An Innovative, Centrifugation-free Method to Prepare Human Platelet Mediator Concentrates Showing Activities Comparable to Platelet-rich Plasma
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Index: WOUNDS 2011;23(6):171–182
Abstract: Slow-healing wounds benefit considerably from treatment with platelet-rich plasma (PrP). The drawback of using PrP is its laborious preparation, which requires expensive technical equipment (centrifuges) and well-trained personnel. Methods. The authors’ new method overcomes these issues and provides the practitioner with an innovative tool to freshly prepare a platelet mediator mix with PrP’s known biological activities, but is much simpler to obtain. This is achieved by employing the sedimentation of a blood sample at regular gravity (no centrifuge necessary) in the presence of an anti-coagulant and a sedimentation accelerator. Thereafter, the supernatant containing the platelets is concentrated on a unique filter, which causes these platelets to release their mediators (different biologically active molecules resembling the substance mix that is released by the platelets upon degranulation). This solution is eluted from the filter, providing a sterile-filtered, enriched fraction of biologically active mediators (TGF-β, PDGF, IGF-1, etc.), most of which are active in wound healing disorders. Results. This preparation triggers in-vitro proliferation of fibroblasts and osteoblasts, the secretion of IL-6 by osteoblasts, and differentiation of fibroblasts into cells with an endothelial morphology resembling cells during angiogenesis. Conclusion. By providing the practioner a sterile concentrate of a whole range of autologous platelet mediators within 1 hour, this new method has the potency to become a substitute of PrP in wound-healing therapy. PMC (platelet mediator concentrate) eases the manufacturing of such preparations, thereby making them not only more widely applicable, but also reducing treatment costs.
Accelerating the healing of chronic wounds is a major goal in human medicine. Major advances have been made in the last 20 years, most of which deal with the development of new types of wound dressing materials.1
Approaches based on the insights modern molecular biology have also provided insights regarding cell-cell communication between the different factors collaborating to correct tissue damage (local tissue cells and the immune system). The introduction of individual recombinant human cytokines, such as PDGF, is a more recent addition to the spectrum of wound healing therapy.2,3 Although the concept of employing genetically engineered preparations is appealing, especially considering their highly defined nature (manufactured under GMP, standardized biological activity, reproducible lot-to-lot quality), the use of either one of these cytokines alone generated disappointing results compared to mixtures of these.4 The latter finding can easily be explained by findings of groups that employed broader genomic or proteomic analyses to elucidate the complexity of the inter-cellular regulatory network that is established during wound healing.5 Therefore, it is evident that neither chronic healing is caused by the lack of a single cytokine, nor will a successful therapy be possible by applying just one of those cytokines, which are central to appropriate wound healing. Yet, an even greater obstacle to broader use of recombinant human mediators in wound healing is the dramatic increase in cost of treatment that usually results.6
One of the first attempts to supply such chronic wounds with more complex, physiological preparations was based on the preparation a blood platelet concentrate, usually by differential centrifugation, now known as platelet-rich plasma (PrP).