title: Chemical modification of extracellular matrix by cold atmospheric plasma-generated reactive species affects chondrogenesis and bone formation.
abstract: The goal of this study was to investigate whether cold plasma generated by dielectric barrier discharge (DBD) modifies extracellular matrices (ECM) to influence chondrogenesis and endochondral ossification. Replacement of cartilage by bone during endochondral ossification is essential in fetal skeletal development, bone growth and fracture healing. Regulation of this process by the ECM occurs through matrix remodelling, involving a variety of cell attachment molecules and growth factors, which influence cell morphology and protein expression. The commercially available ECM, Matrigel, was treated with microsecond or nanosecond pulsed (μsp or nsp, respectively) DBD frequencies conditions at the equivalent frequencies (1 kHz) or power (~1 W). Recombinant human bone morphogenetic protein-2 was added and the mixture subcutaneously injected into mice to simulate ectopic endochondral ossification. Two weeks later, the masses were extracted and analysed by microcomputed tomography. A significant increase in bone formation was observed in Matrigel treated with μsp DBD compared with control, while a significant decrease in bone formation was observed for both nsp treatments. Histological and immunohistochemical analysis showed Matrigel treated with μsp plasma increased the number of invading cells, the amount of vascular endothelial growth factor and chondrogenesis while the opposite was true for Matrigel treated with nsp plasma. In support of the in vivo Matrigel study, 10 T1/2 cells cultured in vitro on μsp DBD-treated type I collagen showed increased expression of adhesion proteins and activation of survival pathways, which decreased with nsp plasma treatments. These results indicate DBD modification of ECM can influence cellular behaviours to accelerate or inhibit chondrogenesis and endochondral ossification. Copyright © 2016 John Wiley & Sons, Ltd.
authors: Department of Orthopaedic Surgery, Thomas Jefferson University, Philadelphia, PA, USA.
authors: Peter
authors: P
authors: Eisenhauer
authors: Department of Orthopaedic Surgery, Thomas Jefferson University, Philadelphia, PA, USA.
authors: Natalie
authors: N
authors: Chernets
authors: Department of Orthopaedic Surgery, Thomas Jefferson University, Philadelphia, PA, USA.
authors: You
authors: Y
authors: Song
authors: Drexel Plasma Institute, Drexel University, Philadelphia, PA, USA.
authors: Danil
authors: D
authors: Dobrynin
authors: Department of Bioengineering, Temple University, Philadelphia, PA, USA.
authors: Nancy
authors: N
authors: Pleshko
authors: School of Biomedical Engineering, Science and Health Systems, Drexel University, Philadelphia, PA, USA.
authors: Marla J
authors: MJ
authors: Steinbeck
authors: Department of Orthopaedic Surgery, Thomas Jefferson University, Philadelphia, PA, USA. theresa.freeman@jefferson.edu.|Drexel Plasma Institute, Drexel University, Philadelphia, PA, USA. theresa.freeman@jefferson.edu.
authors: Theresa A
authors: TA
authors: Freeman
mesh_terms: D000818:Animals; D055396:Bone Morphogenetic Protein 2 / Q000737:chemistry / Q000494:pharmacology; D020219:Chondrogenesis; D005109:Extracellular Matrix / Q000737:chemistry*; D006801:Humans; D051379:Mice; D010012:Osteogenesis; D058626:Plasma Gases / Q000737:chemistry*; D017382:Reactive Oxygen Species / Q000737:chemistry*; D011994:Recombinant Proteins / Q000737:chemistry / Q000494:pharmacology
chemical_list: C525718:BMP2 protein, human; D055396:Bone Morphogenetic Protein 2; D058626:Plasma Gases; D017382:Reactive Oxygen Species; D011994:Recombinant Proteins
keywords: FTIR; bone; chondrogenesis; endochondral ossification; matrigel; plasma