Novel Zinc(II) Complexes [Zn(atc-Et)2] and [Zn(atc-Ph)2]: In Vitro and in Vivo Antiproliferative Studies

Cisplatin and its derivatives are the main metallodrugs used in cancer therapy. However, low selectivity, toxicity and drug resistance are associated with their use. The zinc(II) (Zn^II) thiosemicarbazone complexes [Zn(atc-Et)₂] (1) and [Zn(atc-Ph)₂] (2) (atc-R: monovalent anion of 2-acetylpyridine N4-R-thiosemicarbazone) were synthesized and fully characterized in the solid state and in solution via elemental analysis, Fourier transform infrared (FTIR), ultraviolet-visible (UV-Vis) and proton nuclear magnetic resonance (¹H NMR) spectroscopy, conductometry and single-crystal X-ray diffraction. The cytotoxicity of these complexes was evaluated in the HepG2, HeLa, MDA-MB-231, K-562, DU 145 and MRC-5 cancer cell lines. The strongest antiproliferative results were observed in MDA-MB-231 and HepG2 cells, in which these complexes displayed significant selective toxicity (3.1 and 3.6, respectively) compared with their effects on normal MRC-5 cells. In vivo studies were performed using an alternative model (Artemia salina L.) to assure the safety of these complexes, and the results were confirmed using a conventional model (BALB/c mice). Finally, tests of oral bioavailability showed maximum plasma concentrations of 3029.50 µg/L and 1191.95 µg/L for complexes 1 and 2, respectively. According to all obtained results, both compounds could be considered as prospective antiproliferative agents that warrant further research.     

In Vitro and In Vivo Characterization of Biodegradable Poly(organophosphazenes) for Biomedical Applications

The need and the growing interest in polymers as biomaterials have led to the synthesis of new polymers with a variety of physico-chemical properties. Biomedical application of such materials not only depends on their physical properties but also on biocompatibility and biodegradability. Polyphosphazenes are a family of ‘hybrid inorganic–organic polymers’ with inorganic elements in the backbone and organic side-groups. The polyphosphazenes constitute a family of greatly diverse performance materials with a broad spectrum of properties. The present review focuses on the biodegradable polyphosphazenes, their biocompatibility, and degradation behavior both in vitro and in vivo. This review also covers the use of biodegradable polyphosphazenes as controlled release devices.     

Influence of Soybean Oil Blending with Polylactic Acid (PLA) Films: In Vitro and In Vivo Evaluation

Due to the great interest in oil‐based polymers, which are prepared from renewable resources, different forms and amounts of soybean oil‐based PLA films were prepared and evaluated for their potential usage as a medical biomaterial. Soybean oil, epoxidized soybean oil and auto‐oxidized soybean oil were blended with PLA and PLA/oil films with appropriate oil amounts [2, 7, 14 and 20% (w/w)] were obtained by solvent casting. Thermal stability and plasticization effect were determined by adjusting oil amounts and type. Epoxidized soybean oil blended films showed the smallest increase in elongation breaks (13–20%) and the highest decrease in thermal decomposition temperatures (364–327 °C) compared to other oil blended films. In vitro quantitative and qualitative cytotoxicity results showed no reactivity (grade 0) for the L929 cells treated with 14% (w/w) oil blended PLA films. In vivo irritation and implantation tests concluded that 14% (w/w) oil blended PLA films were non‐irritant. No erythema, no oedema reactions, no traumatic necrosis and foreign debris were observed. Thus, along with superior biocompatibility, PLA/oil films can replace petroleum‐based products for several biomedical uses.     

