TY - JOUR TI - Proteomic Analysis and Cell Viability of Nine Amnion, Chorion, Umbilical Cord, and Amniotic Fluid–Derived Products AU - Becktell, Liliya AU - Matuska, Andrea M. AU - Hon, Stephanie AU - Delco, Michelle L. AU - Cole, Brian J. AU - Begum, Laila AU - Zhang, Sheng AU - Fortier, Lisa A. T2 - CARTILAGE AB - ObjectiveAmnion products are used in various musculoskeletal surgeries and as injections for joint pain with conflicting reports of cell viability and protein contents. The objective of this study was to determine the full proteome and examine cell viability in 9 commercial amnion products using an unbiased bottom-up shotgun proteomics approach and confocal microscopy.DesignProducts were subjected to liquid chromatography?tandem mass spectrometry (LC-MS/MS) analysis and searched against a UniProt Homo sapiens database. Relative protein abundance was determined for each sample. Based on proteomics results, lumican was measured by enzyme-linked immunosorbent assay (ELISA) and Western blot analysis was performed for interleukin-1 receptor antagonist (IL-1Ra) and tissue inhibitor of matrix metalloproteinase-2 (TIMP-2). Cell viability was determined by calcein AM (live) and ethidium homodimer (dead) staining and confocal microscopy.ResultsProteomic analysis revealed 919 proteins in the nine products. Proteins were primarily collagens, keratin, and albumin. Lumican, a small leucine-rich proteoglycan (SLRP) was found in all samples. Western blot analysis for IL-1Ra and TIMP-2 indicated presence of both proteins, with nonspecific antibody binding also present in all samples. No live cells were identified in any product.ConclusionsSeveral novel proteins were identified through proteomics that might impart the beneficial effects of amnion products, including SLRPs, collagens, and regulators of fibroblast activity. IL-1Ra and TIMP-2 were identified, but concentrations measured by ELISA may be falsely increased due to nonspecific antibody binding. The concept that the amnion tissues provide live cells to aid in tissue regeneration cannot be supported by the findings of this study. DA - 2020/12/24/ PY - 2020 DO - 10.1177/1947603520976767 DP - SAGE Journals SP - 1947603520976767 J2 - CARTILAGE LA - en SN - 1947-6035 UR - https://doi.org/10.1177/1947603520976767 Y2 - 2021/01/06/23:19:18 KW - amnion KW - amniotic fluid KW - chorion KW - lumican KW - umbilical cord ER -