Abstract
Photobiomodulation (PBM), as well as plant extracts of Calendula officinalis (Calen), Hypericum perforatum (Hyper), and Echinacea purpurea (Echi-p), have been used to accelerate wound healing. However, the use of homeopathic preparations of these medicinal plants, in combination with PBM, is unknown. The objective of this study was to investigate the combined wound healing potential of these therapies in vitro. Various cell models were created in vitro in commercially available human skin fibroblasts (WS1). PBM was administered using a diode laser at a wavelength of 660 nm and an energy density of 5 J/cm2. For homeopathic treatment, a 5% ethanolic complex containing Calen, Hyper, and Echi-p in 3cH potency was added to the culture medium. For combination therapy, cells were first treated with the homeopathic remedy and then irradiated. The controls included non-treated/irradiated cells and an alcohol control group. Cellular morphology was visualized 0, 24, and 48 h post-treatment. Post-treatment, cells were incubated for 48 h, and the levels of pro-inflammatory cytokines, interleukin-6 (IL-6), and tumor necrosis factor-alpha (TNF-α) were determined by ELISA, cellular viability was determined by the Trypan Blue exclusion assay and adenosine triphosphate (ATP) was determined by the luminescence assay, and cytotoxicity was determined by the lactate dehydrogenase (LDH) membrane integrity assay. Morphological changes showed that, individually, PBM and the homeopathic complex hastened cellular migration in diabetic wounded cells, and combining these therapies had an even greater effect. Cellular viability remained unchanged in both treatments, but the combination therapy increased ATP (p = 0.05). Cytotoxicity was decreased by the homeopathic complex (p = 0.01) and the combination therapy (p = 0.004). TNF-α levels remained unchanged with PBM and the homeopathic complex as independent treatment interventions but combining them increased the TNF-α levels in diabetic models (p = 0.01). PBM decreased IL-6 levels, indicating an anti-inflammatory effect (p = 0.007). These cellular changes indicated that combining PBM and a 3cH homeopathic solution of Calen, Hyper, and Echi-p is promising for treating diabetic foot ulcers.