A novel autologous cell-based therapy to promote diabetic wound healing.
A novel autologous cell-based therapy to promote diabetic wound healing.
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DOI：
10.1097/sla.0b013e31826a9064
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发表时间：
2012-10
影响因子：
9
通讯作者：
Velazquez OC
中科院分区：
文献类型：
--
作者：
Castilla DM;Liu ZJ;Tian R;Li Y;Livingstone AS;Velazquez OC
我们先前已经表明，基质衍生因子-1 α（SDF-1α）在糖尿病皮肤伤口中下调，并且直接应用重组SDF-1α可增加伤口闭合率、新血管形成和内皮祖细胞（EPC）募集。然而，外源性SDF-1α的伤口水平增加导致这种促血管生成趋化因子的全身水平升高，这引起了对肿瘤发生和炎症的担忧。我们现在试图测试一种新的，更安全的基于细胞的治疗（CBT）的有效性，该治疗采用具有SDF-1 α的离体致敏骨髓干细胞（BMDSC）。我们还阐明了这种新方法加速糖尿病伤口愈合的作用机制。将来自糖尿病Leprdb/db小鼠的未分级BMDSC与SDF-1α（100 ng/mL）或BSA（对照）孵育20 h。将预处理的BMDSC（1×106）皮下注射到Leprdb/db小鼠（n=8/组）的全层皮肤伤口中。通过连续摄影、DiI灌注、共聚焦显微镜和免疫组化评估伤口闭合率、毛细血管密度和EPC募集。通过PCRAray和免疫印迹法评价可能介导SDF-1α-致敏的BMDSC促愈合/促血管生成作用的分子靶点的表达。在小鼠伤口愈合模型中测试潜在介质的生物学功能。ELISA法检测血清SDF-1α水平。SDF-1α致敏的BMDSC显著促进伤口愈合（p<.0001）、新生血管形成（p=.0028）和EPC募集（p=.0059）。基因/蛋白质表达研究证明EphRB 4和纤溶酶原作为下游靶标的上调可能介导促愈合和促血管生成应答。离体BMDSC活化和随后将细胞接种到伤口中不会增加全身SDF-1α水平。我们报告了一种新的CBT，它在促进2型糖尿病小鼠模型的愈合和新生血管形成方面非常有效。此外，我们确定了新的分子靶点，这可能对推进伤口愈合领域很重要。
We have previously shown that stromal-derived-factor-1 α(SDF-1α) is down-regulated within diabetic cutaneous wounds, and that direct application of recombinant SDF-1α increases wound closure rates, neovascularization and endothelial progenitor cell(s)(EPC) recruitment. However, increased wound levels of exogenous SDF-1α results in elevated systemic levels of this pro-angiogenic chemokine that raises concerns for tumorgenesis and inflammation. We now seek to test the efficacy of a novel, safer cell-based therapy (CBT) employing ex-vivo primed bone marrow stem cells (BMDSC) with SDF-1 α. We also elucidate the mechanism of action of this new approach for accelerating diabetic wound healing. Unfractionated BMDSC from diabetic Leprdb/db mice were incubated for 20h with SDF-1α(100ng/mL) or BSA(control). Pre-treated-BMDSC (1×106) were injected subcutaneously into full-thickness skin wounds in Leprdb/db mice (n=8/group). Wound closure rates, capillary density and recruitment of EPC were assessed with serial photography, DiI-perfusion, confocal microscopy and immunohistochemistry. Expression of molecular targets, which may mediate pro-healing/pro-angiogenic effects of SDF-1α-primed-BMDSC was evaluated by PCRArray and immunoblotting assay. The biological function of a potential mediator was tested in a mouse wound healing model. Serum SDF-1α levels were measured with ELISA. SDF-1α-primed-BMDSC significantly promote wound healing (p<.0001), neovascularization (p=.0028) and EPC recruitment(p=.0059). Gene/ protein expression studies demonstrate up-regulation of EphRB4 and Plasminogen as downstream targets potentially mediating the pro-healing and pro-angiogenic responses. Ex-vivo BMDSC activation and subsequent inoculation of cells into wounds does not increase systemic SDF-1α levels. We report a novel CBT that is highly effective in promoting healing and neovascularization in a murine model of Type 2 Diabetes. Furthermore, we identify new molecular targets that may be important for advancing the field of wound healing.