Molecular imaging of orthotopic prostate cancer with nanobubble ultrasound contrast agents targeted to PSMA.

Ultrasound imaging is routinely used to guide prostate biopsies, yet delineation of tumors within the prostate gland is extremely challenging, even with microbubble (MB) contrast. A more effective ultrasound protocol is needed that can effectively localize malignancies for targeted biopsy or aid in patient selection and treatment planning for organ-sparing focal therapy. This study focused on evaluating the application of a novel nanobubble ultrasound contrast agent targeted to the prostate specific membrane antigen (PSMA-targeted NBs) in ultrasound imaging of prostate cancer (PCa) in vivo using a clinically relevant orthotopic tumor model in nude mice. Our results demonstrated that PSMA-targeted NBs had increased extravasation and retention in PSMA-expressing orthotopic mouse tumors. These processes are reflected in significantly different time intensity curve (TIC) and several kinetic parameters for targeted versus non-targeted NBs or LUMASON MBs. These, may in turn, lead to improved image-based detection and diagnosis of PCa in the future.

Time-intensity-curve Analysis and Tumor Extravasation of Nanobubble Ultrasound Contrast Agents.

Our group recently presented a simple strategy using the non-ionic surfactant, Pluronic, as a size control excipient to produce nanobubbles in the 100-nm range, which exhibited stability and echogenicity on par with clinically available microbubbles. The objective of the present study was to evaluate biodistribution and extravasation of the Pluronic-stabilized lipid nanobubbles compared with microbubbles in 2 experimental tumor models in mice. Standard lipid-stabilized perfluoropropane bubbles (Pluronic L10) and lipid-stabilized perfluoropropane nanobubbles were intravenously injected into mice bearing either an orthotopic mouse breast cancer (BC4 T1) or subcutaneous mouse ovarian cancer (OVCAR-3) through the tail vein to perform perfusion dynamic studies. No significant differences between the nanobubble and microbubble groups were observed in the peak enhancement of the 3 tested regions (tumor, liver and kidney). However, the decay rates of nanobubble in the tumor and kidney of BC4 T1-bearing mice, as well as in mice with OVRCAR-3 tumors were significantly slower than those of the microbubble. To quantify extravasation, fluorescently labeled bubbles were intravenously injected into mice bearing the same tumors. Histologic analysis showed that nanobubbles were retained in tumor tissue to a greater extent compared with microbubbles in both tumor models at the 3-h time point. Our results demonstrate unique nanobubble behavior compared with microbubbles and support augmented application of these agents in ultrasound molecular imaging and drug delivery beyond the tumor vasculature.

Effect of Bubble Concentration on the in Vitro and in Vivo Performance of Highly Stable Lipid Shell-Stabilized Micro- and Nanoscale Ultrasound Contrast Agents.

Ultrasound (US) is a widely used diagnostic imaging tool because it is inexpensive, safe, portable, and broadly accessible. Ultrasound contrast agents (UCAs) are employed to enhance backscatter echo and improve imaging contrast. The most frequently utilized UCAs are echogenic bubbles made with a phospholipid or protein-stabilized hydrophobic gas core. While clinically utilized, applications of UCAs are often limited by rapid signal decay (<5 min) in vivo under typical ultrasound imaging protocols. Here, we report on a formulation of lipid shell-stabilized perfluoropropane (C3F8) microbubbles and nanobubbles with a significantly prolonged in vivo stability. Microbubbles (875 ± 280 nm) of the target size were prepared by utilizing a multiple-step centrifugation cycle, while nanobubbles (299 ± 189 nm) were isolated from the activated vial using a single centrifugation step. To provide in-depth acoustic characterization of the new construct we evaluated the effect of size and concentration on their in vitro and in vivo performance. In vitro and in vivo characterization were carried out for a range of bubble concentrations normalized by total gas volume quantified via headspace gas chromatography/mass spectrometry (GC/MS). In vitro characterization revealed that nanobubbles at different concentrations are more consistently stable over time with the highest and lowest dilutions (50-fold decrease) only differing in US signal after 8 min exposure by 10.34%, while for microbubbles the difference was 86.46%. As expected, due to the difference in hydrodynamic diameter and scattering cross section difference, nanobubbles showed lower overall initial signal intensity. In vivo experiments showed that both microbubbles and nanobubbles with similar initial peak signal intensity are comparably stable over time with 66.8% and 60.6% remaining signal after 30 min, respectively. This study demonstrates that bubble concentration has significant effects on the persistence of both microbubbles and nanobubbles in vitro and in vivo, but the effects are more pronounced in larger bubbles. These effects should be taken into account when selecting the appropriate bubble parameters for future imaging applications.

