Conditioned Medium from Periodontal Ligament Stem Cells Enhances Periodontal Regeneration
Conditioned Medium from Periodontal Ligament Stem Cells Enhances Periodontal Regeneration
复制标题
DOI：
10.1089/ten.tea.2016.0274
复制
发表时间：
2017-05-01
影响因子：
4.1
通讯作者：
Morita, Ikuo
中科院分区：
文献类型：
--
作者：
Nagata, Mizuki;Iwasaki, Kengo;Morita, Ikuo
牙周病是成人最常见的传染病之一，其特征是牙齿支持组织的破坏。间充质干细胞（MSC）包括中胚层起源的干细胞群体，其已被研究并用于细胞治疗。然而，由于在各种疾病模型中MSC移植后细胞存活率较低，MSC的旁分泌功能作为再生机制受到越来越多的关注。本研究的目的是调查移植的条件培养基（CM）的再生潜力，从培养的牙周膜干细胞（PDLSCs），成人干细胞群体的牙齿支持组织，使用大鼠牙周缺损模型。使用超滤从PDLSC和成纤维细胞收集无细胞CM，并移植到手术创建的牙周缺损中。抗体芯片检测CM蛋白含量。新的牙周组织的形成进行了分析，使用微型计算机断层扫描和组织切片。PDLSC-CM移植以浓度依赖性方式增强牙周组织再生，而成纤维细胞-CM没有显示出任何再生功能。蛋白质组学分析显示，PDLSC-CM中含有细胞外基质蛋白、酶、血管生成因子、生长因子和细胞因子。此外，PDLSC-CM移植导致愈合牙周组织中肿瘤坏死因子-α（TNF-α）的mRNA水平降低。此外，我们发现PDLSC-CM抑制了用IFN-γ刺激的单核细胞/巨噬细胞系RAW细胞中TNF-α的mRNA水平。我们的研究结果表明，PDLSC-CM增强牙周再生抑制炎症反应，通过TNF-α的生产，和PDLSC-CM的移植可能是一种新的方法牙周再生治疗。
Periodontal disease is one of the most common infectious diseases in adults and is characterized by the destruction of tooth-supporting tissues. Mesenchymal stem cells (MSCs) comprise the mesoderm-originating stem cell population, which has been studied and used for cell therapy. However, because of the lower rate of cell survival after MSC transplantation in various disease models, paracrine functions of MSCs have been receiving increased attention as a regenerative mechanism. The aim of this study was to investigate the regenerative potential of transplanted conditioned medium (CM) obtained from cultured periodontal ligament stem cells (PDLSCs), the adult stem cell population in tooth-supporting tissues, using a rat periodontal defect model. Cell-free CM was collected from PDLSCs and fibroblasts, using ultrafiltration and transplanted into surgically created periodontal defects. Protein content of CM was examined by antibody arrays. Formation of new periodontal tissues was analyzed using microcomputed tomography and histological sections. PDLSC-CM transplantation enhanced periodontal tissue regeneration in a concentration-dependent manner, whereas fibroblast-CM did not show any regenerative function. Proteomic analysis revealed that extracellular matrix proteins, enzymes, angiogenic factors, growth factors and cytokines were contained in PDLSC-CM. Furthermore, PDLSC-CM transplantation resulted in the decreased mRNA level of tumor necrosis factor-alpha (TNF-alpha) in healing periodontal tissues. In addition, we found that PDLSC-CM suppressed the mRNA level of TNF-alpha in the monocyte/macrophage cell line, RAW cells, stimulated with IFN-gamma. Our findings suggested that PDLSC-CM enhanced periodontal regeneration by suppressing the inflammatory response through TNF-alpha production, and transplantation of PDLSC-CM could be a novel approach for periodontal regenerative therapy.