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Biological Activities of Frankincense Essential Oil in Human Dermal Fibroblasts

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Although frankincense essential oil (FREO¹) has become increasingly popular in skin care, research on its biological activities in human skin cells is scarce, if not completely absent. In the current study, we explored the biological activities of FREO in pre-inflamed human dermal fibroblasts by analyzing the levels of 17 important protein biomarkers pertinent to inflammation and tissue remodeling. FREO exhibited robust anti-proliferative activity in these skin cells. It also significantly inhibited collagen III, interferon gamma-induced protein 10, and intracellular cell adhesion molecule 1. We also studied its effect in regulating genome-wide gene expression. FREO robustly modulated global gene expression. Furthermore, Ingenuity® Pathway Analysis showed that FREO affected many important signaling pathways that are closely related to inflammation, immune response, and tissue remodeling. This study provides the first evidence of the biological activities of FREO in human dermal fibroblasts. Consistent with existing studies in other models, the current study suggests that FREO possesses promising potential to modulate the biological processes of inflammation and tissue remodeling in human skin.
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Biological Activities of Frankincense Essential Oil in Human Dermal Fibroblasts
Xuesheng Han, Damian Rodriguez, Tory L. Parker
PII: S2214-0085(17)30002-0
DOI: 10.1016/j.biopen.2017.01.003
Reference: BIOPEN 34
To appear in: Biochimie Open
Received Date: 12 December 2016
Revised Date: 25 January 2017
Accepted Date: 27 January 2017
Please cite this article as: X. Han, D. Rodriguez, T.L Parker, Biological Activities of Frankincense
Essential Oil in Human Dermal Fibroblasts, Biochimie Open (2017), doi: 10.1016/j.biopen.2017.01.003.
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Title: Biological Activities of Frankincense Essential Oil in Human Dermal Fibroblasts
Authors and Affiliations:
Xuesheng Han
, Damian Rodriguez, and Tory L Parker
dōTERRA International, LLC, 389 S. 1300 W. Pleasant Grove, UT 84062, USA
Corresponding author: Xuesheng Han, dōTERRA International, LLC, 389 S. 1300 W. Pleasant
Grove, UT 84062, USA. Email: lhan@doterra.com.
Running title: Effect of Frankincense essential oil in human skin cells
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Abstract
Although frankincense essential oil (FREO
1
) has become increasingly popular in skin care,
research on its biological activities in human skin cells is scarce, if not completely absent. In the
current study, we explored the biological activities of FREO in pre-inflamed human dermal
fibroblasts by analyzing the levels of 17 important protein biomarkers pertinent to
inflammation and tissue remodeling. FREO exhibited robust anti-proliferative activity in these
skin cells. It also significantly inhibited collagen III, interferon gamma-induced protein 10, and
intracellular cell adhesion molecule 1. We also studied its effect in regulating genome-wide
gene expression. FREO robustly modulated global gene expression. Furthermore, Ingenuity®
Pathway Analysis showed that FREO affected many important signaling pathways that are
closely related to inflammation, immune response, and tissue remodeling. This study provides
the first evidence of the biological activities of FREO in human dermal fibroblasts. Consistent
with existing studies in other models, the current study suggests that FREO possesses promising
potential to modulate the biological processes of inflammation and tissue remodeling in human
skin.
Keywords: inflammation; immune response; tissue remodeling; alpha-pinene; anti-
proliferation; skin health
1
FREO, frankincense essential oil
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1. Introduction
Frankincense is a resin obtained from trees of the genus Boswellia. Historically, frankincense
whole resin, its extract, and essential oil have been extensively used for a number of health
purposes in Chinese and Ayurvedic medicine. FREO has been traditionally used for its anti-
inflammatory property. Recently, FREO has become increasingly popular for promoting skin
health. However, a literature search showed no published study of the biological activities of
FREO in human skin cells.
