Platelet-Rich Plasma for Regenerative Medicine Associated Treatments in Bottlenose Dolphins (Tursiops truncatus)
Dolphins exhibit an extraordinary capacity to heal deep soft tissue injuries, such as those following shark bites.1 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 proposed the use of a biological-based therapy for dolphin wound healing by the application of platelet-rich plasma (PRP), which has been described to accelerate the healing process in other mammals.2 First, we collected blood samples from six different dolphins and developed a specific and simple protocol, which concentrates platelets greater than two times that of whole blood. Whole blood samples were collected in sodium citrate to prevent clotting and were centrifuged at various revolutions and durations. After centrifugation, the lower half of the serum was considered PRP and collected for use in these experiments. A single centrifugation for 3 minutes at 900 rpm resulted in the best condition, whereas human or canine protocols for PRP concentration (1870 rpm or 460 g/8 min) were not applicable for dolphin platelet concentration. By flow cytometry (FACS) analysis, dolphin platelets showed reactivity to human platelet cell-surface marker CD41. Furthermore, lymphocyte contamination was negligible after centrifugation for 3 minutes at 900 rpm. Analysis by electron microscopy revealed that normal platelet structure and morphology was adequately preserved under the mentioned centrifugation protocol. Dolphin platelets were larger in size and contained more granules than human platelets, potentially explaining the need to reduce the duration and speed of centrifugation compared to other terrestrial mammal concentration protocols. In vitro assays applying PRP over mesenchymal cell cultures significantly promoted cell division and proliferation demonstrating tissue-repairing induction potential. In summary, these data show a simple and well-defined protocol for efficient PRP isolation, which would constitute the basis of a practical, low-cost and easily accessible biological treatment for dolphin wound-healing and tissue regeneration.
The authors would like to thank all the Oceanogràfic Biology Department, for their support in animal training, dedication and commitment in providing best care to the animals. This research was funded by the Fundación García-Cugat and by the Instituto de Salud Carlos III (FISS_PI13/00319).
* Presenting author
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2. Gentile P, Orlandi A, Scioli MG, Di Pasquali C, Bocchini I, Cervelli V. Concise review: adipose-derived stromal vascular fraction cells and platelet-rich plasma: basic and clinical implications for tissue engineering therapies in regenerative surgery. Stem Cells Transl Med. 2012;1:230–236.