64 ANTI-AGEING


and a loss of ECM-synthesizing capacity in these cells’ concomitant with a phenotype of matrix resorption.12,14 Some studies illustrate the importance of measuring collagenase, elastase, and tyrosinase activities in cosmetic formulations to evaluate the potential anti-ageing effects of ingredients. Given that lipoxygenase is involved in the synthesis of inflammatory mediators (leukotrienes), its inhibition by these natural compounds directly targets a mechanism of ‘inflammaging’, which is relevant for soothing and anti-ageing formulations. The evaluation of the antioxidant and anti-


lipoxygenase (LOX) activity of several essential oils and scientifically reinforces the cosmetic value of LOX as a marker of anti-inflammatory activity in natural botanical active ingredient formulations.7,13,16


Natural cosmetic market Natural extract cosmetics are highly favoured by consumers for their safety and efficacy, aligning with their preferences for natural, organic, efficient and sustainable skin care concepts. The market for natural anti-ageing products is also shaped by strong sociological trends. Consumers increasingly associate ageing gracefully with wellness, vitality, and self-care rather than hiding age. There is a growing demand for authenticity and


transparency, with people favouring botanical, plant-based, and sustainably sourced ingredients. Social trends emphasizing clean beauty and reduced chemical exposure drive the shift toward natural solutions. Trust and credibility are critical: consumers


expect scientific validation of efficacy while avoiding synthetic or controversial additives. Generational differences also play a role—younger consumers adopt prevention-oriented routines early, while older consumers seek gentle but effective natural alternatives.2


Dual in vitro study As skin ageing is a multifactorial process, involving both the degradation of the extracellular matrix and the progressive decline in the synthesis of structural proteins. Angelica Root Essential from Natix was evaluated through two complementary approaches: ■ In vitro enzymatic inhibition assays, assessing its ability to inhibit key enzymes involved in skin ageing. ■ Cell-based studies on human dermal fibroblasts (NHDFs), to determine its capacity to stimulate the synthesis of structural proteins (pro- collagen I and elastin).


Enzymatic inhibition: strong firming and soothing potential Experimental conditions Spectrophotometric assays were performed to quantify the inhibition of enzymes known to drive skin ageing: Inhibition is measured and quantified by spectrophotometry and expressed as a percentage. For each test, inhibitory activity is compared to positive controls: pure molecule (30 ppm) and cosmetic active QC (500 ppm).


PERSONAL CARE MAGAZINE January 2026 COL1A1 (ELISA) - Standard curve (48h) 2.0 1.5 1.0 0.5


Y = 0.0007557*X + 0.03624 R2 = 0.9989


0 0 500 1000 1500 COL1A1 (pg/mL)


Figure 2: Collagen type 1 alpha 1 standard curve. Non-linear regression model of the absorbance values obtained by the serial dilutions of collagen type I standard, provided by the manufacturer of Human Pro-Collagen I alpha 1 ELISA Kit subjected to the indicated protocol


These findings point to two complementary mechanisms of action that are highly relevant for skin health. First, the strong inhibition of elastase demonstrates the ingredient’s ability to protect the skin’s structural proteins, supporting firmness and visibly tightening the skin. Second, the pronounced inhibition of


lipoxygenase indicates a capacity to reduce inflammatory pathways, thereby calming irritation and discomfort. Together, these effects contribute to both improved skin resilience and comfort.


Stimulation of key proteins: collagen and elastin To complement the enzymatic approach, the effects of Angelica root essential oil were studied on normal human dermal fibroblasts (NHDFs). Using NHDFs in efficacy testing provides a biologically relevant model that closely mimics human skin physiology, allowing assessment of the effects of cosmetic ingredients on cellular processes, such as ECM (extra-cellular matrix) protein synthesis, wound healing, and anti-ageing properties, among others. This approach enhances the accuracy and


relevance of in vitro testing, offering insights into how products may perform on real human skin, while also aligning with ethical and regulatory requirements that favour human-relevant, non- animal testing methods.


Experimental conditions NHDFs were treated with the Angelica root essential oil at the non-cytotoxic dose 0.001%, for 48 hours. Controls cells without Angelica root essential oil were also tested. After the incubation period, cell culture


supernatants were harvested. The cell-depleted supernatants were then used to measure the levels of secreted collagen I and elastin using the Human Pro-Collagen I alpha 1 ELISA Kit and Human Elastin ELISA Kit (Abcam). The obtained concentration values of collagen I and elastin were corrected versus cell viability


in the corresponding wells, estimated by the reduction of MTT to MTT-formazan solubilized in DMSO (OD 570 nm). This correction was applied to reflect the amount of secreted protein per viable cell, so that slightly cytotoxic or proliferative effects of the products, as well as differences in cell seeding between wells do not mask the real effect of the tested product on the parameters studied. The ELISA method allows to calculate protein


concentration through interpolation using a standard curve made by dilutions of known concentrations of the specific protein in the sample diluent, provided by the kit’s manufacturer. The raw levels of collagen I (pg/mL) and elastin


(ng/mL) were normalized to the raw levels of cell viability (MTT, OD 570 nm) for each technical replicate. Afterwards, protein levels, cell viability, and protein levels relative to cell viability were normalized and graphically represented (%) in bar graphs, as mean ± standard error of the mean (SEM). Protein levels are represented as data


normalized versus untreated control, to show the capacity of the samples to induce the synthesis of each specific ECM component.


Results Cell viability The treatment with Angelica oil at 0.001% during 48 hours in NHDFs did not show any significant effects on cell viability, compared to the untreated control. Secreted proteins normalized to cell viability:


results indicated the treatment with Angelica oil during 48 hours at 0.001% provoked a statistically significant improvement on pro-collagen type I alpha 1 by 16.7%, together with a non-significant increase in elastin synthesis by 20.7%, compared to the untreated control.


Conclusion


These findings demonstrate the oil’s ability to sustain the synthesis of major dermal structural proteins, essential for skin firmness and elasticity.


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