Yulei Chang

An upconversion nanoparticle - Zinc phthalocyanine based nanophotosensitizer for photodynamic therapy

Recent advances in NIR triggering upconversion-based photodynamic therapy have led to substantial improvements in upconversion-based nanophotosensitizers. How to obtain the high efficiency of singlet oxygen generation under low 980 nm radiation dosage still remains a challenge. A highly efficient nanophotosensitizer, denoted as UCNPs-ZnPc, was constructed for photodynamic therapy, which is based on near infrared (NIR) light upconversion nanoparticle (UCNP) and Zn(II)-phthalocyanine (ZnPc) photosensitizer (PS). The high (1)O2 production efficiency came from the enhancement of the 660 nm upconversion emission of NaYF4:Yb(3+), Er(3+) UCNP with 25% Yb(3+) doping, covalent assemblage of UCNP and ZnPc which significantly shortened the distance and enhanced the energy transfer between the two. The high (1)O2 production led to a secure and efficient PDT treatment, as evidenced by the in vivo test where UCNPs-ZnPc of 50 mg per kg body weight was locally injected into the liver tumor in mice, a low 980 nm radiation dose of 351 J/cm(2) (0.39 W/cm(2)) and short irradiation duration of 15 min were sufficient to perform image-guided PDT and caused the liver tumor inhibitory ratio of approximately 80.1%. Histological analysis revealed no pathological changes and inflammatory response in heart, lung, kidney, liver or spleen.

In vivo 808 nm image-guided photodynamic therapy based on an upconversion theranostic nanoplatform.

A new strategy for efficient in vivo image-guided photodynamic therapy (PDT) has been demonstrated utilizing a ligand-exchange constructed upconversion-C60 nanophotosensitizer. This theranostic platform is superior to the currently reported nanophotosensitizers in (i) directly bonding photosensitizer C60 to the surface of upconversion nanoparticles (UCNPs) by a smart ligand-exchange strategy, which greatly shortened the energy transfer distance and enhanced the (1)O2 production, resulting in the improvement of the therapeutic effect; (ii) realizing in vivo NIR 808 nm image-guided PDT with both excitation (980 nm) and emission (808 nm) light falling in the biological window of tissues, which minimized auto-fluorescence, reduced light scatting and improved the imaging contrast and depth, and thus guaranteed noninvasive diagnostic accuracy. In vivo and ex vivo tests demonstrated its favorable bio-distribution, tumor-selectivity and high therapeutic efficacy. Owing to the effective ligand exchange strategy and the excellent intrinsic photophysical properties of C60, (1)O2 production yield was improved, suggesting that a low 980 nm irradiation dosage (351 J cm(-2)) and a short treatment time (15 min) were sufficient to perform NIR (980 nm) to NIR (808 nm) image-guided PDT. Our work enriches the understanding of UCNP-based PDT nanophotosensitizers and highlights their potential use in future NIR image-guided noninvasive deep cancer therapy.

ABT737 enhances cholangiocarcinoma sensitivity to cisplatin through regulation of mitochondrial dynamics.

Cholangiocarcinoma responses weakly to cisplatin. Mitochondrial dynamics participate in the response to various stresses, and mainly involve mitophagy and mitochondrial fusion and fission. Bcl-2 family proteins play critical roles in orchestrating mitochondrial dynamics, and are involved in the resistance to cisplatin. Here we reported that ABT737, combined with cisplatin, can promote cholangiocarcinoma cells to undergo apoptosis. We found that the combined treatment decreased the Mcl-1 pro-survival form and increased Bak. Cells undergoing cisplatin treatment showed hyperfused mitochondria, whereas fragmentation was dominant in the mitochondria of cells exposed to the combined treatment, with higher Fis1 levels, decreased Mfn2 and OPA1 levels, increased ratio of Drp1 60kD to 80kD form, and more Drp1 located on mitochondria. More p62 aggregates were observed in cells with fragmented mitochondria, and they gradually translocated to mitochondria. Mitophagy was induced by the combined treatment. Knockdown p62 decreased the Drp1 ratio, increased Tom20, and increased cell viability. Our data indicated that mitochondrial dynamics play an important role in the response of cholangiocarcinoma to cisplatin. ABT737 might enhance cholangiocarcinoma sensitivity to cisplatin through regulation of mitochondrial dynamics and the balance within Bcl-2 family proteins. Furthermore, p62 seems to be critical in the regulation of mitochondrial dynamics.

