旋轉袖肌腱病變：超音波影像與分子表現的關聯

Rotator Cuff Tendinopathy: Association between Ultrasound Images and Molecular Expression

邱宜姮(Yi-Heng Chiu)

指導教授 ： 趙遠宏

國立臺灣大學/醫學院/物理治療學研究所/碩士(2021年)

https://doi.org/10.6342/NTU202103177

摘要

背景與目的：肩袖肌腱病變 (rotator cuff tendinopathy) 是常見的肩部疾病。先前的研究指出旋轉袖肌腱病變的組織與分子變化包括肌腱組織內細胞數量增多、細胞型態變圓、胞外基質排列紊亂，以及肌腱基質、基質酵素、肌腱特定的轉錄因子和細胞因子基因表現增加。超音波影像檢查是一項可以用來檢測肌腱病理變化的非侵入性工具，先前的研究使用空間頻率分析 (spatial frequency analysis) 量化斑點樣型 (speckle pattern)，可反映肌腱內膠原纖維束排列的品質。空間頻率峰值半徑 (peak spatial frequency radius) 定義為二維空間頻譜中的原點與空間頻率峰值之間的距離，可以表示肌腱中膠原纖維束的排列一致性，文獻指出空間頻率峰值半徑用以檢測肌腱病變是具有高預測率的參數之一，然而，空間頻率峰值半徑仍缺乏從組織和分子層面證據證實肌腱超音波影像呈現的病理變化。本研究旨在探討棘上肌肌腱超音波影像分析與旋轉袖肌腱病變患者基因表現之間的相關性，並且透過組織切片驗證超音波影像分析代表的肌腱特徵。
實驗設計：本篇研究為觀察型、橫斷相關性研究。
材料與方法：本研究招募旋轉袖肌腱撕裂的患者為實驗組，健康成年人為對照組，二組受試者皆進行棘上肌腱超音波影像檢查，並使用空間頻率峰值半徑量化肌腱品質。實驗組接受旋轉袖肌腱修補手術，術中收集撕裂肌腱殘端進一步分析。檢體使用蘇木精-伊紅染色與馬森三色染色觀察肌腱組織型態，使用 CurveAlign 對馬森三色染色組織影像定量膠原纖維的排列一致程度。並分離檢體細胞體外培養，進行基因和蛋白質分析。
結果：實驗組棘上肌腱超音波影像的空間頻率峰值半徑顯著低於年輕健康成人 (實驗組：1.093 ± 0.319 mm-1; 對照組：1.590 ± 0.525 mm-1 ; p = 0.006)。實驗組Scx 基因與空間頻率峰值半徑呈顯著中度負相關 (rs = -0.621, p = 0.01)。組織染色結果發現相較於高空間頻率峰值半徑的實驗組受試者，具有較低空間頻率峰值半徑的受試者之組織染色呈現較差的膠原纖維排列和肌腱結構，有顯著較低的排列係數 (p = 0.026)，Scx 基因的表現顯著較高 (p = 0.026)，Scleraixs 蛋白質的分泌較多。
結論：空間頻率峰值半徑是量化棘上肌肌腱超音波影像斑點樣型合適的參數，空間頻率峰值半徑越低，肌腱中膠原纖維束的排列越差。此外，退化較嚴重的棘上肌肌腱的品質較差，肌腱內膠原纖維排列紊亂，可能是受到 Scx 的調節所致。
臨床意義：探究空間頻率峰值半徑與肌腱分子變化的相關性，可以協助臨床人員解釋肌腱超音波影像的微觀特徵與及早偵測肌腱病變。

關鍵字

旋轉袖肌腱病變 ； 超音波影像 ； 空間頻譜分析 ； 膠原纖維排列 ； 分子生物

並列摘要

Background and purpose: Rotator cuff tendinopathy is a common shoulder disorder. Previous studies indicated that changes include increased cellularity, rounded cell, disorganized extracellular matrix, and increased tendon matrix, matrix enzyme, specific transcription factor, and cytokine gene expression. Ultrasonography is a non-invasive tool to detect the pathology in the tendon. Previous studies introduce the spatial frequency analysis to quantify the speckle pattern, which indicates the quality of the fascicle alignment in tendons. Peak spatial frequency radius (PSFR), defined as the distance between the origin and the peak spatial frequency in 2-D spatial frequency spectrum, can represent the alignment consistency of the collagen bundle in the tendon. PSFR was reported as one of the highest predictive parameters in detecting tendinopathy. However, PSFR still lacks the evidence in tissue and cellular level to confirm the pathological change in the presentation of ultrasonography of a tendon. This study aims to determine the correlation between ultrasonography analysis of the supraspinatus tendon and gene expression in patients with rotator cuff tendinopathy. We would verify the feature of ultrasonography with tissue section in addition.
Study design: This study is an observational and cross-sectional correlation study.
Material and method: We recruited healthy adults and patients with rotator cuff tear into our study. We took the ultrasonography of the supraspinatus tendon in both healthy adults and patients, using PSFR to quantify the quality of the tendon. After patients received rotator cuff repair surgery, stumps of the torn tendon were collected for further analysis. We applied H E and Masson trichrome stain on the specimen. The histological images were quantified using CurveAlign to identify the collagen fiber alignment. We executed gene and protein analysis on the cells derived from the specimen and cultured them in vitro.
Result: PSFR of the supraspinatus tendon ultrasonography in patients was significantly lower than the value in healthy young adults (patients: 1.093 ± 0.319 mm-1; healthy adults: 1.590 ± 0.525 mm-1; p = 0.006). Scx gene showed a significantly moderate negative correlation with PSFR in patients with rotator cuff tear (rs = -0.621, p = 0.01). We also found that patients with lower PSFR showed poor alignment and structure in the histological stain, significantly lower alignment coefficient (p = 0.026), significantly higher expression of Scx gene (p = 0.026), and higher secretion of the Scleraixs protein.
Conclusion: PSFR is a suitable parameter to quantify the speckle pattern of the ultrasonography in the supraspinatus tendon. The lower PSFR, the less organized the collagen bundles in a tendon. Moreover, severer degenerative supraspinatus tendons show poor tendon quality and misaligned collagen fibers, which may result from the regulation of Scx expression.
Clinical relevance: The correlation of PSFR and molecular changes of a tendon can assist clinicians in interpreting the characteristics and early detect tendinopathy by using ultrasonography.

並列關鍵字

Rotator cuff tendinopathy ； Ultrasonography ； Spatial frequency analysis ； Collagen alignment ； Molecular biology

參考文獻

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