Industry News, Cosmetics & Persnoal Cares
Botanical extract for skinlongevity and resilience
Biocogent describes a dual botanical extract that ameliorates the negative impacts of at least four distinct hallmarks of ageing: chronic inflammation, (inflammaging), altered intercellular communication, epigenetic alterations, and diminished proteostasis. These benefits appear to be promulgated by the material’s activity focusing on the dermal-epidermal junction
Modern cosmetic scientists have expended considerable time and effort to elucidate why some people may visibly age quicker or slower than others despite being the same age chronologically. These investigations have built towards an understanding of the collective phenomena responsible for ‘biological ageing’, the cumulative effects of which either accelerate the deterioration or maintain the resiliency of youthful skin.
Colloquially speaking, living forever is not sufficient for current consumers unless they also “age well”. For that desire to be met, cosmetic care active ingredients must both prevent the onset of the physical features of ageing as well as and restore and sustain skin resiliency.
The design, development, and launch of active materials that address such lofty goals continues to be an extremely complicated endeavour requiring a comprehensive understanding of why skin loses its elasticity and strength over time, why hyperpigmentation develops, and of course, why wrinkles form. Many scientific studies have illustrated that the prominent fibres of the extracellular matrix (ECM), such as collagen and elastin, diminish in abundance as we get older.1
Increasing evidence supports that the dermal-epidermal junction (DEJ) is a critical interface at which many of the factors involved in sustaining skin elasticity and longevity converge.2,3 At this site, the epidermis (outer layer of the skin) and dermis (deeper connective tissue layer) physically interact through a meshwork-like basement membrane that precisely coordinates the organized arrangement of proteins, glycoproteins, and cells.
Indeed, this is the site where the dermis and epidermis are mechanically anchored to each other, which facilitates barrier function, nutrient exchange, and signal transduction between cell types. Furthermore, the DEJ represents the niche for epidermal stem cells. It is therefore no surprise that thinning of the DEJ with age leads to reductions in the mechanical and functional resiliency of the skin.
The hallmarks of ageing
Twelve ‘Hallmarks of Ageing’ have been established to explain the observed effects of ‘biological ageing’ relative to simple ‘chronological ageing.
These so-called hallmarks currently include, but potentially are not limited to: 1) dysbiosis of the skin microbiome; 2) chronic low-level inflammation (‘inflammaging’); 3) alterations in intercellular communication; 4) exhaustion of local stem cell populations; 5) increased incidence of cellular senescence; 6) accumulating mitochondrial dysfunction; 7) dysregulated nutritional sensing; 8) disabled macroautophagy; 9) diminished proteostasis; 10) epigenetic alterations; 11) attrition of the telomeres; and 12) cumulative genomic instability (Figure 1).
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These all play a role in how we differentiate chronological ageing from biological ageing, and explain why some people appear to be ageing better than others. These discoveries have provided the impetus for the development of active ingredients that can stymie if not fully reverse the negative impacts of these various ageing factors.4
However, none of these trends are singularly responsible for biological ageing, but collectively can accelerate the ageing process. Many of the existing solutions developed have addressed these damages individually but there are continued efforts to find countermeasures that are effective at more than one aspect of the ageing process.
Yet imagine the rejuvenating potential of a substance that could assert influence over multiple facets of the ’12 Hallmarks of Ageing’. Herein, we describe in vitro, ex vivo, and in vivo evidence of a novel plant-based ingredient that functions as a countermeasure to no less than four of these hallmarks.
Botanical solutions
In addition to devising clever means of countering the facets of ageing, the modern consumer has shown a preference for naturally derived active ingredients. Thus, finding ameliorative substances derived from botanical sources that can address the desires of well-ageing and skin longevity are highly sought after.
Here, we introduce a novel ingredient that is co-extracted from two different plant sources: Rumex acetosella and Pinus massoniana (Figure 2) that synergize in a combinatorial extract to impact multiple features of skin health.
