Abstract：
OBJECTIVE: This study aimed to investigate the effect of human adipose tissue derived stromal vascular fraction (AD-SVF) and mesenchymal stem cells (AD-MSCs) on blood flow recovery and neovascularisation in a rat hindlimb ischaemia model.
METHODS: SVF was isolated using an automated centrifugal system, and AD-MSCs were obtained from adherent cultures of SVF cells. Rats were divided into four groups of six rats each: non-ischaemia (Group 1); saline treated ischaemia (Group 2); SVF treated ischaemia (Group 3); and AD-MSC treated ischaemia (Group 4). Unilateral hindlimb ischaemia was induced in Sprague-Dawley rats via femoral artery ligation. Saline, SVF, or AD-MSCs were injected intramuscularly into the adductor muscle intra-operatively. Cell viability was calculated as the percentage of live cells relative to total cell number. Blood flow improvement, muscle fibre injury, and angiogenic properties were validated using thermal imaging and histological assessment.
RESULTS: The viabilities of SVF and AD-MSCs were 83.3% and 96.7%, respectively. Group 1 exhibited no significant temperature difference between hindlimbs, indicating a lack of blood flow changes. The temperature gradient gradually decreased in SVF and AD-MSC treated rats compared with saline treated rats. In addition, only normal muscle fibres with peripherally located nuclei were observed in Group 1. Groups 3 and 4 exhibited significantly fewer centrally located nuclei, indicating less muscle damage compared with Group 2. Regarding angiogenic properties, CD31 staining of endothelial cells showed similar patterns among all groups, whereas expression of vascular endothelial growth factor, as a crucial angiogenesis factor, was enhanced in the SVF and AD-MSC treated groups.
CONCLUSIONS: SVF and AD-MSCs improved blood flow and neovascularisation in a rat hindlimb ischaemia model, suggesting their potential ability to promote angiogenesis. Further extensive research is warranted to explore their potential applications in the treatment of severe lower extremity arterial disease.
METHODS: SVF was isolated using an automated centrifugal system, and AD-MSCs were obtained from adherent cultures of SVF cells. Rats were divided into four groups of six rats each: non-ischaemia (Group 1); saline treated ischaemia (Group 2); SVF treated ischaemia (Group 3); and AD-MSC treated ischaemia (Group 4). Unilateral hindlimb ischaemia was induced in Sprague-Dawley rats via femoral artery ligation. Saline, SVF, or AD-MSCs were injected intramuscularly into the adductor muscle intra-operatively. Cell viability was calculated as the percentage of live cells relative to total cell number. Blood flow improvement, muscle fibre injury, and angiogenic properties were validated using thermal imaging and histological assessment.
RESULTS: The viabilities of SVF and AD-MSCs were 83.3% and 96.7%, respectively. Group 1 exhibited no significant temperature difference between hindlimbs, indicating a lack of blood flow changes. The temperature gradient gradually decreased in SVF and AD-MSC treated rats compared with saline treated rats. In addition, only normal muscle fibres with peripherally located nuclei were observed in Group 1. Groups 3 and 4 exhibited significantly fewer centrally located nuclei, indicating less muscle damage compared with Group 2. Regarding angiogenic properties, CD31 staining of endothelial cells showed similar patterns among all groups, whereas expression of vascular endothelial growth factor, as a crucial angiogenesis factor, was enhanced in the SVF and AD-MSC treated groups.
CONCLUSIONS: SVF and AD-MSCs improved blood flow and neovascularisation in a rat hindlimb ischaemia model, suggesting their potential ability to promote angiogenesis. Further extensive research is warranted to explore their potential applications in the treatment of severe lower extremity arterial disease.
摘要：
目的：本研究旨在研究人脂肪组织来源的基质血管分数（AD-SVF）和间充质干细胞（AD-MSCs）对大鼠后肢缺血模型血流恢复和新生血管形成的影响。
方法：使用自动离心系统分离SVF，从SVF细胞的贴壁培养物中获得AD-MSC。将大鼠分成四组,每组六只大鼠:非缺血(组1);盐水处理的缺血(组2);SVF处理的缺血(组3);和AD-MSC处理的缺血(组4)。通过股动脉结扎在Sprague-Dawley大鼠中诱发单侧后肢缺血。盐水,SVF,或AD-MSC在术中肌肉注射到内收肌中。细胞活力计算为活细胞相对于总细胞数的百分比。血流改善，肌肉纤维损伤，和血管生成特性使用热成像和组织学评估进行验证。
结果：SVF和AD-MSCs的存活率分别为83.3%和96.7%，分别。第1组后肢之间没有明显的温度差，表明缺乏血流变化。与盐水处理的大鼠相比,在SVF和AD-MSC处理的大鼠中温度梯度逐渐降低。此外,在第1组中，仅观察到具有外周核的正常肌纤维。第3组和第4组显示出明显较少的位于中央的核，表明与第2组相比肌肉损伤较少。关于血管生成特性，CD31染色的内皮细胞在所有组中显示相似的模式，而血管内皮生长因子的表达，作为重要的血管生成因子，在SVF和AD-MSC治疗组中增强。
结论：SVF和AD-MSCs改善了大鼠后肢缺血模型的血流和新生血管形成，表明了它们促进血管生成的潜在能力。需要进一步广泛的研究来探索其在严重下肢动脉疾病治疗中的潜在应用。
方法：使用自动离心系统分离SVF，从SVF细胞的贴壁培养物中获得AD-MSC。将大鼠分成四组,每组六只大鼠:非缺血(组1);盐水处理的缺血(组2);SVF处理的缺血(组3);和AD-MSC处理的缺血(组4)。通过股动脉结扎在Sprague-Dawley大鼠中诱发单侧后肢缺血。盐水,SVF,或AD-MSC在术中肌肉注射到内收肌中。细胞活力计算为活细胞相对于总细胞数的百分比。血流改善，肌肉纤维损伤，和血管生成特性使用热成像和组织学评估进行验证。
结果：SVF和AD-MSCs的存活率分别为83.3%和96.7%，分别。第1组后肢之间没有明显的温度差，表明缺乏血流变化。与盐水处理的大鼠相比,在SVF和AD-MSC处理的大鼠中温度梯度逐渐降低。此外,在第1组中，仅观察到具有外周核的正常肌纤维。第3组和第4组显示出明显较少的位于中央的核，表明与第2组相比肌肉损伤较少。关于血管生成特性，CD31染色的内皮细胞在所有组中显示相似的模式，而血管内皮生长因子的表达，作为重要的血管生成因子，在SVF和AD-MSC治疗组中增强。
结论：SVF和AD-MSCs改善了大鼠后肢缺血模型的血流和新生血管形成，表明了它们促进血管生成的潜在能力。需要进一步广泛的研究来探索其在严重下肢动脉疾病治疗中的潜在应用。