In this study, we explored the biological activities of a commercially available FREO in
human dermal fibroblasts in vitro. We first studied the effect of FREO on the levels of 17
important biomarkers related to inflammation, immune response, and tissue remodeling in the
skin cells. Then, we studied the effect of FREO on the expression levels of 21,224 genes, using
genome-wide analysis of the same cells. The results showed that FREO was biologically active
and significantly affected expression of these biomarkers and genes.
2. Materials and Methods
All experiments were conducted in a BioMAP HDF3CGF system, a cell culture of human
dermal fibroblasts that is designed to model chronic inflammation and fibrosis in a robust and
reproducible way. The system consists of three components: a cell type, stimuli to create the
disease environment, and set of biomarker (protein) readouts to examine how treatments
affect that disease environment [1].
2.1. Cell cultures
Primary human neonatal foreskin fibroblasts (HDFn) were obtained as previously described
[2]. HDFn were plated in low serum conditions, 24-h before stimulation with cytokines. Cell
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culture and stimulation conditions for HDF3CGF assays have been described in detail
elsewhere, and were performed in a 96-well format [2,3].
2.2. Protein-based readouts
Direct ELISA was used to measure the biomarker levels of cell-associated and cell
membrane targets. Soluble factors from supernatants were quantified using HTRF® detection,
bead-based multiplex immunoassay, or capture ELISA. Overt adverse effects of the test agents
on cell proliferation and viability (i.e., cytotoxicity) were measured using SRB
2
assay. For
proliferation assays, cells were cultured and then assayed after 72 h, which was optimized for
the HDF3CGF system. Detailed information has been described elsewhere [2]. Measurements
were performed in triplicate wells. See Table S1 in Supplementary Materials for a glossary of
the biomarkers used in this study.
2.3. RNA isolation
Total RNA was isolated from cell lysates using the Zymo Quick-RNA™ MiniPrep kit (Zymo
Research Corporation, Irvine, CA), according to manufacturer’s instructions. RNA concentration
was determined using NanoDrop ND-2000 (Thermo Fisher Scientific). RNA quality was assessed
with a Bioanalyzer 2100 (Agilent Technologies, Santa Clara, CA) and an Agilent RNA 6000 Nano
Kit. All samples had an A260/A280 ratio between 1.9 and 2.1, and an RNA Integrity Number
score greater than 8.0.
2.4. Microarray analysis for genome-wide gene expression
A 0.003% (v/v) concentration of FREO was tested for its effect on expression of 21,224
genes in the HDF3CGF system after 24-h treatment. Samples for microarray analysis were
2
SRB, sulforhodamine B
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processed by Asuragen, Inc. (Austin, TX), according to the company’s standard operating
procedures. Biotin-labeled cRNA was prepared from 200 ng of total RNA with an Illumina®
TotalPrep™ RNA Amplification kit (Thermo Fisher Scientific) and one round of amplification. The
cRNA yields were quantified via UV spectroscopy, and the distribution of transcript sizes was
assessed using the Agilent Bioanalyzer 2100. Labeled cRNA (750 ng) was used to probe Illumina
Human HT-12 v4 Expression BeadChips (Illumina, Inc., San Diego, CA). Hybridizing, washing,
staining with streptavidin-conjugated Cyanine-3, and scanning of the Illumina arrays was
performed according to the manufacturer’s instructions. Illumina BeadScan software was used
to produce the data files for each array; raw data were extracted using Illumina BeadStudio
software.
Raw data were uploaded into R [3] and analyzed for quality-control metrics using the
beadarray package [4]. Data were normalized using quantile normalization [5], then re-
annotated and filtered to remove probes that were non-specific or mapped to intronic or
intragenic regions [6]. The remaining probe sets comprised the data set for the remainder of
the analysis. Fold-change expression for each value was calculated as the log
2
ratio of FREO to
vehicle control. These fold-change values were uploaded to Ingenuity® Pathway Analysis (IPA®
3
,
QIAGEN, Redwood City, CA, www.qiagen.com/ingenuity) to generate the network and pathway
analyses.
