CAJ | 학술논문

背景：骨髓间充质干细胞对骨和软骨在细胞水平的发育和再生至关重要，其数量缺失和功能受损被广泛认为是骨关节炎病因之一。目的：探讨骨关节炎患者骨髓间充质干细胞功能状态与病情进展的关系。方法：2013年7月至2014年10月间收集3组病例，每组10例，分别为对照组、轻度骨关节炎患者组、重度骨关节炎患者组。获取3组患者股骨或胫骨骨髓5 mL分离培养骨髓间充质干细胞，培养至3代时采用CCK-8法检测各组患者骨髓间充质干细胞的增殖能力，成软骨诱导14 d后行甲苯胺蓝染色鉴定，Real time PCR法检测对照组诱导后软骨细胞特征基因Aggrecan、Col2A1的表达。结果与结论：骨髓间充质干细胞体外培养后贴壁生长，胞浆含有较大的颗粒，核卵圆形，细胞呈多角形、梭形，有多个大小不一的突起，细胞分裂相多见。对照组增殖能力最强，重度骨关节炎患者组增殖能力最弱，但均在培养1周后进入平台期。成软骨诱导14 d后细胞呈多角形、类圆形改变，紫红色异染颗粒分布于胞浆外围。对照组诱导后软骨细胞特征基因 Aggrecan、Col2A1有高表达趋势。结果表明骨关节炎患者骨髓间充质干细胞功能状态与病情严重程度成负相关，其功能状态影响骨关节炎患者的病情进展。
배경：골수간충질간세포대골화연골재세포수평적발육화재생지관중요，기수량결실화공능수손피엄범인위시골관절염병인지일。목적：탐토골관절염환자골수간충질간세포공능상태여병정진전적관계。방법：2013년7월지2014년10월간수집3조병례，매조10례，분별위대조조、경도골관절염환자조、중도골관절염환자조。획취3조환자고골혹경골골수5 mL분리배양골수간충질간세포，배양지3대시채용CCK-8법검측각조환자골수간충질간세포적증식능력，성연골유도14 d후행갑분알람염색감정，Real time PCR법검측대조조유도후연골세포특정기인Aggrecan、Col2A1적표체。결과여결론：골수간충질간세포체외배양후첩벽생장，포장함유교대적과립，핵란원형，세포정다각형、사형，유다개대소불일적돌기，세포분렬상다견。대조조증식능력최강，중도골관절염환자조증식능력최약，단균재배양1주후진입평태기。성연골유도14 d후세포정다각형、류원형개변，자홍색이염과립분포우포장외위。대조조유도후연골세포특정기인 Aggrecan、Col2A1유고표체추세。결과표명골관절염환자골수간충질간세포공능상태여병정엄중정도성부상관，기공능상태영향골관절염환자적병정진전。
BACKGROUND:Bone marrow mesenchymal stem cels are crucial for bone and cartilage development and regeneration at a celular level. Insufficient quantity and functional impairment of bone marrow mesenchymal stem cels is widely considered to be one of osteoarthritis causes. OBJECTIVE: To explore the relationship between the functional status of bone marrow mesenchymal stem cels and disease progression in osteoarthritis patients.METHODS: Thirty patients with osteoarthritis were enroled from July 2013 to October 2014, and divided into control, mild osteoarthritis, and severe osteoarthritis groups, with 10 cases in each group. 5 mL bone marrow from the femur or tibia was extracted from each patient to isolate and culture bone marrow mesenchymal stem cels. Proliferation ability of cels at passage 3 was detected using cel counting kit-8; toluidine blue staining was performed at 14 days after chondrogenic induction; real-time PCR was used to detect the mRNA expression of Aggrecan and Col2A1 in the control group after chondrogenic induction. RESULTS AND CONCLUSION:Afterin vitro culture, bone marrow mesenchymal stem cels grew adherently in polygonal and fusiform shape with multiple processes at uniform size. The cytoplasm contained larger particles and the nuclei were ovoid. Most of cels were in cel division phase. The proliferation ability was strongest in the control group and weakest in the severe osteoarthritis group. Cels from the three groups were al at plateau phase after 1 week culture. At 14 days after chondrogenic induction, the cels were polygonal and quasi-circular, and purple metachromatic granules distributed outside of the cytoplasm. The expression of Aggrecan and Col2A1 in the control group displayed an overexpression trend. These findings indicate that the functional status of bone marrow mesenchymal stem cels from osteoarthritis patients is negatively correlated with the severity of disease, which can influence the disease progression in osteoarthritis patients.