簡易檢索 / 詳目顯示

回結果列表
研究生： 胡庭斳
Hu, Ting-Chin
論文名稱： 機械刺激模擬腕隧道症候群發病機制及可行治療方法研究
Study of the mechanism of pathogenesis and feasible treatment in mechanical stimulation simulated carpal tunnel syndrome
指導教授： 楊岱樺
Yang, Tai-Hua
學位類別： 碩士
Master
系所名稱： 工學院 - 生物醫學工程學系
Department of BioMedical Engineering
論文出版年： 2021
畢業學年度： 109
語文別： 英文
論文頁數： 94
中文關鍵詞： 腕隧道症候群 、滑膜下結締組織 、屈指淺肌 、發炎 、上皮間質轉化路徑 、纖維化 、類固醇治療
外文關鍵詞： Carpal tunnel syndrome (CTS), Subsynovial connective tissue (SSCT), Flexor digitorum superficialis III (FDS III), Inflammation, Epithelial mesenchymal transition, Fibrosis, Steroid treatment
相關次數： 點閱：187  下載：0 
分享至:
查詢本校圖書館目錄 查詢臺灣博碩士論文知識加值系統 勘誤回報

    • 腕隧道症候群(Carpal tunnel syndrome, CTS)為常見之周邊神經病變，長期反覆手腕運動，致使腕部滑膜下結締組織(Subsynovial connective tissue, SSCT)增殖壓迫正中神經，出現麻木、感覺異常、疼痛等。臨床治療方式為末期手術及局部注射類固醇，減少發炎及壓迫。然而，最近研究顯示慢性患者未發現發炎因子且在超音波與組織學相關研究中發現腕部組織纖維化，藉此本實驗驗證細胞體外模型在不同時間與頻率下拉伸，及動物體內模型在不同時間點，是否給予類固醇治療下的變化。冀望能了解CTS之發炎反應調節及致使纖維化之機轉。
    本實驗建立體外及體內動物模型模擬CTS。體外模型，大鼠纖維母細胞NIH3T3於不同時間及頻率重複拉伸，藉即時聚合酶連鎖反應、西方墨點法及免疫螢光染色測定發炎狀態，予以類固醇對比給藥前後差異；再者確認纖維化，測定相關蛋白表現，探討纖維化與發炎之相關性。體內模型，於大鼠腕部植入微量吸管，模擬該疾病腕部SSCT壓迫正中神經於4、8、12周檢測給藥有無腕管內部壓力，並進行組織染色及生物力學檢測比較不同時間下之差異。
    體外模型中顯示，早期階段發炎及纖維化相關因子表現增加，TGF-β、IL1β、TNF-α、CXCL12、CXCL4、MMP9及TIMP1，尤以2.0Hz第三天拉伸下更具顯著性意義，且類固醇治療發炎因子表現有下調趨勢。動物模型中，早期階段TGF-β、CXCL12上升，力學測試中也顯示力學性質變異；類固醇治療則可抑制發炎表現。該結果分析於One way-ANOVA以及SPSS軟體中發炎及纖維化機制與類固醇治療具有顯著性差異。
    藉此，本研究實驗證實CTS實有發炎反應參與其中，且與後續纖維化形成有關。此外該發炎反應可藉由給予類固醇有效達到緩解，與實驗結果一致，或許可以做為未來CTS患者早期發現治療之依據，進而減緩中、後期不可逆之腕部傷害。

    Carpal tunnel syndrome (CTS) a common peripheral neuropathy, which subsynovial connective tissue (SSCT) proliferating and compressing median nerve inducing numbness, paresthesia, and pain. Clinical treatments are surgery and steroids injection to reduce the inflammation. However, the studies indicated CTS was a non-inflammatory fibrotic disease. To investigate whether carpal tunnel syndrome regulates the inflammation and fibrosis, verifying in animal and cell model at different time points and frequency, and common steroid treated. In vitro, qPCR, Western blot and immunofluorescence to confirm inflammatory status and fibrotic genes, and steroids applied to confirm if fibrosis related to inflammation. In vivo models, we implanted tips into wrist, the difference was detected at different time points at 4, 8, and 12 weeks, including pressure, tissue and tendons. In vitro, inflammatory and fibrotic factors increased and downregulated after drug treatment, including TGF-β, IL1β, TNF-α, CXCL12, CXCL4, MMP9 and TIMP1, especially under the 2.0Hz stretching on the day 3. The animal model, TGF-β and CXCL12 both increased in early stage and decreased in steroid treatment. The results are analyzed in One way-ANOVA and SPSS software. Our experiment confirmed CTS participates inflammation involved fibrosis. The inflammatory response can alleviate by steroids, was consistent with our results. It may be used as a treatment basis for early detection and early treatment of CTS patients in the future, thereby reducing irreversible wrist injuries in the middle and late stages.

