Platinum(II) ring-fused chlorins as efficient theranostic agents: Dyes for tumor-imaging and photodynamic therapy of cancer.

The discovery of Pt-chlorin-type theranostic agents is described. Luminescent Pt(II) 4,5,6,7-tetrahydropyrazolo[1,5-a]pyridine-fused chlorins, with different degrees of hydrophilicity, have been synthesized and their in vitro photocytotoxicity against human melanoma, oesophageal and bladder carcinomas was studied. A di(hydroxymethyl)-substituted chlorin was identified as a privileged molecule to explore imaging-guided photodynamic therapy. In addition to the high activity as PDT agent and absence of cytotoxicity per se, this molecule showed the ideal photophysical and photochemical properties. In vivo studies using a A375 melanoma mouse model, proved the extraordinary properties of this chlorin as a luminescent probe and the ability to impair tumor growth, making image guided treatment and follow up a possibility.

Doping ß-TCP as a Strategy for Enhancing the Regenerative Potential of Composite ß-TCP-Alkali-Free Bioactive Glass Bone Grafts. Experimental Study in Rats.

The present work aims at evaluating the potential gains derived from partially replacing calcium in resorbable ß-tricalcium phosphate (ß-TCP) by two different molar percentages of strontium (5, 10) and zinc (1, 2), concomitantly with a fixed molar percentage (0.5) of manganese. Synthetic granular composite bone filling grafts consisting of doped ß-TCP and an alkali-free bioactive glass were prepared and implanted in ~4 mm diameter bone defects drilled in the calvaria of Wistar rats used as animal models. The animals were sacrificed after 9 weeks of implantation and the calvaria was excised. Non-manipulated bone was used as positive control, while empty defects were used as a negative control group. The von Kossa staining revealed an enhanced new bone formation with increasing doping levels, supporting the therapeutic effects exerted by the doping elements. The percentage of newly formed bone was similar when the defects were filled with autologous bone, BG (previous results) or 3TCP2/7BG, which indicates that the latter two are excellent candidates for replacement of autologous bone as bone regeneration material. This finding confirms that doping with suitable doses of therapeutic ions is a good strategy towards transposing the bone graft materials to biomedical applications in humans.

The in vivo performance of an alkali-free bioactive glass for bone grafting, FastOs® BG, assessed with an ovine model.

Although bioactive glasses are successfully used as bone substitutes, recent studies have revealed that the high alkali content in these glasses leads to fast in vivo degradation rates that may not match the rate of new bone ingrowth. This prompted us to design and develop novel bioactive glasses that are devoid of alkali but still demonstrate high bioactivity in vitro. This article describes the in vivo performance of an alkali-free bioactive glass with the following composition (Wt %): 13.03 MgO-33.98 CaO-13.37 P2 O5 -38.84 SiO2 -0.77 CaF2 (labelled as FastOs® BG). An animal model was used to assess the in vivo performance of FastOs® BG, using 45S5 Bioglass® as control. The evaluation was performed through implantation of FastOs® BG and 45S5 Bioglass® , during one month, in femoral bone defects in sheep. Subcutaneous implantation of both glasses was also performed in order to assess tissue response through a standardized method. Histological and scanning electron microscopy assessment of retrieved subcutaneous and bone samples demonstrated that FastOs® BG is biocompatible, osteoconductive, that it can be osteointegrated, and that it is more slowly resorbed than 45S5 Bioglass® . These features suggest that FastOs® BG is a potential candidate for bone grafting. © 2015 Wiley Periodicals, Inc. J Biomed Mater Res Part B: Appl Biomater, 105B: 30-38, 2017.

Osteogenic capacity of alkali-free bioactive glasses. In vitro studies.

