關節內注射透明質酸鈉鹽結合骨髓間葉幹細胞治療兔膝關節軟骨再生的研究

鍾佩君

doi:10.6342/NTU.2015.00473

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關節內注射透明質酸鈉鹽結合骨髓間葉幹細胞治療兔膝關節軟骨再生的研究

Intra-articular injection Sodium Hyaluronate Combined with Bone Marrow Stem Cell for Articular Cartilage Regeneration of Stifle Joint in Rabbits Model.

鍾佩君 , Masters　　Advisor：郭宗甫　　Co-advisor ：鄭益謙

繁體中文 DOI： 10.6342/NTU.2015.00473

骨髓間葉幹細胞；透明質酸；組織工程；關節軟骨再生；骨關節炎； Hyaluronan ； Mesenchymal stem cells from bone marrow ； Tissue engineering ； Articular cartilage regeneration ； Osteoarthritis

Abstract │ Reference (111) │ Altmetrics

Abstract 〈TOP〉

膝蓋關節軟骨的受損一直被認為是導致骨關節炎(Osteoarthritis, OA)加速發病的一個重要的原因，因為軟骨沒有血管，所以限制了其內在的癒合和再生能力。因此，醫療上需要更新穎的修復療法來治療骨關節炎，並使軟骨再生(Cartilage regeneration)。在組織工程研究中，發展出促進關節軟骨再生的材料是重要的。
    透明質酸(Hyaluronan, HA)，又稱為玻尿酸，是維持軟骨細胞外基質物理化學特性的關鍵分子之一。為關節滑液的主要成份，並發現能增加關節滑液的粘彈性，也是關節軟骨的重要組成部份，它能夠在軟骨細胞的周圍行保護的功能。
    骨髓間葉幹細胞(Mesenchymal stem cells derived from bone marrow or bone marrow stem cells, BMSC)是多能性幹細胞，具有分化成多種細胞類型的能力，包括：成骨細胞、軟骨細胞，和脂肪細胞。幹細胞(stem cell)有兩種特性，一是自我更新能力，二是分化的能力。
    本實驗目的為關節內注射透明質酸鈉鹽和骨髓間葉幹細胞，已驗證是否能夠達到治療膝蓋骨關節炎和關節軟骨再生修復的療效。
    在這項研究中，使用紐西蘭大白兔作為實驗動物，將實驗分為四組，每一組各六隻，總共使用二十四隻動物。第一組為對照組;第二組為注射骨髓間葉幹細胞3×106 cells /0.5ml PBS;第三組為注射透明質酸鈉鹽0.1ml/kg；第四組為注射透明質酸鈉鹽0.1ml/kg混合骨髓間葉幹細胞3×106cells/0.2ml PBS。
    手術在無菌操作下進行，製造出右後肢股骨關節內髁軟骨缺損，軟骨缺損範圍為直徑3mm、深度2mm的圓柱形傷口，縫合關閉傷口。手術後一週傷口癒合拆線，同時於手術部位的關節囊內注射實驗材質，隔一週後再次注射實驗材質一次，並且於術後十四週犧牲實驗動物，取下膝關節內髁進行軟骨修復狀況的評估。
    研究的結果顯示出，在兔軟骨缺損模式中，移植透明質酸結合骨髓間葉幹細胞，配合組織工程(Tissue engineering)的原理，可以達到良好的組織修復再生能力，並且減緩手術誘導的骨關節炎的發展過程，以及可能應用在兔子OA的早期治療。
    在治療組和對照組之間能觀察出統計學上的顯著性差異，最接近的正常的軟骨組織結構的第四組(HA+BMSC)，隨後則是第三組(HA)。這些實驗結果也能提供未來進一步支持組合療法的使用，例如幹細胞療法在其他疾病的應用上，以及對於骨關節炎的預防和治療，可以提供實質上的參考。

Parallel Abstract 〈TOP〉

Articular cartilage does not contain blood vessels, so it does not have the ability to regenerate and repair. Therefore, in tissue engineering studies, materials that promote articular cartilage regeneration are very important. Sodium hyaluronate is one of the critical molecules for the maintenance of the physico-chemical characteristics of cartilage extracellular matrix. Mesenchymal stem cells are multipotent stem cells and can differentiate into a variety of cell types, involving: adipocytes, chondrocytes, and osteoblasts. We want to know the effect of intra-articular injection of hyaluronan plus bone marrow stem cells (BMSC) promote cartilage regeneration in the treatment of osteoarthritis. 
    In this study, an injectable sodium hyaluronate combine bone marrow stem cells (BMSC) was used for articular cartilage regeneration. Female 16-week-old New Zealand white rabbits were divided into four groups for treatment (n=6 per group): (1) control group - no treatment; (2) injected BMSC 3×106 cells/0.5 ml PBS; (3) injected sodium hyaluronate 0.1ml/kg; (4) injected sodium hyaluronate 0.1ml/kg combine BMSC 3×106 cells/0.2 ml PBS. 
    Each animal underwent surgery to create full thickness cartilage defects on the articular surface of the medial femoral condyle of right stifle joint. The defects were 3 mm diameter and 2 mm depth. At 1 and 2 weeks after surgical, experiment material was injected into the right articular capsule of each rabbit in the transplanted group. At 14 weeks after surgical, the regenerated cartilage was examined using the International Cartilage Repair Society (ICRS) Macroscopic Score and the ICRS Visual Histological Score.
    A statistically significant difference between the experimental and control groups was observed. The forth group closest to the normal cartilage tissue structure, then followed by the third group. It is a promising therapeutic approach to joint damage. The combination of BMSC and HA yield good results and are beginning to gain recognition in regenerative medicine.

Reference ( 111 ) 〈TOP〉

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連結：
Alford, J. W. (2005). "Cartilage Restoration, Part 1: Basic Science, Historical Perspective, Patient Evaluation, and Treatment Options." Am J Sports Med 33(2): 295-306.
連結：
Armentano, I., E. Fortunati, S. Mattioli, N. Rescignano and J. M. Kenny (2013). "Biodegradable composite scaffolds: a strategy to modulate stem cell behaviour." Recent Pat Drug Deliv Formul 7(1): 9-17.
連結：
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連結：
Brody, L. T. (2014). "Knee osteoarthritis: Clinical connections to articular cartilage structure and function." Phys Ther Sport. S1466-853X(14)00106-0.
連結：

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