A facile and general route to synthesize silica-coated SERS tags with the enhanced signal intensity.

Silica-coated SERS tags have been attracting greater attention in recent years. However, the reported methods to synthesize these tags are tedious, and often subjected to the limited signal intensity. Here, we report a facile and general method to prepare the silica-coated Ag SERS tags with the enhanced signal intensity by no introducing the primers. This approach mainly depends on the colloidal stability of the Ag NPs in alcohol solution. By decreasing the concentration of salt in Ag NP solution, the citrate-stabilized Ag NPs can be well dispersed in alcohol solution. Based on this, the Ag SERS tags can be directly coated with thickness-controlled and homogeneous silica shells. This approach is highly reproducible for silica shell growth and signal intensity, not depending on the properties of Raman molecules, proved by 7 kinds of the Raman molecules. Moreover, this kind of SERS tags coated with silica hold the stronger SERS signals than the traditional method due to no interference from the priming molecules.

A SERS nano-tag-based fiber-optic strategy for in situ immunoassay in unprocessed whole blood

Assay technologies capable of detecting biomarker concentrations in unprocessed whole blood samples are fundamental for applications in medical diagnostics. SERS nano-tags integrated fiber-optic biosensor (FOB) was realized for the first time for in situ immunoassay in whole blood. The reliability and sensitivity of this method rely, in a large extent, on the quality and properties of the SERS nano-tags. The constructed silica-coated Ag SERS nano-tags as labels were used in a rapid and specific in situ FOB immune sensor to detect alpha fetoprotein (AFP) in unprocessed blood samples. Preliminary results of in vivo and in situ dynamic observation of AFP of whole blood in wistar rat highlight the power of this new method.

One-step in situ solid-substrate-based whole blood immunoassay based on FRET between upconversion and gold nanoparticles

Despite their general clinical applications, current fluorescence-based immunoassays are confronted with serious challenges, e.g. the advance serum/ plasma separation and the tedious washing process in current heterogeneous approaches, and aggregation of particles, low sensitivity and the narrow linear range in homogeneous approaches. In this paper, these urgent problems were solved in a novel one-step in situ immunoassay of whole blood samples by combining the traditional fluorescence resonance energy transfer (FRET) technology (between upconversion nanoparticles (UCNPs) and gold nanoparticles (GNPs)) and the solid-substrate based immunoassay technology. The low detection limits of goat IgG (gIgG) as 0.042μg/mL in buffers, 0.51μg/mL in 20-fold diluted whole blood samples and a wide linear range from 0.75μg/mL to 60μg/mL in blood samples were achieved. To the best of our knowledge, it is the first one-step in situ solid-substrate-based immunoassay of whole blood samples with large linear detection range. This development provides a promising platform for a rapid and sensitive immunoassay of various bio-molecules directly in whole blood without tedious separation, washing steps and aggregation problems.

Accurate Quantitative Sensing of Intracellular pH based on Self-ratiometric Upconversion Luminescent Nanoprobe.

Accurate quantitation of intracellular pH (pHi) is of great importance in revealing the cellular activities and early warning of diseases. A series of fluorescence-based nano-bioprobes composed of different nanoparticles or/and dye pairs have already been developed for pHi sensing. Till now, biological auto-fluorescence background upon UV-Vis excitation and severe photo-bleaching of dyes are the two main factors impeding the accurate quantitative detection of pHi. Herein, we have developed a self-ratiometric luminescence nanoprobe based on förster resonant energy transfer (FRET) for probing pHi, in which pH-sensitive fluorescein isothiocyanate (FITC) and upconversion nanoparticles (UCNPs) were served as energy acceptor and donor, respectively. Under 980 nm excitation, upconversion emission bands at 475 nm and 645 nm of NaYF4:Yb3+, Tm3+ UCNPs were used as pHi response and self-ratiometric reference signal, respectively. This direct quantitative sensing approach has circumvented the traditional software-based subsequent processing of images which may lead to relatively large uncertainty of the results. Due to efficient FRET and fluorescence background free, a highly-sensitive and accurate sensing has been achieved, featured by 3.56 per unit change in pHi value 3.0–7.0 with deviation less than 0.43. This approach shall facilitate the researches in pHi related areas and development of the intracellular drug delivery systems.

