Health Sciences Research Centre (CICS-UBI), University of Beira Interior, Av. Infante D. Henrique, 6200-506, Covilhã, Portugal; Faculty of Health Sciences, University of Beira Interior, Av. Infante D. Henrique, 6200-506, Covilhã, Portugal.
Health Sciences Research Centre (CICS-UBI), University of Beira Interior, Av. Infante D. Henrique, 6200-506, Covilhã, Portugal.
J Ethnopharmacol. 2023 Feb 10;302(Pt A):115830. doi: 10.1016/j.jep.2022.115830. Epub 2022 Oct 13.
Thymus mastichina (L.) L. (TM) and Cistus ladanifer L. (CL) are two Portuguese autochthonous species with traditional skin application in folk medicine. TM is majorly known for its antiseptic and wound healing properties, as an external anti-inflammatory agent and for its application in folk cosmetics and hygiene products. Its use in acne vulgaris has also been reported. CL is traditionally used in remedies for wounds, ulcers and other skin ailments such as psoriasis and eczema. Its application has been found useful due to its anti-inflammatory, astringent, wound healing and antiseptic properties.
With this work, we aimed to investigate relevant bioactivities related with the traditional application of TM and CL essential oils (EOs) and hydrolates (by-products of EO production) in skin ailments. Specifically their in vitro antioxidant, anti-inflammatory, cytotoxic, wound healing and antimicrobial properties were evaluated. The chemical composition of both EOs and respective hydrolates was also characterized.
Chemical characterization of EOs and hydrolates was performed by GC-FID and GC-MS. Cellular biocompatibility was evaluated using the MTT assay in macrophages (RAW 264.7) and fibroblasts (L929) cell lines. Anti-inflammatory activity was investigated by studying nitric oxide (NO) production by macrophages with Griess reagent. Wound healing potential was evaluated with the scratch-wound assay. The antioxidant potential was studied by the DPPH scavenging method. Antimicrobial activity was evaluated by broth microdilution assay against relevant microbial strains and skin pathogens, namely Staphylococcus aureus, Staphylococcus epidermidis, Cutibacterium acnes, Pseudomonas aeruginosa, Escherichia coli, Candida albicans and Aspergillus brasiliensis.
The major compounds present in TM and CL EOs were 1,8-cineole and α-pinene, respectively. 1,8-cineole and E-pinocarveol were the major compounds in the correspondent hydrolates. CL EO presented the highest anti-inflammatory potential [EC = 0.002% (v/v)], still with significant cytotoxicity [IC = 0.012% (v/v)]. TM preparations presented anti-inflammatory potential, also presenting higher biocompatibility. The same profile was present on fibroblasts regarding biocompatibility of the tested preparations. CL EO and hydrolate increased fibroblasts' migration by 155.7% and 148.4%, respectively. TM hydrolate presented a milder activity than CL hydrolate, but wound healing potential was still present, increasing cell migration by 125.1%. All preparations presented poor antioxidant capacity. CL EO presented higher antimicrobial activity, with MICs ranging from 0.06% (v/v) to 2% (v/v), against different microorganisms.
Anti-inflammatory and skin repairing potential were present for CL preparations. TM hydrolate presented an interesting biocompatible profile on both cell lines, also presenting anti-inflammatory potential. Furthermore, EOs from both species presented antimicrobial activity against a panel of different microorganisms. These in vitro bioactivities support some of their traditional skin applications, specifically regarding their antiseptic, wound healing and anti-inflammatory uses.
岩玫瑰（L.）L.（TM）和乳香（L.）是葡萄牙两种具有传统皮肤应用的本土物种，在民间医学中具有防腐和伤口愈合的特性，作为外用消炎药，并用于民间化妆品和卫生产品。也有报道称其可用于治疗寻常痤疮。CL 传统上用于治疗伤口、溃疡和其他皮肤病，如银屑病和湿疹。由于其具有抗炎、收敛、伤口愈合和抗菌特性，因此其应用被发现是有用的。
通过这项工作，我们旨在研究 TM 和 CL 精油（EOs）及其水蒸馏物（EO 生产的副产物）在皮肤疾病中的传统应用相关的相关生物活性。具体来说，评估了它们的体外抗氧化、抗炎、细胞毒性、伤口愈合和抗菌特性。还对两种精油及其相应水蒸馏物的化学成分进行了表征。
通过 GC-FID 和 GC-MS 对 EO 和水蒸馏物进行化学表征。使用 MTT 测定法在巨噬细胞（RAW 264.7）和成纤维细胞（L929）细胞系中评估细胞生物相容性。通过考马斯亮蓝法研究巨噬细胞中一氧化氮（NO）的产生来研究抗炎活性。通过划痕实验评估伤口愈合潜力。通过 DPPH 清除法研究抗氧化潜力。通过肉汤微量稀释法评估对相关微生物菌株和皮肤病原体（金黄色葡萄球菌、表皮葡萄球菌、痤疮丙酸杆菌、铜绿假单胞菌、大肠杆菌、白色念珠菌和巴西曲霉）的抗菌活性。
TM 和 CL EO 中的主要化合物分别为 1,8-桉叶素和α-蒎烯。相应的水蒸馏物中的主要化合物是 1,8-桉叶醇和 E-对伞花烃。CL EO 表现出最高的抗炎潜力[EC=0.002%（v/v）]，仍然具有显著的细胞毒性[IC=0.012%（v/v）]。TM 制剂表现出抗炎潜力，同时也具有更高的细胞相容性。对于测试的制剂，成纤维细胞也具有相同的特性。CL EO 和水蒸馏物分别使成纤维细胞迁移增加了 155.7%和 148.4%。TM 水蒸馏物的活性比 CL 水蒸馏物温和，但仍具有伤口愈合潜力，使细胞迁移增加了 125.1%。所有制剂的抗氧化能力都很差。CL EO 表现出更高的抗菌活性，MIC 范围为 0.06%（v/v）至 2%（v/v），对不同的微生物有效。
CL 制剂具有抗炎和皮肤修复潜力。TM 水蒸馏物在两种细胞系上均呈现出有趣的细胞相容性特征，同时也具有抗炎潜力。此外，两种物种的 EO 对一系列不同的微生物均具有抗菌活性。这些体外生物活性支持它们在皮肤方面的一些传统应用，特别是在防腐、伤口愈合和抗炎方面的应用。