Sink or float? Characterization of shell-stabilized bulk nanobubbles using a resonant mass measurement technique.

Nano-sized shell-stabilized gas bubbles have applications in various fields ranging from environmental science to biomedical engineering. A resonant mass measurement (RMM) technique is demonstrated here as a new and only method capable of simultaneously measuring the size and concentration of buoyant and non-buoyant particles in a nanobubble sample used as a next-generation ultrasound contrast agent.

3D Printing of Photocurable Cellulose Nanocrystal Composite for Fabrication of Complex Architectures via Stereolithography.

The advantages of 3D printing on cost, speed, accuracy, and flexibility have attracted several new applications in various industries especially in the field of medicine where customized solutions are highly demanded. Although this modern fabrication technique offers several benefits, it also poses critical challenges in materials development suitable for industry use. Proliferation of polymers in biomedical application has been severely limited by their inherently weak mechanical properties despite their other excellent attributes. Earlier works on 3D printing of polymers focus mainly on biocompatibility and cellular viability and lack a close attention to produce robust specimens. Prized for superior mechanical strength and inherent stiffness, cellulose nanocrystal (CNC) from abaca plant is incorporated to provide the necessary toughness for 3D printable biopolymer. Hence, this work demonstrates 3D printing of CNC-filled biomaterial with significant improvement in mechanical and surface properties. These findings may potentially pave the way for an alternative option in providing innovative and cost-effective patient-specific solutions to various fields in medical industry. To the best of our knowledge, this work presents the first successful demonstration of 3D printing of CNC nanocomposite hydrogel via stereolithography (SL) forming a complex architecture with enhanced material properties potentially suited for tissue engineering.

Grafted carbazole-assisted electrodeposition of graphene oxide.

The electrodeposition of graphene oxide (GO) by covalently linked electroactive monomer, carbazole (Cbz) is first demonstrated herein. This is based on the electropolymerization and electrodeposition of covalently linked Cbz units when a potential is applied. During the electrochemical process, the Cbz groups electropolymerize and carry the GO nanosheets as it electrodeposits on the substrate. Moreover, the GO-Cbz sheets selectively deposit onto the conducting regions of the substrate, which demonstrates its promise for the fabrication of electropatterned graphene-based devices. In addition, GO-Cbz is a promising material for the fabrication of nanocomposite coatings for anticorrosion application. In as little as 1 wt % GO-Cbz loading, a protection efficiency as high as 95.4% was achieved.

Superhydrophobic colloidally textured polythiophene film as superior anticorrosion coating.

In this paper, we demonstrated for the first time the use of electrodeposited superhydrophobic conducting polythiophene coating to effectively protect the underlying steel substrate from corrosion attack: by first preventing water from being absorbed onto the coating, thus preventing the corrosive chemicals and corrosion products from diffusing through the coating, and second by causing an anodic shift in the corrosion potential as it galvanically couples to the metal substrate. Standard electrochemical measurements revealed the steel coated with antiwetting nanostructured polythiophene film, which was immersed in chloride solution of different pH and temperature for up to 7 days, is very well protected from corrosion evidenced by protection efficiency of greater than 95%. Fabrication of the dual properties superhydrophobic anticorrosion nanostructured conducting polymer coating follows a two-step coating procedure that is very simple and can be used to coat any metallic surface.