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R. acetosella, more commonly known as field sorrel or red sorrel, has been established in the archives of herbal medicine to have several health benefits. Native to multiple regions of the northern hemisphere, reports suggest that extracts of this plant can alleviate conditions as varied as skin erythema (redness), eczema, and psoriasis.5
Furthermore, antimicrobial activity has been attributed to topical mixtures containing R. acetosella potentially suggesting the capacity for microbiome modulation that could address the ageing hallmark of microbial dysbiosis.
As with R. acetosella, P. massoniana also has a storied history detailing multiple ameliorative impacts. Frequently called ‘Masson’s Pine’, the beneficial health effects of extracts of this plant have been chronicled in traditional Chinese medicine (TCM). Indigenous to much of the Asian continent, TCM suggests that solutions infused with Masson’s Pine exert antioxidant, anti-inflammatory, and anti-diabetic activities.6,7
The combination of such distinctive properties from these two botanical sources promises a truly powerful active ingredient.
Through a proprietary high-yield extraction process, the two botanical sources were co-extracted into a material containing approximately 59 compounds that were consistently liberated from batch-to-batch. The mass spectrometry chromatograms that detected this panoply of molecules became the signature ‘fingerprint’ of the co-extract of R. acetosella and P. massoniana. These compounds included a variety of phytochemicals that are unable to be synthesized commercially. Truly mother nature at her finest.
Pathway to longevity — in vitro and ex vivo EVIDENCE
R. acetosella and P. massoniana have both been investigated individually, but here we explored the possibility of synergizing their activity in a combined co-extracted material. The botanical co-extraction product was tested on monolayers of normal human dermal fibroblasts (NHDF cells). The cells were treated with the material at various concentrations over the course of three days.
At that time, cells were harvested, lysed, and analysed by Western blot probing for changes in the abundance of proteins considered to be critical to skin resiliency such as the much discussed collagen fibres.8
The first finding presented from the in vitro experiments showed a boost in the expression of two specific collagen genes in skin: COL3A1 and COL4A1 (Figure 3A). Type III collagen (COL3A1) represents a fibrillar collagen that coalesces with type I collagen fibres to construct a formidable lattice work that buttresses the skin against physical insults.
Expression of the COL3A1 gene diminishes over an individual’s life span, which is often accelerated from extended UV exposure. In contrast, the type IV collagen gene (COL4A1) product is not fibrillar in nature, but forms a supportive meshwork at the basement membrane of the DEJ.
From these early results, we considered that this material could potentially reverse loss of proteostasis, one of the key hallmarks of ageing where the normal homeostatic levels of key protein factors are disrupted to the point of inducing structural instability in the ECM and at the DEJ. To that end, the relative abundance of other critical protein molecules was tested in similar fashion.
Similar to the collagen protein levels, there was also a dose-dependent increase in elastin (ELN) protein expression (Figure 3B). As their name suggests, these proteins are vital contributors to the elasticity of the skin, allowing for it to be stretched and return to its original shape much like the action of a rubber band.9
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They are first secreted from dermal fibroblast cells in the form of soluble tropoelastin that is subsequently crosslinked by lysyl oxidase enzymes into insoluble, durable fibre complexes. The induction of their synthesis represents a critical step to restoring and maintaining skin resiliency to frequent stretching and recoiling.
In addition to boosting important fibre proteins in the ECM, the co-extraction product augmented the expression of three enzymes belonging to the lysyl oxidase like (LOXL) family of enzymes: LOXL 1, 2, and 3 (Figure 4). These enzymes are responsible for crosslinking both collagen and elastin fibres, which is an essential activity for sustaining the structural and tensile strength of skin tissue.
Additionally, it has been observed that LOXL levels diminish with age, contributing to increases in skin fragility and wrinkle formation.10,11 Thus, a boost in their expression levels alongside the augmented levels of collagen and elastin will collectively enhance youthful features of skin.