The high alkali content bioactive glasses commonly used to regenerate bone in dental, orthopedic, and maxillofacial surgeries induce some cytotoxicity in surrounding tissues. The present study aims the ability of some alkali-free bioactive glasses compositions, recently developed by our research group, to stimulate human mesenchymal stem cells (hMSCs) differentiation into osteoblasts in comparison to 45S5 Bioglass® . The obtained results demonstrated that alkali-free bioactive glasses possess higher stimulating towards differentiation of hMSCs in comparison to the control 45S5 Bioglass® . The von Kossa assay demonstrated that all bioactive glasses studied were able to induce the appearance of calcium deposits even when the cells were cultured in DMEM, proving that these biomaterials per se induce hMSCs cell differentiation. It was also observed that in both cell culture medium used (DMEM, and osteogenesis differentiation medium) alkali-free bioactive glasses clearly induced the appearance of more calcium deposits than the 45S5 Bioglass® , indicating their greater ability to induce cell differentiation. In summary, these results indicate that alkali-free bioactive glasses are a safe and valid alternative to 45S5 Bioglass® . © 2016 Wiley Periodicals, Inc. J Biomed Mater Res Part B: Appl Biomater, 105B: 2360-2365, 2017.

Fluorine-18 Fluorodeoxyglucose Uptake in Hepatocellular Carcinoma: Correlation with Glucose Transporters and p53 Expression.

INTRODUCTION: Hepatocellular Carcinoma (HCC) is one of most lethal cancers worldwide. The prognosis is very poor and therapeutic options are limited. The aim of this study was to determine the correlation of the [(18)F]FDG uptake profile of three HCC cell lines with p53 and glucose transporters (GLUTs) 1, 2, 3, 5 and 12 expression and with the glucose level present in the cell culture medium. METHODS: Cell lines used are HepG2 (wp53), HuH7 (overexpress p53) and Hep3B2.1-7 (p53null). An immunocytochemical analysis was performed to evaluate p53 expression. Through uptake studies were analyzed the [(18)F]FDG uptake profiles of all cell lines under study. The expression of GLUTs were quantified by flow cytometry. The [(18)F]FDG uptake studies GLUTs expression analysis were performed on cells that grew in a high and low glucose medium in order to determine the effect of glucose concentration on GLUTs expression and on [(18)F]FDG uptake. RESULTS: Immunocytochemical analysis confirmed the p53 expression profiles of all cell lines. It was found out that for all cell lines, [(18)F]FDG uptake is higher when cells grow in low glucose medium, however, the glucose level doesn't affect mostly the GLUTs expression. The Hep3B2.1-7 (p53null) is always the one that have higher [(18)F]FDG uptake. It was found that not always GLUT1 and GLUT3 are the most expressed by these cell lines. CONCLUSIONS: Our results shown that the p53 expression influences [(18)F]FDG uptake. This suggests that [(18)F]FDG may be used in HCC diagnosis, and may even provide some information about the genetic profile of the tumor.

Influence of P53 on the radiotherapy response of hepatocellular carcinoma.

BACKGROUND/AIMS: Hepatocellular carcinoma (HCC) is one of the most common cancers worldwide, and it has a poor prognosis and few therapeutic options. Radiotherapy is one of the most effective forms of cancer treatment, and P53 protein is one of the key molecules determining how a cell responds to radiotherapy. The aim of this study was to determine the therapeutic efficacy of iodine-131 in three human HCC cell lines. METHODS: Western blotting was used to measure P53 expression. The effects of radiotherapy with iodine-131 were assessed by using the clonogenic assay to evaluate cell survival. Flow cytometry was carried out to examine the effects of iodine-131 on cell death, oxidative stress, reduced intracellular glutathione expression, the mitochondrial membrane potential, and the cell cycle. RESULTS: The P53 protein was not expressed in Hep3B2.1-7 cells, was expressed at normal levels in HepG2 cells, and was overexpressed in HuH7 cells. P53 expression in the HuH7 and HepG2 cell lines increased after internal and external irradiation with iodine-131. Irradiation induced a decrease in cell survival and led to a decrease in cell viability in all of the cell lines studied, accompanied by cell death via late apoptosis/necrosis and necrosis. Irradiation with 131-iodine induced mostly cell-cycle arrest in the G0/G1 phase. CONCLUSIONS: These results suggest that P53 plays a key role in the radiotherapy response of HCC.

