Comprehensive characterizations of nanoparticle biodistribution following systemic injection in mice.

Various nanoparticle (NP) properties such as shape and surface charge have been studied in an attempt to enhance the efficacy of NPs in biomedical applications. When trying to undermine the precise biodistribution of NPs within the target organs, the analytical method becomes the determining factor in measuring the precise quantity of distributed NPs. High performance liquid chromatography (HPLC) represents a more powerful tool in quantifying NP biodistribution compared to conventional analytical methods such as an in vivo imaging system (IVIS). This, in part, is due to better curve linearity offered by HPLC than IVIS. Furthermore, HPLC enables us to fully analyze each gram of NPs present in the organs without compromising the signals and the depth-related sensitivity as is the case in IVIS measurements. In addition, we found that changing physiological conditions improved large NP (200-500 nm) distribution in brain tissue. These results reveal the importance of selecting analytic tools and physiological environment when characterizing NP biodistribution for future nanoscale toxicology, therapeutics and diagnostics.

Functionalized nanoparticles provide early cardioprotection after acute myocardial infarction.

Recent developments in nanotechnology have created considerable potential toward diagnosis and cancer therapy. In contrast, the use of nanotechnology in tissue repair or regeneration remains largely unexplored. We hypothesized that intramyocardial injection of insulin-like growth factor (IGF)-1-complexed poly(D,L-lactide-co-glycolide) (PLGA) nanoparticles (PLGA-IGF-1 NPs) increases IGF-1 retention, induces Akt phosphorylation, and provides early cardioprotection after acute myocardial infarction (MI). We synthesized 3 different sizes of PLGA particles (60 nm, 200 nm, and 1 µm) which were complexed with IGF-1 using electrostatic force to preserve the biological function of IGF-1. Afterward, we injected PLGA-IGF-1 NPs in the heart after MI directly. Compared with the other two larger particles, the 60 nm-sized PLGA-IGF-1 NPs carried more IGF-1 and induced more Akt phosphorylation in cultured cardiomyocytes. PLGA-IGF-1 NPs also prolonged Akt activation in cardiomyocytes up to 24h and prevented cardiomyocyte apoptosis induced by doxorubicin in a dose-dependent manner. In vivo, PLGA-IGF-1 NP treatment significantly retained more IGF-1 in the myocardium than the IGF-1 alone treatment at 2, 6, 8, and 24 h. Akt phosphorylation was detected in cardiomyocytes 24h post-MI only in hearts receiving PLGA-IGF-1 NP treatment, but not in hearts receiving injection of PBS, IGF-1 or PLGA NPs. Importantly, a single intramyocardial injection of PLGA-IGF-1 NPs was sufficient to prevent cardiomyocyte apoptosis (P<0.001), reduce infarct size (P<0.05), and improve left ventricle ejection fraction (P<0.01) 21 days after experimental MI in mice. Our results not only demonstrate the potential of nanoparticle-based technology as a new approach to treating MI, but also have significant implications for translation of this technology into clinical therapy for ischemic cardiovascular diseases.

The enhancement of endothelial cell therapy for angiogenesis in hindlimb ischemia using hyaluronan.

Growing evidence shows that injection of hyaluronan (HA) benefits ischemic injury in animals. On the other hand, cell therapy is an emerging approach to treat occlusive arterial diseases, although the low retention rate of cells after direct injection remains a major concern. Here, we tested whether injection of HA along with endothelial cells promotes the retention and growth of transplanted cells, thus improving therapeutic angiogenesis in a mouse model of hindlimb ischemia (HI). In culture, HA improved human umbilical vein endothelial cell (HUVEC) proliferation proportional to HA concentration and protected HUVECs from apoptosis. Subsequently, in immunocompromised mice HI was induced by femoral artery ligation and treatments were given 24h later. At 4 weeks, injection of HA along with HUVECs had a greater effect for restoring blood perfusion and salvaging the ischemic limb compared to injection of HA or HUVECs alone. In addition, angiogenesis and arteriogenesis were significantly increased by HA+HUVECs injection. Lastly, HA+HUVECs injection resulted in the retention of more cells than HUVECs alone, and allowed their engraftment into the vasculature of the ischemic limb. These results suggest that this combined approach can be translated into a clinical therapy for peripheral artery occlusive disease.

Detection of triglyceride using an iridium nano-particle catalyst based amperometric biosensor.

The detection and quantification of triglyceride (TG) using an iridium nano-particle modified carbon based biosensor was successfully carried out in this study. The detection procedures were based on the electrochemical detection of enzymatically produced NADH. TG was hydrolyzed by lipase and the glycerol produced was catalytically oxidized by NAD-dependent glycerol dehydrogenase producing NADH in a solution containing NAD(+). Glyceryl tributyrate, a short chain triglyceride, was chosen as the substrate for the evaluation of this TG biosensor in bovine serum and human serum. A linear response to glyceryl tributyrate in the concentration range of 0 to 10 mM and a sensitivity of 7.5 nA mM(-1) in bovine serum and 7.0 nA mM(-1) in human serum were observed experimentally. The potential interference of species such as uric acid (UA) and ascorbic acid (AA) was assessed. The incorporation of a selected surfactant and an increase in the incubation temperature appeared to enhance the performance of this biosensor. The conditions for the determination of TG levels in bovine serum using this biosensor were optimized, with sunflower seed oil being used as an analyte to simulate the detection of TG in blood. The experimental results demonstrated that this iridium nano-particle modified working electrode based biosensor provided a relatively simple means for the accurate determination of TG in serum.