    摘要 I Abstract II 誌謝 III Content IV List of tables VII List of figures IX Abbreviation XII Chapter 1 Introduction 1 1.1 Anatomy of Carpal tunnel syndrome 1 1.2 The population of Carpal tunnel syndrome 2 1.3 The pathophysiology and etiology of Carpal tunnel syndrome 3 1.4 Clinical diagnosis and treatment 3 1.5 Healing process in Carpal tunnel syndrome 5 1.5.1 Early stage 5 1.5.2 Cellular condition 6 1.5.2.1 Macrophages 6 1.5.2.2 Fibroblasts 6 1.6 The inflammatory markers in Carpal tunnel syndrome 6 1.6.1 TGF-Beta 6 1.6.2 TNF-alpha and IL1-Beta 7 1.6.3 TIMP1 and MMP9 8 1.6.4 CXCR4 and CXCL12 8 1.7 The markers of fibrosis in Carpal tunnel syndrome via EMT pathway 9 1.7.1 ERK and Phos-ERK 10 1.7.2 E-cadherin and N-cadherin 10 1.8 Research model 11 1.8.1 In vitro 11 1.8.2 In-vivo 12 1.9 Motivation and Purpose 12 Chapter 2 Materials and Methods 14 2.1 Materials 14 2.1.1 Cell line 14 2.1.2 SD rat 14 2.2 Methods 15 2.2.1 In-vitro cell line experimental preparation 15 2.2.2 The collection and purification of conditioned medium from the cultured the stretching cell 16 2.2.3 Cell proliferation and Cytotoxicity assay 17 2.2.4 The mRNA level of stretching cell detection 18 2.2.5The protein level of stretching cell detection 21 2.2.6 Immunofluorescence (IF) 24 2.2.7 Steroids medicine treatment on cell model 25 2.2.8 In-vivo 26 2.2.9 Data analysis 32 2.2.10 Statistics 32 Chapter 3 Results 33 3.1 Established the in-vitro model in NIH-3T3 33 3.1.1 The survival rate NIH-3T3 33 3.1.2 The inflammatory dependent and frequency dependent measured by qPCR 34 3.1.3 The condition of time and frequency dependent tested by western blotting 41 3.1.4 The cytotoxicity of NIH-3T3 54 3.1.5 The effect of dexamethasone in-vitro 55 3.1.6 The morphology of stretched NIH-3T3 under the repetitive force with differ Frequencies 64 3.2 The animal model with operated 68 3.2.1 The efficacy of dexamethasone treated with animal model and the tip was inserted into the trans carpal tunnel 68 3.2.2 The mechanical characters of FDS III tendon 71 3.2.3 The reaction occurred in SSCT measuring by H＆E and IHC staining 74 Chapter 4 Discussion and limitation 81 Chapter 5 Conclusion and future work 87 Chapter 6 References 89

    1. Li, C., et al., Mutations in COMP cause familial carpal tunnel syndrome. Nature Communications, 2020. 11(1): p. 3642.
    2. Tse, C.T.F. and P.J. Keir, External Compression and Partial Ischemia Decrease Human Finger Flexor Tendon and Subsynovial Connective Tissue Relative Motion. J Orthop Res, 2020. 38(5): p. 1038-1044.
