A preliminary study on the potential of manuka honey and platelet-rich plasma in wound healing

Abstract

Aim. The purpose of this study was to determine the in vitro response of cells critical to the wound healing process in culture media supplemented with a lyophilized preparation rich in growth factors (PRGF) and Manuka honey. Materials and Methods. This study utilized cell culture media supplemented with PRGF, as well as whole Manuka honey and the medical-grade Medihoney (MH), a Manuka honey product. The response of human fibroblasts (hDF), macrophages, and endothelial cells (hPMEC) was evaluated, with respect to cell proliferation, chemotaxis, collagen matrix production, and angiogenic potential, when subjected to culture with media containing PRGF, MH, Manuka honey, and a combination of PRGF and MH. Results. All three cell types demonstrated increases in cellular activity in the presence of PRGF, with further increases in activity seen in the presence of PRGF+MH. hDFs proved to be the most positively responsive cells, as they experienced enhanced proliferation, collagen matrix production, and migration into an in vitro wound healing model with the PRGF+MH-supplemented media. Conclusion. This preliminary in vitro study is the first to evaluate the combination of PRGF and Manuka honey, two products with the potential to increase regeneration individually, as a combined product to enhance dermal regeneration.

Figure 1

(a) Fluorescence results from TGF-β western blot for acid- and honey-activated PRP. All ratios are PRP : MH by volume. (b) Quantification of fluorescence from TGF-β Western blot.

Figure 2

Results of mean hDF (a), hPMEC (b), and macrophage (c) proliferation from MTS assays at days 1, 4, and 7. Manuka honey results are not shown for hDF and hPMEC proliferation as they were not statistically different from MH.

Figure 3

Results of the macrophage inflammation response studies, with mean TNF-α (a), IL-10 (b), and VEGF (c) from days 1, 4, and 7. Dashed lines indicate the minimum levels of detection of 0.031, 0.005, and 0.023 ng/mL for the TNF-α IL-10, and VEGF ELISAs, respectively.

Figure 4

Results of macrophage (a) and hPMEC (b) chemotaxis from MTS assay quantification in both the top insert and bottom well. The results of cell chemotaxis from the addition of Manuka honey are not included as they were not significantly different from those of MH. Higher honey concentrations (5, 10, and 20% v/v) were excluded as they resulted in cell death.

Figure 5

Light microscopy images of the in vitro wound healing assay taken at time 0, 18, and 30 hr after wounding (a). Graph of the mean % area healed for each of the test media at 6, 12, 18, and 30 hr after wounding (b).

Figure 6

Results of the hydroxyproline assay performed on retained media on days 1, 7, 14, 21, and 28.

Figure 7

Representative light microscopy images of the in vitro bead angiogenesis assay performed with hPMECs cultured on Cytodex beads. The 0.1% MH, 0.1 mg/mL PRGF, 1 mg/mL PRGF, PRGF+MH, and control are shown on days 1, 4, and 6 as those test medias resulted in maximum sprout formation (a). Mean sprout densities (b, left) and mean sprout lengths (b, right) are shown for MH, PRGF, and control medias. The results of Manuka honey supplemented media are not included as they were not significantly different from those of MH. Higher honey concentrations (5, 10, and 20% v/v) were excluded from the sprout density and sprout length graphs as they resulted in cell death.