Evaluation of Physical-Chemical Properties and Biocompatibility of in-situ Polymerizable Vitreous

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Project title

Code/計畫編號

NSC99-2314-B075-004-MY2

Translated Name/計畫中文名

玻尿酸/Pluronic F127可原位聚合人工玻璃體之物化特性與生物相容性之評估

Funding Organization/主管機關

National Science and Technology Council

Co-Investigator(s)/共同執行人

陳克華(計畫主持人)

Department/Unit

Department of Ophthalmology, Taipei Veterans General Hospital

Website

https://www.grb.gov.tw/search/planDetail?id=2126028

Year

2010

Start date/計畫起

01-08-2010

Expected Completion/計畫迄

31-07-2010

Co-Investigator(s)

Yung-Kai Lin

Bugetid/研究經費

900千元

ResearchField/研究領域

臨床醫學
醫學工程

第一年：化學修飾玻尿酸/pluronic F127水膠之物化特性與製備可原位聚合人工玻璃體之評估在白內障盛行之後，玻璃體-視網膜病變目前仍是目前導致失明的主要原因。自1960年以降，由於矽膠油具有高度安定性、透明度及高介面張力等特性，因此普遍被應用於玻璃體-視網膜手術過程中來填補視網膜的玻璃體取代物。然而，由於矽膠油的高度疏水特性導致其無法適當填補體液/視網膜間之介面，低比重特性導致在術後6個月內需手術取出並反覆注射以避免造成對生命威脅之併發症。目前部分玻璃體取代物由玻尿酸或膠原蛋白製備，這些天然高分子可提供良好的生物相容性，但在玻璃體腔內會迅速液化水解且填補效果不佳。有鑑於此，申請者提出一種智慧型原位聚合玻璃體取代物，其利用經化學改質之玻尿酸與Pluronic F-127所組成。經設計之人工玻璃體具有在不同溫度下會產生溶液-膠體轉換之特性，在眼外(室溫)下呈現液態並可以注射針筒吸取，待注入眼內(體溫)時會形成半固態的水膠。此種水膠相較於矽膠油有較高的親水性、安定性及生物安定性。本研究案將藉由分析不同配方之化學修飾玻尿酸/ pluronic F-127水膠之化學特性、黏彈性、光學特性及生物降解性並探索最佳化組成配方。第二年化學修飾玻尿酸/pluronic F127水膠製備可原位聚合人工玻璃體之生物相容性評估申請者初步資料顯示，高黏度矽油具有細胞毒性，且造成角膜內皮細胞 (human corneal endothelial cells, hCEs)凋亡現象，但矽油應用做為人工玻璃體是否亦會對視網膜色素上皮細胞 (human retinal pigment epithelial cells, hRPEs) 造成同樣效應有待探討。本計畫擬將hCEs 與hRPEs培養於嵌入式培養皿中，以模擬人類玻璃體/視網膜介面，再灌注不同配方組合之可原位聚合人工玻璃體及不同黏度矽油並進一步觀察存活率 (MTS bioassay)、細胞型態、Live/dead staining、凋亡分析 (apoptosis assay)。待分析完成後將選擇三組生物相容性較佳之可原位聚合人工玻璃體與矽油進行動物實驗。兔隻左眼注入不同人工玻璃體 (0.7-1.5 mL)並於術後一週、一個月、三個月與六個月觀察視網膜電圖(electroretinogram, ERG)、組織學評估、Slit Lamp與眼內壓 (Intraocular pressure, IOP)，藉助動物實驗結果，將可確認本計畫所開發之人工玻璃體與眼內組織之交互作用並作為臨床應用之參考。 First year Evaluation of physical-chemical properties of in-situ polymerizable vitreous substitute composed of modified hyaluronic acid and pluronic F127 Vitreouretinal pathologies still remain the significant leading cause of blindness, after cataract throughout the world. Silicone oil has been used in vitreoretinal surgery since 1960s for it suitable properties of stability, transparency and high interfacial surface energy of silicone oil at the tamponade/aqueous/retinal interface. However, the hydrophobic properties of silicone oil lead to a poor contact with the retinal and aqueous fluids, low density and long period (6 months) leads to life-threatening complication therefore necessitate removing. Although the hyaluronic acid and collagen exhibits good tissue biocompatibility, their solution has a short longevity in vitreous cavity and exhibits a poor tamponade effect. The proposer will be fabricates a smart in-situ polymerizable vitreous substituent composed of chemical modified HA/ pluronic F127. It is reveals a unique solution-gel transformation at different temperature that is liquid (room temperature) and injectable outside of eye, but which form a semi-solid hydrogel (physiological temperature) inside the eye. This hydrogel would be more hydrophilic, persistent and providing higher biocompatibility than silicone oil. The chemical, viscoelestic, optical properties and biodegradability will be conducted to investigating the optimal formulation for chemical modified HA/ pluronic F127 hydrogel. Second year Evaluation of biocompability of in-situ polymerizable vitreous substitute composed of modified hyaluronic acid and pluronic F127 Based on the preliminary data suggested that the silicone oil was induced cytotoxic and apoptotic effects for corneal endothelial cell (CEs). However, cytotoxic and apoptotic effects of silicone oil for retinal pigment epithelial cells (RPEs) and interactions between silicone oil/retinal tissue still remained to investigate. The in vitro test of silicone oil and vitreous substitutes were conducted using CEs and RPEs cultivation, the RPEs and CEs will be seed on micro-porous inserts to observing the morphological properties, viability (MTS bioassay), apoptosis assay and live/dead staining. Three lowest cytotoxicity formulations and silicone oil will be injected into left eye of New Zealand half-lop rabbits to subject in vivo experiment. The electroretinogram (ERG), histological evaluation, slit lamp and intraocular pressure (IOP) will be perform at postoperative 1 week, 1 month, 3 month and 6 month. Both in vitro and in vivo biocompatibility would be confirming the optimum formulation of vitreous substitute, interactions between vitreous substitute and retinal tissue and provide information for clinical applications.

Keyword(s)

人工玻璃體
玻尿酸
水膠
可注射性
Artificial vitreous substitute
Hyaluronic acid
hydrogel; injectable

DSpace CRIS

Evaluation of Physical-Chemical Properties and Biocompatibility of in-situ Polymerizable Vitreous

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