Search

Objective Recent innovations in biology are boosting gene and cell therapy, but monitoring the response to these treatments is difficult. The purpose of this study was to find an MRI-reporter gene that can be used to monitor gene or cell therapy and that can be delivered without a viral vector, as viral vector delivery methods can result in long-term complications. Materials and methods CMV promoter-human organic anion transporting polypeptide 1B3 (CMV-hOATP1B3) cDNA or CMV-blank DNA (control) was transfected into HEK293 cells using Lipofectamine. OATP1B3 expression was confirmed by western blotting and confocal microscopy. In vitro cell phantoms were made using transfected HEK293 cells cultured in various concentrations of gadoxetic acid for 24 hours, and images of the phantoms were made with a 9.4T micro-MRI. In vivo xenograft tumors were made by implanting HEK293 cells transfected with CMV-hOATP1B3 (n = 4) or CMV-blank (n = 4) in 8-week-old male nude mice, and MRI was performed before and after intravenous injection of gadoxetic acid (1.2 μL/g). Results Western blot and confocal microscopy after immunofluorescence staining revealed that only CMV-hOATP1B3-transfected HEK293 cells produced abundant OATP1B3, which localized at the cell membrane. OATP1B3 expression levels remained high through the 25th subculture cycle, but decreased substantially by the 50th subculture cycle. MRI of cell phantoms showed that only the CMV-hOATP1B3-transfected cells produced a significant contrast enhancement effect. In vivo MRI of xenograft tumors revealed that only CMV-hOATP1B3-transfected HEK293 tumors demonstrated a T1 contrast effect, which lasted for at least 5 hours. Conclusion The human endogenous OATP1B3 gene can be non-virally delivered into cells to induce transient OATP1B3 expression, leading to gadoxetic acid-mediated enhancement on MRI. These results indicate that hOATP1B3 can serve as an MRI-reporter gene while minimizing the risk of long-term complications.

In recent days, while taller and more massive structures such as huge bridges and super skyscrapers have been welcomed, the structural stabilization in design and construction have been gradually limited due to the major weakness of current concrete which is relatively heavier when compared with its strength. To improve the weakness of the current concrete, The lightweight concrete with light weight and high strength should be used; however, not many researchers in Korea have studied on the lightweight concrete. Generally, artificial lightweight aggregate produced through high-temperature-plasticization has a possibility of its body-expansion with many bubbles. Therefore, depending on the size of aggregate, the effects of bubbles on the specific weight and strength of the lightweight concrete should be studied. In this study, considering grain-size, the mix design of the artificial lightweight aggregate produced through the high-temperature-plasticization and the body-expansion of waste and clay from the fire power plant in Korea was conducted. The experiment to analyze the variation in specific weight and strength of the lightweight concrete was followed. From these experiments, the optimized grain-size ratio of the artificial lightweight aggregate for the enhancement of high-strength from the lightweight concrete was revealed.