2.5. Reagents
FREO (lot number 143504A, dōTERRA International LLC, Pleasant Grove, UT, USA) was
diluted in DMSO to 8X the specified concentrations (final DMSO concentration in culture media
3
IPA, Ingenuity® Pathway Analysis
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was no more than 0.1% (v/v)); 25 µL of each 8X solution was added to the cell culture to a final
volume of 200 µL. DMSO (0.1% (v/v)) served as the vehicle control. Gas chromatography–mass
spectrometry analysis of FREO indicated that its major chemical constitutes (i.e., >5%) were
alpha-pinene (57%), limonene (8%), and caprylyl acetate (7%).
3. Results
3.1. Bioactivity profile of FREO in pre-inflamed human dermal fibroblasts
Four different concentrations (0.003, 0.001, 0.00033, and 0.00011% (v/v)) of FREO were
initially tested for biological activity in the dermal fibroblasts. None of the four concentrations
was overtly cytotoxic, and, therefore, the highest concentration (i.e., 0.003%) was analyzed
further. FREO showed significant anti-proliferative activity in dermal fibroblasts. Biomarkers
were designated if FREO values were significantly different (p < 0.05) from vehicle controls,
with an effect size of at least 10% (more than 0.05 log ratio units) (Figure 1). The level of a
tissue remodeling biomarker, collagen III, decreased in response to FREO. FREO significantly
reduced levels of
interferon gamma-induced protein 10 (
IP-10
4
) and
intracellular cell adhesion
molecule 1 (
ICAM-1
5
), both important inflammatory biomarkers. FREO also slightly lowered the
levels of PAI-I, serine proteinase inhibitor and inhibitor of tissue plasminogen activator (tPA)
and urokinase (uPA), which is involved in tissue remodeling.
4
IP-10, interferon gamma-induced protein 10
5
ICAM-1, intracellular cell adhesion molecule 1
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Figure 1. The bioactivity profile of FREO (0.003% (v/v) in DMSO) in BioMAP System HDF3CGF. X-
axis denotes protein-based biomarker readouts. Y-axis denotes the relative expression levels of
biomarkers compared to vehicle control values, in log form. Vehicle control values are shaded
in gray, denoting the 95% confidence level. A * indicates a biomarker designated with “key
activity,” i.e., biomarker values were significantly different (p < 0.05) from vehicle controls, with
an effect size of at least 10% (more than 0.05 log ratio units).
3.2. Effects of FREO on gene expression: a genome-wide total RNA expression assay
To further explore the effect of 0.003% (v/v) FREO on human skin cells, we analyzed its
effect on the RNA expression of 21,224 genes. The results show a robust effect of FREO on
regulating human genes, with many genes being upregulated and many others being
downregulated. Among the top 83 regulated genes (absolute value of the fold-change ratio of
gene expression over vehicle control ≥ 1.5) by FREO, 42 were upregulated, and 41 were
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downregulated (Table S2). IPA analysis showed that the bioactivity of FREO significantly
overlapped with many canonical pathways (Figure 2) from the literature-validated database
(IPA®, QIAGEN, Redwood City, CA, www.qiagen.com/ingenuity). Many of these signaling
pathways are closely related to the biological processes of inflammation, immune response,
and tissue remodeling in human cells. See Supplementary Materials for more details.
Figure 2. Top 20 canonical pathways matching FREO’s bioactivity profile of gene expression in
the HDF3CGF system, produced via Ingenuity® Pathway Analysis (IPA®, QIAGEN, Redwood City,
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CA, www.qiagen.com/ingenuity). Each p-value is calculated with the right-tailed Fisher's Exact
Test. The p-value measures the likelihood that the observed association between a specific
pathway and the dataset is due to random chance. The smaller p value (the bigger - log (p-
value), indicated by the black bars) the pathway has, the more significantly it matches with the
bioactivity of FREO. A ratio, indicated by each gray bar, is calculated by taking the number of
genes from the FREO dataset that participate in a canonical pathway, and dividing it by the total
number of genes in that pathway.
