Quiescent platelets stimulate angiogenesis and diabetic wound repair.
Quiescent platelets stimulate angiogenesis and diabetic wound repair.
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DOI：
10.1016/j.jss.2008.09.010
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发表时间：
2010-05-01
期刊：
影响因子：
--
通讯作者：
Orgill DP
中科院分区：
文献类型：
--
作者：
Pietramaggiori G;Scherer SS;Mathews JC;Gennaoui T;Lancerotto L;Ragno G;Valeri CR;Orgill DP
血小板参与止血、伤口愈合和肿瘤生长。尽管富血小板血浆（PRP）已在外科手术中使用多年，但其作用机制和应用方法仍缺乏表征。将通过血小板去除术获得的单个单位的人血小板在血浆中稀释并分成三个等体积。一个体积作为新鲜血小板（RT）在室温下储存，另一个体积通过在-80 °C下储存（FZ）冷冻，第三个体积在-80 °C下用6%DMSO冷冻（FZ 6）。血浆（PL）用作对照。使用流式细胞术，检测血小板的血小板糖蛋白GPIb和膜联蛋白V结合，分别作为存活和活化标志物。通过血栓弹性测定法评估止血功能。在体内，在db/db小鼠（n=10只/组）的1 cm 2全层伤口上局部施用血小板，并在显微镜和肉眼下对愈合进行分期。所有血小板制剂均显示出止血能力。RT血小板GPIb阳性（非活化静止血小板）和刺激血管生成3倍，细胞增殖4倍，在体内。FZ血小板膜联蛋白V阳性，表明活化的血小板，在体内，增加只有伤口肉芽。FZ 6血小板含有30%的非活化-静止和50%的活化血小板，并在体内刺激肉芽形成、血管生成、细胞增殖和促进再上皮化。血小板显示出诱导止血和伤口愈合的独特机制。静止血小板是体内诱导血管生成所必需的。血小板在室温下储存，用6%DMSO冷冻并储存在−80 °C下，在糖尿病小鼠中实现了最佳的伤口愈合。
Platelets partake in hemostasis, wound healing and tumor growth. Although platelet-rich-plasma (PRP) has been used in surgery for several years, its mechanism of action and application methods are still poorly characterized. A single unit of human platelets obtained by plateletpheresis was diluted in plasma and divided into three equal volumes. One volume was stored at room temperature as fresh platelets (RT), another volume frozen by storage at −80 °C (FZ) and the third volume frozen at −80 °C with 6% DMSO (FZ6). Plasma (PL) was used as control. Using flow cytometry, platelets were tested for platelet glycoprotein GPIb and annexin V binding, as survival and activation markers, respectively. Hemostatic function was assessed by thromboelastometry. In vivo, platelets were topically applied on 1 cm2, full-thickness wounds on db/db mice (n=10/group) and healing was staged microscopically and macroscopically. All platelet preparations showed hemostatic ability. RT platelets were GPIb positive (nonactivated-quiescent platelets) and stimulated angiogenesis by 3-fold, and cell proliferation by 4-fold in vivo. FZ platelets were positive for annexin V, indicating activated platelets and, in vivo, increased only wound granulation. FZ6 platelets contained 30% nonactivated-quiescent and 50% activated platelets and stimulated granulation, angiogenesis, cell proliferation and promoted re-epithelialization in vivo. Platelets showed distinct mechanisms to induce hemostasis and wound healing. Quiescent platelets are required to induced angiogenesis in vivo. Platelets stored at room temperature and frozen with 6% DMSO and stored at −80 °C achieved optimal wound healing in diabetic mice.
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