Tenocyte cell density, migration, and extracellular matrix deposition with amniotic suspension allograft.
Tenocyte cell density, migration, and extracellular matrix deposition with amniotic suspension allograft.
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DOI：
10.1002/jor.24173
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发表时间：
2019-03
期刊：
影响因子：
--
通讯作者：
Mowry KC
中科院分区：
文献类型：
--
作者：
Kimmerling KA;McQuilling JP;Staples MC;Mowry KC
同种异体羊膜悬浮移植物来源于胎盘组织，含有颗粒状羊膜和羊水细胞。最近，ASA和其他胎盘来源的同种异体移植物已被用于骨科应用，包括肌腱疾病和肌腱损伤。本研究的目的是确定ASA对腱细胞密度、迁移和对炎症刺激的反应的潜在影响。使用AlamarBlue在多个时间点上测量腱细胞密度，而使用Boyden小室实验来确定迁移。使用BioColor试剂盒测量ECM标志物的沉积。用定量聚合酶链式反应和ELISA检测促炎症因子β和肿瘤坏死因子α刺激后细胞因子和生长因子的基因表达和蛋白产生。条件培养液(CM)由ASA制成，用于本研究的所有检测。在体外，ASA CM处理显著促进了腱细胞密度和迁移的增加，与测定介质对照组相比。ASA CM还增加了细胞外基质(ECM)蛋白的沉积，包括胶原、弹性蛋白和SGAG。在炎症刺激和ASA CM治疗后，肌腱细胞下调IL-8基因的表达，IL-8是一种促炎细胞因子，通常在肌腱愈合的炎症阶段升高。此外，与对照组相比，经AsA CM处理的腱细胞转化生长因子-β1的蛋白水平显著降低。这项研究评估了ASA及其对腱细胞的影响；具体地说，ASA治疗导致细胞密度增加，更强劲的迁移和基质沉积，以及一些炎症靶点的改变。©2018作者。《整形外科研究杂志》由威利期刊公司代表整形外科研究学会出版。J Orthop Res 37：412-420,2019年。
Amniotic suspension allografts (ASA), derived from placental tissues, contain particulated amniotic membrane and amniotic fluid cells. Recently, ASA and other placental‐derived allografts have been used in orthopaedic applications, including tendinopathies and tendon injuries. The purpose of this study was to determine the potential effects of ASA on tenocyte cell density, migration, and responses to inflammatory stimuli. Tenocyte cell density was measured using AlamarBlue over multiple time points, while migration was determined using a Boyden chamber assay. Deposition of ECM markers were measured using BioColor kits. Gene expression and protein production of cytokines and growth factors following stimulus with pro‐inflammatory IL‐1β and TNF‐α was measured using qPCR and ELISAs. Conditioned media (CM) was made from ASA and used for all assays in this study. In vitro, ASA CM treatment significantly promoted tenocyte increases in cell density and migration compared to assay media controls. ASA CM also increased the deposition of extracellular matrix (ECM) proteins, including collagen, elastin, and sGAG. Following inflammatory stimulation and treatment with ASA CM, tenocytes downregulated IL‐8 gene expression, a pro‐inflammatory cytokine normally elevated during the inflammatory phase of tendon healing. Additionally, tenocytes treated with ASA CM had significantly lower protein levels of TGF‐β1 compared to controls. This study evaluated ASA and its effect on tenocytes; specifically, treatment with ASA resulted in increased cell density, more robust migration and matrix deposition, and some alteration of inflammatory targets. © 2018 The Authors. Journal of Orthopaedic Research® Published by Wiley Periodicals, Inc. on behalf of Orthopaedic Research Society. J Orthop Res 37:412–420, 2019.
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