Quantitative analysis of collagen after intense pulsed light and erbium-doped yttrium aluminum garnet laser treatment on rabbit skin.

OBJECTIVE: To investigate the effects of intense pulsed light (IPL) and ablative 2.940 nm erbium-doped yttrium aluminum garnet (Er: YAG) laser on dynamic changes in collagen by quantitative analysis of type I collagen (Col I) and type III collagen (Col III), transforming growth factor (TGF), and matrix metalloproteinases (MMPs) in the dermis of rabbits. METHODS: Backs of ten rabbits were divided into four treatment areas: IPL normal energy group, IPL high-energy group, erbium laser normal energy group, and erbium laser high-energy group. HE staining was performed immediately after the first treatment and two weeks after the first treatment. Col I, Col III, TGF-ß1, and MMP-1 were collected by real-time PCR at baseline, 2 weeks after each session, and three months after the entire treatment monthly. RESULTS: HE staining showed that collagen fibers in the superficial layer of the dermis in the four treatment groups were increased and thickened to different degrees. Real-time PCR showed that statistically differences were noted in each checkpoint before and after treatments in the four groups, whereas no significant difference in the change in the four biomarkers was found among the four treatment groups. CONCLUSION: Both the IPL and Er: YAG laser can effectively upregulate collagens, but in this experiment, there was no significant difference in the therapeutic effect among the four irradiation groups. Moreover, the high-energy level group tended to bring more serious epidermal injury.

Study on cytotoxicity, cellular uptake and elimination of rare-earth-doped upconversion nanoparticles in human hepatocellular carcinoma cells.

The growing use of rare-earth doped upconversion nanoparticles (UCNPs) has caused increasing concern about their biosafety. Here, to understand the toxicity of UCNPs and their mechanism in HepG2 cells, we systematically study the cytotoxicity, uptake and elimination behaviors of three types of UCNPs combined multiple cytotoxicity evaluation means with inductively coupled plasma mass spectrometry (ICP-MS) detection. Sodium yttrium fluoride, doped with 18% (molar ratio) ytterbium and 2% erbium (NaYF4: Yb3+, Er3+) was selected as the model UCNPs with two sizes (35 and 55 nm), and the poly(acrylic acid) and polyethylenimine were selected as the representatives of negative and positive surface coating of UCNPs, respectively. UCNPs were found to induce cytotoxicity in time- and dose-dependent manners, which might be mediated by reactive oxygen species generation and oxidative stress. Apoptosis, inflammation, and metabolic process were enhanced after cells exposed to 200 mg/L UCNPs for 48 h. Increase in the protein levels of cleaved caspased-9, cleaved caspase-3 and Bax and decrease in the anti-apoptotic protein, Bcl-2 suggested that the mitochondria mediated pathway was involved in UCNP-induced apoptosis. With the aid of ICP-MS, it demonstrated that the cytotoxicity was associated with internalized amount of UCNPs, which largely relied on their surface properties rather than size in the tested range. By comparing UCNPs with Y3+ ions, it demonstrated that NPs properties played a nonnegligible role in the cytotoxicity of UCNPs. These findings provide new insights for fundamental understanding of cytotoxicity of UCNPs and may contribute to more rational use of these materials in the future.

αvß3-Isoform specific erbium complexes highly specific for bladder cancer imaging and photodynamic therapy.

We have synthesized a bifunctional erbium-porphyrin tumor imaging and PDT agent (Er-R3) that is capable of killing bladder cancer cells via its selective binding to the integrin αvß3 isoform overexpressed on the cell membrane.

Noninvasive and Reversible Cell Adhesion and Detachment via Single-Wavelength Near-Infrared Laser Mediated Photoisomerization.

