Abstract:
Platelet-rich plasma (PRP) is the cell free plasma, which has an enriched concentration of platelets
and clotting factors with the ability to enhance the natural healing process. PRP is often used by
physicians in an office setting to accelerate the healing of a variety of sports related injuries, chronic
wounds and enhance skin rejuvenation. PRP mimics the wound healing cascade by enhancing the
recruitment, proliferation and differentiation of cells involved in tissue regeneration. Although PRP is
used to enhance healing, the efficacy thereof is debated as no clear-cut set of parameters is available
that device manufacturers and protocols should follow. The lack of uniformity in the PRP preparation
methods results in differing PRP volume, platelet contents and unavoidably platelet-derived growth
factors. Therefore, the aim of this study was to develop a simple and rapid method for preparing
autologous PRP in an office setting using a tabletop centrifuge for point-of-care use. The simplified preparation procedure involved a single centrifugation step of 18 ml of whole blood, which sufficiently
enriched the platelet content in the PRP fraction. As activated platelets express and release growth
factors and cytokines that mediate the different phases of the wound healing cascade, the extracted
PRP fraction was activated with an ethanol, calcium chloride (CaCl2) and platelet poor plasma (PPP)
preparation in glass containers, without the collection of additional blood as required in some
protocols. The activated PRP formed a fibrin clot, trapping the degranulating platelets and its released
growth factors. The concentration of TGF- 1 obtained from the fibrin clot was 45.49 ± 3.80 ng/ml, in
range with the available literature. During the in vitro studies, the extracted PRP by the developed
method was able to significantly induce cell proliferation in a dose dependent manner. Cells
enumerated with the crystal violet assay indicated that the cells treaded with 5% or 10% PRP
significantly increased the percentage of viable cells to 165-176% and 156-158%, when compared to
the positive controls. Cells enumerated with the MTT-assay indicated that the cells treaded with 5% or
10% PRP increased the percentage of viable cells to 79-91% and 87-105% which is comparable to
that of the positive control. Data from the cellular proliferation assays indicate that sufficient plateletderived
growth factors had been obtained with the preparation procedure. Furthermore, data from the
in vivo studies indicated that the extracted PRP was able to augment soft tissue regeneration and
bone formation. Treatment with the activated PRP resulted in symptom reduction and accelerated
healing of various injuries. The simplified preparation and the use of the provided study product packaged in a kit developed during this study will enable physicians to easily obtain autologous PRP,
in an office setting for point-of-care use, with the ability to induce tissue regeneration.