In Vitro and in Vivo Anti-Hyperglycemic Effects of Omija (Schizandra chinensis) Fruit

The entrocytes of the small intestine can only absorb monosaccharides such as glucose and fructose from our diet. The intestinal absorption of dietary carbohydrates such as maltose and sucrose is carried out by a group of α-glucosidases. Inhibition of these enzymes can significantly decrease the postprandial increase of blood glucose level after a mixed carbohydrate diet. Therefore, the inhibitory activity of Omija (Schizandra chinensis) extract against rat intestinal α-glucosidase and porcine pancreatic α-amylase were investigated in vitro and in vivo. The in vitro inhibitory activities of water extract of Omija pulp/skin (OPE) on α-glucosidase and α-amylase were potent when compared to Omija seeds extract (OSE). The postprandial blood glucose lowering effect of Omija extracts was compared to a known type 2 diabetes drug (Acarbose), a strong α-glucosidase inhibitor in the Sprague-Dawley (SD) rat model. In rats fed on sucrose, OPE significantly reduced the blood glucose increase after sucrose loading. Furthermore, the oxygen radical absorbance capacity (ORAC) of OSE and OPE was evaluated. OPE had higher peroxyl radical absorbing activity than OSE. These results suggest that Omija, which has high ORAC value with α-glucosidase inhibitory activity and blood glucose lowering effect, could be physiologically useful for treatment of diabetes, although clinical trials are needed.     

In Vitro and in Vivo Models of Non-Alcoholic Fatty Liver Disease (NAFLD)

By now, non-alcoholic fatty liver disease (NAFLD) is considered to be among the most common liver diseases world-wide. NAFLD encompasses a broad spectrum of pathological conditions ranging from simple steatosis to steatohepatitis, fibrosis and finally even cirrhosis; however, only a minority of patients progress to end-stages of the disease, and the course of the disease progression to the later stages seems to be slow, developing progressively over several years. Key risk factors including overweight, insulin resistance, a sedentary life-style and an altered dietary pattern, as well as genetic factors and disturbances of the intestinal barrier function have been identified in recent years. Despite intense research efforts that lead to the identification of these risk factors, knowledge about disease initiation and molecular mechanisms involved in progression is still limited. This review summarizes diet-induced and genetic animal models, as well as cell culture models commonly used in recent years to add to the understanding of the mechanisms involved in NAFLD, also referring to their advantages and disadvantages.     

Increased In Vitro and In Vivo Digestibility of Soy Proteins by Chemical Modification of Disulfide Bonds

To improve protein digestibility of aqueously extracted soy proteins, an effective chemical treatment under mild conditions is needed. Soy proteins, including storage protein glycinin and antinutritional factors such as trypsin inhibitors, are rich in disulfide bonds. Reduction of these disulfide bonds by incubating soy proteins with sodium sulfite and sodium metabisulfite at 55 °C showed no net increase of free sulfhydryl groups after dialysis to remove the residual reducing agent. However, the in vitro digestibility measured by trypsin hydrolysis using the pH-Stat method was significantly increased. Sodium metabisulfite (SMBS) was more effective in increasing in vitro digestibility than sodium sulfite at the same molar concentration. The digestibility of soy protein treated by 0.5 mmol SMBS/g soy flour at 55 °C was more than doubled compared to that of the control without reduction treatment. Large-scale testing of soy proteins treated with SMBS for an in vivo animal feeding study showed similar in vitro digestibility by trypsin, e.g., the degree of hydrolysis of the treated sample was 8.5% compared to 1.6% of the control. These soy proteins were further evaluated using a chick growth model. The protein efficiency ratio (PER) increased by 57% when the chicks fed SMBS-treated soy were compared to the chicks fed raw soy flour. SMBS-fed chicks did not display any pancreatic hypertrophy compared to those fed with raw soy control. These results indicate that there is great potential to use safe chemicals and mild temperature to inactivate the antinutritional factors in soybeans and thus improve digestibility of soy proteins that are extracted with low-temperature aqueous process.     

配合物{[Co（HFlu）2Cl2】·2C2H5OH}n的合成、结构及体外抗真菌活性

新合成了1种钴-氟康唑配合物{[Co（HFlu）2Cl2】·2C2H5OH}n,利用X-射线单晶衍射仪等对配合物进行了结构分析,并研究了该配合物的体外抗真菌活性,标题化合物对丝状真菌的抗菌效果明显优于氟康唑,对酵母菌的体外抗真菌活性与氟康唑相当,     

In Vitro/In Vivo Toxicity Evaluation and Quantification of Iron Oxide Nanoparticles