    3. Yoshii, Y., et al., The effect of wrist position on the relative motion of tendon, nerve, and subsynovial connective tissue within the carpal tunnel in a human cadaver model. J Orthop Res, 2008. 26(8): p. 1153-8.
    4. Soubeyrand, M., et al., Anatomy of the median nerve and its clinical applications. Hand Surg Rehabil, 2020. 39(1): p. 2-18.
    5. Robbins, H., ANATOMICAL STUDY OF THE MEDIAN NERVE IN THE CARPAL TUNNEL AND ETIOLOGIES OF THE CARPAL-TUNNEL SYNDROME. J Bone Joint Surg Am, 1963. 45: p. 953-66.
    6. Katz, J.N. and B.P. Simmons, Carpal Tunnel Syndrome. New England Journal of Medicine, 2002. 346(23): p. 1807-1812.
    7. Atroshi, I., et al., Prevalence of Carpal Tunnel Syndrome in a General Population. JAMA, 1999. 282(2): p. 153-158.
    8. Lee, S.K., et al., The Influence of Transverse Carpal Ligament Thickness on Treatment Decisions for Idiopathic Mild to Moderate Carpal Tunnel Syndrome. Annals of Plastic Surgery, 2020. 85(2): p. 127-134.
    9. Aboonq, M.S., Pathophysiology of carpal tunnel syndrome. Neurosciences (Riyadh), 2015. 20(1): p. 4-9.
    10. Almasi-Doghaee, M., et al., Carpal compression, Phalen's and Tinel's test: Which one is more suitable for carpal tunnel syndrome? Iran J Neurol, 2016. 15(3): p. 173-4.
    11. Jayaraman, S. and T.P. Naidich, The carpal tunnel: ultrasound display of normal imaging anatomy and pathology. Neuroimaging Clin N Am, 2004. 14(1): p. 103-13, viii.
    12. Lozano-Calderón, S., S. Anthony, and D. Ring, The quality and strength of evidence for etiology: example of carpal tunnel syndrome. J Hand Surg Am, 2008. 33(4): p. 525-38.
    13. Tat, J., K.E. Wilson, and P.J. Keir, Pathological changes in the subsynovial connective tissue increase with self-reported carpal tunnel syndrome symptoms. Clin Biomech (Bristol, Avon), 2015. 30(4): p. 360-5.
    14. Chtompel, Y., Atlas of Common Pain Syndromes, Fourth Edition. Canadian Journal of Anesthesia/Journal canadien d'anesthésie, 2018. 65(11): p. 1277-1278.
    15. Raihan, H., et al., Orthotic Treatment Overview of Carpal Tunnel Syndrome. 2020.
    16. Zhang, D., et al., Accuracy of Provocative Tests for Carpal Tunnel Syndrome. Journal of Hand Surgery Global Online, 2020. 2(3): p. 121-125.
    17. GOLDING, D.N., D.M. ROSE, and K. SELVARAJAH, CLINICAL TESTS FOR CARPAL TUNNEL SYNDROME: AN EVALUATION. Rheumatology, 1986. 25(4): p. 388-390.
    18. O'Connor, D., S. Marshall, and N. Massy-Westropp, Non-surgical treatment (other than steroid injection) for carpal tunnel syndrome. Cochrane Database Syst Rev, 2003. 2003(1): p. Cd003219.
    19. Yang, T.-H., et al., Triamcinolone Acetonide affects TGF-β signaling regulation of fibrosis in idiopathic carpal tunnel syndrome. BMC Musculoskeletal Disorders, 2018. 19(1): p. 342.
