SPORT MASSAGE OIL

OZONIZED OIL EVOO + CBD

RELAX · REVITALIZE · RECOVER · HYDRATE

Relaxing and revitalizing muscle oil with

EVOO ozonated oil + CBD
It promotes relaxation and muscle rest, providing vitality
and contributing to muscle recovery.

SCIENTIFIC ENDORSEMENT: The tests carried out suggest that the

use of ozonated oil during sports massage increases the elimination of
lactate in the blood, improves performance and reduces the perception
of fatigue in cyclists in 3 Wingate anaerobic test. Anti-
inflammatory.

RECOVERED MOISTURIZER NO GREASY FINISH ALL SKIN TYPES LAB TESTED CRUELTY FREE + VEGAN

INSTRUCTIONS FOR USE: Apply to the skin with an intense massage,

to help with muscle relaxation and rest, until completely absorbed.
Producing a cold effect, followed by a heat effect, after a few minutes. Vegan product, free of
ingredients of animal origin
WARNINGS: Store in a cool, dry place, away from sunlight. Tested Product
Keep out of the reach of children. Topical use. Avoid contact with eyes. Dermatologically

INGREDIENTS:
*OZONIZED OLIVE OIL,
**PRUNUS AMYGDALUS
DULCIS OIL, ***MENTHOL,
****LAVENDULA
ANGUSTIFOLIA OIL,
LINALOOL,
*****CANNABIDIOL,
GLYCINE SOY OIL,
TOCOPHEROL,
BETA-SITOSTEROL,
LIMONENE, SQUALENE,
GERANIOL
*90% Organic Extra Virgin Olive Oil ozonated PeroxiBiokey® Plug&Play (600IP). Ingredient manufactured cold, respecting the molecular structure of fatty acids, contains
cellularly available oxygen that activates the skin's energy. ENERGIZING AND ANTIOXIDANT
**7.5% Almond Oil. It is an oil with high skin tolerance that provides fatty acids in the form of triglycerides. The main fatty acid is oleic acid, which has emollient properties,
favors massage and penetration through the skin. The result is greater ease of absorption of the active ingredients in the formulations. Its content of unsaturated fatty
acids helps to restore the oil phase of the epicutaneous emulsion, thus reinforcing the barrier function, helping to maintain hydrated and nourished skin. MOISTURIZING AND
EMOLIENT
***2% Menthol. Applied to the skin, it has a rubefacient action, subsequently giving a sensation of cold and then manifesting a local anesthetic action. COLD AND SOOTHING
EFFECT
****0.6% Lavender essential oil. It has antimicrobial, antioxidant and anti-inflammatory action. It is suitable for use on sensitive and/or irritated skin. ANTIOXIDANT AND
SOOTHING
*****0.1% CBD (Cannabidiol). It is one of the main components of the hemp plant, it has no psychoactive or dependent effects. It is considered a great ally for muscles
and skin. It acts as an antioxidant, antiseborrheic, conditioner and skin protector. Helps relieve stress and reduce skin irritation. CONDITIONER AND PROTECTOR

Ozonized EVOO: Contains Triolein - a natural structural compound of the skin. Contains natural Azelaic Acid.

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CELLULAR NUTRITION WITH OXYGEN
BIOTECHNOLOGY APPLIED TO THE INGREDIENT PEROXIBIOKEY®

Nutritious source of fatty acids and triglycerides

that return natural moisture to the skin. PEROXIBIOKEY®
Unique manufacturing
Fills the skin spaces produced by dry skin.
process through controlled diffusion
of filtered ozone gas on organic
Extraordinary cuticle moisturizer. Extra Virgin Olive Oil (EVOO)
Helps reduce and soften marks (such as scars (production respectful of
and stretch marks). natural resources, contributing to
the sustainability of the environment
Contains triolein (structural fatty acids of skin cells)
and the agricultural
and natural azelaic acid, which contribute to skin
environment), obtained at low
repair.
temperature and under conditions
Suitable for all skin types stable and controlled,
avoiding the degradation of the ing

MUSCLE RECOVERY
WITH OZONIZED OIL
Reducing lactate production or increasing its elimination rate has been an objective
pursued by many researchers and is related to the hypothesis of the relationship between
fatigue and lactate level (Fitts and Holloszy, 1976).
The use of ozonated oil in sports massage can lead to greater lactate
elimination and improved sports performance. The effects of
passive rest and sports massage with and without
ozonated oil on performance, heart rate and
level of fatigue and blood lactate elimination in
professional cyclists after testing the Wingate
anaerobic test were examined.

