淫羊藿苷含药血清促进3种细胞共培养体系中软骨细胞增殖和干细胞成软骨分化

被引:1
|
作者
刘琪 [1 ,2 ]
李林臻 [1 ,2 ]
李玉生 [1 ,2 ]
焦泓焯 [1 ,2 ]
杨程 [1 ,2 ]
张君涛 [1 ,2 ]
机构
[1] 天津中医药大学第一附属医院
[2] 国家中医针灸临床医学研究中心
关键词
淫羊藿苷含药血清; 软骨细胞; 骨髓间充质干细胞; 滑膜间充质干细胞; 共培养; 细胞增殖; Ⅱ型胶原; 成软骨分化;
D O I
暂无
中图分类号
R285 [中药药理学];
学科分类号
摘要
背景:关节软骨损伤修复能力十分有限,组织工程技术为修复受损软骨提供了新的治疗方案,其中软骨细胞、骨髓间充质干细胞和滑膜间充质干细胞之间的相互影响和诱导作用是自体软骨损伤愈合的基础。目的:构建软骨细胞-骨髓间充质干细胞-滑膜间充质干细胞共培养体系用以模拟软骨细胞体内微环境,并探究其最佳细胞接种比例,同时观察淫羊藿苷含药血清对该体系中软骨细胞增殖和干细胞成软骨分化的影响。方法:提取、培养、鉴定大鼠膝关节软骨细胞、骨髓间充质干细胞和滑膜间充质干细胞,按不同细胞接种比例构建软骨细胞-骨髓间充质干细胞-滑膜间充质干细胞非接触共培养体系,共培养72 h后观察软骨细胞增殖活性和表型能力,选择综合效应最佳的共培养体系;用淫羊藿苷溶液(0.25 mg/m L)灌胃新西兰大白兔制备淫羊藿苷含药血清,对照组共培养体系用含体积分数为10%胎牛血清、1%双抗的高糖DMEM培养液培养,实验组共培养体系在此基础之上加入体积分数为10%淫羊藿苷含药血清进行干预,24,48 h后检测两组软骨细胞增殖活性和Ⅱ型胶原表达,14 d后采用免疫荧光染色检测骨髓间充质干细胞、滑膜间充质干细胞成软骨分化情况。结果与结论:(1)3种细胞以不同比例共培养时均可正常贴壁生长,当软骨细胞、骨髓间充质干细胞、滑膜间充质干细胞接种比例为2∶1∶1时,共培养体系中软骨细胞表现出最佳增殖活性和表型能力;(2)与对照组相比,实验组培养24 h后软骨细胞增殖活性和Ⅱ型胶原表达显著升高(P<0.01),48 h后两组仍有差异(P<0.05),培养14 d后两组骨髓间充质干细胞和滑膜间充质干细胞出现明显的软骨分化,部分细胞呈现圆形或椭圆形,胞浆Ⅱ型胶原免疫荧光染色为阳性,实验组荧光强度明显高于对照组(P<0.01);(3)结果表明,以非接触共培养方法可以成功建立软骨细胞-骨髓间充质干细胞-滑膜间充质干细胞共培养体系且细胞比例为2∶1∶1时软骨细胞增殖活性和表型能力最佳;同时淫羊藿苷含药血清具有促进该体系中软骨细胞增殖及骨髓间充质干细胞、滑膜间充质干细胞成软骨分化的作用。
引用
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页码:1371 / 1379
页数:9
相关论文
共 44 条
  • [1] Rehabilitation of Soccer Players' Knee Injuries: Cartilage Reconstruction, Anterior Cruciate Ligament Surgery, and Intensive Recovery-A Pilot Study
    Kacprzak, Bartlomiej
    Rosinska, Karolina
    [J]. JOURNAL OF CLINICAL MEDICINE, 2023, 12 (21)
  • [2] Icariin-conditioned serum combined with chitosan attenuates cartilage injury in rabbit knees with osteochondral defect
    Zhang, Juntao
    Fan, Fangyang
    Zhang, Chao
    Liu, Aifeng
    Shang, Man
    Meng, Lin
    [J]. JOURNAL OF ORTHOPAEDIC SURGERY AND RESEARCH, 2023, 18 (01)
  • [3] Icariin: A Promising Natural Product in Biomedicine and Tissue Engineering
    Seyedi, Zahra
    Amiri, Mohammad Sadegh
    Mohammadzadeh, Vahideh
    Hashemzadeh, Alireza
    Haddad-Mashadrizeh, Aliakbar
    Mashreghi, Mohammad
    Qayoomian, Mohsen
    Hashemzadeh, Mohammad Reza
    Simal-Gandara, Jesus
    Taghavizadeh Yazdi, Mohammad Ehsan
    [J]. JOURNAL OF FUNCTIONAL BIOMATERIALS, 2023, 14 (01)
  • [4] Regeneration of articular cartilage defects: Therapeutic strategies and perspectives..[J].Guo Xueqiang;Xi Lingling;Yu Mengyuan;Fan Zhenlin;Wang Weiyun;Ju Andong;Liang Zhuo;Zhou Guangdong;Ren Wenjie.Journal of tissue engineering.2023,
  • [5] Cartilage Extracellular Matrix-derived Matrikines in Osteoarthritis..[J].Rapp Anna E;Zaucke Frank.American journal of physiology. Cell physiology.2022, 2
  • [6] Icariin-loaded hydrogel with concurrent chondrogenesis and anti-inflammatory properties for promoting cartilage regeneration in a large animal model.[J].Xu Songshan;Zhao Shaohua;Jian Yanpeng;Shao Xinwei;Han Dandan;Zhang Fan;Liang Chen;Liu Weijie;Fan Jun;Yang Zhikui;Zhou Jinge;Zhang Wenqiang;Wang Yigong.Frontiers in Cell and Developmental Biology.2022,
  • [7] The synovial microenvironment suppresses chondrocyte hypertrophy and promotes articular chondrocyte differentiation
    Chau, Michael
    Dou, Zelong
    Baroncelli, Marta
    Landman, Ellie B.
    Bendre, Ameya
    Kanekiyo, Masaru
    Gkourogianni, Alexandra
    Barnes, Kevin
    Ottosson, Lars
    Nilsson, Ola
    [J]. NPJ REGENERATIVE MEDICINE, 2022, 7 (01)
  • [8] Chondrocytes In Vitro Systems Allowing Study of OA.[J].Bednarczyk Ewa.International Journal of Molecular Sciences.2022, 18
  • [9] Traditional Chinese medicine promotes bone regeneration in bone tissue engineering
    Gao, Zheng-Rong
    Feng, Yun-Zhi
    Zhao, Ya-Qiong
    Zhao, Jie
    Zhou, Ying-Hui
    Ye, Qin
    Chen, Yun
    Tan, Li
    Zhang, Shao-Hui
    Feng, Yao
    Hu, Jing
    Ou-Yang, Ze-Yue
    Dusenge, Marie Aimee
    Guo, Yue
    [J]. CHINESE MEDICINE, 2022, 17 (01)
  • [10] Patchouli Alcohol Inhibits D-Gal Induced Oxidative Stress and Ameliorates the Quality of Aging Cartilage via Activating the Nrf2/HO-1 Pathway in Mice.[J].Chen Ming;Wen Haiyan;Zhou Siqi;Yan Xinxin;Li Haohuan.Oxidative Medicine and Cellular Longevity.2022,