| Summary: | 碩士 === 國立東華大學 === 物理學系 === 99 === In recent years, nanodiamond (ND) has been used in various biomedical applications due to its good natural fluorescence, strong Raman signals and superb compatibility with bio organism and non-toxicity with biological cell. It can be used to connect anti-cancer drug. The future goal is to use in a living system to carry and deliver anti-cancer drugs.
A biomarker or drug carrier should be tested in animal models or clinical trials. The first problem of trials that it may cause the blood to experience a variety of responses, especially red blood cells (RBC), so far, there are only very few ND interaction with RBC studies. In this work, we used confocal Raman spectroscopy and confocal fluorescence microscopy to study the RBC and RBC interaction of ND. We used fluorescence images and Raman signals to obtain NDs interaction with a single RBC. Using NDs and RBC’s characteristic Raman signals to identify determined whether RBC is damaged or experience unexpected side effects. The experimental results may have implementation for ND in applications on biological system.
This study is divided into two parts: the first part, we used different particle sizes of NDs and carboxylated NDs to interact with a single RBC using confocal Raman spectroscopy and to decide the impact of ND on the oxygen carrying function of RBC, and used Raman spectroscopy to perform quantitative analysis on the RBC oxygenation, changes over time. The second part includes discussions in different concentrations of ND on the oxygen carrying capacity of RBC, using RBC’s oxygenation (R-state) and deoxygenation (T-state) state by observing the Raman peaks’ changes, combine with confocal fluorescence microscopy fluorescent images to observe high concentration NDs coated on membrane of RBCs to see if ND affect the oxygen capacity of RBC.
From the Raman spectroscopy and fluorescence imaging, ND did not change the structure of hemoglobin within RBC. NDs would not cause RBC hemolysis, but the NDs could cause RBCs aggregation. When high concentrations NDs were added, aggregation of the RBC would form more obviously. The aggregation caused small time delay in the re-oxygenation of RBC. Therefor an appropriate concentration of NDs as dosage or biological target vector is feasible. However, due to the complexity of the physiological state of the blood, further research is needed to explore the possibility of the immune response of ND on the blood system before ND can be applied in animal or human models.
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