4. Discussion
4.1. The anti-inflammatory and immune-modulating properties of FREO
Inflammation is a protective response that involves immune cells, blood vessels, and
molecular mediators. The purpose of inflammation is to eliminate the initial cause of cell injury,
remove necrotic cells and tissues damaged from the injury and inflammatory process, and
initiate tissue repair. Chronic inflammation may lead to a variety of diseases, such as hay fever,
periodontitis, atherosclerosis, rheumatoid arthritis, and cancer [7,8].
FREO significantly reduced the levels of IP-10 and ICAM-1, important pro-inflammatory
biomarkers, suggesting its anti-inflammatory potential. Alpha-pinene, the top constituent of
FREO, is widely recognized as the major anti-inflammatory component of FREO. Gayathri et al.
showed that alpha-pinene showed anti-inflammatory properties in human peripheral blood
mononuclear cells and mouse macrophages through inhibition of tumor necrosis factor-α,
interleukin-1β, nitric oxide, and mitogen activated protein kinases [9]. An in vitro study showed
that isolated alpha-pinene had the ability to reduce the expression of pro-inflammatory
cytokines [10]. Another study found that alpha-pinene inhibited the nuclear translocation of
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NF-kB induced by
lipopolysaccharide
in THP-1 cells, explaining its benefits in the treatment of
upper and lower airway diseases [11].
Recent research has also provided evidence that alpha-pinene has some immune-enhancing
properties, particularly regarding enhanced T-cell activity. In two related studies, the effects on
human immune function of essential oils from trees were investigated [12,13]. In both studies,
it was found that exposure to alpha-pinene increased T-cell activity and decreased stress
hormone levels. Consistent with these studies, microarray results of current study showed that
FREO affected some important inflammation- and immune-related signaling pathways. Gene
expression of many cytokines and other important players in inflammation and immune
responses was significantly inhibited in pre-stimulated, inflamed skin cells, indicating that FREO
has potential immune modulating properties.
4.2. Potential roles of FREO in the wound healing process
Wound healing is an intricate process, where the skin or other body tissue repairs itself
after injury. This process is composed of several phases: blood clotting (hemostasis),
inflammation, growth of new tissue (proliferation), and remodeling of tissue (maturation) [14].
The wound healing process is not only complex, but also fragile, and it is susceptible to
disruption, leading to the formation of non-healing, chronic wounds [15]. Collagen III is
secreted by fibroblasts during the wound remodeling or repairing process, prior to the
deposition of collagen I. FREO dramatically lowered the level of collagen III, and therefore, it
would likely improve healing by reducing the chance of scar formation or wound persistence.
Additionally, the robust, anti-proliferative activity of FREO in skin cells could also contribute to
better wound healing.
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The significant, anti-proliferative activity of FREO observed in this study may have important
implications in skin and other cells. An in vitro study showed that FREO induced cell death in J82
bladder cancer cells via NRF-2-mediated oxidative stress [16]. FREO and/or its top constituent,
alpha-pinene, has also been shown to be anti-proliferative and pro-apoptotic toward several
other types of cancer cells [17–21]. Further research into FREO’s activities in cancer cells will
better our understanding of its biological mechanism of action.
5. Conclusions
To our knowledge, this was the first study of the biological activities of FREO in human
dermal fibroblasts. FREO was significantly anti-proliferative to these cells. FREO significantly
inhibited the production of collagen III, IP-10, and ICAM-1. Genome-wide gene expression
analysis showed that FREO modulated global gene expression. It also robustly affected signaling
pathways which are relevant to inflammation and tissue remodeling.
Acknowledgement
This study was funded by dōTERRA Intl (UT, USA). We would like to thank Editage (www.editage
.com) for English language editing.
Conflicts of Interest
X.H., D.R., and T.P. are employees of dōTERRA.