    20. Kluemper, C.T., et al., Flexor Tenosynovectomy for Recurrent Carpal Tunnel Syndrome: A Retrospective Case Series of 108 Hands. HAND, 2021. 16(1): p. 18-24.
    21. Koh, T.J. and L.A. DiPietro, Inflammation and wound healing: the role of the macrophage. Expert Rev Mol Med, 2011. 13: p. e23.
    22. Wang, J.C., et al., Efficacy of Combined Ultrasound-Guided Steroid Injection and Splinting in Patients With Carpal Tunnel Syndrome: A Randomized Controlled Trial. Arch Phys Med Rehabil, 2017. 98(5): p. 947-956.
    23. Gressner, A.M., et al., Roles of TGF-beta in hepatic fibrosis. Front Biosci, 2002. 7: p. d793-807.
    24. Chikenji, T., et al., Transforming growth factor-β (TGF-β) expression is increased in the subsynovial connective tissues of patients with idiopathic carpal tunnel syndrome. J Orthop Res, 2014. 32(1): p. 116-22.
    25. Freeland, A.E., et al., Biochemical evaluation of serum and flexor tenosynovium in carpal tunnel syndrome. Microsurgery, 2002. 22(8): p. 378-85.
    26. Kulshrestha, R., et al., Beneficial effects of N-acetylcysteine on protease-antiprotease balance in attenuating bleomycin-induced pulmonary fibrosis in rats. Iran J Basic Med Sci, 2020. 23(3): p. 396-405.
    27. Chou, D.H., W. Lee, and C.A. McCulloch, TNF-alpha inactivation of collagen receptors: implications for fibroblast function and fibrosis. J Immunol, 1996. 156(11): p. 4354-62.
    28. Gu, S., et al., TNF Alpha Attenuates Bleomycin-Induced Pulmonary Fibrosis Via TNFR1 Signaling and Neutrophil Recruitment, in A62. LUNG FIBROSIS: INSIGHTS FROM THE BENCH. 2020, American Thoracic Society. p. A2242-A2242.
    29. Roman-Blas, J.A., D.G. Stokes, and S.A. Jimenez, Modulation of TGF-beta signaling by proinflammatory cytokines in articular chondrocytes. Osteoarthritis Cartilage, 2007. 15(12): p. 1367-77.
    30. Masola, V., et al., In vitro effects of interleukin (IL)-1 beta inhibition on the epithelial-to-mesenchymal transition (EMT) of renal tubular and hepatic stellate cells. J Transl Med, 2019. 17(1): p. 12.
    31. Kolb, M., et al., Transient expression of IL-1beta induces acute lung injury and chronic repair leading to pulmonary fibrosis. J Clin Invest, 2001. 107(12): p. 1529-36.
    32. Lachowski, D., et al., Matrix stiffness modulates the activity of MMP-9 and TIMP-1 in hepatic stellate cells to perpetuate fibrosis. Sci Rep, 2019. 9(1): p. 7299.
    33. Dong, J. and Q. Ma, TIMP1 promotes multi-walled carbon nanotube-induced lung fibrosis by stimulating fibroblast activation and proliferation. Nanotoxicology, 2017. 11(1): p. 41-51.
    34. Medeiros, T., et al., Liver fibrosis improvement in chronic hepatitis C after direct acting-antivirals is accompanied by reduced profibrogenic biomarkers-a role for MMP-9/TIMP-1. Dig Liver Dis, 2020. 52(10): p. 1170-1177.
    35. Seeland, U., et al., Myocardial fibrosis in transforming growth factor-beta(1) (TGF-beta(1)) transgenic mice is associated with inhibition of interstitial collagenase. Eur J Clin Invest, 2002. 32(5): p. 295-303.
    36. Zhao, F.Y., et al., G-CSF Inhibits Pulmonary Fibrosis by Promoting BMSC Homing to the Lungs via SDF-1/CXCR4 Chemotaxis. Sci Rep, 2020. 10(1): p. 10515.
