LAUSR

Abstract Exposure to solar ultraviolet (UV) radiation is a major contributing factor to the increasing number of skin cancer cases. Interest has grown to use plant extracts as natural ingredients in cosmetic formulations due to their photoprotective effect, antioxidant and anti-inflammatory activity, as well as other biological activities. The aim of this study was to evaluate the biological activity of two South African plant extracts, Helichrysum odoratissimum (L.) Sweet. and Buddleja saligna Willd., and to successfully incorporate these extracts into sunscreen formulations (o/w emulsions) due to their reported biological activity. Ethanolic extracts were prepared from the leaves and stems of H. odoratissimum and B. saligna and evaluated for their antioxidant activity, mutagenic potential and antiproliferative activity against human dermal fibroblasts (MRHF). The extracts were further characterized using gas chromatography-mass spectrometry (GC-MS). Thereafter, the extracts were incorporated into separate sunscreen formulations to evaluate the in vivo dermal irritancy potential, in vivo sun protection factor, in vitro UVA protection, photostability and long term stability of the formulation, to confirm that by incorporating the extracts, the stability or photoprotective effect of the sunscreen formulation was not reduced and that these formulation were considered safe for topical application. Three separate sunscreen formulations were prepared; the base sunscreen formulation (formulation A), the base sunscreen formulation containing B. saligna (formulation B) and H. odoratissimum (formulation C) respectively. Both extracts showed significant radical scavenging activity using the 2,2-diphenyl-1-picrylhydrazyl (DPPH) assay with a fifty percent inhibitory concentration (IC50) of 5.13 ± 0.07 and 8.16 ± 0.34 µg/mL for H. odoratissimum and B. saligna respectively. No mutagenic activity was observed when the extracts were tested in the Ames assay using Salmonella typhimurium (TA98 and TA100). The PrestoBlue® cell viability assay was used to determine the antiproliferative activity of the extracts against MRHF cells, both extracts showed an IC50 value >90 µg/mL. Photoprotective activity was measured using in vivo sun protection factor (SPF) test method according to South African (SANS 1557) and International (ISO 24444) standards as well as the in vitro UVA SPF testing procedure (ISO 24443). The SPF results showed that the formulations had broad-spectrum UV protection with SPF values of 15.8±0.41, 16.1±0.66 and 16.0±0.49 and UVAPF values of 6.47±0.06, 6.45±0.06 and 6.47±0.07 for formulation A, B and C respectively. Furthermore, the formulations remained stable under normal and extreme conditions and the plant extracts did not affect the photoprotective effect of the sunscreen formulations and contributed towards the formulations stability. Additionally, each of the formulations were photostable, whereas the formulations with the addition of the extracts showed an incremental increase in photostability when compared to the base formulation. Both these extracts have been previously reported to display antiproliferative activity against skin cancer cell lines (previously published data), with an IC50 value of 31.80 ± 0.35 µg/mL (human malignant melanoma, UCT-MEL-1) for B. saligna and IC50 values of 15.50 ± 0.20 (human epidermoid carcinoma, A431) and 55.50 ± 6.60 µg/mL (human malignant melanoma, A375) for H. odoratissimum, contributing towards the medicinal benefit of using these extracts as ingredients into sunscreen formulations. Therefore, Helichrysum odoratissimum and Buddleja saligna could be considered as useful and viable additives to sunscreen formulations due to their reported biological activity.