以可注射性富含血小板纖維素釋放液混合骨髓幹細胞治療兔膝關節軟骨再生之研究

A novel method for rabbit stifle joint cartilage regeneration with injectable platelet-rich fibrin releasate and bone marrow stem cell

曾嘉川(Chia-Chuan Tseng)

指導教授 ： 郭宗甫

國立臺灣大學/獸醫專業學院/獸醫學研究所/碩士(2014年)

https://doi.org/10.6342/NTU.2014.10572

摘要

中文摘要
骨關節炎(osteoarthritis)，又稱退化性關節病(degenerative joint disease)，是一種非炎症性的退化性關節軟骨病變，造成細胞外基質嚴重流失發展成軟骨出現裂隙、侵蝕、軟骨下硬化甚至出現骨贅，最後導致關節功能喪失。由於成熟關節軟骨沒有血管分佈，缺乏提供軟骨再生及修補的能力，因此組織工程學應用於關節軟骨再生的研究日漸重要。
成熟間葉幹細胞可以於不同的成熟組織中發現並分離出來，其中包括:骨髓、周邊血液、臍帶血、脂肪、滑液囊膜、牙周韌帶、皮膚或其他固型間葉組織。其有能力分化成結締組織系的細胞，包括:硬骨、軟骨、脂肪和肌肉組織，在不同培養條件中被誘導分化成不同組織的細胞，因此使其臨床應用性大為增加。
此外，在醫學研究中發現血小板並非僅有凝血及止血的功能，對於組織修復和再生過程中，可幫助調節體內新陳代謝，促進組織的修復和癒合。隨著科技的發展，現在可以天然的取得血小板濃縮產物，富含血小板纖維素釋放液( Platelet-rich fibrin releasate)含有多種生長因子及細胞激素對於軟骨細胞的增生與分化很有幫助。
本研究利用可注射性富含血小板纖維素釋放液混合骨髓幹細胞達到關節軟骨組織再生，實驗分為四組，每一組各含紐西蘭白兔六隻，第一組為對照組;第二組為注射富含血小板纖維素釋放液1ml;第三組為注射富含血小板纖維素釋放液0.8ml混合骨髓幹細胞3×106cells
/0.2ml medium;第四組為注射骨髓幹細胞3×106cells/1ml medium。在經體內試驗十四周後，外觀復原程度依照新生軟骨覆蓋程度、新生軟骨的顏色、缺損邊緣及軟骨表面進行評分，實驗組與對照組間有顯著差異，以第三組外觀的復原程度最接近正常軟骨面，其次為第二組。組織學下復原程度依照軟骨表層、基質、細胞分佈、細胞族群的存活率及軟骨下硬骨與軟骨礦物質化的程度進行評分，實驗組與對照組間有顯著差異，以第三組的組織復原程度最接近正常軟骨組織，其次為第二組。綜合以上結果發現在關節軟骨經過組織工程手術後，無論是在外觀形態表現及組織學下細胞生長量以及細胞外基質都有顯著的再生結果。

關鍵字

可注射性富含血小板纖維素釋放液 ； 骨髓幹細胞 ； 組織工程 ； 關節軟骨再生

並列摘要

Abstracts
Osteoarthritis, is a degenerative joint disease accompanied with non-inflammatory articular cartilage lesion, losing of extracellular matrix and develop into cracks, erosion, subchondral sclerosis, osteophytes and loss of joint function eventually. Unlike other connective tissues, articular cartilage does not contain blood vessels so it does not have the ability to regenerate and repair; therefore, materials that promote articular cartilage regeneration are important in tissue engineering studies.
Mesenchymal stem cells (MSCs) can be found and isolated in different tissue including bone marrow, peripheral blood, umbilical cord blood, fat, synovial capsule, periodontal ligament, skin or solid mesenchymal tissues. The ability to transform into connective tissue system including hard bone, cartilage, fat and muscle tissue, which were cultured in several conditions that may induced to different tissues.
Furthermore, functions of the platelets in medical research are not only coagulation and hemostasis, in the process of tissue repair and regeneration, the platelets can help regulate body metabolism, and promote tissue repair and healing. With the development of technology, we can get the concentrated platelet, platelet-rich fibrin releasate , which contains a variety of growth factor that maybe promote proliferation and differentiation of chondrocytes.
In this study, an injectable platelet-rich fibrin releasate combine bone marrow stem cells was used for articular cartilage regeneration. New Zealand white rabbits were divided into four groups for treatment (n=6 per group): (1) control group; (2) injected platelet-rich fibrin releasate 1ml; (3) injected platelet-rich fibrin releasate 0.8 ml combine bone marrow stem cells 3×106 cells/0.2 ml medium; (4) injected bone marrow stem cells 3×106 cells/1 ml medium. After fourteen weeks, the bone appearance of healing was evaluated including coverage, color, cartilage surface defect edge and surface smooth. Histological investigation including cartilage surface, matrix, cell distribution, cell population viability, subchondral bone and cartilage mineralization. A statistically significant difference between the experimental and control groups was observed. The third group closest to the normal cartilage tissue structure, then followed by the second group. It was concluded that the promising experimental data are beginning to emerge in support of the use of platelet-rich fibrin relesate combine bone marrow stem cells for regenerative applications.

並列關鍵字

injectable platelet-rich fibrin releasate ； bone marrow stem cells ； tissue engineering ； articular cartilage regen

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