From the Department of Plastic Surgery, Tongji Hospital, Tongji Medical College, Huazhong University of Science and Technology.
Research Center of Plastic Surgery Hospital, Chinese Academy of Medical Sciences and Peking Union Medical College.
Plast Reconstr Surg. 2023 Jul 1;152(1):98-107. doi: 10.1097/PRS.0000000000010145. Epub 2023 Jan 2.
Wound healing is a widespread health problem that imposes a financial burden on health systems. Cell therapy with genetically modified adipose-derived stem cells (ADSCs) is a promising strategy for dysregulated wound repair. E2F transcription factor 1 (E2F1) is a bidirectional regulator of cytokines. Here, the authors aimed to investigate the impact and potential mechanism of E2F1 -/- ADSCs in promoting the wound healing process.
Forty-five C57BL/6 mice (specific pathogen-free, male) with 10-mm full-thickness wounds were randomly treated with subcutaneous injection of 2 × 10 6 wild-type ADSCs, 2 × 10 6 E2F1 -/- ADSCs, or phosphate-buffered saline. The wound closure rate was monitored at days 0, 3, 7, 10, and 14 after treatment. The collagen synthesis, angiogenesis, and wound contraction were calculated by Masson, immunohistochemistry, and immunofluorescent staining (CD31 and KI67), Western blotting (α-smooth muscle actin, collagen I, vascular endothelial growth factor, and transforming growth factor-β1) separately at day 14. In vitro, the conditioned media (CM) of wild-type ADSCs and E2F1 -/- ADSCs were collected to evaluate the impact on proliferation, migration, and angiogenesis.
In vivo, the E2F1 -/- ADSC group exhibited increased healing rate, proliferating vessels, and collagen synthesis compared with control at day 14 ( P < 0.05). Moreover, E2F1 -/- ADSCs showed enhanced vascular endothelial growth factor and transforming growth factor-β1 expression in the wound site and CM, and the CM from E2F1 -/- ADSCs promoted the proliferation, migration, and tube formation of co-cultured cells in vitro ( P < 0.05).
The E2F1 -/- ADSCs exhibited a strong paracrine ability to improve the vascularization process and collagen deposition, thereby accelerating wound healing in the rodent model.
These findings show that targeting transcription factor E2F1 could regulate the paracrine function of ADSCs, developing E2F1-modified ADSCs as an effective therapeutic option for wound healing and regeneration.
伤口愈合是一个普遍存在的健康问题，给卫生系统带来了经济负担。用基因修饰的脂肪来源干细胞（ADSCs）进行细胞治疗是一种有前途的调控失调伤口修复的策略。E2F 转录因子 1（E2F1）是细胞因子的双向调节剂。在这里，作者旨在研究 E2F1-/-ADSCs 促进伤口愈合过程的影响和潜在机制。
45 只 C57BL/6 雄性无特定病原体小鼠（SPF）的 10-mm 全层伤口被随机接受 2×10 6 个野生型 ADSCs、2×10 6 个 E2F1-/-ADSCs 或磷酸盐缓冲盐水的皮下注射。在治疗后第 0、3、7、10 和 14 天监测伤口闭合率。第 14 天分别通过 Masson、免疫组织化学和免疫荧光染色（CD31 和 KI67）计算胶原蛋白合成、血管生成和伤口收缩，Western blot （α-平滑肌肌动蛋白、I 型胶原蛋白、血管内皮生长因子和转化生长因子-β1）。体外收集野生型 ADSCs 和 E2F1-/-ADSCs 的条件培养基（CM），以评估对增殖、迁移和血管生成的影响。
体内，E2F1-/-ADSC 组在第 14 天的愈合率、增殖血管和胶原蛋白合成均高于对照组（P<0.05）。此外，E2F1-/-ADSCs 在伤口部位和 CM 中显示出增强的血管内皮生长因子和转化生长因子-β1表达，CM 从 E2F1-/-ADSCs 促进共培养细胞的增殖、迁移和管形成体外（P<0.05）。
E2F1-/-ADSCs 表现出强大的旁分泌能力，可改善血管生成过程和胶原蛋白沉积，从而加速啮齿动物模型中的伤口愈合。
这些发现表明，靶向转录因子 E2F1 可以调节 ADSCs 的旁分泌功能，将 E2F1 修饰的 ADSCs 作为治疗伤口愈合和再生的有效治疗选择。
