumor cells in vivo (typically fewer than 10 cells/mL for an active tumor) among billions of erythrocytes in the blood is nearly impossible. In this paper, a newly developed technique, magnetomotive photoacoustic (mmPA) imaging, which can greatly increase the sensitivity and specificity of sensing targeted cells or molecular interactions, is reviewed. Its primary advantage is suppression of background signals through magnetic enrichment/manipulation with simultaneous PA detection of magnetic contrast agent targeted objects. Results from phantom and in vitro studies demonstrate the capability of mmPA imaging to differentiate regions targeted with magnetic nanoparticles from the background, and to trap and sensitively detect targeted cells at a concentration of a single cell per milliliter in a flow system mimicking a human peripheral artery. This technique provides an example of the ways in which molecular imaging can potentially enable robust molecular diagnosis and treatment, and accelerate the translation of molecular medicine into the clinic.     

LyP-1-conjugated Fe3O4 nanoparticles suppress tumor growth by magnetic induction hyperthermia

To find a promising drug carrier to suppress tumor using magnetic induction hyperthermia (MIH) and targeted therapy, two superparamagnetic iron oxide nanoparticles (SPIONs) coated with polyethylene glycol (PEG) and LyP-1, respectively, were prepared and compared. The particle size ranges of PEG-SPIONs and LyP-1-SPIONs were 10–15 nm, and 15–20 nm, respectively. In FTIR spectra, PEG-SPIONs and LyP-1-SPIONs had strong peaks between 575 and 1630 cm^?1. Specifically, the PEG-SPIONs mainly has peaks in 581 and 1630 cm^?1. The LyP-1-SPIONs mainly had peaks in 575, 1050 and 1625 cm^?1. The contents of Fe₃O₄ in the PEG-SPIONs and LyP-1-SPIONs were about 94.24 and 89.26%, respectively. The iron contents in the MCF-7 and CT-26 cells were 33.1 ± 1.8 and 27.9 ± 0.95 pg, respectively, after co-incubation with LyP-1-SPIONs for 8 h. The LyP-1-SPIONs accumulated in the nucleus of MCF-7 cells while PEG-SPIONs in cytoplasma. In vitro, after 30 days we can found the tumor almost stopped to grow in Group LyP-1-SPIONs. LyP-1-SPIONs are promising in treating cancer as they accumulated in the nucleus of MCF-7 cells which expressed p32 and almost stopped tumor growth by combined MIH and targeted therapy.     

Nuclear polymorphism in osteosarcomas as a prognostic factor for the effect of chemotherapy

Summary A current strategy for osteosarcoma treatment is neo-adjuvant chemotherapy prior to the resection of the tumour. It appears that some tumours respond very well to the cytostatic therapy, while others show little or no effect. It is desirable to be able predict the response of the tumour before starting chemotherapy.16 biopsy specimens from patients with osteosarcoma who had been treated according to the protocol of the study COSS-80 and COSS-82 were examined. 100 tumour cells from each biopsy have been measured by an electronic interactive image analysis system (IBAS II; Kontron/ ZEISS). After completion of chemotherapy en bloc resection of the tumour was performed. The entire surgical specimen was completely examined at two levels by means of undecalcified sections, and assigned a grade for the effect of chemotherapy analogous to the grading of Salzer-Kuntschik et al. (1983). The quantitative analysis of tumour cell nuclei revealed two different patterns of nuclear sizes, which were correlated significantly with the chemotherapy response (P<0.002).Tumour cell nuclei of well responders were significantly larger and showed a greater variance in size (mean value 66 + 41 µm²), than those of poor responders (mean value 38 + 18 µm²).We conclude from our results that quantitative analysis and classification of nuclear size of osteosarcoma cells may be useful for predicting chemotherapy response in patients with osteosarcoma.Institut of Pathology (Head: Prof. Dr. G. Seifert) Offprint requests to: G. Delling at the above address     

Molecular targeted therapies for diffuse large B‐cell lymphoma based on apoptosis profiles

Diffuse large B‐cell lymphoma (DLBCL) is the most common type of adult non‐Hodgkin lymphoma and is treated with chemotherapy in combination with rituximab. Despite this aggressive therapy, the disease is fatal in 30–40% of patients. Inhibition of the apoptosis signalling pathways is strongly related to response to chemotherapy and eventual clinical outcome. In order to survive, lymphoma cells depend on disruption of the apoptosis pathway by mutations in apoptosis inducing genes or by continuous expression of anti‐apoptotic proteins. The development of molecules targeting these apoptosis inhibitors provides a very promising opportunity to specifically target tumour cells without toxicity to non‐malignant cells in DLBCL patients. Sensitivity for most of these antagonists can be predicted based on biological markers, suggesting the possibility of pre‐defining patients who will most likely benefit from these targeted therapies. Experimental therapies aimed at restoring the upstream apoptosis pathway or targeting apoptosis inhibitors are currently being tested in clinical trials and are expected to be effective particularly in chemotherapy‐refractory DLBCL, providing hope for patients who are refractory to current therapies. Copyright © 2009 Pathological Society of Great Britain and Ireland. Published by John Wiley & Sons, Ltd.     

