目的: 研究人脂肪间充质干细胞(human adipose-derived mesenchymal stem cells，hASCs)外泌体对骨质疏松小鼠骨髓间充质干细胞(bone marrow mesenchymal stem cells，BMSCs)成骨分化的影响，以及对雌激素缺乏引起的骨质疏松的预防效果。方法: 采用超速离心法提取hASCs分泌的外泌体，然后提取骨质疏松小鼠BMSCs，并将这些细胞暴露于含有hASCs外泌体的成骨诱导培养基中作为实验组，同时使用不含hASCs外泌体的成骨诱导培养基对BMSCs进行成骨诱导作为对照组，通过碱性磷酸酶(alkaline phosphatase, ALP)染色和定量分析以及实时荧光定量聚合酶链反应(quantitative reverse transcription polymerase chain reaction，qPCR)检测评估hASCs外泌体对BMSCs成骨分化的影响。将标记有荧光信号的hASCs外泌体通过尾静脉注射入小鼠体内，观察外泌体的体内分布情况。切除小鼠双侧卵巢构建小鼠雌激素缺乏模型，术后两周将小鼠分为3组：实验组为雌激素缺乏小鼠接受hASCs外泌体注射；阴性对照组为雌激素缺乏小鼠接受磷酸缓冲盐溶液(phosphate buffered saline，PBS)注射；阳性对照组为假手术(Sham)小鼠接受PBS注射，每3天注射1次，共注射8次后收集小鼠股骨，拍摄显微CT(micro computed tomography，micro-CT)，计算骨密度并进行骨形态学分析。结果: 使用超速离心法成功提取hASCs外泌体。hASCs外泌体使骨质疏松小鼠BMSCs的ALP染色加深，ALP活性增强，成骨相关基因表达上调。注射hASCs外泌体的雌激素缺乏小鼠较注射PBS的雌激素缺乏小鼠股骨骨小梁更为致密，骨密度增大，并且与Sham组相比并未出现明显的骨密度降低。结论: hASCs外泌体不仅能在体外促进骨质疏松小鼠BMSCs的成骨分化，还能在体内有效预防去势小鼠骨质疏松的发生，这些发现为hASCs外泌体作为一种新型的生物治疗手段在骨质疏松症的预防中的应用提供了科学依据。

Objective: To investigate the effect of human adipose-derived mesenchymal stem cells (hASCs) exosomes on osteogenic differentiation of bone marrow mesenchymal stem cells (BMSCs) extracted from osteoporotic mice, and to evaluate the effect of hASCs exosomes on preventing bone loss induced by estrogen deficiency. Methods: hASCs exosomes were extracted by ultracentrifugation. The osteoporotic mice were established by bilateral ovariectomy (OVX). BMSCs were isolated from osteo-porotic mice and cultured for further analysis. In the experimental group, these BMSCs were exposed to an osteogenic induction medium supplemented with hASCs exosomes to evaluate their potential effects on osteogenesis. In contrast, the control group was treated with the same osteogenic induction medium, but without the addition of hASCs exosomes, to serve as a baseline comparison for the study. To comprehensively assess the osteogenic differentiation of BMSCs influenced by hASCs exosomes, alkaline phosphatase (ALP) staining, ALP activity quantitative analysis and quantitative reverse transcription polymerase chain reaction (qPCR) were performed. These evaluations provided critical insights into the role of hASCs exosomes in promoting osteoblast differentiation and bone formation in osteoporotic conditions. The fluorescence labeled hASCs exosomes were injected via the tail vein to observe the biodistribution of exosomes. Two weeks after OVX, the mice were divided into three groups: The experimental group consisted of estrogen-deficient mice receiving hASCs exosome injections; the negative control group consisted of estrogen-deficient mice receiving phosphate-buffered saline (PBS) injections; and the positive control group consisted of mice that underwent Sham surgery and received PBS injections.The injections were administered once every 3 days, for a total of 8 injections. Afterward, the femurs were collected from the mice, and micro-computed tomography (micro-CT) was performed to measure bone mineral density and conduct bone morphometric analysis. Results: hASCs exosomes were successfully extracted using ultracentrifugation. After the induction by hASCs exosomes, ALP staining and ALP activity in the BMSCs extracted from osteoporotic mice were significantly enhanced, the expression of osteogenesis related genes in BMSCs were significantly up-regulated. More trabecular bone and higher bone mineral density were observed in estrogen-deficient mice injected with hASCs exosomes compared with estrogen-deficient mice injected with PBS, and there was no significant decrease in bone mineral density compared with the Sham operation group. Conclusion: hASCs exosomes promoted the osteogenic differentiation of BMSCs extracted from osteoporotic mice. hASCs exosomes prevented bone loss induced by estrogen deficiency.