Development and characterization of an all-solid-state potentiometric biosensor array microfluidic device for multiple ion analysis.

A microfluidic device with an all-solid-state potentiometric biosensor array was developed using microfabrication technology. The sensor array included a pH indicator, and potassium and calcium ion-selective microelectrodes. The pH indicator was an iridium oxide thin film modified platinum microelectrode and the iridium oxide was deposited by an electrochemical method. The potassium and calcium ion-selective microelectrodes were platinum coated with silicon rubber based ion-selective membranes with respectively potassium (valinomycin) and calcium (ETH 1001) ionophores. The detection system was integrated with a micro-pneumatic pump which can continuously drive fluids into the microchannel through sensors at flow rates ranging from 52.4 microl min(-1) to 7.67 microl min(-1). The sensor array microfluidic device showed near-Nernstian responses with slopes of 62.62 mV +/- 2.5 mV pH(-1), 53.76 mV +/- 3 mV -log[K+](-1) and 25.77 mV +/- 2 mV -log[Ca2+](-1) at 25 degrees C +/- 5 degrees C, and a linear response within the pH range of 2-10, with potassium and calcium concentrations between 0.1 M and 10(-6) M. In this study the device provided a convenient way to measure the concentration of hydrogen, potassium and calcium ions, which are important physiological parameters.

Fabrication of a planar-form screen-printed solid electrolyte modified Ag/AgCl reference electrode for application in a potentiometric biosensor.

This study features the fabrication of a planar-form, solid electrolyte modified, (PSEM) Ag/AgCl reference electrode using a screen-printing method. The PSEM Ag/AgCl reference electrode uses agar gel as the inner electrolyte and chloroprene rubber for the liquid junction and insulator. These common low-cost materials and the simple fabrication processes involved render the proposed reference electrode an ideal candidate for cost-efficient mass production. It is shown that the developed reference electrode is insensitive to most of the physiologically important ionic species, including Na+, K+, Li+, Ca2+, NH4+, and Cl-, under continuous measurement conditions. Moreover, as with conventional commercial reference electrodes, the proposed reference electrode exhibits a reversible response, which is maintained until the agar gel dries out. The PSEM Ag/AgCl reference electrode is integrated with an iridium oxide modified Pt-based pH indicator electrode to form a chip-type pH biosensor. The performance of this biosensor is consistent with that obtained from a pH meter based on a macroscopic commercial Ag/AgCl reference electrode. The experimental results confirm that the proposed biosensor is capable of providing precise pH measurements of various real samples. Accordingly, the PSEM Ag/AgCl reference electrode presented in this study provides a viable alternative to the macroscopic Ag/AgCl reference electrode used in many conventional chip-based pH sensors.

Microfluidic pH-sensing chips integrated with pneumatic fluid-control devices.

This paper presents a microfluidic chip capable of performing precise continuous pH measurements in an automatic mode. The chip is fabricated using micro-electro-mechanical-systems (MEMS)-based techniques and incorporates polydimethylsiloxane (PDMS) microstructures, pH-sensing electrodes and pneumatic fluid-control devices. Through its enhanced microchannel design and use of pneumatic fluid-control devices, the microfluidic chip reduces the dead volume of the sample and increases the pumping rate. The maximum pumping rate of the developed micro-pump is 28 microL/min at an air pressure of 10 psi and a driving frequency of 10 Hz. The total sample volume consumed in each sensing operation is just 0.515 microL. As a result, the developed chip reduces the sample volume compared to conventional large-scale pH-sensing systems. The microfluidic chip employs the electrochemical sensing method to conduct precise pH level measurements. The sensing electrodes are fabricated by sputtering a layer of SiO(2)-LiO(2)-BaO-TiO(2)-La(2)O(3) (SLBTLO) onto platinum (Pt) electrodes and the pH value of the sample is evaluated by measuring the potential difference between the sensing electrodes and a reference electrode. Additionally, the integration of the microfluidic chip with a pneumatic fluid-control device facilitates automatic sample injection and a continuous sensing operation. The developed system provides a valuable tool with which to examine pH values in a wide range of biomedical and industrial applications.

Telemetric electrochemical sensor.

A telemetric system was designed and constructed to sense pH and ethanol variation in aqueous solutions. The measured signals were transferred by software digitally and transmitted wirelessly by the telemeter, personal digital assistant (PDA), through the General Packet Radio Service (GPRS) protocol. The pH sensing electrode was designed to measure a chemical potential induced by a proton concentration gradient on the electrode's surface which exhibits internal Donnon diffusion behavior, and a linear relationship between the electrical potential and pH was found. The result shows that the wireless sensing system allowed not only long-term usage and long-distance transmission but also with high accuracy (e.g. S.D. less than +/-2%). The telemetric system can also be modified to measure ethanol concentration in aqueous solution amperometrically. It was found that the sensitivity of that ex situ measurements matched those of in field measurements with negligible deviation, less than 4%.