    37. Derlin, T., et al., Clinical Molecular Imaging of Pulmonary CXCR4 Expression to Predict Outcome of Pirfenidone Treatment in Idiopathic Pulmonary Fibrosis. Chest, 2021. 159(3): p. 1094-1106.
    38. Zheng, G., et al., Disruption of E-cadherin by matrix metalloproteinase directly mediates epithelial-mesenchymal transition downstream of transforming growth factor-beta1 in renal tubular epithelial cells. Am J Pathol, 2009. 175(2): p. 580-91.
    39. López-Novoa, J.M. and M.A. Nieto, Inflammation and EMT: an alliance towards organ fibrosis and cancer progression. EMBO molecular medicine, 2009. 1(6-7): p. 303-314.
    40. Foglia, B., et al., ERK Pathway in Activated, Myofibroblast-Like, Hepatic Stellate Cells: A Critical Signaling Crossroad Sustaining Liver Fibrosis. Int J Mol Sci, 2019. 20(11).
    41. Jeng, K.S., et al., Liver Fibrosis and Inflammation under the Control of ERK2. Int J Mol Sci, 2020. 21(11).
    42. Rout-Pitt, N., et al., Epithelial mesenchymal transition (EMT): a universal process in lung diseases with implications for cystic fibrosis pathophysiology. Respir Res, 2018. 19(1): p. 136.
    43. Loh, C.Y., et al., The E-Cadherin and N-Cadherin Switch in Epithelial-to-Mesenchymal Transition: Signaling, Therapeutic Implications, and Challenges. Cells, 2019. 8(10).
    44. Piazzini, D.B., et al., A systematic review of conservative treatment of carpal tunnel syndrome. Clinical Rehabilitation, 2007. 21(4): p. 299-314.
    45. Yamanaka, Y., et al., Blocking fibrotic signaling in fibroblasts from patients with carpal tunnel syndrome. J Cell Physiol, 2018. 233(3): p. 2067-2074.
    46. Hsieh, J.-L., et al., Estrogen and mechanical loading-related regulation of estrogen receptor-β and apoptosis in tendinopathy. PLOS ONE, 2018. 13(10): p. e0204603.
    47. Toyoshima, Y., et al., A non–invasive technique for evaluating carpal tunnel pressure with ultrasound vibro–elastography for patients with carpal tunnel syndrome: A pilot clinical study. Journal of Biomechanics, 2021. 116: p. 110228.
    48. Kubo, K., et al., The quantitative evaluation of the relationship between the forces applied to the palm and carpal tunnel pressure. Journal of Biomechanics, 2018. 66: p. 170-174.
    49. Festen-Schrier, V.J.M.M. and P.C. Amadio, The biomechanics of subsynovial connective tissue in health and its role in carpal tunnel syndrome. Journal of Electromyography and Kinesiology, 2018. 38: p. 232-239.
    50. Park, J.-J., J.-G. Choi, and B.-C. Son, Carpal Tunnel Syndrome Caused by Bifid Median Nerve in Association with Anomalous Course of the Flexor Digitorum Superficialis Muscle at the Wrist. Nerve, 2017. 3(1): p. 21-23.
    51. Tat, J., K.E. Wilson, and P.J. Keir, Pathological changes in the subsynovial connective tissue increase with self-reported carpal tunnel syndrome symptoms. Clinical Biomechanics, 2015. 30(4): p. 360-365.
    52. Chu, C.Y., et al., Induction of chemokine receptor CXCR4 expression by transforming growth factor-β1 in human basal cell carcinoma cells. J Dermatol Sci, 2013. 72(2): p. 123-33.
    53. Antoniou, K.M., et al., Expression analysis of angiogenic growth factors and biological axis CXCL12/CXCR4 axis in idiopathic pulmonary fibrosis. Connect Tissue Res, 2010. 51(1): p. 71-80.
