Chih-Hsien Lee

Synthesis, Characterization, and Bioconjugation of Fluorescent Gold Nanoclusters toward Biological Labeling Applications

Synthesis of ultrasmall water-soluble fluorescent gold nanoclusters is reported. The clusters have a decent quantum yield, high colloidal stability, and can be readily conjugated with biological molecules. Specific staining of cells and nonspecific uptake by living cells is demonstrated.

Fluorescent Gold Nanoclusters as a Biocompatible Marker for In Vitro and In Vivo Tracking of Endothelial Cells

We have been investigating the fluorescent property and biocompatibility of novel fluorescent gold nanoclusters (FANC) in human aortic endothelial cells (HAEC) and endothelial progenitor cells (EPC). FANC (50-1000 nmol/L) was delivered into cells via the liposome complex. The fluorescence lasted for at least 28 days with a half-life of 9 days in vitro. Examination of 12 transcripts regulating the essential function of endothelial cells after a 72 h delivery showed that only the vascular cell adhesion molecule 1 and the vascular endothelial cadherin were down-regulated at high concentration (500 nmol/L). In addition, no activation of caspase 3 or proliferating cell nuclear antigens was detected. 3-[4,5-Dimethylthiazol-2-yl]-2,5- diphenyltetrazolium bromide (MTT) assay demonstrated that, unlike the markedly suppressed viability in cells treated with quantum dots, FANC had minimal effect on the viability, unless above 500 nmol/L, at which level a minor reduction of viability mainly caused by liposome was found. Tube formation assay showed no impaired angiogenesis in the EPC treated with FANC. In vivo study using hindlimb ischemic mice with an intramuscular injection of FANC-labeled human EPC showed that the cells preserved an angiogenic potential and exhibited traceable signals after 21 days. These findings demonstrated that FANC is a promising biocompatible fluorescent probe.

Recombination dynamics of photoluminescence in thiol-protected gold nanoclusters

Recombination dynamics of photoluminescence (PL) in Au nanoclusters (NCs) with different capping molecules were studied with time-resolved PL. Based on the emission-energy of carrier lifetimes; we suggest that the fast and slow PL decay of Au NCs originates from recombination of the linear Au–S bond and the staple motif, respectively. The effect of carrier localization in Au NCs was found to depend on the capping molecules. The zero-dimensionality of carriers in Au NCs was demonstrated by the temperature dependence of the time-resolved PL.

Synthesis and surface modification of highly fluorescent gold nanoclusters and their exploitation for cellular labeling

We introduce a general approach to make colorful fluorescent gold nanoclusters which are protected by dihydrolipoic acid, mercaptoundecanoic acid and polyethylenimine. The fluorescent gold nanoclusters can perform a variety of bioconjugation processes such as PEGylation, biotinylation as well as forming complex nanobioconjugates with streptavidins. The brightening effects under proper surface modification are also reported. The clusters have a decent quantum yield, high colloidal stability, and can be readily conjugated with biological molecules. Nonspecific uptake by human aortic endothelial cells is demonstrated.

Impact of dihydrolipoic acid on mouse embryonic stem cells and related regulatory mechanisms

α‐Lipoic acid (LA) is a thiol with antioxidant properties that protects against oxidative stress‐induced apoptosis. LA is absorbed from the diet, taken up by cells and tissues, and subsequently reduced to dihydrolipoic acid (DHLA). Recently, DHLA has been used as the hydrophilic nanomaterial preparations, and therefore, determination of its bio‐safety profile is essential. In this article, we show that DHLA (50–100 μM) induces apoptotic processes in mouse embryonic stem cells (ESC‐B5), but exerts no injury effects at treatment dosages below 50 μM. Higher concentrations of DHLA (50–100 μM) directly increased the reactive oxygen species (ROS) content in ESC‐B5 cells, along with a significant increase in cytoplasmic free calcium and nitric oxide (NO) levels, loss of mitochondrial membrane potential (MMP), activation of caspases‐9 and −3, and cell death. Pretreatment with NO scavengers suppressed the apoptotic biochemical changes induced by 100 μM DHLA and promoted the gene expression levels of p53 and p21 involved in apoptotic signaling. Our results collectively indicate that DHLA at concentrations of 50–100 μM triggers apoptosis of ESC‐B5 cells, which involves both ROS and NO. Importantly, at doses of less than 50 μM (0–25 μM), DHLA does not exert hazardous effects on ESC‐B5 cell properties, including viability, development and differentiation. These results provide important information in terms of dosage safety and biocompatibility of DHLA to facilitate its further use as a precursor for biomaterial preparation.