| Gout, one kind of osteoarthritis (OA), is a serious disease that endangers human’s health. Gout has existed since ancient time and now has become more serious, especially in coastal areas. When MSU crystals that caused by metabolic disorder deposit in joints, then gout will come up as the pain and swelling of joints, thus making the patient suffer a lot, even can’t walk. It often takes several weeks even months for the patient to recover by himself. However, when bad weather comes or unhealthy life style continues, gout will attack repeatly, making the patients extremely anguished again and again. In clinic therapy, NSAIDs and other drugs are usually used to treat gout, such as colchicine and other Chinese medicine. In short-term therapy, taking these drugs can be effective, but the side effects of these medicines cannot be ignored. And if the drugs stop, gout will easily recurrent attack. In this case, patients have to take more drugs and therefore leading to a vicious cycle. So how to cure gout and its recurrence effectively and safely is a great challenge.Based on the aforementioned, this work is aimed to choose a kind of medicine that could impactfully treat gout with little side effects and develop a new drug carrier to overcome gout recurrence. For this purpose, in this work luteolin was selected as model drug and then electrospun with PCL and gelatin into composite nanofibers. Raising the ratio of PCL in the fibers could extend release time of the fibers. When PCL is 100%, the release time could be up to about three weeks, which is very promising in the therapy of recurrent gout attacks. The detailed research methods and conclusions are as follows:First, luteolin-loaded PCL/gelatin (5:5) and PCL/gelatin (9:1) nanofibers and luteolin-loaded PCL microfibers were fabricated via electrospinning. During the process of preparing spinning solution, adding HAc could solve the phase separation of PCL and gelatin.At the same time, pre-dissolving luteolin into tiny amount of DMF could improve luteolin’s final solubility in the total spinning solution.Then, in vitro and in vivo gout models were set up. In vitro, LPS stimulated RAW 264.7, which could simulate flammation, is a classic model; Besides, in vitro, XOD reaction in 37℃ could simulate the process of enzyme metabolism of gout, and the absorbance @ 295 nm could represent the amount of purine trione. In vivo, gout model was set up by injecting MSU into New Zealand Rabbits’ joints. After 24 hours, the flammation would be obvious and the synovium membranes were as gout sites and researched. What’s more, at day 0, day 7 and day 14, injecting MSU into joints could simulate recurrent gout attacks.Finally, drug loaded fibers were tested in these models. Luteolin-loaded PCL/gelatin (5:5) were tested both in in vitro model and in vivo model. Luteolin-loaded PCL microfibers were tested in recurrent gout model. In the model of LPS-RAW 264.7, the release solution of nanofibers on different days were co-cultured with LPS-RAW 264.7, it could be found that the metabolic activity was obviously inhibited and the flammation-related cytokines were decreased. In the model of XOD reaction, reaction rate and speed were inhibited by co-cultured with release solution of different days. In vivo, when gout was treated by implanting PG-L (5:5), the results revealed that the flammation was obviously relieved. In long-term recurrent gout model, compared with injection group and blank group, PCL-L showed the "once implanted, long-term effective" ability, and this result shows that modern technology could relieve ancient pain. |
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