| 摘要: | 研究背景:藍光是高能可見光，近年人們使用手機等電子產品時間增加，讓眼睛持續暴露在藍光之下，藍光引起的視網膜病變與發炎反應有關，使視網膜藍光損傷問題越來越受到關注。而一種抗氧化/發炎藥物TGF-β活化激?1(TAK1)之抑制劑 (5z-7- oxozeaenol, Oxo）具有降低活性氧與抑制發炎等效果。該研究選用Oxo來治療藍光引起的視網膜問題，視網膜位於後眼部，這使藥物遞送和治療功效面臨許多挑戰。奈米顆粒作為藥物載體來治療眼部疾病是一個良好選擇，其具有保護藥物、增加藥物停留眼部組織及緩慢釋放藥物的優勢，提升治療效果，上述優點是用於治療後眼部的理想選擇。本研究選用天然高分子明膠作為奈米載體材料。其中，透明質酸（Hyaluronic acid, HA）能順利通過玻璃體，增強細胞吞噬能力，具有標靶視網膜上皮細胞的功能。
目的:本研究採用明膠奈米顆粒(GNP)作為藥物載體，在給藥系統中攜帶抗發炎藥5Z-7-oxozeatenol (Oxo)作為奈米藥物(GNP-Oxo)，並於表面披覆HA(GNP-Oxo-HA)以研究其是否可有效治療因藍光而受損的視網膜。
材料與方法:使用粒徑分析儀與穿透式電子顯微鏡檢測奈米顆粒大小、表面電位與顆粒型態檢測， 由奈米顆粒追蹤分析觀察奈米顆粒在水溶液中的分布情形，並檢測奈米顆粒的藥物釋放曲線。在體外檢測中，奈米顆粒與視網膜色素上皮細胞共同培養，使用流式細胞儀檢測細胞吞噬情形，並觀察藍光誘導前後之細胞活性與ROS含量；在動物試驗中，使用C57BL/6小鼠，並建立藍光誘導損傷模型誘導小使視網膜損傷後藉由玻璃體注射給予藥物治療，後續檢測使用眼底攝影與螢光素眼底血管成像(FFA)、眼科光學同調斷層掃描(OCT)與視網膜電位圖(ERG)進行追蹤，後續犧牲後使用蘇木精-伊紅 (H&E)組織染色觀察。
結果:選用pH8明膠奈米顆粒搭載Oxo並包覆HA奈米顆粒尺寸與表面電荷為166.27 nm與-47.3 mV，包覆率為84.33%，並具有良好型態與在水溶液中有良好分布，在藥物釋放曲線奈米顆粒展現緩釋效果，包覆HA後具有更好的效果。在細胞測試中，透過細胞活性得知Oxo對細胞安全濃度為0.1 μg/ mL，在藍光誘導後損傷細胞後能提高細胞存活率，從ROS測試得知Oxo能有效抑制細胞內ROS，GNP-Oxo-HA具有更好效果。動物實驗中，經照射藍光損傷小鼠視網膜後，就由觀察視網膜厚度與內核層(ONL)厚度得知Oxo藥物能減緩藍光所帶來的損傷，GNP-Oxo-HA具有更好的治療效果。
結論:已知Oxo能有效治療或減緩因藍光所帶來的視網膜損傷，使用明膠奈米顆粒是能夠高效率將藥物傳輸到視網膜的一種載體，除此之外，結合透明質酸性質，提高了在後眼部的傳輸與治療效率，對於這樣的結果有利於改善臨床上治療後眼部疾病。
Background: Blue light is high-energy visible light. In recent years, people have spent more time using electronic products such as mobile phones, so that the eyes are continuously exposed to blue light. Retinopathy caused by blue light is related to inflammation, and the problem of blue light damage to the retina has attracted more attention. And an anti-oxidant/inflammatory drug TGF-β-activated kinase 1 (TAK1) inhibitor (5z-7-oxozeaenol, Oxo) has the effect of reducing reactive oxygen species and inhibiting inflammation. The study selected Oxo to treat retinal problems caused by blue light. The study selected Oxo to treat retinal problems caused by blue light. This is located posterior segment, which poses many challenges for drug delivery and therapeutic efficacy. Nanoparticles are a good choice as a drug carrier to treat eye diseases. They protect the drug, increase retention of the drug in the eye tissue, and slowly releasing the drug to improve the therapeutic effect. This study selected natural polymer gelatin as the nano-carrier material because of the above advantages. Among them, hyaluronic acid (HA) can pass through the vitreous body smoothly, enhance cell phagocytosis, and target retinal epithelial cells.
Aim: This study will used gelatin nanoparticles (GNP) loaded with 5Z-7-Oxozeaenol (GNP-Oxo) and coated with hyaluronic acid (GNP-Oxo-HA) to study its efficacy in the treatment of blue light-damaged retina.
Material and methods: Use dynamic light scattering (DLS) and transmission electron microscopy (TEM) to detect nanoparticle size,zeta potential and morphology use nanoparticle tracking analysis (NTA) to observe the distribution of nanoparticles in aqueous solution. and detect the drug release profile of the nanoparticles. In vitro , nanoparticles were confirmed with retinal pigment epithelial cells (RPE), cell activity and ROS content were observed before and after blue light induction;In vivo, C57BL/6 mice were used, and create blue light damage model induces mice retinal ,after drug treatment by intravitreal injection , and observe the damage by Fundus photography, fluorescein fundus angiography (FFA), Ophthalmological optical coherence tomography (OCT) and Electroretinogram (ERG) then, after treat 14 day sacrifice , using hematoxylin-eosin (H&E) histological staining.
Result: The pH8 gelatin nanoparticles were selected to carry Oxo and coated with HA. The size and surface charge of nanoparticles were 166.27 nm and -47.3 mV, the entrapment efficacy rate was 84.33%, and they had good shape and good distribution in aqueous solution. nanoparticles show a slow release effect, and the effect is better after coating with HA. In vitro , it is known from the cell activity that the safe concentration of Oxo to the cells is 0.1 μg/mL, and can be improved after damaged by blue light induced. the ROS assay, it is known that Oxo can effectively inhibit intracellular ROS, GNP-Oxo-HA has a better effect. In vivo, after the retina of mice was damaged by blue light induced, it was known from the observation of retinal thickness and outer nuclear layer (ONL) thickness that Oxo drugs can slow down the damage caused by blue light, and GNP-Oxo-HA has a better therapeutic effect.
Conclusion: Oxo can effectively treat or slow down the retinal damage caused by blue light. The use of gelatin nanoparticles is a carrier that can efficiently deliver drugs to the retina. In addition, combined with the properties of hyaluronic acid, it improves The transmission and treatment efficiency in the posterior eye, for such results, is beneficial to improve the clinical treatment of posterior ocular diseases. |
