Nanocomposite powders of hydroxyapatite-graphene oxide for biological applications

The repair of bone fractures is a clinical challenge for patients with impaired healing, such as osteoporosis. Currently, different strategies have been developed to design new biomaterials, enhancing their interactions with biological systems and conducting the cellular behavior in the desired direction to help fracture healing. In the present work, hydroxyapatite-graphene oxide (HA-GO) nanocomposites were produced and the morphological and physicochemical influences of the addition of 0.5 wt%, 1.0 wt% and 1.5 wt% of GO to HA were observed. FEG-SEM and TEM analyses of HA-GO nanocomposites showed HA nanoparticles adhered to the surface of the GO sheets, suggesting an effective method to form nanostructured graphene-based biomaterials. As confirmation, physicochemical analyses by Raman, FTIR and TGA demonstrated a strong affinity between HA and GO, according to the increase of concentration from 0.5 wt% to 1.5 wt% GO in the HA-GO nanocomposites. Also, in order to evaluate the HA-GO nanocomposites behavior under biological microenvironment, in vitro bioactivity and indirect cytotoxicity tests were performed. FEG-SEM analyses confirmed the positive results for the bioactivity properties of HA-GO nanocomposite and indirect cytotoxicity demonstrated that even with a decrease in the hDPSCs viability and proliferation, when increasing to 1.5 wt% of GO concentration, high level of cell viability was exhibited by HA-GO nanocomposites. These biological results suggested the 0.5 wt% HA-GO nanocomposite as a potential bioactive bone graft and a promising biomaterial for bone tissue regeneration, when compared to the pure HA.     

Hemocompatibility,cytotoxicity, and genotoxicity of poly(methylmethacrylate)/nanohydroxyapatite nanocomposites synthesized by melt blending method

The aim of this study was to show the hemocompatibility, cytotoxicity, and genotoxicity of nanocomposites that were synthesized with different molecular weights of poly(methyl methacrylate) (PMMA) and different concentrations of nanohydroxyapatite (nHAp). Different techniques to characterize the nanocomposites were used. The cytotoxicity and genotoxic effects of the polymers and nanocomposites on human lymphocytes were determined by acid phosphatase assay, viability test, and comet assay. Moreover, hemocompatibility test was performed. It was found that all of the PMMA/nHAp nanocomposites are highly hemocompatible and biocompatible, none of the nanocomposites showed a cytotoxic effect, and nHAp addition decreased the genotoxicity.     

Facile fabrication of hollow mesoporous bioactive glass spheres: From structural behaviour to in vitro biology evaluation

Bone defects accompanied by infection or inflammation can significantly delay the healing process. To simultaneously achieve controlled release of local antibiotics for infection control and bone healing, bone-implantable delivery systems have been considered as a promising strategy. This study aims to improve drug loading capacity of bone-implantable delivery systems by introducing hollow structure mesoporous bioactive glass nanospheres (HMBGs) through a sol–gel process. Particularly, such core–shell bimodal-porous structured nanoparticles were prepared through a sacrificing template using cetyltrimethylammonium bromide (CTAB) as surfactant. It was found that varying the amount of CTAB during the synthesis process is a simple and effective approach for tuning the particle size, morphology, and structure of HMBGs. For in vitro drug release, HMBGs could sustain storage and release of vancomycin hydrochloride (VAN) via diffusion-controlled mechanism, thereby inhibiting the bacteria growth in the subsequent bacterial study. Moreover, HMBGs incorporated with VAN provided a biomimetic microenvironment favored by cell adhesion and proliferation. These findings support the compatibility of HMBG nanoparticles with antibiotics and their potential application in the treatment of infectious bone defects.     

