Platelet-rich plasma and adipose-derived mesenchymal stem cells for regenerative medicine-associated treatments in bottlenose dolphins (Tursiops truncatus)

• Published in: PloS one Vol. 9; no. 9; p. e108439
• Main Authors: Griffeth, Richard J, García-Párraga, Daniel, Mellado-López, Maravillas, Crespo-Picazo, Jose Luis, Soriano-Navarro, Mario, Martinez-Romero, Alicia, Moreno-Manzano, Victoria
• Format: Journal Article
• Language: English
• Published: United States Public Library of Science 24.09.2014; Public Library of Science (PLoS)
• Subjects: Adipose tissue; Angiogenesis; Animals; Aquatic mammals; Biological treatment; Biology and Life Sciences; Blood; Blood & organ donations; Blood platelets; Bottle-Nosed Dolphin - blood; Cadavers; Cell culture; Cell cycle; Cell Differentiation; Cell Proliferation; Cell surface; Cells, Cultured; Centrifugation; Cetacea; Dolphins; Dolphins & porpoises; Electron microscopy; Engineering and Technology; Female; Flow cytometry; Growth factors; Humans; Infections; Male; Marine animals; Marine mammals; Marine organisms; Medicine and Health Sciences; Mesenchymal stem cells; Mesenchymal Stromal Cells - physiology; Mesenchyme; Plasma; Platelet-derived growth factor; Platelet-derived growth factor BB; Platelet-Rich Plasma - immunology; Platelet-Rich Plasma - physiology; Platelets; Regeneration; Regeneration (physiology); Regenerative Medicine; Research and Analysis Methods; Side effects; Stem cell transplantation; Stem cells; Studies; Surface markers; Tissue engineering; Transforming Growth Factor beta - metabolism; Transplants & implants; Tursiops truncatus; Ulcers; Vascular endothelial growth factor; Vascular Endothelial Growth Factor A - metabolism; Veterinary medicine; Wound Healing
• ISSN: 1932-6203; 1932-6203
• DOI: 10.1371/journal.pone.0108439

Dolphins exhibit an extraordinary capacity to heal deep soft tissue injuries. Nevertheless, accelerated wound healing in wild or captive dolphins would minimize infection and other side effects associated with open wounds in marine animals. Here, we propose the use of a biological-based therapy for wound healing in dolphins by the application of platelet-rich plasma (PRP). Blood samples were collected from 9 different dolphins and a specific and simple protocol which concentrates platelets greater than two times that of whole blood was developed. As opposed to a commonly employed human protocol for PRP preparation, a single centrifugation for 3 minutes at 900 rpm resulted in the best condition for the concentration of dolphin platelets. By FACS analysis, dolphin platelets showed reactivity to platelet cell-surface marker CD41. Analysis by electron microscopy revealed that dolphin platelets were larger in size than human platelets. These findings may explain the need to reduce the duration and speed of centrifugation of whole blood from dolphins to obtain a 2-fold increase and maintain proper morphology of the platelets. For the first time, levels of several growth factors from activated dolphin platelets were quantified. Compared to humans, concentrations of PDGF-BB were not different, while TGFβ and VEGF-A were significantly lower in dolphins. Additionally, adipose tissue was obtained from cadaveric dolphins found along the Spanish Mediterranean coast, and adipose-derived mesenchymal stem cells (ASCs) were successfully isolated, amplified, and characterized. When dolphin ASCs were treated with 2.5 or 5% dolphin PRP they exhibited significant increased proliferation and improved phagocytotic activity, indicating that in culture, PRP may improve the regenerative capacity of ASCs. Taken together, we show an effective and well-defined protocol for efficient PRP isolation. This protocol alone or in combination with ASCs, may constitute the basis of a biological treatment for wound-healing and tissue regeneration in dolphins.