Increasing biomedical applications of iron oxide nanoparticles (IONPs) in academic and commercial settings have alarmed the scientific community about the safety and assessment of toxicity profiles of IONPs. The great amount of diversity found in the cytotoxic measurements of IONPs points toward the necessity of careful characterization and quantification of IONPs. The present document discusses the major developments related to in vitro and in vivo toxicity assessment of IONPs and its relationship with the physicochemical parameters of IONPs. Major discussion is included on the current spectrophotometric and imaging based techniques used for quantifying, and studying the clearance and biodistribution of IONPs. Several invasive and non-invasive quantification techniques along with the pitfalls are discussed in detail. Finally, critical guidelines are provided to optimize the design of IONPs to minimize the toxicity.     

Water-Soluble Ruthenium(III)-Dimethyl Sulfoxide Complexes: Chemical Behaviour and Pharmaceutical Properties

In this paper we report a review of the results obtained in the last few years by our group in the development of ruthenium(III) complexes characterized by the presence of sulfoxide ligands and endowed with antitumor properties. In particular, we will focus on ruthenates of general formula Na[trans-RuCl(4)(R(1)R(2)SO)(L)], where R(1)R(2)SO = dimethylsulfoxide (DMSO) or tetramethylenesulfoxide (TMSO) and L = nitrogen donor ligand. The chemical behavior of these complexes has been studied by means of spectroscopic techniques both in slightly acidic distilled water and in phosphate buffered solution at physiological pH. The influence of biological reductants on the chemical behavior is also described. The antitumor properties have been investigated on a number of experimental tumors. Out of the effects observed, notheworthy appears the capability of the tested ruthenates to control the metastatic dissemination of solid metastasizing tumors. The analysis of the antimetastatic action, made in particular on the MCa mammary carcinoma of CBA mouse, has demonstrated a therapeutic value for these complexes which are able to significantly prolong the survival time of the treated animals. The antimetastatic effect is not attributable to a specific cytotoxicity for metastatic tumor cells although in vitro experiments on pBR322 double stranded DNA has shown that the test ruthenates bind to the macromolecule, causing breaks corresponding to almost all bases, except than thymine, and are able to cause interstrand bonds, depending on the nature of the complex being tested, some of which results active as cisplatin itself.     

In Vitro Activity of Copper(II) Complexes,Loaded or Unloaded into a Nanostructured Lipid System,against Mycobacterium tuberculosis

Tuberculosis (TB) is an infectious disease caused mainly by the bacillus Mycobacterium tuberculosis (Mtb), presenting 9.5 million new cases and 1.5 million deaths in 2014. The aim of this study was to evaluate a nanostructured lipid system (NLS) composed of 10% phase oil (cholesterol), 10% surfactant (soy phosphatidylcholine, sodium oleate), and Eumulgin^® HRE 40 ([castor oil polyoxyl-40-hydrogenated] in a proportion of 3:6:8), and an 80% aqueous phase (phosphate buffer pH = 7.4) as a tactic to enhance the in vitro anti-Mtb activity of the copper(II) complexes [CuCl₂(INH)₂]·H₂O (1), [Cu(NCS)₂(INH)₂]·5H₂O (2) and [Cu(NCO)₂(INH)₂]·4H₂O (3). The Cu(II) complex-loaded NLS displayed sizes ranging from 169.5 ± 0.7095 to 211.1 ± 0.8963 nm, polydispersity index (PDI) varying from 0.135 ± 0.0130 to 0.236 ± 0.00100, and zeta potential ranging from −0.00690 ± 0.0896 to −8.43 ± 1.63 mV. Rheological analysis showed that the formulations behave as non-Newtonian fluids of the pseudoplastic and viscoelastic type. Antimycobacterial activities of the free complexes and NLS-loaded complexes against Mtb H₃₇Rv ATCC 27294 were evaluated by the REMA methodology, and the selectivity index (SI) was calculated using the cytotoxicity index (IC₅₀) against Vero (ATCC^® CCL-81), J774A.1 (ATCC^® TIB-67), and MRC-5 (ATCC^® CCL-171) cell lines. The data suggest that the incorporation of the complexes into NLS improved the inhibitory action against Mtb by 52-, 27-, and 4.7-fold and the SI values by 173-, 43-, and 7-fold for the compounds 1, 2 and 3, respectively. The incorporation of the complexes 1, 2 and 3 into the NLS also resulted in a significant decrease of toxicity towards an alternative model (Artemia salina L.). These findings suggest that the NLS may be considered as a platform for incorporation of metallic complexes aimed at the treatment of TB.     

