Acellular cartilage extracellular matrix homogenate combined with oxymatrine on treatment of osteoarthritis in rats

doi:10.12307/2025.042

Abstract

Abstract: BACKGROUND: Acellular cartilage extracellular matrix is effective in the treatment of osteoarthritis, but its efficacy is limited when applied alone. Oxymatrine alleviates interleukin-1β-induced chondrocyte apoptosis and extracellular matrix degradation.
OBJECTIVE: To observe the effect of different doses of oxymatrine combined with acellular cartilage extracellular matrix injection in knee cavity on osteoarthritis in rats.
METHODS: The femoral cartilage of SD rats was obtained. The acellular cartilage extracellular matrix was prepared by physical, chemical and enzyme methods. Fifty SD rats were selected and divided into sham operation group, osteoarthritis group, simple material group, low-dose drug group, and high-dose drug group by random number table method, with 10 rats in each group. The latter four groups were treated with modified Hulth method to establish the rat model of osteoarthritis. After successful modeling, acellular cartilage extracellular matrix homogenate was injected into the knee cavity of rats in the simple material group. Acellular cartilage extracellular matrix homogenate containing 50, 100 µg oxymatrine was injected into the knee cavity of rats in the low-dose drug group and the high-dose drug group, respectively. Samples were taken 4 weeks after injection for relevant detection.
RESULTS AND CONCLUSION: (1) Compared with the sham operation group, the concentrations of interleukin 1β, tumor necrosis factor ɑ, and malondialdehyde in the joint effusion were increased (P < 0.05), and the concentration of superoxide dismutase in the joint effusion was decreased (P < 0.05) in the osteoarthritis group. Compared with osteoarthritis group, the levels of interleukin 1β, tumor necrosis factor ɑ, and malondialdehyde in joint effusion were decreased (P < 0.05), while the level of superoxide dismutase was increased (P < 0.05) in the low-dose drug group and high-dose drug group, and the changes were more significant in high-dose drug group. (2) TUNEL staining showed that compared with sham operation group, apoptotic chondrocytes increased in osteoarthritis group. Compared with the osteoarthritis group, the apoptotic chondrocytes decreased in the simple material group, the low-dose drug group, and the high-dose drug group, and the decrease was more significant in the high-dose drug group. (3) Hematoxylin-eosin staining and immunohistochemical staining showed that the knee cartilage was seriously degraded, the expression of type II collagen was decreased (P < 0.05), and the expression of matrix metalloproteinase-9 was increased (P < 0.05) in the osteoarthritis group. Compared with the osteoarthritis group, the knee cartilage degeneration of rats in the simple material group, the low-dose drug group, and the high-dose drug group was significantly improved; the expression of type II collagen was increased (P < 0.05) and the expression of matrix metalloproteinase 9 was decreased (P < 0.05), and the improvement was most significant in the high-dose drug group. (4) Western blot assay showed that compared with sham operation group, the expression of Nrf2, HO-1, and Bcl-2 protein in cartilage tissue decreased (P < 0.05); expression levels of Cleaved-caspase-3, Cleaved-caspase-9, and Bax were increased (P < 0.05) in the osteoarthritis group. Compared with the osteoarthritis group, the protein expression levels of Nrf2, HO-1, and Bcl-2 were increased (P < 0.05), and the protein expressions of Cleaved-caspase-3, Cleaved-caspase-9, and Bax were decreased (P < 0.05) in the low-dose and high-dose drug groups. The improvement was more significant in the high-dose drug group. (5) In conclusion, intracavicular injection of acellular cartilage extracellular matrix and oxymatrine can promote the synthesis of extracellular matrix, inhibit inflammation and oxidative stress, and suppress chondrocyte apoptosis, and play a therapeutic role in osteoarthritis, which may be related to the activation of Nrf2/HO-1 pathway.

Key words: osteoarthritis, acellular cartilage extracellular matrix, oxymatrine, chondrocyte, extracellular matrix

CLC Number:

Wu Fuzhang, Zhang Pengli, Zhang Zhenhua, He Yongbing, Sun Heyan. Acellular cartilage extracellular matrix homogenate combined with oxymatrine on treatment of osteoarthritis in rats[J]. Chinese Journal of Tissue Engineering Research, 2025, 29(13): 2675-2682.

Figures/Tables 10

2.2 实验动物数量分析术后无大鼠死亡，未出现膝关节感染、伤口感染与皮肤坏死等情况。骨关节炎造模大鼠造模后膝关节肿胀、活动受限与跛行等。50只大鼠全部进入结果分析。 2.3 各组大鼠关节软骨病理形态变化苏木精-伊红染色显示，假手术组大鼠膝关节表面软骨组织结构完整，软骨细胞大小及形态正常，各层次排列整齐且分布均匀；骨关节炎组大鼠膝关节出现严重退变，关节表面软骨结构遭到完全破坏，出现裂隙且深达软骨下骨，软骨细胞大小及形态异常，细胞萎缩、排列紊乱且无层次，软骨细胞数量显著减少；单纯材料组大鼠膝关节软骨存在退变表现，但病变未累及软骨下骨，软骨细胞形态较正常，软骨细胞数量较骨关节炎组增多；低剂量药物组、高剂量药物组大鼠膝关节软骨退变程度较轻，软骨细胞形态正常，软骨细胞数量较单纯材料组增多，其中高剂量药物组改善效果更明显，见图2。免疫组化染色显示，假手术组大鼠关节软骨组织中Ⅱ型胶原呈强阳性表达，基质金属蛋白酶9呈弱阳性表达；与假手术组相比，骨关节炎组大鼠关节软骨组织中Ⅱ型胶原阳性表达显著减弱，基质金属蛋白酶9阳性表达显著增强；与骨关节炎组相比，单纯材料组大鼠关节软骨组织中Ⅱ型胶原阳性表达增强，基质金属蛋白酶9阳性表达减弱；与单纯材料组相比，低剂量药物组、高剂量药物组大鼠关节软骨组织中Ⅱ型胶原阳性表达进一步增强，基质金属蛋白酶9阳性表达进一步减弱；相较于低剂量药物组，高剂量药物组大鼠关节软骨组织中Ⅱ型胶原阳性表达增强，基质金属蛋白酶9阳性表达减弱，见图3，4。5组大鼠关节软骨组织中Ⅱ型胶原与基质金属蛋白酶9表达灰度值，见表2。"

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