Cholangiocarcinoma: from molecular biology to treatment.

Cholangiocarcinoma is a rare tumor originating in the bile ducts, which, according to their anatomical location, is classified as intrahepatic, extrahepatic and hilar. Nevertheless, incidence rates have increased markedly in recent decades. With respect to tumor biology, several genetic alterations correlated with resistance to chemotherapy and radiotherapy have been identified. Here, we highlight changes in KRAS and TP53 genes that are normally associated with a more aggressive phenotype. Also IL-6 and some proteins of the BCL-2 family appear to be involved in the resistance that the cholangiocarcinoma presents toward conventional therapies. With regard to diagnosis, tumor markers most commonly used are CEA and CA 19-9, and although its use isolated appears controversial, their combined value has been increasingly advocated. In imaging terms, various methods are needed, such as abdominal ultrasound, computed tomography and cholangiopancreatography. Regarding therapy, surgical modalities are the only ones that offer chance of cure; however, due to late diagnosis, most patients cannot take advantage of them. Thus, the majority of patients are directed to other therapeutic modalities like chemotherapy, which, in this context, assumes a purely palliative role. Thus, it becomes urgent to investigate new therapeutic options for this highly aggressive type of tumor.

Positron emission tomography diagnostic imaging in multidrug-resistant hepatocellular carcinoma: focus on 2-deoxy-2-(18F)Fluoro-D-Glucose.

Hepatocellular carcinoma (HCC) is one of the most lethal cancers worldwide. Surgical resection and liver transplantation are still the best options for treatment. Nevertheless, as the number of patients who may benefit from these therapies is limited, alternative therapies have been developed, including chemotherapy. However, partly due to the expression of multidrug resistance (MDR) proteins, it has been found that HCC is a highly chemoresistant tumor. The major family of MDR proteins is the ATP-binding cassette (ABC) transporter superfamily, which includes P-glycoprotein (Pgp) and MDR-associated protein 1 (MRP1). Positron emission tomography using the radiolabeled analog of glucose, 2-deoxy-2-((18)F)fluoro-D-glucose ([(18)F]FDG), has been used in diagnostic imaging of various types of tumors. Clinical studies are inconsistent but experimental studies have shown that [(18)F]FDG uptake is associated with tumor grade and is inversely proportional to Pgp expression in HCC. These studies unveil that [(18)F]FDG can be a substrate of Pgp, although that relationship remains unclear. This review sums up the relationship between MDR expression in HCC, and [(18)F]FDG uptake by tumor cells, showing that this radiopharmaceutical may provide a useful tool for the study of chemoresistance in HCC, and that the use of this marker may contribute to the therapeutic choice on this highly aggressive tumor.

Targeting triple-negative breast cancer cells with 6,7-bis(hydroxymethyl)-1H,3H-pyrrolo[1,2-c]thiazoles.

Further studies on 6,7-bis(hydroxymethyl)-1H,3H-pyrrolo[1,2-c]thiazoles as anticancer agents against breast cancer are reported, allowing to demonstrate the potential of these compounds for the therapy of the triple-negative breast cancer, the most challenging tumors in clinical practice. These compounds were assayed for their in vitro cytotoxicity on several human breast cancer cell lines (MCF7, HCC1954 and HCC1806 cell lines). Particularly interesting were the results obtained for 4-hydroxyphenyl substituted derivative, which proved to be the most promising compound regarding HCC1806 cell line, a triple-negative breast cancer. The effects of the two most active compounds on cell survival, viability, cell cycle, DNA damage and expression of proteins related to cell death pathways were studied. The reported results consolidate the potential of 6,7-bis(hydroxymethyl)-1H,3H-pyrrolo[1,2-c]thiazoles for the therapy of breast cancer, particularly the triple-negative.