Tenocyte cell density, migration, and extracellular matrix deposition with amniotic suspension allograft

Abstract

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.

Species variations in tenocytes’ response to inflammation require careful selection of animal models for tendon research

“…We used IL-1β and TNF-α, two hallmark cytokines of inflammation in tendons, which are associated with tendon injury and tendinopathy in vivo and in vitro, to induce disease-relevant inflammation 28 , 30 , 33 , 45 , 89 – 97 . IL-1 activates the NF-κB pathway in tenocytes, induces the production of inflammatory mediators including COX2 and IL6, and matrix remodelling factors such as MMP1, MMP3 and MMP13 28 , 30 , 89 – 91 .…”

Treatment with Human Amniotic Suspension Allograft Improves Tendon Healing in a Rat Model of Collagenase-Induced Tendinopathy

“…Additionally, several groups have evaluated the effect of cytokines and growth factors on tendon repair [19,20,21]. Two studies used conditioned media from human amniotic mesenchymal stem cells (AMSCs) or amniotic membrane progenitor cells (AMPCs) at the site of Achilles tendon transection in rats; they found increased collagen production and biomechanical properties following treatment [20,21].…”

Species variations in tenocytes’ response to inflammation require careful selection of animal models for tendon research

“…We used IL-1β and TNF-α, two hallmark cytokines of inflammation in tendons, which are associated with tendon injury and tendinopathy in vivo and in vitro, to induce disease-relevant inflammation. 28,30,33,45,[89][90][91][92][93][94][95][96][97] IL-1 activates the NF-κB pathway in tenocytes, induces the production of inflammatory mediators including COX2 and IL6, and matrix remodelling factors such as MMP1, MMP3 and MMP13. 28,30,[89][90][91] It can even cause loss of the tenocyte phenotype, which is associated with decreased expression of tendon-related genes, e.g.…”