SKIN CARE Botanical Co-Extraction Inhibition of LOXL1-3 Expression
200 180 160 140 120 100 80 60 40 20 0
-20 1 ng /ml TGFβ1 0.1 Concentration of Active (% v/v)
Figure 4: Induction of LOXL 1-3. Normal human dermal fibroblasts (NHDF) were treated with the botanical co-extracted material for three days after which the cells were harvested, lysed, and examined by Western blot probing with antibodies specific for LOXL enzymes 1, 2 and 3. Band intensity was quantified using Image J software
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
140 120 100 80 60 40 20 0
-20
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). 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
Inhibition of Inflammatory Cytokines Botanical Co-Extraction
*** *** ** *** *** ** *** *** *** *** *** **
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
0.0003% (v/v) ■ 0.003% (v/v) ■ 0.03% (v/v) ■ *** *** 0.2 0.3 LOXL1 ■ LOXL2 ■ LOXL3 ■
59
GM-CSF
IL-6
IL-8
IP-10
MCP-1
MCP-2
MIP-1β
RANTES **p<0.01 ***p<0.001
Figure 5: Inhibition of 8 different inflammatory mediators. The supernatants from the cell cultures of normal human keratinocytes stimulated with IFN-g/MDP in the presence/absence of the botanical co-extract were analyzed by ELISA to detect and quantify any changes in the levels of different inflammatory molecules
www.personalcaremagazine.com September 2025 PERSONAL CARE
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