Applied Microbiology Research Center, Systems Biology and Poisonings Institute, Baqiyatallah University of Medical Sciences, Tehran, Iran.
Islamic Azad university of sciences and researches, Tehran, Iran.
Bioorg Chem. 2022 Jul;124:105797. doi: 10.1016/j.bioorg.2022.105797. Epub 2022 Apr 8.
Mesenchymal stem cells (MSCs) therapeutically intensify wound-healing. Meanwhile, probiotics are capable to inhibit pathogenic bacteria by secreting antimicrobial and antioxidant agents. They ameliorate more the wounds, by MSCs' synergistic effect.
This way, MSCs exposed to probiotics metabolites (the extracts of Lactobacillus plantarum and Lactobacillus casei), become more viable, and the optimum doses of probiotics metabolites were determined by Trypan blue and MTT assays. Then, preconditioned MSCs (pre-MSCs) faced again the oxidative stress (100 µM HO), and their survival rate was assessed by MTT assay. Conditioned medium (CM) prepared from pre-MSCs and the expression rate of antioxidant proteins evaluated by Real-time PCR. The antimicrobial activity of the samples was evaluated using the MIC method. Then mice wound models were treated with the mixture of a basal ointment and each CM of pre-MSCs. The proliferation and viability of MSCs improved by the probiotic's metabolites preconditioning (P ≤ 0.05). pre-MSCs exhibited significant up-regulated in antioxidant genes expression (P ≤ 0.001). Also, probiotics metabolites presented a significant free radical scavenging effect (P ≤ 0.001). CMs of pre-MSCs showed considerably increased antibacterial activity (P ≤ 0.05). Experimental groups compared with the controls revealed that wound contraction was more rapid in groups that received CMs of LP-MSCs, LC-MSCs, MSCs, and then finally mixed probiotics metabolites, respectively.
These results provide a novel insight into the probiotics extract effect on the improvement of MSCs' potential for wound healing procedures.
间充质干细胞(MSCs)可增强伤口愈合的治疗效果。同时,益生菌通过分泌抗菌和抗氧化剂来抑制病原菌。它们通过与 MSCs 的协同作用使伤口得到更好的改善。
暴露于益生菌代谢物(植物乳杆菌和干酪乳杆菌的提取物)的 MSCs 变得更具活力,并且通过台盼蓝和 MTT 测定法确定了益生菌代谢物的最佳剂量。然后,预处理的 MSCs(预 MSC)再次面临氧化应激(100µM HO),并通过 MTT 测定法评估其存活率。通过实时 PCR 评估来自预 MSC 的条件培养基(CM)中抗氧化蛋白的表达率。使用 MIC 法评估样品的抗菌活性。然后,使用基础软膏和每种预 MSC 的 CM 混合物处理小鼠伤口模型。益生菌代谢物预处理可提高 MSCs 的增殖和活力(P≤0.05)。预 MSC 表现出抗氧化基因表达的显著上调(P≤0.001)。此外,益生菌代谢物具有显著的自由基清除作用(P≤0.001)。预 MSC 的 CM 表现出相当大的抗菌活性增强(P≤0.05)。实验组与对照组相比,接受 LP-MSCs、LC-MSCs、MSCs 的 CM 的组的伤口收缩速度更快,最后是混合益生菌代谢物的组。
这些结果为益生菌提取物对改善 MSC 伤口愈合潜力的作用提供了新的见解。