Dynamically regulating cell-molecule interactions is fundamental to a variety of biological and biomedical applications. Herein, for the first time, by utilizing spiropyran conjugated multishell upconversion nanoparticles (UCNPs) as a new generation of single-wavelength near-infrared (NIR)-controlled photoswitch, we report a simple yet versatile strategy for controlling cell adhesion/detachment reversibly and noninvasively. Specifically, the two-way isomerization of the photoswitch was merely dependent on the excitation power density of the 980 nm laser. At high power density, the ring-opening was prominent, whereas its reverse ring-closing process occurred upon irradiation by the same laser but with the lower power density. Such transformations made the interactions between spiropyran and cell surface protein fibronectin switchable, thus leading to reversible cell adhesion and detachment. Moreover, efficient adhesion-and-detachment of cells could be realized even after 10 cycles. Most importantly, the utilization of NIR not only showed little damage toward cells, but also improved penetration depth. Our work showed promising potential for in vivo dynamically manipulating cell-molecule interactions and biological process.

Transport of NaYF4:Er3+, Yb3+ up-converting nanoparticles into HeLa cells.

An effective, simple and practically useful method to incorporate fluorescent nanoparticles inside live biological cells was developed. The internalization time and concentration dependence of a frequently used liposomal transfection factor (Lipofectamine 2000) was studied. A user friendly, one-step technique to obtain water and organic solvent soluble Er(3+) and Yb(3+) doped NaYF4 nanoparticles coated with polyvinylpyrrolidone was obtained. Structural analysis of the nanoparticles confirmed the formation of nanocrystals of the desired sizes and spectral properties. The internalization of NaYF4 nanoparticles in HeLa cervical cancer cells was determined at different nanoparticle concentrations and for incubation periods from 3 to 24 h. The images revealed a redistribution of nanoparticles inside the cell, which increases with incubation time and concentration levels, and depends on the presence of the transfection factor. The study identifies, for the first time, factors responsible for an effective endocytosis of the up-converting nanoparticles to HeLa cells. Thus, the method could be applied to investigate a wide range of future 'smart' theranostic agents. Nanoparticles incorporated into the liposomes appear to be very promising fluorescent probes for imaging real-time cellular dynamics.

[Research of biocompatibility of nanoparticles with fluorescent domen Er/Yb in the neutrophilic granulocytes system].

Fluorescent tags are extensively used for the diagnostic, labeling and marking in vivo and in vitro systems. In this study, fluorescent nanoparticles with Er/Yb lightning center were tested on neutrophilic granulocytes. The main purpose was to idenfity possible toxic effect. The negative impact of fluorophores on the metabolism of neutrophilic and their enzyme systems, on the receptor-mediated cell responses and on the rigidity of cell membranes was shown. The viability of neutrophils (estimated with the use of propidium iodide) after 2 hours of incubation with the fluorescent nanoparticles in concentrations of 10(-4) and 10(-3) mM was 27.0 +/- 6.6 and 19.07 +/- 3.34 %, respectively.

Spectroscopic studies on the interaction between tryptophan-erbium(III) complex and herring sperm DNA.

By means of UV and fluorescence spectra, the binding ratios between Er(III)-Trp and DNA in physiological pH environment (pH 7.40) were determined as n(Trp):n(Er(III))=3:1 and (n)ER(III)(Trp)(3):(n)(DNA) = 2:1, and the apparent molar absorptivity of epsilon(Er(III)-Trp-DNA) is 4.33 x 10(5) L mol(-1)cm(-1) which was confirmed by molar ratio method. The binding constants at different temperatures K(B25 degrees C)(theta)=1.93 x 10(4)L mol(-1) and K(B37 degrees C)(theta)=5.28 x 10(3)L mol(-1) were obtained by double reciprocal method. Thermodynamic function computation demonstrates that Delta(r)H(m)(theta) is the primary driving power of the interaction between Er(III)(Trp)(3) and DNA. By combination analysis of the Scatchard method and CD spectrometry, we suggested that the interaction mode between Er(III)(Trp)(3) complex and herring sperm DNA is groove and intercalation bindings.

X-ray studies reveal lanthanide binding sites at the A/B5 interface of E. coli heat labile enterotoxin.

The crystal structure determination of heat labile enterotoxin (LT) bound to two different lanthanide ions, erbium and samarium, revealed two distinct ion binding sites in the interface of the A subunit and the B pentamer of the toxin. One of the interface sites is conserved in the very similar cholera toxin sequence. These sites may be potential calcium binding sites. Erbium and samarium binding causes a change in the structure of LT: a rotation of the A1 subunit of up to two degrees relative to the B pentamer.