Topical application of ozone during sports

massage appears to be a promising way
to reduce recovery time and attenuate
some components of muscle
fatigue. In particular, it could be useful
to introduce sports massage with ozonated
oil in physical activities that require repeated
performances, such as cycling, boxing,
athletics... in which massage is commonly
used as a recovery methodology¹.

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ANTI-INFLAMMATORY PROCESS OF THE
AC EITEDEOLI VA OZO NI ZAD O
To elucidate the mechanism of anti-inflammatory action of ozonated olive oil, we
investigated the effects of ozonated olive oil on the generation of PGE2 and the expression
of cyclooxygenase-2 (COX-2) in macrophages stimulated by lipopolysaccharide
(LPS) - such as THP-1 cells. Olive oil did not affect cell viability or morphology
within the concentration used. LPS-induced COX-2 expression and PGE2 generation in
macrophage-like THP-1 cells were not affected by olive oil itself. However,
ozonated olive oil inhibited both reactions in a concentration-dependent manner.
Furthermore, ozonated olive oil inhibited the phosphorylation of IÿB, which
is necessary for the activation of nuclear factor kappa B (NFÿB) and subsequent
transcription of the COX-2 gene in LPS-stimulated macrophages. These results
demonstrate that ozonated olive oil suppresses LPS-induced PGE2 generation in
macrophage-like THP-1 cells by inhibiting gene transcription through suppression of the
IÿBÿ/NFÿB pathway.

Ozonated olive oil significantly inhibits PGE2 overproduction and COX-2 expression in
LPS-stimulated macrophage-like THP-1 cells. It was shown that the effect of ozonated olive
oil on the COX-2/PGE2 pathway is the consequence of the inactivation of NFÿB through
the inhibition of IÿB phosphorylation
ÿ as shown in Fig.5. Furthermore, ozonated olive oil can suppress the generation of LPS-
induced proinflammatory cytokines such as TNF-ÿ and IL-1ÿ 2.

LPS
Toll-like receptor

cell membrane MyD88

ozonide IRAQ
Ras

inhibition

IkBa TRAF6

NFkB IKKa TAB2 MEK MEKK

TAK1
Q phosphorylation

IkBa
NFkB
ERK JNK
p38

degradation

arachidonic acid AP-1

Q
COX-2
COX-2 gene Binding region

prostaglandin translation
AP-1

inflammation, pain, fever, etc.

COX-2 gene transcription

Fig.5

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PAIN RELIEF
MUSCULAR
After intense sports training, it is possible to notice sore
muscles. This happens in elite athletes in phases
of over-training, pre-season or
after sports events and also in those people who are
not used to eccentric exercises (active movement
of a muscle while it is lengthening under load),
and can lead to injuries to muscle fibers.

To recover more quickly from these injuries,

inflammatory substances should be eliminated from
our body and damaged parts should be rebuilt. During
this process, the athlete could suffer muscle pain. This
condition can persist for four to five days 5.

Many people stretch their muscle groups before and after training to prevent
or reduce muscle tenderness. However, evidence suggests that it does not produce any
reduction in delayed onset muscle soreness in healthy adults 6. Therefore, in order to
ensure better regeneration after practicing sports, CBD could be very interesting
for athletes and doctors. sports.

The human body has an endocannabinoid system (ECS), composed of endogenous ligands
(endocannabinoids) and receptors. These receptors also work with cannabidiol, an active
component found in the cannabis plant. Through this interaction,
the
cannabidiol has demonstrated the ability to
produce many healing properties and
soothing. The anti-inflammatory properties of CBD,
given the inhibition of pro-inflammatory NF-kB
signaling[4], this means that due to its anti-
inflammatory potential, muscle regeneration
can progress faster, and muscle pain
can be prevented.

Many studies have shown a high

concentration of endocannabinoid
receptors (CB1/CB2) in the skin and, because
of this, cannabinoids such as CBD can be
easily absorbed through it. The interaction
results in
suppression of cell inflammation
resident and infiltrating immune cells 5.

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SPECIFIC BIBLIOGRAPHICAL REFERENCES:
1. Paoli A, Bianco A, Battaglia G, Bellafiore M, Grainer A, Marcolin G, Cardoso CC, Dall'aglio R, Palma A. Sports massage with ozonised oil or non-ozonised oil: Comparative effects on recovery parameters after maximal
effort in cyclists. Phys Ther Sport. 2013 Nov;14(4):240-5. doi: 10.1016/j.ptsp.2012.11.004. Epub 2013 Apr 24. PMID: 23623301.

