Objective: To observe the expressions of transforming growth factor β 1 (TGF-β 1) and basic fibroblast growth factor (bFGF) induced membrane by Masquelet technique in rats treated with glycoside of short-horned epimedium Herb, and to explore the effect of glycoside of short-horned epimedium Herb on Masquelet induced membrane.
Methods: Sixty 3-month-old male Wistar rats were randomly divided into 3 groups with 20 rats in each group; a tibial bone defect (6 mm in length) model was established. The blank group (group A) was not treated; the control group (group B) and the experimental group (group C) were filled with vancomycin antibiotic bone cement in the drawing stage, and the bone cement was completely solidified. Group C was given perfused flavonoids glycoside of short-horned epimedium Herb (10 μmol/L) by gavage once a day (0.3 mL) from 1 day after operation, and groups A and B were given the same amount of normal saline by gavage. After operation, the recovery and wound healing of experimental animals were observed; at 4 weeks after operation, X-ray film was taken to observe the recovery of bone defect of proximal tibia; at 6 weeks after operation, the bone defect was observed, and the morphological changes and vascularization degree of granulation tissue and induction membrane tissue were observed; the expressions of TGF-β 1 and bFGF were observed by immunohistochemistry staining and ELISA detection.
Results: The bone defect models of the 3 groups were established successfully, and there was no abnormality after operation. The incisions healed by first intention after operation. At 4 weeks after operation, X-ray films of proximal tibial defect showed that there was obvious space in group A, while bone cement was filled and Kirschner wire fixation was good in groups B and C. At 6 weeks after operation, the gross observation showed that the granulation tissue was filled in the defect area in group A; transparent membrane was formed in groups B and C, and microvessels were seen in some areas, and the microvessels in group C were significantly more than those in group B. Immunohistochemical staining showed that the expressions of TGF-β 1 and bFGF were negative in group A, but they were expressed in groups B and C, and the expressions of TGF-β 1 and bFGF in group B were significantly less than those in group C. ELISA detection showed that the expressions of TGF-β 1 and bFGF in group C were significantly higher than those in groups A and B ( P<0.05), but there was no significant difference between groups A and B ( P>0.05).
Conclusion: Glycoside of short-horned epimedium Herb can significantly increase the expressions of TGF-β 1 and bFGF, accelerate the process of osteogenesis, and contribute to bone shaping and reconstruction.
目的: 观察淫羊藿苷对 Masquelet 技术治疗大鼠胫骨骨缺损后诱导膜内血管化及其相关生物活性因子 TGF-β 1 和 bFGF 表达的影响，探讨淫羊藿苷在 Masquelet 技术修复骨缺损中的作用。.
方法: 将 60 只 3 月龄 Wistar 雄性大鼠随机分为 3 组，每组 20 只，建立右后肢 6 mm 长胫骨骨缺损模型。空白组（A 组）骨缺损处不作处理；对照组（B 组）和实验组（C 组）将处于拉丝期的去甲万古霉素抗生素骨水泥填充于骨缺损处，待骨水泥完全凝固。C 组从术后第 1 天开始每天给予淫羊藿总黄酮注射液稀释液（10 μmol/L）灌胃 1 次（0.3 mL），A、B 组予以等量生理盐水灌胃。观察实验动物术后恢复及切口愈合情况；术后 4 周摄 X 线片观察胫骨近端骨缺损处恢复情况；术后 6 周大体观察骨缺损处情况，并取材观察肉芽组织及诱导膜组织形态变化及其血管化程度，免疫组织化学染色观察及 ELISA 检测 TGF-β 1、bFGF 表达。.
结果: 3 组大鼠骨缺损模型均造模成功，术后无异常，切口均Ⅰ期愈合。术后 4 周胫骨近端骨缺损处 X 线片示 A 组有明显空隙，B、C 组为骨水泥充填、克氏针固定良好。术后 6 周大体观察示，A 组可见缺损区被肉芽组织充填；B、C 组可见透明薄膜形成，部分区域可见微血管，且 C 组薄膜及微血管明显多于 B 组。免疫组织化学染色示 A 组 TGF-β 1、bFGF 表达均为阴性，B、C 组均有表达且 B 组表达明显少于 C 组。ELISA 检测示，C 组 TGF-β 1、bFGF 表达水平显著高于 A、B 组（ P<0.05）；A、B 组间比较差异均无统计学意义（ P>0.05）。.
结论: 淫羊藿苷在大鼠胫骨骨缺损模型形成诱导膜时可显著提高 TGF-β 1 及 bFGF 的表达，加速成骨过程，有利于骨质塑形与改建。.
Keywords: Bone defect; Masquelet technique; basic fibroblast growth factor; glycoside of short-horned epimedium Herb; osteogenic activity; transforming growth factor β1.