Organogenesis Inc., 2641 Rocky Ridge Lane, Birmingham, AL 35216.
Milestone Research Organization, 4901 Morena Blvd, Suite 132, San Diego, California, 92117.
J Orthop Res. 2019 Feb;37(2):412-420. doi: 10.1002/jor.24173. Epub 2018 Nov 30.
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.
羊膜悬浮同种异体移植物（ASA）来源于胎盘组织，包含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，包括腱病和肌腱损伤。本研究的目的是确定 ASA 对肌腱细胞密度、迁移和对炎症刺激的反应的潜在影响。使用 AlamarBlue 在多个时间点测量肌腱细胞密度，而使用 Boyden 室测定法测定迁移。使用 BioColor 试剂盒测量细胞外基质（ECM）标志物的沉积。使用 qPCR 和 ELISA 测量刺激促炎细胞因子 IL-1β 和 TNF-α 后细胞因子和生长因子的基因表达和蛋白产生。使用 ASA 制备条件培养基（CM）并用于本研究中的所有测定。在体外，与测定培养基对照相比，ASA CM 处理显着促进了肌腱细胞密度和迁移的增加。ASA CM 还增加了细胞外基质（ECM）蛋白的沉积，包括胶原蛋白、弹性蛋白和 sGAG。在炎症刺激和 ASA CM 治疗后，肌腱细胞下调了 IL-8 基因表达，这是一种在肌腱愈合的炎症阶段通常升高的促炎细胞因子。此外，与对照相比，用 ASA CM 处理的肌腱细胞 TGF-β1 的蛋白水平显着降低。本研究评估了 ASA 及其对肌腱细胞的影响；具体而言，ASA 处理导致细胞密度增加、迁移更加强壮、基质沉积增加，以及一些炎症靶点的改变。© 2018 作者。发表于 Wiley Periodicals, Inc. 代表骨科研究协会的《骨科研究杂志》。J Orthop Res 37:412-420, 2019。