2. Ugazio E, Tullio V, Binello A, Tagliapietra S, Dosio F. Ozonated Oils as Antimicrobial Systems in Topical Applications. Their Characterization, Current Applications, and Advances in Improved Delivery Techniques.
Molecules. 2020 Jan 14;25(2):334. doi: 10.3390/molecules25020334. PMID: 31947580; PMCID: PMC7024311.

3. Rojas-Valverde D, Fallas-Campos A. Cannabidiol in sports: insights on how CBD could improve performance and recovery. Front Pharmacol. 2023 Sep 22;14:1210202. doi: 10.3389/fphar.2023.1210202.
PMID: 37808192; PMCID: PMC10556669.

4. Peters EN, Yardley H, Harrison A, Eglit GML, Antonio J, Turcotte C, Bonn-Miller MO. A randomized, double-blind, placebo-controlled, repeated-dose pilot study of the safety, tolerability, and preliminary effects of a cannabidiol
(CBD)- and cannabigerol (CBG)-based beverage powder to support recovery from delayed onset muscle soreness ( DOMS). J Int Soc Sports Nutr. 2023 Dec;20(1):2280113. doi: 10.1080/15502783.2023.2280113. Epub
2023 Nov 10. PMID: 37947792; PMCID: PMC10653658.

5. Proske, U. (2005). Muscle tenderness from exercise: mechanisms? The Journal of Physiology, 564(1), 1–1. doi:10.1113/jphysiol.2005.085514

6. Herbert, RD, de Noronha, M., & Kamper, SJ (2011). Stretching to prevent or reduce muscle soreness after exercise. Cochrane Database of Systematic Reviews. doi:10.1002/14651858.cd004577.pub3

7. Bíró, Tamás et al. “The endocannabinoid system of the skin in health and disease: novel perspectives and therapeutic opportunities.” Trends in pharmacological sciences vol. 30.8 (2009): 411-20. doi:10.1016/
j.tips.2009.05.004

KEYBIOLOGICAL R&D
1. KeyBiological Dept. R&D, PeroxibioKey molecular characterization, “Molecular characterization of the products obtained from the ozonation of olive and sunflower oils by establishing the exact molecular
mass of the major components of both oils using high-resolution mass spectrometry. (QTof)”
SUMMARY: Two ozonized samples with different degrees of ozonation are analyzed to establish how it affects the chemical composition of the final product. In addition, the ozonolysis starting materials are also
analyzed to aid the interpretation of the acquired data.
CONCLUSIONS:
9-oxononanoic acid (8) and azaleic acid (9) were found in the ozonated oils analyzed, as cleavage products of linoleic and oleic acid. This observation is in agreement with a previous study published by Tortini et. Alabama.
where the authors found these fatty acid derivatives in ozonated sunflower oil using GC-MS.
Many derivatives of triacylglycerols were proposed as products of the ozonolysis reaction. These fatty acids are part of the structural compound of skin cells and have not been reported in ozonated oils so far.

2. KeyBiological Dept. R&D, Cell viability in murine macrophages and fibroblasts. “Determine the cell viability of murine macrophages and fibroblasts (RAW 264.7 and 3T3 L1 – MBX cell lines) in the presence of different
samples of PeroxibioKey ozonated oil using a colorimetric assay”
SUMMARY: Given that cell cultures grow in aqueous-based culture media, for the cell viability test it was necessary to evaluate different sample dilution methods, with the aim of obtaining homogeneous solutions compatible
with cell cultures: Dilution in PBS with ethanol (EtOH) at 5% and Dilution in dimethyl sulfoxide (DMSO) and culture medium.
CONCLUSIONS:
The analysis of cell viability in fibroblasts (3T3 L1-MBX), the oils tested do not significantly affect the survival of the cell line.

3. KeyBiological Dept. R&D, Efficacy Research, “Investigation of the Efficacy of PeroxibioKey Ozonated Oil Samples”
SUMMARY: The suspension study will be carried out for each of the four samples, according to concentration and in triplicate, against Escherichia coli and Staphylococcus aureus. This test will allow us to quantitatively know
the bactericidal efficacy of the samples. The results will be expressed as a percentage of cell death.
CONCLUSIONS:
The objective is to demonstrate, by performing a laboratory test, an observation that had been made in the ozonated oil samples: that these samples have antimicrobial activity.

