Kuroda Kazunari, Kabata Tamon, Hayashi Katsuhiro, Maeda Toru, Kajino Yoshitomo, Iwai Shintaro, Fujita Kenji, Hasegawa Kazuhiro, Inoue Daisuke, Sugimoto Naotoshi, Tsuchiya Hiroyuki
Department of Orthopaedic Surgery, Graduate School of Medical Science, Kanazawa University, 13-1, Takara-machi, Kanazawa, 920-8641, Japan.
Department of Physiology, Graduate School of Medical Science, Kanazawa University, Kanazawa, Japan.
BMC Musculoskelet Disord. 2015 Sep 3;16:236. doi: 10.1186/s12891-015-0701-4.
This study aimed to determine whether intra-articularly injected adipose-derived stem cells (ADSCs) inhibited articular cartilage degeneration during osteoarthritis (OA) development in a rabbit anterior cruciate ligament transection (ACLT) model. The paracrine effects of ADSCs on chondrocytes were investigated using a co-culture system.
ACLT was performed on both knee joints of 12 rabbits. ADSCs were isolated from the subcutaneous adipose tissue. ADSCs with hyaluronic acid were intra-articularly injected into the left knee, and hyaluronic acid was injected into the right knee. The knees were compared macroscopically, histologically, and immunohistochemically at 8 and 12 weeks. In addition, cell viability was determined using co-culture system of ADSCs and chondrocytes.
Macroscopically, osteoarthritis progression was milder in the ADSC-treated knees than in the control knees 8 weeks after ACLT. Histologically, control knees showed obvious erosions in both the medial and lateral condyles at 8 weeks, while cartilage was predominantly retained in the ADSC-treated knees. At 12 weeks, the ADSC-treated knees showed a slight suppression of cartilage degeneration, unlike the control knees. Immunohistochemically, MMP-13 expression was less in the ADSC-treated cartilage than in the control knees. The cell viability of chondrocytes co-cultured with ADSCs was higher than that of chondrocytes cultured alone. TNF-alpha-induced apoptotic stimulation was similar between the two groups.
Intra-articularly injected ADSCs inhibited cartilage degeneration progression by homing to the synovium and secreting a liquid factor having chondro-protective effects such as chondrocyte proliferation and cartilage matrix protection.
本研究旨在确定在兔前交叉韧带切断(ACLT)模型中,关节内注射脂肪来源干细胞(ADSCs)是否能在骨关节炎(OA)发展过程中抑制关节软骨退变。使用共培养系统研究了ADSCs对软骨细胞的旁分泌作用。
对12只兔的双侧膝关节进行ACLT手术。从皮下脂肪组织中分离ADSCs。将含透明质酸的ADSCs关节内注射到左膝,右膝注射透明质酸。在第8周和第12周对膝关节进行宏观、组织学和免疫组织化学比较。此外,使用ADSCs与软骨细胞的共培养系统测定细胞活力。
宏观上,ACLT术后8周,ADSC治疗组膝关节的骨关节炎进展比对照组膝关节更轻。组织学上,对照组膝关节在8周时内外侧髁均出现明显侵蚀,而ADSC治疗组膝关节软骨基本得以保留。在12周时,与对照组膝关节不同,ADSC治疗组膝关节的软骨退变有轻微抑制。免疫组织化学显示,ADSC治疗组软骨中MMP - 13表达低于对照组膝关节。与ADSCs共培养的软骨细胞的细胞活力高于单独培养的软骨细胞。两组之间TNF - α诱导的凋亡刺激相似。
关节内注射的ADSCs通过归巢至滑膜并分泌具有软骨保护作用的液体因子(如软骨细胞增殖和软骨基质保护)来抑制软骨退变进展。