All-in-one low-intensity pulsed ultrasound stimulation system using piezoelectric micromachined ultrasonic transducer (pMUT) arrays for targeted cell stimulation

A novel cell-stimulation system was fabricated using 10 × 29 piezoelectric micromachined ultrasonic transducer (pMUT) arrays for targeted ultrasonic cell stimulation. Both the diameter of a single pMUT element and the edge-to-edge gap were 120 μm, and the size of a pMUT array was 2.27 × 6.84 mm, to be placed at the bottom of a Transwell. The measured resonance frequency of a single pMUT element was 1.48 ± 0.13 MHz and the measured acoustic intensity of the pMUT array was 0.15 ± 0.03 MPa at 1 mm away from the transducer. A pMUT array was mounted on a print circuit board (PCB), which was designed in accordance with the size of a 12-well Transwell. The Transwell was placed on the PCB and wire bonding was performed to electrically connect the PCB and pMUT arrays. After wiring, the PCB and pMUT arrays were coated with 2.6-μm thick parylene-C to ensure biocompatibility and waterproofing. PC12 cells were used for ultrasonic cell stimulation tests to examine the proposed all-in-one low-intensity pulsed ultrasound stimulation system. Various stimulation times and duty cycles were used simultaneously for cell proliferation in a confined cell culture environment. All stimulation groups showed increased cell proliferation rates, in the range 138–166%, versus the proliferation rate of the control group.     

Stereocomplex micelle efficiently transports Doxorubicin for enhanced lymphoma suppression in vivo

The application of Doxorubicin (DOX) in the chemotherapy for lymphoma is seriously hampered by the side effects of DOX, especially the cardiotoxicity and nephrotoxicity. Nanoscale micelle as a promising drug delivery system has gained more and more interest in malignancy chemotherapy. In this study, we successfully fabricated DOX-loaded stereocomplex micelle (SCM/DOX) from the equimolar mixture of the enantiomeric four-armed poly(ethylene glycol)-polylactide (PDM and PLM) copolymers. The SCM/DOX showed proper hydrodynamic size of ~90 nm and slow DOX release in phosphate-buffered saline at pH 7.4. The antitumor activities of DOX, PDM/DOX, PLM/DOX, and SCM/DOX toward lymphoma cells were tested in vitro and in vivo. Our data demonstrated that the SCM/DOX more effectively inhibited the cell proliferation than PDM/DOX, PLM/DOX, and free DOX in vitro. In the in vivo antitumor test, the SCM/DOX more effectively inhibited the growth of EL4 lymphoma, too. In addition, the body weight loss caused by SCM/DOX was alleviated than DOX. More importantly, the cardiotoxicity, nephrotoxicity, and hepatotoxicity caused by DOX in mice were obviously attenuated compared to the free DOX treatment group. Taken together, all the results indicated that the SCM/DOX could inhibit the growth of EL4 lymphoma cells and attenuate the toxicity of DOX more efficiently, which suggested SCM/DOX was promising for the prevention and treatment of lymphoma.     

Improved antifilarial activity of ivermectin in chitosan–alginate nanoparticles against human lymphatic filarial parasite, Brugia malayi

The current antifilarial treatments are not up to the mark partly due to deep location of filarial parasites in the human lymphatic system. We report here on the improvement in the antifilarial activity of ivermectin (IVM) using chitosan–alginate nanoparticles prepared by modified complex coacervation method. The nanoparticles were spherical having 155 nm size and 4.56 and 75.67 % loading and entrapment efficiency respectively for IVM. The delivery system maintained the sustained release and significantly augmented the microfilaricidal (MIF) activity at a single low dose (200 μg/kg body weight, subcutaneously) in contrast to much higher dose of free ivermectin (400 μg/kg body weight, subcutaneously) against human lymphatic filariid, Brugia malayi in rodent host, Mastomys coucha. To substantiate increase in MIF activity, pharmacokinetics study was designed on Wistar rats which revealed a greater peak plasma concentration (45.3?±?1.79 ng/ml), area under the concentration curve (298?±?38.7 ng d/ml) and extended mean residence time (23.4?±?8.56 days)of IVM in chitosan–alginate nanoparticles. Administration of 25 mg/kg of diethylcarbamazine following nanoparticle therapy significantly improved the MIF and macrofilaricidal action of encapsulated drug and was considered superior in this study.