Azam Maryam, Ghufran Hafiz, Butt Hira, Mehmood Azra, Ashfaq Ramla, Ilyas Asad M, Ahmad Muhammad R, Riazuddin Sheikh
National Centre of Excellence in Molecular Biology, 87-West Canal Bank Road, University of the Punjab, Lahore, Pakistan.
Burns Trauma. 2021 Sep 11;9:tkab021. doi: 10.1093/burnst/tkab021. eCollection 2021.
Following recent findings from our group that curcumin preconditioning augments the therapeutic efficacy of adipose-derived stem cells in the healing of diabetic wounds in rats, we aimed to investigate the regenerative effects of curcumin preconditioned adipose-derived mesenchymal stem cells (ASCs) for better recovery of acid inflicted burns in this study.
ASCs were preconditioned with 5 μM curcumin for 24 hours and assessed for proliferation, migration, paracrine release potential and gene expression comparative to naïve ASCs. Subsequently, the healing capacity of curcumin preconditioned ASCs (Cur-ASCs) versus naïve ASCs was examined using acidic wounds in rats. For this, acid inflicted burns of 20 mm in diameter were made on the back of male Wistar rats. Then, 2 × 10 cells of Cur-ASCs and naïve ASCs were intradermally injected in the wound periphery (n = 6) for comparison with an untreated saline control. Post-transplantation, wounds were macroscopically analysed and photographed to evaluate the percentage of wound closure and period of re-epithelization. Healed wound biopsies were excised and used for histological evaluation and expression analysis of wound healing markers at molecular level by quantitative PCR and western blotting.
We found that Cur-ASCs exhibited greater proliferation, migration and paracrine potential . Further, Cur-ASCs showed more effective recovery than naïve ASCs as exhibited by gross morphology, faster wound closure and earlier re-epithelialization. Masson's trichrome and hematoxylin and eosin staining demonstrated the improved architecture of the healing burns, as evidenced by reduced infiltration of inflammatory cells, compact collagen and marked granulation in Cur-ASC treated rats. Corroborating these findings, molecular assessment showed significantly reduced expressions of pro-inflammatory factors (interleukin-1 beta, interleukin-6, tumor necrosis factor alpha) a with striking upsurge of an oxidative marker (superoxide dismutase 1), pro-angiogenic factors (vascular endothelial growth factor, hepatocyte growth factor, hypoxia-inducible factor-1 alpha) and collagen markers (transforming growth factor beta 1, fibroblast growth factor-2, collagen type 1 alpha 1), verifying that Cur-ASCs modulate the regulation of pro-inflammatory and healing markers at burn sites.
Treatment with Cur-ASCs resulted in faster re-epithelization of acid inflicted burns compared to the treatment with naïve ASCs. Based on observed findings, we suggest the transplantation of Cur-ASCs is a valuable therapy for the potent clinical management of acidic burns.
我们团队最近发现姜黄素预处理可增强脂肪干细胞对大鼠糖尿病伤口愈合的治疗效果，在此研究中，我们旨在探讨姜黄素预处理的脂肪间充质干细胞（ASCs）的再生作用，以促进酸性烧伤更好地恢复。
用5μM姜黄素对ASCs进行24小时预处理，并与未处理的ASCs比较评估其增殖、迁移、旁分泌释放潜能及基因表达。随后，利用大鼠酸性伤口检测姜黄素预处理的ASCs（Cur-ASCs）与未处理的ASCs的愈合能力。为此，在雄性Wistar大鼠背部制作直径为20mm的酸性烧伤。然后，将2×10个Cur-ASCs和未处理的ASCs皮内注射到伤口周边（n = 6），与未处理的生理盐水对照组进行比较。移植后，对伤口进行宏观分析和拍照，以评估伤口闭合百分比和再上皮化时间。切除愈合伤口的活检组织，用于组织学评估，并通过定量PCR和蛋白质印迹在分子水平分析伤口愈合标志物的表达。
我们发现Cur-ASCs表现出更强的增殖、迁移和旁分泌潜能。此外，Cur-ASCs比未处理的ASCs显示出更有效的恢复，表现为大体形态、更快的伤口闭合和更早的再上皮化。Masson三色染色和苏木精-伊红染色显示愈合烧伤的结构得到改善，Cur-ASCs处理的大鼠炎症细胞浸润减少、胶原致密和肉芽明显，证明了这一点。证实这些发现的分子评估显示促炎因子（白细胞介素-1β、白细胞介素-6、肿瘤坏死因子-α）的表达显著降低，而氧化标志物（超氧化物歧化酶1）、促血管生成因子（血管内皮生长因子、肝细胞生长因子、缺氧诱导因子-1α）和胶原标志物（转化生长因子-β1、成纤维细胞生长因子-2、Ⅰ型胶原α1）显著上调，证实Cur-ASCs可调节烧伤部位促炎和愈合标志物的调控。
与未处理的ASCs治疗相比，Cur-ASCs治疗可使酸性烧伤的再上皮化更快。基于观察结果，我们认为Cur-ASCs移植是酸性烧伤有效临床管理的一种有价值的治疗方法。