In Vivo X-Ray Imaging of Phosphor-Doped PDMS and Phosphor-Doped Aerogel Biomaterials

Noninvasive rapid in vivo imaging and detection of biomedical implants is a critical part of the design and implementation of smart implants. Thermographic phosphors offer a precise and remotely accessible sensing method that has been utilized here. We present the first in vivo X-ray images of La₂O₂S:Eu-doped crosslinked silica aerogels and polydimethylsiloxane (PDMS) with increasing dopant concentrations. Results show that native PDMS and crosslinked silica aerogel do not show noticeable attenuation of X-rays while image analysis yields values of the absorption coefficient of 0.014 for the doped aerogel and a range of 0.015–0.017 for the doped PDMS.     

Toxicity, Spectroscopic Characterization and Electrochemical Behaviour of New Macrocclic Complexes of Lead(II) and Palladium(II) Metals

Tetraazamacrocyclie complexes of lead and palladium have been synthesized by the template process using the bis(benzil)ethylenediamine precursor. The tetradentate macrocycle (maL) reacts with PbCl(2), PdCl(2) and different diamines in a 1:1:1 molar ratio in methanol to give several solid complexes of the types [Pb(maL)(R)Cl(2)] and [Pd(maL)(R)]Cl(2) (where R = 2,6-diaminopyridine or 1,2-phenylenediamine). The macrocycle and its metal complexes have been characterized by elemental analysis, molecular weight determinations, molar conductivity, IR, (1)H NMR, (13)C NMR, electronic, mass and electrochemical studies. The macrocyclic ligand coordinates through the four azomethine nitrogen atoms which are bridged by benzil moieties. IR spectra suggest that the pyridine nitrogen is not coordinating. The palladium complexes exhibit tetracoordinated square-planar geometry, whereas a hexacoordinated octahedral geometry is suggested for lead complexes. The macrocycle along with its complexes have been screened in vitro against a number of pathogenic fungi and bacteria to assess their growth inhibiting potential.     

Neuroprotective Effects of Pre-Treament with l-Carnitine and Acetyl-l-Carnitine on Ischemic Injury In Vivo and In Vitro

The therapeutic effect of stroke is hampered by the lack of neuroprotective drugs against ischemic insults beyond the acute phase. Carnitine plays important roles in mitochondrial metabolism and in modulating the ratio of coenzyme A (CoA)/acyl-CoA. Here, we investigate the neuroprotective effects of l-carnitine (LC) and Acetyl-l-carnitine (ALC) pre-treatment on ischemic insults under the same experimental conditions. We used a transient middle cerebral artery occlusion (MCAO) model to evaluate the protective roles of LC and ALC in acute focal cerebral ischemia in vivo and to understand the possible mechanisms using model of PC12 cell cultures in vitro. Results showed that ALC, but not LC, decreased infarction size in SD rats after MCAO in vivo. However, both LC and ALC pretreatment reduced oxygen-glucose deprivation (OGD)-induced cell injury and decreased OGD-induced cell apoptosis and death in vitro; at the same time, both of them increased the activities of super oxide dismutase (SOD) and ATPase, and decreased the concentration of malondialdehyde (MDA) in vitro. Thus, our findings suggested that LC and ALC pre-treatment are highly effective in the prevention of neuronal cell against ischemic injury in vitro, however, only ALC has the protective effect on neuronal cell injury after ischemia in vivo.     