Subsequently, a cellular-based examination of the anti-inflammatory potential of this material was examined. Normal human keratinocytes were stimulated with interferon-gamma (IFN-γ) and muramyl dipeptide (MDP) to induce an inflammatory response promulgated by several skin-relevant inflammatory mediators.
The supernatants from these cell culture reactions were collected and analysed by ELISA to quantify the concentrations of the different inflammatory molecules.
Relative to controls, the IFN-γ/MDP stimulus was co-administered with varying concentrations of the dual extract to explore if this material could possibly function as a counteragent to chronic inflammation, another hallmark of ageing.12 In this situation, consistent low-level inflammation gradually erodes the integrity of skin tissue — a phenomenon now referred to as “inflammaging”.
The outcomes observed with this experiment were nothing less than stunning: the botanical co-extracted material down-regulated the expression levels of at least eight different inflammatory mediators, including: interleukin-6 (IL-6), IL-8, granulocyte-macrophage colony-stimulating factor (GM-CSF), IFN-γ induced protein-10 (IP-10), monocyte chemoattractant protein-1 (MCP-1), MCP-2, macrophage inflammatory protein-1 beta (MIP-1β), and regulated activation normal T-cell expressed and secreted (RANTES) factor (Figure 5).
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Follow-up studies also confirmed dose-dependent inhibition of both phospholipase A2 (PLA2) and cyclooxygenase 2 (COX-2). These results confirmed that the material could indeed be considered as remedy for skin inflammaging conditions and would likely have a soothing effect.
The exciting results generated from the different in vitro studies led to an ex vivo investigation using human skin explants. Similar to the in vitro research, the dual botanical co-extract was incubated at various doses on three-dimensional tissue for approximately nine days.
After which, RNA was harvested and purified from tissue lysates that was then subject to reverse transcriptase quantitative polymerase chain reaction (or what is commonly called RT-qPCR) against a panel of skin-relevant genes. The results corroborated much of the prior work that had been done with similar genes being up-regulated in their expression (data not shown).
This also detected additional factors important to the ECM and DEJ, including other collagens and crosslinking factors as well as tissue remodeling proteins. This reinforcing data also suggested that the dual botanical extract could address another hallmark of ageing: epigenetic alterations.
Pathway to longevity — in vivo evidence
The accumulated data from both the in vitro and ex vivo investigations supported the commissioning of a clinical study to evaluate the benefits of a finished formulation containing the botanical co-extracted material at 3% (or a placebo) on a panel of 60 participants aged 30-65 that applied it to their face twice daily for 12 weeks.
At baseline, four weeks, eight weeks, and 12 weeks, the participants were examined using Elastimeter and Cutometer instruments that quantitatively assessed their skin elasticity and firmness, respectively.
Both the Elastimeter and Cutometer results demonstrated a trendline of continual improvement with the co-extract starting at the four-week timepoint that was sustained through to the conclusion of the clinical at 12 weeks (Figure 6).
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The instrument data was further supported by expert grading that showed progressive increases in skin smoothness, texture, and visible firmness with concomitant reductions in forehead lines, crow’s feet, under eye lines, and glabella lines.
Unexpectedly, there was an observed recuperation in overall skin tone. These macroscopic augmentations in youthful skin appearance are highlighted in the participant panel images shown below (Figure 7).
Conclusion
Discovering natural solutions to act as countermeasures to multiple facets of biological ageing is a significant undertaking that sometimes results in truly exciting findings. Furthermore, it is exceedingly rare to identify planted-based substances that are able to not only address one but many aspects of the skin ageing process.
Herein, we have elaborated upon a dual botanical extract that ameliorates the negative impacts of at least four distinct hallmarks of ageing: chronic inflammation (inflammaging), altered intercellular communication, epigenetic alterations, and diminished proteostasis. These benefits appear to be promulgated by the material’s activity focusing on the DEJ, one of the most critical interfaces in the skin architecture.
The combined laboratory and clinical data strongly supports that this combinatorial substance will assert a rejuvenating impact on skin tissue, with increased elasticity and resiliency.