目的 研究間充質(zhì)干細胞注射液（MSCsI）對手術(shù)法誘導(dǎo)的大鼠膝骨關(guān)節(jié)炎的治療作用。方法 自70只大鼠中隨機取8只為假手術(shù)組，其余62只動物均進行右側(cè)膝關(guān)節(jié)單側(cè)手術(shù)造模。術(shù)后4周，進行X光檢查并進行K-L評分，選擇模型成功、狀態(tài)良好的模型動物40只，根據(jù)K-L評分隨機分為模型組、玻璃酸鈉（陽性藥，每關(guān)節(jié)腔0.5 mg玻璃酸鈉注射液）組和MSCsI低、中、高劑量（每關(guān)節(jié)腔1.5×10⁵、5.0×10⁵、1.5×10⁶個細胞）組，每組8只。分組后次日開始，玻璃酸鈉組每周1次，其余各組每2周1次，關(guān)節(jié)腔內(nèi)注射給藥，持續(xù)5周，假手術(shù)組與模型組給予等體積0.9%氯化鈉溶液。使用X射線機檢測大鼠膝關(guān)節(jié)病變；給藥后記錄大鼠行為學(xué)及痛閾變化；剖殺時記錄關(guān)節(jié)軟骨大體評分；試劑盒法檢測血清中白細胞介素-1β（IL-1β）、白細胞介素-6（IL-6）、腫瘤壞死因子-α（TNF-α）、轉(zhuǎn)化生長因子β（TGF-β）、II型膠原C端肽（CTX-Ⅱ）、前列腺素E₂（PGE₂）水平；取膝關(guān)節(jié)組織進行病理切片，HE染色、甲苯胺藍染色觀察組織損傷。結(jié)果 與模型組相比，MSCsI高劑量顯著降低大鼠X光評分（P<0.05）、降低大鼠關(guān)節(jié)軟骨大體觀察評分（P<0.05）；給藥4周后，MSCsI低、中、高劑量顯著降低大鼠行為學(xué)評分（P<0.05、0.01），中、高劑量顯著升高大鼠痛閾值（P<0.01）；MSCsI低劑量顯著降低血清中IL-1β水平、升高TGF-β水平（P<0.05），中劑量顯著降低血清中IL-1β、IL-6、CTX-Ⅱ水平、升高TGF-β水平（P<0.05、0.01），高劑量顯著降低血清中IL-1β、IL-6、CTX-Ⅱ、PGE₂水平、升高TGF-β水平（P<0.05、0.01）；HE染色結(jié)果顯示，MSCsI各劑量能有效減輕模型大鼠組織病理學(xué)改變，滑膜、軟骨及軟骨下骨病變較輕；甲苯胺藍染色顯示，MSCsI各劑量可減小模型大鼠軟骨基質(zhì)損傷。結(jié)論 MSCsI每2周1次關(guān)節(jié)腔內(nèi)注射對手術(shù)誘導(dǎo)的大鼠膝骨關(guān)節(jié)炎有明顯的治療作用。

Objective To evaluate the efficacy of Mesenchymal Stem Cells Injection (MSCsI) on knee osteoarthritis induced by surgery in rats. Methods Eight out of 70 rats were randomly selected as the sham-operated group, while the remaining 62 animals underwent unilateral surgical modeling of the right knee joint. Four weeks after surgery, X-ray examination and K-L score were performed. Forty model animals with successful and good condition were selected and randomly divided into model group, sodium hyaluronate (positive drug, 0.5 mg sodium hyaluronate injection per joint cavity) group, and MSCsI low, medium, and high doses (1.5×10⁵, 5.0×10⁵, 1.5×10⁶ per joint cavity) group based on K-L score, with eight in each group. Starting from the day after grouping, the sodium hyaluronate group was given once a week, while the other groups were given intra-articular injection once every two weeks for five weeks. The sham-operated group and model group were given an equal volume of 0.9% sodium chloride solution. The knee lesions of rats were detected by X-ray machine. The behavior and pain threshold of rats were recorded after administration. The gross observation score of articular cartilage was recorded during dissection. Serum levels of interleukin-1β (IL-1β), interleukin-6 (IL-6), tumor necrosis factor-α (TNF-α), transforming growth factor-β (TGF-β), C-terminal peptide of type II collagen (CTX-Ⅱ) and prostaglandin E₂ (PGE₂) were detected by kits. The knee tissue was taken for pathological section, and the tissue injury was observed by HE staining and toluidine blue staining. Results Compared to model group, MSCsI 1.5×10⁶ cells significantly reduced the X-ray score of rats (P< 0.05) and the gross observation score of rat articular cartilage (P< 0.05). After four weeks of administration, MSCsI 1.5×10⁵, 5.0×10⁵, and 1.5×10⁶ cells significantly reduced rat behavioral scores (P< 0.05, 0.01), 5.0×10⁵, 1.5×10⁶ cells significantly increased the pain threshold in rats (P< 0.01). MSCsI 1.5×10⁵ cells significantly reduce IL-1β in serum level, elevated TGF-β level (P< 0.05), 5.0×10⁵ cells significantly reduce IL-1β, IL-6, CTX-Ⅱ levels in serum, elevated TGF-β level (P< 0.05, 0.01), 1.5×10⁶ cells significantly reduce IL-1β, IL-6, CTX-Ⅱ, PGE2 levels in serum, elevated TGF-β levels (P< 0.05, 0.01). The HE staining results showed that each dose of MSCsI could effectively alleviate the histopathological changes in the model rats, with mild synovial, cartilage, and subchondral bone lesions. Toluidine blue staining showed that various doses of MSCsI can reduce cartilage matrix damage in model rats. Conclusion Intra-articular injection of stem cells once every two weeks has a significant therapeutic effect on surgically induced knee osteoarthritis in rats.

R965