Chung-Hao Wang

Photothermolysis of glioblastoma stem-like cells targeted by carbon nanotubes conjugated with CD133 monoclonal antibody

UNLABELLED CD133(+) cells in glioblastoma (GBM) display cancer stem cell-like properties and have been considered as the culprit of tumor recurrence, justifying exploration of potential therapeutic modalities targeting CD133(+) cancer stem-like cells (CSCs). For photothermolysis studies, GBM-CD133(+) and GBM-CD133(-) cells mixed with various ratios were challenged with single-walled carbon nanotubes (SWNTs) conjugated with CD133 monoclonal antibody (anti-CD133) and then irradiated with near-infrared laser light. Results show that GBM-CD133(+) cells were selectively targeted and eradicated, whereas GBM-CD133(-) cells remained viable. In addition, in vitro tumorigenic and self-renewal capability of GBM-CD133(+) treated with localized hyperthermia was significantly blocked. Furthermore, GBM-CD133(+) cells pretreated with anti-CD133-SWNTs and irradiated by near-infrared laser 2 days after xenotransplantation in nude mice did not exhibit sustainability of CSC features for tumor growth. Taken altogether, our studies demonstrated that anti-CD133-SWNTs have the potential to be utilized as a thermal-coupling agent to effectively target and destroy GBM CSCs in vitro and in vivo. FROM THE CLINICAL EDITOR Glioblastoma remains one of the most notorious cancer from the standpoint of recurrence and overall resistance to therapy. CD133+ stem cells occur among GBM cells, and may be responsible for the huge recurrence risk. This paper discusses a targeted elimination method of these cells, which may enable more efficient therapy in an effort to minimize or prevent recurrence.

In vitro photothermal destruction of neuroblastoma cells using carbon nanotubes conjugated with GD2 monoclonal antibody

Despite aggressive multimodality therapy, most neuroblastoma-bearing patients relapse and survival rate remains poor. Exploration of alternative therapeutic modalities is needed. Carbon nanotubes (CNTs), revealing optical absorbance in the near-infrared region, warrant their merits in photothermal therapy. In order to specifically target disialoganglioside (GD2) overexpressed on the surface of neuroblastoma stNB-V1 cells, GD2 monoclonal antibody (anti-GD2) was conjugated to acidified CNTs. To examine the fate of anti-GD2 bound CNTs after incubation with stNB-V1 cells, rhodamine B was labeled on carboxylated CNTs functionalized with and without anti-GD2. Our results illustrated that anti-GD2-linked CNTs were extensively internalized by neuroblastoma cells via GD2-mediated endocytosis. In addition, we showed that anti-GD2 bound CNTs were not ingested by PC12 cells without GD2 expression. After anti-GD2 conjugated CNTs were incubated with neuroblastoma cells for 6 h and endocytosed by the cells, CNT-laden neuroblastoma cells were further irradiated with an 808 nm near-infrared (NIR) laser with intensity ramping from 0.6 to 6 W cm(-2) for 10 min which was then maintained at 6 W cm(-2) for an additional 5 min. Post-NIR laser exposure, and after being examined by calcein-AM dye, stNB-V1 cells were all found to undergo necrosis, while non-GD2 expressing PC12 cells all remained viable. Based on the in vitro study, CNTs bound with anti-GD2 have the potential to be utilized as a therapeutic thermal coupling agent that generates heat sufficient to selectively kill neuroblastoma cells under NIR laser light exposure.

Photothermal Ablation of Stem-Cell Like Glioblastoma Using Carbon Nanotubes Functionalized with Anti-CD133

Despite aggressive multimodality therapy, most glioblastoma-bearing patients relapse and survival rate remains poor. Exploration of alternative therapeutic modalities is needed. Biologically modified materials with intrinsic optical properties, such as multi-walled carbon nanotubes (MWNTs) and gold nanoparticles, have been recently extensively explored for their potential use in biomedical applications. MWNTs revealing strong optical absorbance in the nearinfrared region warrant their merits in photothermal therapy. In order to specific target CD133 over-expressed on the surface of glioblastoma cell line GBM S1R1, anti-CD133 monoclonal antibody was employed by conjugated to acidified MWNTs pre-modified with N-hydroxysuccinimide (NHS). In order to visualize anti-CD133 tagged MWNTs ingested by GBM S1R1 cells, fluorescent dye rhadoamine B was labeled onto MWNTs conjugated with anti-CD133. In comparison with DAOY cells, with negative CD133 expression, confocal images and flow cytometric analysis were showed. After CD133-grafted MWNTs were added into cell culture, GBM S1R1 glioblastoma cells and DAOY cells were irradiated separately with 808 nm near-infrared (NIR) laser for 15 minutes under intensity of 6 W/cm2. Our results showed prominent photothermal damage of GBM S1R1 cells, while less necrosis of CD133-negative DAOY cells.