References
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[1] Berg EL, Yang J, Melrose J, Nguyen D, Privat S, Rosler E, et al. Chemical target and pathway
toxicity mechanisms defined in primary human cell systems. J Pharmacol Toxicol Methods
2010;61:3–15. doi:10.1016/j.vascn.2009.10.001.
[2] Bergamini G, Bell K, Shimamura S, Werner T, Cansfield A, Müller K, et al. A selective
inhibitor reveals PI3Kγ dependence of T(H)17 cell differentiation. Nat Chem Biol
2012;8:576–82. doi:10.1038/nchembio.957.
[3] R Development Core Team. R: A Language and Environment for Statistical Computing.
Vienna, Austria: The R Foundation for Statistical Computing: 2011.
[4] Dunning MJ, Smith ML, Ritchie ME, Tavaré S. beadarray: R classes and methods for
Illumina bead-based data. Bioinforma Oxf Engl 2007;23:2183–4.
doi:10.1093/bioinformatics/btm311.
[5] Bolstad BM, Irizarry RA, Astrand M, Speed TP. A comparison of normalization methods for
high density oligonucleotide array data based on variance and bias. Bioinforma Oxf Engl
2003;19:185–93.
[6] Barbosa-Morais NL, Dunning MJ, Samarajiwa SA, Darot JFJ, Ritchie ME, Lynch AG, et al. A
re-annotation pipeline for Illumina BeadArrays: improving the interpretation of gene
expression data. Nucleic Acids Res 2010;38:e17. doi:10.1093/nar/gkp942.
[7] Coussens LM, Werb Z. Inflammation and cancer. Nature 2002;420:860–7.
doi:10.1038/nature01322.
[8] Eming SA, Krieg T, Davidson JM. Inflammation in wound repair: molecular and cellular
mechanisms. J Invest Dermatol 2007;127:514–25. doi:10.1038/sj.jid.5700701.
[9] Gayathri B, Manjula N, Vinaykumar KS, Lakshmi BS, Balakrishnan A. Pure compound from
Boswellia serrata extract exhibits anti-inflammatory property in human PBMCs and mouse
macrophages through inhibition of TNFalpha, IL-1beta, NO and MAP kinases. Int
Immunopharmacol 2007;7:473–82. doi:10.1016/j.intimp.2006.12.003.
[10] Choi I-Y, Lim JH, Hwang S, Lee J-C, Cho G-S, Kim W-K. Anti-ischemic and anti-inflammatory
activity of (S)-cis-verbenol. Free Radic Res 2010;44:541–51.
doi:10.3109/10715761003667562.
[11] Zhou J, Tang F, Mao G, Bian R. Effect of alpha-pinene on nuclear translocation of NF-kappa
B in THP-1 cells. Acta Pharmacol Sin 2004;25:480–4.
[12] Li Q, Kobayashi M, Wakayama Y, Inagaki H, Katsumata M, Hirata Y, et al. Effect of
phytoncide from trees on human natural killer cell function. Int J Immunopathol
Pharmacol 2009;22:951–9.
[13] Li Q, Kobayashi M, Inagaki H, Hirata Y, Li YJ, Hirata K, et al. A day trip to a forest park
increases human natural killer activity and the expression of anti-cancer proteins in male
subjects. J Biol Regul Homeost Agents 2010;24:157–65.
[14] Stadelmann WK, Digenis AG, Tobin GR. Physiology and healing dynamics of chronic
cutaneous wounds. Am J Surg 1998;176:26S–38S.
[15] Diegelmann RF, Evans MC. Wound healing: an overview of acute, fibrotic and delayed
healing. Front Biosci 2004:283–9.
[16] Dozmorov MG, Yang Q, Wu W, Wren J, Suhail MM, Woolley CL, et al. Differential effects of
selective frankincense (Ru Xiang) essential oil versus non-selective sandalwood (Tan Xiang)
essential oil on cultured bladder cancer cells: a microarray and bioinformatics study. Chin
Med 2014;9:18. doi:10.1186/1749-8546-9-18.