    54. Moghtaderi, A.R., N. Moghtaderi, and A. Loghmani, Evaluating the effectiveness of local dexamethasone injection in pregnant women with carpal tunnel syndrome. J Res Med Sci, 2011. 16(5): p. 687-90.
    55. Eroğlu, A., et al., Recurrent carpal tunnel syndrome: Evaluation and treatment of the possible causes. World journal of clinical cases, 2018. 6(10): p. 365-372.
    56. Karimi, N., S. Abedian, and F. Darvari, Serum levels of inflammatory cytokines in patients with idiopathic carpal tunnel syndrome. International Journal of Neuroscience, 2020. 131: p. 1-8.
    57. Farias Zuniga, A., et al., Ultrasound examination predicts 6-month progression in carpal tunnel syndrome patients. J Orthop Res, 2021. 39(3): p. 609-618.
    58. Farias Zuniga, A., et al., Blood flow velocity but not tendon mechanics relates to nerve function in carpal tunnel syndrome patients. J Neurol Sci, 2020. 411: p. 116694.
    59. Güneş, M. and H. Büyükgöl, Correlation of neutrophil/lymphocyte and platelet/lymphocyte ratios with the severity of idiopathic carpal tunnel syndrome. Muscle Nerve, 2020. 61(3): p. 369-374.
    60. Sharma, D., A.S. Jaggi, and A. Bali, Clinical evidence and mechanisms of growth factors in idiopathic and diabetes-induced carpal tunnel syndrome. European Journal of Pharmacology, 2018. 837: p. 156-163.
    61. Kang, Y.-M., et al., Relaxin Modulates the Expression of MMPs and TIMPs in Fibroblasts of Patients with Carpal Tunnel Syndrome. ymj, 2017. 58(2): p. 415-422.
    62. Isles, H.M., et al., The CXCL12/CXCR4 Signaling Axis Retains Neutrophils at Inflammatory Sites in Zebrafish. Frontiers in Immunology, 2019. 10(1784).
    63. Borgschulze, A., et al., New insights into the Tissue Homeostasis between TIMP and MMP in Dupuytren’s Disease. Cell Biol (Henderson, NV) 6, 2017. 2: p. 2.
    64. Takasu, S., et al., Serum hyaluronic acid and interleukin-6 as possible markers of carpal tunnel syndrome in chronic hemodialysis patients. Artif Organs, 1994. 18(6): p. 420-4.
    65. Suarez‐Carmona, M., et al., EMT and inflammation: inseparable actors of cancer progression. Molecular oncology, 2017. 11(7): p. 805-823.
    66. Xie, L., et al., Activation of the Erk Pathway Is Required for TGF-β1-Induced EMT In Vitro. Neoplasia, 2004. 6(5): p. 603-610.
    67. Song, Y., et al., Adipose-derived stem cells ameliorate renal interstitial fibrosis through inhibition of EMT and inflammatory response via TGF-β1 signaling pathway. International Immunopharmacology, 2017. 44: p. 115-122.
    68. Borthwick, L., et al., Inflammation and epithelial to mesenchymal transition in lung transplant recipients: role in dysregulated epithelial wound repair. American Journal of Transplantation, 2010. 10(3): p. 498-509.
    69. Schrier, V., A. Vrieze, and P.C. Amadio, Subsynovial connective tissue development in the rabbit carpal tunnel. Vet Med Sci, 2020. 6(4): p. 1025-1033.
    70. Yoshii, Y., W.-L. Tung, and T. Ishii, Measurement of Median Nerve Strain and Applied Pressure for the Diagnosis of Carpal Tunnel Syndrome. Ultrasound in Medicine & Biology, 2017. 43(6): p. 1205-1209.
    71. Seradge, H., Y.C. Jia, and W. Owens, In vivo measurement of carpal tunnel pressure in the functioning hand. J Hand Surg Am, 1995. 20(5): p. 855-9.

    下載圖示
    2026-10-15公開
    QR CODE