Anti-Neuroblastoma Activity of Gold Nanorods Bound with GD2 Monoclonal Antibody under Near-Infrared Laser Irradiation

High-risk neuroblastoma is one of the most common deaths in pediatric oncology. Current treatment of this disease involves a coordinated sequence of chemotherapy, surgery, and radiation. Further advances in therapy will require the targeting of tumor cells in a more selective and efficient way so that survival can be improved without substantially increasing toxicity. To achieve tumor-selective delivery, disialoganglioside (GD2) expressed by almost all neuroblastoma tumors represents a potential molecular target that can be exploited for tumor-selective delivery. In this study, GD2 monoclonal antibody (anti-GD2) was conjugated to gold nanorods (GNRs) which are one of anisotropic nanomaterials that can absorb near-infrared (NIR) laser light and convert it to energy for photothermolysis of tumor cells. Thiolated chitosan, due to its biocompatibility, was used to replace cetyltrimethylammonium bromide (CTAB) originally used in the synthesis of gold nanorods. In order to specifically target GD2 overexpressed on the surface of neuroblastoma stNB-V1 cells, anti-GD2 was conjugated to chitosan modified GNRs (CGNRs). To examine the fate of CGNRs conjugated with anti-GD2 after incubation with neuroblastoma cells, rhadoamine B was labeled on CGNRs functionalized with anti-GD2. Our results illustrated that anti-GD2-conjugated CGNRs were extensively endocytosed by GD2+ stNB-V1 neuroblastoma cells via antibody-mediated endocytosis. In addition, we showed that anti-GD2 bound CGNRs were not internalized by GD2− SH-SY5Y neuroblastoma cells. After anti-GD2-linked CGNRs were incubated with neuroblatoma cells for six hours, the treated cells were further irradiated with 808 nm NIR laser. Post-NIR laser exposure, when examined by calcein-AM dye, stNB-V1 cells all underwent necrosis, while non-GD2 expressing SH-SY5Y cells all remained viable. Based on the in vitro study, CGNRs bound with anti-GD2 has the potential to be utilized as a therapeutic thermal coupling agent that generates heat sufficient to selectively kill neuroblastoma cells under NIR laser light exposure.

Designing a stronger interface through graded structures in amorphous/nanocrystalline ZrCu/Cu multilayered films.

Many multilayered nano-structures appear to fail due to brittle matter along the interfaces. In order to toughen them, in this study, the microstructure and interface strength of multilayered thin films consisting of amorphous ZrCu and nanocrystalline Cu (with sharp or graded interfaces) are examined and analyzed. The interface possesses a gradient nature in terms of composition, nanocrystalline phase size and volume fraction. The bending results extracted from the nano-scaled cantilever bending samples demonstrate that multilayered films with graded interfaces would have a much higher interface bending strength/strain/modulus, and an overall improvement upgrade of more than 50%. The simple graded interface design of multilayered thin films with improved mechanical properties can offer much more promising performance in structural and functional applications for MEMS or optical coating.

Rapid antiviral assay using QD-tagged fish virus as imaging nanoprobe.

Development of rapid antiviral assays can expedite the process of screening potential agents against viral pathogens. In the present study, fluorescent quantum dots (QDs) incorporated with infectious pancreatic necrosis virus (IPNV) were used as imaging nanoprobes to detect the threshold amount of poly I:C (an interferon inducer) required to induce zebrafish cells into an antiviral state against IPNV. QD-IPNV hybrids were formed by colloidal clustering of negatively charged QDs and IPNV, using the cationic polymer polybrene (50 μg/mL). To test the screening potential of the QD-IPNV hybrids for anti-IPNV drug candidates, zebrafish ZF4 cells primed with the immunostimulant poly I:C at concentrations of 1, 5, and 10 μg/mL for 6h were used as a model system. After poly I:C treatment, cells were exposed to the QD-IPNV hybrids for 6h at a multiplicity of infection (MOI) of 5. The anti-IPNV effectiveness of poly I:C was assessed via fluorescence intensity of the QDs. Our results showed that ZF4 cells primed with poly I:C at 10 μg/mL were highly protected from IPNV challenge (i.e., no detection of QD fluorescence). In summary, a rapid and efficient cell-based imaging platform has been developed for assessing the anti-IPNV activity of poly I:C on ZF4 cells using QD-IPNV hybrids. This approach may be applied to a wider range of fish species and fish pathogenic viruses.

Gold Nanorods Modified with Chitosan As Photothermal Agents

Gold nanorods (GNRs) are one of the nanomaterials, which could absorb near-infrared (NIR) light and convert to heat for the application on photothermal therapy. According to literature, Cetyltrimethylammonium bromide (CTAB) is one of the widely used surfactant for the synthesis of GRNs. However, due to cell toxicity of CTAB, it is necessary to modify the surface of gold nanorods for cell-related studies. In this study, thiolated chitosan was employed to replace CTAB on gold nanorods due to its biocompatibility. The absorption spectra ranged from visible to NIR wavelength were tuned by changing the aspect ratio of chitosan-tagged GNRs. We further conjugated chitosan/GNRs with disiloganglioside (GD2) monoclonal antibody which can lead to the functionalized nanomaterials endocytosed into stNB-V1 neuroblastoma cells. After exposure with NIR laser at 808 nm, photothermal destruction of stNB-V1 cells was clearly demonstrated by calcein AM staining.