研究⽺膜萃取物懸浮點眼劑以及細胞外基質替代物對於⼤⿏實驗誘發眼⾓膜傷⼝的癒合效果

Investigation of topical amniotic membrane extract suspension and extracellular matrix substitute eyedrops on corneal wound healing in an induced rat corneal wounding model

李晧(Hao Lee)

指導教授 ： 林中天

國立臺灣大學/獸醫專業學院/臨床動物醫學研究所/碩士(2022年)

https://doi.org/10.6342/NTU202201658

摘要

角膜潰瘍是一個在獸醫臨床眼科之中非常常見的疾病，除了會造成病患不適之外，角膜潰瘍也可能會導致嚴重的併發症。嚴重的角膜基質潰瘍會需要手術的介入，來幫助受傷的角膜恢復結構的穩定性以及促進角膜的癒合過程。然而，如果病患有生理方面的異常無法進行麻醉手術，或是有經費方面的考量的情況，就會需要以積極的內科點眼藥水的方式治療。此研究的目標是希望可以調查在使用羊膜懸浮點眼劑以及細胞外基質替代物，在治療手術誘導的大鼠深層角膜潰瘍動物模式的治療效果。
有24隻老鼠參與實驗並且分成四組，分別為控制組（生理食鹽水一天三次，共六隻）、EyeQ組（羊膜萃取物懸浮液一天三次，共六隻）、EMS組（細胞外基質替代物每兩天一次，共六隻）以及EyeQ+EMS組（羊膜萃取物懸浮液一天三次；細胞外基質替代物每兩天一次，共六隻）。實驗使用角膜層切術在大鼠的角膜上製造出深層潰瘍。角膜基質的癒合過程會藉由臨床的檢查、光學同調斷層掃描、組織病理學以及免疫組織化學染色的方式來評估。
各組之間在手術後的24與48小時的時間內，角膜上皮的癒合百分比並沒有出現顯著的差異。並且角膜基質的厚度在術後的第三天與第七天，各組也沒有出現統計上的顯著差異。然而在術後第七天時，在各組之間的角膜混濁度的評估以及肌纖維母細胞的數量，會發現在EMS組以及EyeQ+EMS組別中與控制組相比有顯著的差異存在。
此研究的結果發現，羊膜萃取物懸浮液與細胞外基質替代物都可以有效的幫助角膜基質的癒合過程。在使用細胞外基質替代物的角膜中，可以觀察到較為顯著的角膜混濁以及較大量的肌纖維母細胞浸潤在角膜基質內。未來需要更多的後續研究來了解，這些眼藥水在臨床的環境下治療犬貓的深層角膜潰瘍時的治療效果。

關鍵字

角膜基質潰瘍 ； 光學同調斷層掃描 ； 肌纖維母細胞 ； 羊膜萃取物懸浮點眼劑 ； 細胞外基質替代物

並列摘要

Corneal ulcer is a common disease in veterinary ophthalmology, which not only causes the patient’s discomfort, but also lead to serious complications. Severe corneal stromal ulcer is often required surgical management to restore corneal structural stability and promote healing process. However, if patient is unable to undergo general anesthesia due to physiological factors or financial restriction, vigorous medical therapy is an alternative. The aim of our study is to investigate the therapeutic response of amniotic membrane suspension and extracellular matrix substitute in surgically induced rat deep stromal ulcer model.
Twenty-four rats were divided into 4 groups: control group (topical normal saline, tid, n=6); EyeQ group (topical amniotic membrane suspension, tid, n=6); EMS group (topical extracellular matrix substitute, q2d, n=6); EyeQ+EMS group (topical amniotic membrane suspension, tid; topical extracellular matrix substitute, q2d, n=6). Lamellar keratectomy was used to induce deep stromal ulcer in the corneas. The evaluation of stromal wound healing process was performed by clinical ophthalmic examinations, spectral domain optical coherence tomography (SD-OCT), histopathology, and immunohistochemistry of markers of corneal wound healing.
There was no significant difference in epithelial healing percentage in both 24 and 48 hours postoperatively between groups (P>0.05). There was no significant difference in stroma thickness between groups (P>0.05) at both day 3 and 7. The corneal opacity grading scores and myofibroblast counts in EMS and EyeQ+EMS groups were significantly higher than those in the control group (P<0.05) at day 7.
The results from our study showed both amniotic membrane suspension and extracellular matrix substitute could improve the corneal stromal healing process. Significant corneal opaque and myofibroblast infiltration were found in the eyes treated with extracellular matrix substitute, which may be related to its therapeutic mechanism. Further study is required to understand these topical treatments to canine and feline deep corneal stromal ulcer under clinical setting.

並列關鍵字

stromal ulcer ； OCT ； myofibroblast ； amniotic membrane suspension ； extracellular matrix substitute

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