Docosahexaenoic Acid (DHA) and Docosapentaenoic Acid (DPAn-6) Algal Oils Reduce Inflammatory Mediators in Human Peripheral Mononuclear Cells In Vitro and Paw Edema In Vivo

The anti-inflammatory activity associated with fish oil has been ascribed to the long-chain polyunsaturated fatty acids (LC-PUFA), predominantly eicosapentaenoic acid (EPA) and docosahexaenoic acid (DHA). Here we examined the anti-inflammatory effects of two DHA-rich algal oils, which contain little EPA, and determined the contribution of the constituent fatty acids, particularly DHA and docosapentaenoic acid (DPAn-6). In vitro, lipopolysaccharide (LPS)-stimulated Interleukin-1 beta (IL-1β) and Tumor Necrosis Factor-alpha (TNF-α) secretion in human peripheral blood mononuclear cells (PBMC) was inhibited with apparent relative potencies of DPAn-6 (most potent) > DHA > EPA. In addition, DPAn-6 decreased intracellular levels of cyclooxygenase-2 (COX-2) and was a potent inhibitor of pro-inflammatory prostaglandin E2 (PGE2) production. DHA/DPAn-6-rich DHA-S™ (DHA-S) algal oil was more effective at reducing edema in rats than DHA-rich DHA-T™ (DHA-T), suggesting that DPAn-6 has anti-inflammatory properties. Further in vivo analyses demonstrated that feeding DPAn-6 alone, provided as an ethyl ester, reduced paw edema to an extent approaching that of indomethacin and enhanced the anti-inflammatory activity of DHA when given in combination. Together, these results demonstrate that DPAn-6 has anti-inflammatory activity and enhances the effect of DHA in vitro and in vivo. Thus, DHA-S algal oil may have potential for use in anti-inflammatory applications.     

基因工程人红细胞生成素对骨髓增生异常综合征患者体内外治疗作用的研究

EPO体外对26例MDS患者CFU－E的作用及体内用于13例患者的治疗结果显示：（1）EPO对MDS患者CFU－E的作用明显低于对照组；（2）EPO可促进患者CFU－E增殖，这种增殖为剂量依赖型，EPO100u／ml时最强；（3）EPO对CFU－E作用与患者自身CFU－E明显相关。自身CFU－E越接近正常者对EPO反应越好；（4）EPO治疗13例MDS有效3例（23％）。以上结果证明，EPO对于部分MDS患者有效，可作为选择治疗MDS药物。     

重组人促红素体内、体外生物学活性差异成因分析

目的分析造成重组人促红素(rHuEPO)体内、体外生物学活性差异的原因。方法按《中国药典》三部(2005版)方法检测rHuEPO样品体内、体外生物学活性,并通过等电聚焦电泳分析、唾液酸含量、纯度、N-糖链糖基化和相对分子质量检测,探讨样品体内、体外生物学活性产生差异的原因。结果体内、体外生物学活性有差异的rHuEPO样品中,存在糖链末端N-乙酰神经氨酸残基不完整的rHuEPO成分。纯度、N-糖链糖基化和相对分子质量检测结果未见异常。结论为提高rHuEPO产品质量,应在生产过程中有效去除N-乙酰神经氨酸残基不完整的rHuEPO。     

Toxicological Assessment of Inhaled Nanoparticles: Role of in Vivo,ex Vivo,in Vitro,and in Silico Studies

The alveolar epithelium of the lung is by far the most permeable epithelial barrier of the human body. The risk for adverse effects by inhaled nanoparticles (NPs) depends on their hazard (negative action on cells and organism) and on exposure (concentration in the inhaled air and pattern of deposition in the lung). With the development of advanced in vitro models, not only in vivo, but also cellular studies can be used for toxicological testing. Advanced in vitro studies use combinations of cells cultured in the air-liquid interface. These cultures are useful for particle uptake and mechanistic studies. Whole-body, nose-only, and lung-only exposures of animals could help to determine retention of NPs in the body. Both approaches also have their limitations; cellular studies cannot mimic the entire organism and data obtained by inhalation exposure of rodents have limitations due to differences in the respiratory system from that of humans. Simulation programs for lung deposition in humans could help to determine the relevance of the biological findings. Combination of biological data generated in different biological models and in silico modeling appears suitable for a realistic estimation of potential risks by inhalation exposure to NPs.