To demonstrate this, a suspension antimicrobial efficacy test was proposed against two bacteria commonly used for this purpose: E.coli and S. aureus.
For a compound to be considered an effective antimicrobial, it must produce a death >99.00% (equivalent to R>2) during its testing. Taking into account the results obtained from the test carried out, it can be concluded that
all the oils tested have activity. antibacterial.

4. KeyBiological Dept. R&D, Minimum Inhibitory Concentration (MIC) Research, “Determine the lowest concentration of PeroxibioKey ozonated oil that, under in vitro test conditions, inhibits the growth of the microorganism in
a defined period of time.
SUMMARY: The MIC evaluation was carried out against Escherichia coli and Staphylococcus aureus, independently, in the presence of decreasing concentrations of the products under study (50.00% / 25.00% /
12.50% / 6.25% / 3.15% /1.56% / 0.78% / 0.39%) after a 24-hour incubation.
CONCLUSIONS:
· The results show that the non-ozonized sample oil does not present antimicrobial activity against Escherichia coli and Staphylococcus aureus, at concentrations greater than 50%.
· PeroxibioKey ozonized oil: 100% organic (600IP) presents activity against S aureus and E coli at a concentration of 50%.
· According to the results obtained, the PeroxibioKey ozonized Oil: 15% Olive + 85% Sunflower (1200IP) presents antimicrobial activity when tested at 25% against S. aureus and E. coli.
· PeroxibioKey Ozonized Oil: 50% Olive + 50% Sunflower (800IP) tested at 12.5% presents antimicrobial activity against S. aureus, however in the case of E. coli the 25% concentration is what inhibits growth of the bacteria.

5. KeyBiological Dept. R&D, Anti-aging activity assay, “Anti-aging activity of PeroxibioKey ozonated oil in aged human skin explants”
SUMMARY: This study was conducted to evaluate the effects of topical application of the compound PeroxibioKey on parameters associated with skin aging in human skin explants.

CONCLUSIONS:
· The study product at both 1% and 3% did not produce a decrease in resazurin reduction or an increase in LDH in human skin explants compared to the healthy control group under the test conditions. Therefore,
PeroxibioKey ozonated oil at the concentrations tested was not toxic to human skin explants under the test conditions.

· Treatment with corticosteroids caused several changes in the parameters measured in the present study related to skin aging: there was a clear reduction in collagen and elastin content. Therefore, corticosteroid treatment
was able to successfully mimic skin aging in this study.
· Topical application of 1% PeroxiBiokey ozonated oil for 10 days on aged skin explants improved the skin's ability to avoid the effects of corticosteroids. Topical application of this product resulted in increased collagen
and elastin production in aged human skin samples.
· Topical application of 3% PeroxiBiokey ozonated oil for 10 days on aged skin explants resulted in an improvement in the skin's ability to avoid the effects of corticosteroids, completely recovering its original appearance and
structure. Topical application of this product resulted in increased collagen and elastin production in aged human skin samples.

6. KeyBiological Dept. R&D. Research on cell viability. Establishment and maintenance of Hep2 cell line (human hepatocytes).
SUMMARY: Using the MTT test, cell viability was determined spectrophotometrically based on mitochondrial activity in living cells.
Abbreviation Composition: A1 Olive + Sunflower MP (50%), A2 OE100-600ECO, A3 Recycled ozonated oil, A5 MixOE50HA50-800.
CONCLUSIONS:
The oils that showed lower viability were A2 and A5 compared to A1 and A3. The results of the study demonstrated that treatment of cells with encapsulated A2 oil (FA2) shows significantly higher viability than cells treated
with non-encapsulated A2 oil (A2). In fact, treatment with the encapsulated oil (FA2) does not affect cell viability compared to the control (FA).

7. KeyBiological Dept. R&D. Safety and skin compatibility trial in healthy adult volunteers.
SUMMARY: Under the supervision of a dermatologist, the product is applied to the patch and fixed on the upper back using an adhesive patch. In parallel, a blank test without product is carried out. Duration and frequency:
single application, for 48 hours.
Evaluation: Clinical observations carried out by a dermatologist approximately 15-30 minutes after removing the patch. Verification of the absence of reaction by the volunteers 24 hours later.

CONCLUSIONS:
Based on the results obtained with the adopted methodology, said product meets the requirements of the skin compatibility test, and can be classified as NON-IRRITANT with VERY GOOD SKIN COMPATIBILITY. The
result obtained is CPI of 0.00

*Information for the exclusive use of professionals, which should not be understood as advertising. Delivery to the general public is prohibited.

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