In vitro cytotoxicity of monticellite based bioactive ceramic powder synthesized from boron derivative waste

The cytotoxicity of monticellite based bioactive ceramic powder, which was synthesized from boron derivative waste has been determined by in vitro assays of MTT, NRU, and JC-1 staining. The toxicity of powder on different mammalian cell lines (3T3-L1, HUVEC, CRL-2120) was evaluated at the concentrations of 10, 100, 200, 400 and 800?µg/mL to justify its potential for biomedical applications. The obtained results showed that monticellite based bioactive ceramic powder possesses not only bioactive feature but also biocompatible characteristic at the concentration range of 10–200?µg/mL. Hence, monticellite based bioactive ceramics have high potential as a bone graft substitute for bone void filling and coating applications.     

Catalytic supercritical water oxidation of tri-n-butyl phosphate: Process optimization by response surface methodology and cytotoxicity assessment

Catalytic supercritical water oxidation (SCWO) of an organophosphate flame retardant, namely tri-n-butyl phosphate (TNBP) was studied. Firstly, copper oxide nanoparticles (NPs) were synthesized in SCW and their properties were characterized by various analyses. Afterwards, their catalytic performance was investigated under different conditions including reaction temperature (400–500 °C), TNBP volume percentage in the feed (1–4%), oxidant ratio (0–2) and reaction time (50–150 min) based on response surface methodology (RSM). The synthesized CuO NPs had an average particle size of 30 nm with a narrow distribution. According to RSM analysis, the reaction temperature and time are the most significant factors; whereas, the impact of the other factors, especially TNBP volume percentage in the feed, was found to be negligible. Overall, excellent performance was achieved under optimal conditions found by the RSM, which was reaction temperature of 500 °C, TNBP volume percentage of 4%, oxidant ratio of 1.5, and reaction time of 90 min. The TOC removal efficiency as an indicator of TNBP degradation was about 99%. Finally, in vitro cell viability assays for the cytotoxicity evaluation of fresh and SCW-treated solution were applied. The results of MTT showed that SCWO converts TNBP into by-product that did not induce any cytotoxicity.     

A study of the mechanism of in vitro cytotoxicity of metal oxide nanoparticles using catfish primary hepatocytes and human HepG2 cells

Nanoparticles (NPs), including nanometal oxides, are being used in diverse applications such as medicine, clothing, cosmetics and food. In order to promote the safe development of nanotechnology, it is essential to assess the potential adverse health consequences associated with human exposure. The liver is a target site for NP toxicity, due to NP accumulation within it after ingestion, inhalation or absorption. The toxicity of nano-ZnO, TiO₂, CuO and Co₃O₄ was investigated using a primary culture of channel catfish hepatocytes and human HepG2 cells as in vitro model systems for assessing the impact of metal oxide NPs on human and environmental health. Some mechanisms of nanotoxicity were determined by using phase contrast inverted microscopy, 3-(4,5-dimethylthiazol-2-yl)-2,5-diphenyltetrazolium bromide (MTT) assays, reactive oxygen species (ROS) assays, and flow cytometric assays. Nano-CuO and ZnO showed significant toxicity in both HepG2 cells and catfish primary hepatocytes. The results demonstrate that HepG2 cells are more sensitive than catfish primary hepatocytes to the toxicity of metal oxide NPs. The overall ranking of the toxicity of metal oxides to the test cells is as follows: TiO₂ < Co₃O₄ < ZnO < CuO. The toxicity is due not only to ROS-induced cell death, but also to damages to cell and mitochondrial membranes.     

Bioanalytical and chemical assessment of the disinfection by-product formation potential: Role of organic matter

