Multimodal VEGF-Targeted Contrast-Enhanced Ultrasound and Photoacoustic Imaging of Rats with Inflammatory Arthritis: Using Dye-VEGF-Antibody-Loaded Microbubbles.

Owing to the heavy health burdens from rheumatoid arthritis, a sensitive and objective imaging method is needed for early diagnosis and accurate evaluation of the disease. We aimed to fabricate vascular epithelial growth factor (VEGF)-targeted microbubbles (MBs) to evaluate the expression levels of VEGF within the inflammatory lesions of rats with adjuvant-induced arthritis (AIA) using a multimodal photoacoustic (PA)/ultrasound (US) imaging system. Fluorescein isothiocyanate-biotin double-labeled vascular endothelial growth factor receptor 2 antibodies and Cy5.5-biotin double-labeled VEGF2 antibodies were added to the avidin-labeled MBs to synthesize VEGF-targeted MBs. The antibodies could specifically bind to the MBs according to the flow cytometry and fluorescence imaging. In vitro experiments on the cellular uptake of the target MBs also validated the interaction of the VEGF antibodies and the MBs. Multimodal contrast-enhanced US (CEUS)/PA imaging was performed in sequence on the inflamed paws of the AIA rats with a single PA/US imaging system after the injection of the targeted MBs. The CEUS and PA signals were then quantified and verified by the pathologic results. A CEUS pattern of fast wash in and slow washout was observed in the AIA rats after injection of targeted MBs. Compared with AIA rats injected with unconnected VEGF antibodies and naked MBs, AIA rats injected with targeted MBs presented a higher peak intensity (p = 0.0079 and 0.0079 respectively) and a longer time to peak (p = 0.0117 and 0.0117, respectively). The PA signals were also significantly enhanced after injection of targeted MBs (p = 0.0112 and 0.0119, respectively), which was in accordance with the pathologic and immunohistochemical results. In conclusion, VEGF-targeted MBs can be used as agents for multimodal CEUS/PA imaging and to detect VEGF expression in the inflammatory lesions of AIA rats in vivo. This strategy may be useful in imaging evaluation of arthritis by identifying inflammation-related molecules in different imaging modes.

Effects of Ca/P molar ratios on regulating biological functions of hybridized carbon nanofibers containing bioactive glass nanoparticles.

Biological functions of hybridized carbon nanofibers (CNFs) depend closely on the incorporated bioactive components. For hybridized CNFs containing bioactive glass (BG) nanoparticles (CNF/BG), chemical compositions of BG nanoparticles might have decisive effects on their cell affinity and osteocompatibility. Herein, three hybridized CNF/BGs were produced by incorporating 68S-type BG nanoparticles with different Ca/P molar ratios (1.0, 1.67 or 2.5) into CNFs via a sol-gel/electrospinning and carbonization method. Structural evolution of these hybridized CNF/BGs was studied in relation to their Ca/P molar ratios. Crystalline wollastonite was found to be the dominant phase at a high feeding Ca/P molar ratio (i.e. 2.5), but weak crystallized hydroxyapatite was the main phase at the low feeding Ca/P molar ratio (i.e. 1.0). These findings were correlated to the biological functions of the resulted CNF/BG hybrids including apatite formation ability in simulated body fluid and osteoblast behaviors in in vitro culture. All the CNF/BG hybrids displayed a strong affinity for inducing apatite deposition, showing insignificant difference after the initial nucleation stage, while they behaved differently in promoting the proliferation and osteogenic differentiation of osteoblasts. The fastest proliferation rate and the highest expression of alkaline phosphatase activity was found on the CNF/BG (Ca/P = 1.0). The results suggested a feasible way to upregulate osteoblast behaviors is by changing the feeding Ca/P molar ratios in the preparation of CNF/BG hybrids for potential bone repairing applications.