MANUS CRIP T
ACCEP TED
ACCEPTED MANUSCRIPT
[17] Matsuo AL, Figueiredo CR, Arruda DC, Pereira FV, Scutti JAB, Massaoka MH, et al. α-Pinene
isolated from Schinus terebinthifolius Raddi (Anacardiaceae) induces apoptosis and
confers antimetastatic protection in a melanoma model. Biochem Biophys Res Commun
2011;411:449–54. doi:10.1016/j.bbrc.2011.06.176.
[18] Suhail MM, Wu W, Cao A, Mondalek FG, Fung K-M, Shih P-T, et al. Boswellia sacra
essential oil induces tumor cell-specific apoptosis and suppresses tumor aggressiveness in
cultured human breast cancer cells. BMC Complement Altern Med 2011;11:129.
doi:10.1186/1472-6882-11-129.
[19] Kusuhara M, Urakami K, Masuda Y, Zangiacomi V, Ishii H, Tai S, et al. Fragrant environment
with α-pinene decreases tumor growth in mice. Biomed Res Tokyo Jpn 2012;33:57–61.
[20] Ni X, Suhail MM, Yang Q, Cao A, Fung K-M, Postier RG, et al. Frankincense essential oil
prepared from hydrodistillation of Boswellia sacra gum resins induces human pancreatic
cancer cell death in cultures and in a xenograft murine model. BMC Complement Altern
Med 2012;12:253. doi:10.1186/1472-6882-12-253.
[21] Chen W, Liu Y, Li M, Mao J, Zhang L, Huang R, et al. Anti-tumor effect of α-pinene on
human hepatoma cell lines through inducing G2/M cell cycle arrest. J Pharmacol Sci
2015;127:332–8. doi:10.1016/j.jphs.2015.01.008.
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Please consider as potential referees:
Zhenhua Liu, Ph.D.
Email: zliu@nutrition.umass.edu
Dr. Liu is an expert in the research of health benefits of natural products, and their impact on
molecular pathways such as Wnt-pathway.
Sreejayan Nair, Ph.D.
Email: Sreejay@uwyo.edu
Dr. Nair is an expert in studying the health benefits of natural compounds, including essential
oils.
Runzhi Lai, Ph.D.
Email:
r.lai@utah.edu
Dr. Lai is an expert in molecular biology and biochemistry.
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Frankincense essential oil
(FREO) was anti-proliferative to human dermal fibroblasts.
FREO significantly inhibited collagen III, interferon gamma-induced protein 10, and
intracellular cell adhesion molecule 1.
FREO robustly modulated global gene expression.
FREO affected many important signaling pathways that are closely related to
inflammation, immune response, and tissue remodeling.

Supplementary resource (1)

... Frankincense has been extensively used for medical purposes, for thousands of years, in traditional Islamic, Chinese and Indian medicine [6,10], in addition to its use as chewing gum, incense or in perfumery [11]. Frankincense products are available as resins, extracts and essential oils. ...
... Various frankincense preparations are marketed as dietary supplements as well as their wide use as valuable ingredients in skin care products [7,11]. Pharmacological research highlighted myriad biological activities of frankincense essential oil (FEO) including antimicrobial, anti-inflammatory, analgesic, anticancer and immunomodulatory activities [6,10,[12][13][14][15]. ...
... Its anti-inflammatory effect has been demonstrated via inhibiting the transactivation of NF-kB through the stabilization of I k Bα [16]. Besides, the oil possessed a promising potential to modulate the biological processes of inflammation and tissue remodeling in human skin [10]. Additionally, FEO-loaded hydrogel wound dressings demonstrated antimicrobial and wound-healing efficacy in rats [17]. ...