Disinfection by-products (DBP) formed from natural organic matter and disinfectants like chlorine and chloramine may cause adverse health effects. Here, we evaluate how the quantity and quality of natural organic matter and other precursors influence the formation of DBPs during chlorination and chloramination using a comprehensive approach including chemical analysis of regulated and emerging DBPs, total organic halogen quantification, organic matter characterisation and bioanalytical tools. In vitro bioassays allow us to assess the hazard potential of DBPs early in the chain of cellular events, when the DBPs react with their molecular target(s) and activate stress response and defence mechanisms. Given the reactive properties of known DBPs, a suite of bioassays targeting reactive modes of toxic action including genotoxicity and sensitive early warning endpoints such as protein damage and oxidative stress were evaluated in addition to cytotoxicity. Coagulated surface water was collected from three different drinking water treatment plants, along with reverse osmosis permeate from a desalination plant, and DBP formation potential was assessed after chlorination and chloramination. While effects were low or below the limit of detection before disinfection, the observed effects and DBP levels increased after disinfection and were generally higher after chlorination than after chloramination, indicating that chlorination forms higher concentrations of DBPs or more potent DBPs in the studied waters. Bacterial cytotoxicity, assessed using the bioluminescence inhibition assay, and induction of the oxidative stress response were the most sensitive endpoints, followed by genotoxicity. Source waters with higher dissolved organic carbon levels induced increased DBP formation and caused greater effects in the endpoints related to DNA damage repair, glutathione conjugation/protein damage and the Nrf2 oxidative stress response pathway after disinfection. Fractionation studies indicated that all molecular weight fractions of organic carbon contributed to the DBP formation potential, with the humic rich fractions forming the greatest amount of DBPs, while the low molecular weight fractions formed more brominated DBPs due to the high bromide to organic carbon ratio. The presence of higher bromide concentrations also led to a higher fraction of brominated DBPs as well as proportionally higher effects. This study demonstrates how a suite of analytical and bioanalytical tools can be used to effectively characterise the precursors and formation potential of DBPs.     

Cytotoxicity mechanism of non-viral carriers polyethylenimine and poly-l-lysine using real time high-content cellular assay

Cationic polymers polyethylenimine (PEI) and poly-l-lysine (PLL) used as non-viral gene/drug delivery vehicles, showed high cytotoxicity but their molecular mechanisms of toxicity have been inadequately understood. Therefore, we tried to investigate the toxicity pathway triggered by these polymers through a high-content cellular imaging technique. The results revealed that PEI induced apoptosis via an intrinsic pathway, whereas PLL showed cytotoxicity through both intrinsic and extrinsic caspase cascade. Both PEI and PLL provide different apoptotic activities on HepG2 cells depending on their molecular weight. The degree of apoptosis of PEI also depends on its structure. The branched PEI showed higher cytotoxicity than linear PEI. This observation was verified through Annexin V-FITC/PI assay and real-time high-content monitoring of cytosolic calcium, mitochondrial membrane disruption, and caspase-3 activation methods. The study therefore provides important implications on the molecular mechanisms of PEI and PLL induced cytotoxicity.     

Mortality response of folate receptor-activated,PEG–functionalized TiO2 nanoparticles for doxorubicin loading with and without ultraviolet irradiation

TiO₂ nanoparticles (NPs) were synthesized by hydrothermal assisted sol–gel technique. In the next step, as-synthesized NPs were modified by poly ethylene glycol (PEG). Then, folic acid (FA) was conjugated to TiO₂–PEG. Finally, Doxorubicin (Dox) as an anticancer drug was loaded on as-prepared TiO₂–PEG–FA nanocarrier. The optimization of TiO₂ and FA concentration and the influence of ultraviolet (UV) irradiation on photocatalytic activity of nanocarrier and Dox loaded carrier were assessed by utilizing the 3-(4,5-dimethylthiazol-2yl)-2,5-diphenyltetrazolium bromide (MTT)-assay method.     

Cytotoxicity induced by nanobacteria and nanohydroxyapatites in human choriocarcinoma cells

We explored the cytotoxic effects of nanobacteria (NB) and nanohydroxyapatites (nHAPs) against human choriocarcinoma cells (JAR) and the mechanisms of action underlying their cytotoxicity. JAR cells were co-cultured with NB and nHAPs for 48 h, and ultrastructural changes were more readily induced by NB than nHAPs. Autophagy in the plasma of JAR cells were observed in the NB group. The rate of apoptosis induced by NB was higher than that for nHAPs. The expression of Bax and FasR proteins in the NB group was stronger than that for the nHAP group. NB probably resulted in autophagic formation. Apoptosis was possibly activated via FasL binding to the FasR signaling pathway.