Cell Sheets from Adipose Tissue MSC Induce Healing of Pressure Ulcer and Prevent Fibrosis via Trigger Effects on Granulation Tissue Growth and Vascularization
Cell Sheets from Adipose Tissue MSC Induce Healing of Pressure Ulcer and Prevent Fibrosis via Trigger Effects on Granulation Tissue Growth and Vascularization
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DOI：
10.3390/ijms21155567
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发表时间：
2020-08-01
影响因子：
5.6
通讯作者：
Makarevich, Pavel
中科院分区：
文献类型：
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作者：
Alexandrushkina, Natalya;Nimiritsky, Peter;Makarevich, Pavel
我们报道了注射多能间充质基质细胞(MSC)、基于MSC的细胞片(CS)和MSC分泌体诱导C57BL/6小鼠皮肤压疮愈合的比较研究。我们发现，脂肪来源的骨髓间充质干细胞移植后，纤维化减轻，皮肤结构及其附属物(毛发和皮腺)恢复。尽管CS在溃疡表面的滞留时间较短(3-7天)，但它引起肉芽组织(GT)结构的深刻变化，增加了其厚度，改变了血管形态，血管密度降低，血管成熟度增加。在GT血管化方面，MSC分泌组也有类似的效果，但在CS组中，这种治疗方法并不能诱导皮肤及其附件的修复。对MSC产生的单层或片状分泌体成分的研究表明，CS产生更多参与周细胞趋化和血管成熟的因子(PDGF-BB、HGF、G-CSF)，而不是发芽诱导剂(VEGF165)。使用RNA测序和基因本体论作图对转录组进行分析，在CS中发现负责胶原结合和GT成熟的蛋白质以及已知的负调控血管萌发的脂肪酸代谢酶的上调。同时，激活毛细血管生长的因素丰富了下调的转录本，这表明在MSC薄片中，旁分泌活性可能转向基质重塑和血管系统的成熟，而不是血管发芽的激活。我们提出了一种可能的旁分泌触发机制，可能对GT的血管化和重塑产生影响。我们的结果表明，在片材内，MSC可能改变其功能状态和影响组织修复的可溶性因子的谱，并诱导更有效的皮肤愈合，倾向于再生和减少瘢痕形成。
We report a comparative study of multipotent mesenchymal stromal cells (MSC) delivered by injection, MSC-based cell sheets (CS) or MSC secretome to induce healing of cutaneous pressure ulcer in C57Bl/6 mice. We found that transplantation of CS from adipose-derived MSC resulted in reduction of fibrosis and recovery of skin structure with its appendages (hair and cutaneous glands). Despite short retention of CS on ulcer surface (3-7 days) it induced profound changes in granulation tissue (GT) structure, increasing its thickness and altering vascularization pattern with reduced blood vessel density and increased maturation of blood vessels. Comparable effects on GT vascularization were induced by MSC secretome, yet this treatment has failed to induce repair of skin with its appendages we observed in the CS group. Study of secretome components produced by MSC in monolayer or sheets revealed that CS produce more factors involved in pericyte chemotaxis and blood vessel maturation (PDGF-BB, HGF, G-CSF) but not sprouting inducer (VEGF165). Analysis of transcriptome using RNA sequencing and Gene Ontology mapping found in CS upregulation of proteins responsible for collagen binding and GT maturation as well as fatty acid metabolism enzymes known to be negative regulators of blood vessel sprouting. At the same time, downregulated transcripts were enriched by factors activating capillary growth, suggesting that in MSC sheets paracrine activity may shift towards matrix remodeling and maturation of vasculature, but not activation of blood vessel sprouting. We proposed a putative paracrine trigger mechanism potentially rendering an impact on GT vascularization and remodeling. Our results suggest that within sheets, MSC may change their functional state and spectrum of soluble factors that influence tissue repair and induce more effective skin healing inclining towards regeneration and reduced scarring.