Platelet-Rich Plasma and Adipose-Derived Mesenchymal Stem Cells for Regenerative Medicine-Associated Treatments in Bottlenose Dolphins (Tursiops truncatus)

Abstract

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.

A phylogenomic analysis of the role and timing of molecular adaptation in the aquatic transition of cetartiodactyl mammals

“…This reduced clotting has been attributed to the contact of blood with water rather than air, as well as the need to sequester blood in stagnant reservoirs while diving [ 56 ]. Moreover, cetaceans show accelerated wound healing, reducing infection and accelerating tissue repair [ 57 ].…”

Clinical management of Candida albicans keratomycosis in a bottlenose dolphin (Tursiops truncatus)

“…In humans, ASCs have been used to promote healing in radiation ischemia and perianal fistulas, and animal models show promise for the treatment of cardiovascular, genitourinary, and musculoskeletal disorders . In dolphins, ASCs have been isolated, cultured, and differentiated into multiple cell lineages, suggesting that they may have similar regenerative capabilities as in other mammalian species . Because the stem cells were derived from the sire of the patient, there was less risk of immune rejection of the cells than if they were derived from another, unrelated animal.…”

“…Blood was collected from the periarterial venous rete at the distal ventral peduncle and processed to collect platelet‐rich plasma (PrP) using a veterinary benchtop kit primarily utilized in equine patients, which included spinning whole blood at 720 g for 15 min (Genesis CS‐2, Vet Stem Pharma, Poway, CA). Recent studies have shown that dolphin platelets have a significantly larger area compared with equine and human platelets, and an ideal centrifugation protocol for dolphin platelet concentration is 106 g (900 rpm) for 3 min, a much lower speed and shorter duration than was recommended with the benchtop kit . Because of the size difference between equine and cetacean platelets, this protocol was not successful in collecting PrP, and fresh plasma was used in all procedures.…”

The use of fat grafting and platelet‐rich plasma for wound healing: A review of the current evidence

“…ADSCs cocultured with PRP secrete low chemokine concentrations and PRP also encourages downregulation of the strongly proinflammatory gene IL1B in ADSCs both of which suggest an anti‐inflammatory effect . ADSCs cultured with PRP from dolphins also show an enhanced ability to phagocytose suggesting a role in the inflammation stage of healing …”

“…84 ADSCs cultured with PRP from dolphins also show an enhanced ability to phagocytose suggesting a role in the inflammation stage of healing. 85 Several in vitro studies have shown that PRP enhances proliferation of ADSCs 39,68,[86][87][88] and it can be used as a safe and reliable alternative to standard expansion media. 89,90 This enhanced proliferative effect on ADSCs does not affect their ability to differentiate 91 and a higher volume of ADSCs within a fat graft has been shown to have a positive effect on graft take and survival.…”