DeWitt Daughtry Family Department of Surgery, University of Miami Miller School of Medicine, Miami, FL.
John P. Hussman Institute for Human Genomics, Dr. John T. Macdonald Foundation Department of Human Genetics, University of Miami Miller School of Medicine, Miami, FL.
Ann Surg. 2023 Sep 1;278(3):383-395. doi: 10.1097/SLA.0000000000005949. Epub 2023 Jun 19.
Here, we report a new method to increase the therapeutic potential of mesenchymal stem/stromal cells (MSCs) for ischemic wound healing. We tested biological effects of MSCs modified with E-selectin, a cell adhesion molecule capable of inducing postnatal neovascularization, on a translational murine model.
Tissue loss significantly worsens the risk of extremity amputation for patients with chronic limb-threatening ischemia. MSC-based therapeutics hold major promise for wound healing and therapeutic angiogenesis, but unmodified MSCs demonstrate only modest benefits.
Bone marrow cells harvested from FVB/ROSA26Sor mTmG donor mice were transduced with E-selectin-green fluorescent protein (GFP)/AAV-DJ or GFP/AAV-DJ (control). Ischemic wounds were created via a 4 mm punch biopsy in the ipsilateral limb after femoral artery ligation in recipient FVB mice and subsequently injected with phosphate-buffered saline or 1×10 6 donor MSC GFP or MSC E-selectin-GFP . Wound closure was monitored daily for 7 postoperative days, and tissues were harvested for molecular and histologic analysis and immunofluorescence. Whole-body DiI perfusion and confocal microscopy were utilized to evaluate wound angiogenesis.
Unmodified MSCs do not express E-selectin, and MSC E-selectin-GFP gain stronger MSC phenotype yet maintain trilineage differentiation and colony-forming capability. MSC E-selectin-GFP therapy accelerates wound healing compared with MSC GFP and phosphate-buffered saline treatment. Engrafted MSC E-selectin-GFP manifest stronger survival and viability in wounds at postoperative day 7. Ischemic wounds treated with MSC E-selectin-GFP exhibit more abundant collagen deposition and enhanced angiogenic response.
We establish a novel method to potentiate regenerative and proangiogenic capability of MSCs by modification with E-selectin/adeno-associated virus. This innovative therapy carries the potential as a platform worthy of future clinical studies.
本研究报告了一种提高间充质干细胞（MSCs）治疗缺血性创面愈合潜力的新方法。我们在转化型小鼠模型中测试了经细胞黏附分子 E-选择素修饰后的 MSCs 的生物学效应，E-选择素可诱导出生后血管新生。
组织缺失会显著增加慢性肢体威胁性缺血患者截肢的风险。基于 MSC 的治疗方法为创面愈合和治疗性血管生成带来了巨大的希望，但未经修饰的 MSC 仅显示出适度的益处。
从 FVB/ROSA26Sor mTmG 供体小鼠的骨髓细胞中提取细胞，用 E-选择素-绿色荧光蛋白（GFP）/AAV-DJ 或 GFP/AAV-DJ（对照）进行转导。受体 FVB 小鼠通过股动脉结扎在其同侧肢体上创建 4mm 穿孔活检以创建缺血性创面，随后用磷酸盐缓冲盐水或 1×10 6 个供体 MSC GFP 或 MSC E-选择素-GFP 进行注射。术后第 7 天每天监测创面闭合情况，并采集组织进行分子和组织学分析及免疫荧光检测。利用 DiI 全身灌注和共聚焦显微镜评估创面血管生成。
未经修饰的 MSC 不表达 E-选择素，而 MSC E-选择素-GFP 获得更强的 MSC 表型，但仍保持三系分化和集落形成能力。与 MSC GFP 和磷酸盐缓冲盐水处理相比，MSC E-选择素-GFP 治疗可加速创面愈合。在术后第 7 天，移植的 MSC E-选择素-GFP 在创面中表现出更强的存活和活力。经 MSC E-选择素-GFP 处理的缺血性创面显示出更多的胶原沉积和增强的血管生成反应。
我们建立了一种通过 E-选择素/腺相关病毒修饰来增强 MSCs 再生和促血管生成能力的新方法。这种创新疗法具有作为未来临床研究值得关注的平台的潜力。