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Frankincense oil has gained increased popularity in skin care, yet its anti-aging effect remains unclear. The current study aimed to investigate the anti-photoaging effect of frankincense (Boswellia papyrifera (Del.) Hochst., Family Burseraceae) essential oil in an in vivo model. The oil was initially extracted by two methods: hydro-distillation (HD) and microwave-assisted hydro-distillation (MAHD). GC/MS analysis revealed the dominance of n-octyl acetate, along with other marker compounds of B. papyrifera including octanol and diterpene components (verticilla 4(20) 7, 11-triene and incensole acetate). Thereafter, preliminary investigation of the anti-collagenase and anti-elastase activities of the extracted oils revealed the superior anti-aging effect of HD-extracted oil (FO), comparable to epigallocatechin gallate. FO was subsequently formulated into solid lipid nanoparticles (FO-SLNs) via high shear homogenization to improve its solubility and skin penetration characteristics prior to in vivo testing. The optimimal formulation prepared with 0.5% FO, and 4% Tween® 80, demonstrated nanosized spherical particles with high entrapment efficiency percentage and sustained release for 8 hours. The anti-photoaging effect of FO and FO-SLNs was then evaluated in UVB-irradiated hairless rats, compared to Vitamin A palmitate as a positive standard. FO and FO-SLNs restored the antioxidant capacity (SOD and CAT) and prohibited inflammatory markers (IL6, NFκB p65) in UVB-irradiated rats via downregulation of MAPK (pERK, pJNK, and pp38) and PI3K/AKT signaling pathways, alongside upregulating TGF-β expression. Subsequently, our treatments induced Procollagen I synthesis and downregulation of MMPs (MMP1, MMP9), where FO-SLNs exhibited superior anti-photoaging effect, compared to FO and Vitamin A, highlighting the use of SLNs as a promising nanocarrier for FO. In particular, FO-SLNs revealed normal epidermal and dermal histological structures, protected against UVβ-induced epidermal thickness and dermal collagen degradation. Our results indicated the potential use of FO-SLNs as a promising topical anti-aging therapy.
... Studies suggest that FEO enhances the proliferation phase by stimulating fibroblast activity and encouraging granulation tissue formation [13]. Additionally, it aids in the remodeling phase by promoting collagen synthesis, improving the tensile strength of the wound, and minimizing scarring. ...
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... However, the cellular and molecular mechanisms involved in the decrease of intracellular infection by P. gingivalis in human GECs had not yet been examined. A previous study showed that frankincense essential oil robustly modulated genome-wide gene expression in human dermal fibroblasts [58]. Given that frankincense may affect the expression of several genes in host cells, we hypothesize that B. serrata extracts may modulate the expression of host surface receptors involved in the recognition, uptake, and immune responses against P. gingivalis. ...
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Frankincense is produced by Boswellia trees, which can be found throughout the Middle East and parts of Africa and Asia. Boswellia serrata extract has been shown to have anti-cancer, anti-inflammatory, and antimicrobial effects. Periodontitis is an oral chronic inflammatory disease that affects nearly half of the US population. We investigated the antimicrobial effects of B. serrata extract on two oral pathogens associated with periodontitis. Using the minimum inhibitory concentration and crystal violet staining methods, we demonstrated that Porphyromonas gingivalis growth and biofilm formation were impaired by treatment with B. serrata extracts. However, the effects on Fusobacterium nucleatum growth and biofilm formation were not significant. Using quantification of colony-forming units and microscopy techniques, we also showed that concentrations of B. serrata that were not toxic for host cells decreased intracellular P. gingivalis infection in human gingival epithelial cells. Our results show antimicrobial activity of a natural product extracted from Boswellia trees (B. serrata) against periodontopathogens. Thus, B. serrata has the potential for preventing and/or treating periodontal diseases. Future studies will identify the molecular components of B. serrata extracts responsible for the beneficial effects.
... Frankincense, a splendidly organic resin extracted from the majestic Boswellia trees of the esteemed Burseraceae family (Han et al., 2017), emanates from none other than the illustrious Boswellia carterii (B. carterii), which thrives in the arid and lofty realms of East Africa, China, and India (Frank et al., 2009). ...
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