Precisely Tailoring Upconversion Dynamics via Energy Migration in Core–Shell Nanostructures

Abstract Upconversion emission dynamics have long been believed to be determined by the activator and its interaction with neighboring sensitizers. Herein this assumption is, however, shown to be invalid for nanostructures. We demonstrate that excitation energy migration greatly affects upconversion emission dynamics. “Dopant ions’ spatial separation” nanostructures are designed as model systems and the intimate link between the random nature of energy migration and upconversion emission time behavior is unraveled by theoretical modelling and confirmed spectroscopically. Based on this new fundamental insight, we have successfully realized fine control of upconversion emission time behavior (either rise or decay process) by tuning the energy migration paths in various specifically designed nanostructures. This result is significant for applications of this type of materials in super resolution spectroscopy, high‐density data storage, anti‐counterfeiting, and biological imaging.

Dependence between cytotoxicity and dynamic subcellular localization of up-conversion nanoparticles with different surface charges

Intensive investigations have been devoted to lanthanide-doped upconversion nanoparticles (UCNPs), which have shown great potential in applications such as biomedical imaging and therapy. Recently, various polymer-coated UCNPs, as novel bioprobes or nanocarriers, have been developed and their cellular uptake behavior and cytotoxicity have been widely studied. However, the interactions between the UCNPs and subcellular organelles are poorly understand, which restrict their applications in biomedicine. Herein, we engineered UCNPs with different surface charges (positive, negative, and neutral) and studied the dependence between cytotoxicity, internalization, and subcellular localization in normal and cancer cell lines. It was observed that UCNPs with positive or neutral charges entered most of the studied cell lines, whereas UCNPs with negative charges internalize mostly inside the cancer cell lines. Moreover, upon entering into the cells, these UCNPs are localized in different cell compartments, e.g. the cytoplasm, mitochondria or lysosomes, depending on their surface charges and incubation time with the cells. It is revealed that the cytotoxicity of the differently charged UCNPs towards the studied cell lines significantly depends on localization in the mitochondria rather than in the lysosomes or cytoplasm. The corresponding changes in the mitochondria structure were visualized showing the likelihood of cell death. These results have enriched our knowledge on the cytotoxicity of UCNPs in organelle and sheds light on the design of organelle-targeted UCNPs in tumor imaging and therapy.

Catalysis-reduction strategy for sensing inorganic and organic mercury based on gold nanoparticles

In view of the high biotoxicity and trace concentration of mercury (Hg) in environmental water, developing simple, ultra-sensitive and highly selective method capable of simultaneous determination of various Hg species has attracted wide attention. Here, we present a novel catalysis-reduction strategy for sensing inorganic and organic mercury in aqueous solution through the cooperative effect of AuNP-catalyzed properties and the formation of gold amalgam. For the first time, a new AuNP-catalyzed-organic reaction has been discovered and directly used for sensing Hg2+, Hg22+ and CH3Hg+ according to the change of the amount of the catalytic product induced by the deposition of Hg atoms on the surface of AuNPs. The detection limit of Hg species is 5.0pM (1 ppt), which is 3 orders of magnitude lower than the U.S. Environmental Protection Agency (EPA) limit value of Hg for drinking water (2 ppb). The high selectivity can be exceptionally achieved by the specific formation of gold amalgam. Moreover, the application for detecting tap water samples further demonstrates that this AuNP-based assay can be an excellent method used for sensing mercury at very low content in the environment.