Nonthermal atmospheric pressure plasma enhances mouse limb bud survival, growth, and elongation.

Authors

Natalie Chernets, Electrical and Computer Engineering Department, Drexel UniversityFollow
Jun Zhang, Department of Orthopedic Surgery, Thomas Jefferson University; Department of Orthopedics, Second Hospital of Jilin UniversityFollow
Marla J Steinbeck, School of Biomedical Engineering, Science and Health Systems, Drexel University; Department of Orthopedic Surgery, Drexel University College of MedicineFollow
Deepa S Kurpad, Department of Orthopedic Surgery, Thomas Jefferson UniversityFollow
Eiki Koyama, Division of Orthopedic Surgery, Children's Hospital of Philadelphia
Gary Friedman, Electrical and Computer Engineering Department, Drexel University
Theresa A. Freeman, Department of Orthopaedic Surgery, Thomas Jefferson UniversityFollow

Document Type

Article

Publication Date

1-1-2015

Comments

This article has been peer reviewed. It was published in: Tissue Engineering - Part A.

Volume 21, Issue 1-2, 1 January 2015, Pages 300-309.

The published version is available at DOI: 10.1089/ten.tea.2014.0039

Copyright © 2015 Mary Ann Liebert, Inc.

Abstract

The enhanced differentiation of mesenchymal cells into chondrocytes or osteoblasts is of paramount importance in tissue engineering and regenerative therapies. A newly emerging body of evidence demonstrates that appendage regeneration is dependent on reactive oxygen species (ROS) production and signaling. Thus, we hypothesized that mesenchymal cell stimulation by nonthermal (NT)-plasma, which produces and induces ROS, would (1) promote skeletal cell differentiation and (2) limb autopod development. Stimulation with a single treatment of NT-plasma enhanced survival, growth, and elongation of mouse limb autopods in an in vitro organ culture system. Noticeable changes included enhanced development of digit length and definition of digit separation. These changes were coordinated with enhanced Wnt signaling in the distal apical epidermal ridge (AER) and presumptive joint regions. Autopod development continued to advance for approximately 144 h in culture, seemingly overcoming the negative culture environment usually observed in this in vitro system. Real-time quantitative polymerase chain reaction analysis confirmed the up-regulation of chondrogenic transcripts. Mechanistically, NT-plasma increased the number of ROS positive cells in the dorsal epithelium, mesenchyme, and the distal tip of each phalange behind the AER, determined using dihydrorhodamine. The importance of ROS production/signaling during development was further demonstrated by the stunting of digital outgrowth when anti-oxidants were applied. Results of this study show NT-plasma initiated and amplified ROS intracellular signaling to enhance development of the autopod. Parallels between development and regeneration suggest that the potential use of NT-plasma could extend to both tissue engineering and clinical applications to enhance fracture healing, trauma repair, and bone fusion.

Recommended Citation

Chernets, Natalie; Zhang, Jun; Steinbeck, Marla J; Kurpad, Deepa S; Koyama, Eiki; Friedman, Gary; and Freeman, Theresa A., "Nonthermal atmospheric pressure plasma enhances mouse limb bud survival, growth, and elongation." (2015). Department of Orthopaedic Surgery Faculty Papers. Paper 82.
https://jdc.jefferson.edu/orthofp/82

PubMed ID

25102046