SLOW RELEASE ESSENTIAL OILS

A NEW GENERATION OF NUTRITIONAL + COSMETIC PROGRAMS CONTAINING LONG ACTING – SLOW RELEASE ESSENTIAL OILS

 

Note: These programs are based on thorough scientific research, their efficacy is pharmacologically & scientifically proved, the key discoveries are patented, published, and this research work received the European PAEXA award & French Academy of Science award.

Prince Louis de Polignac, award by the French Academy of Science

European PAEXA award for in vitro research

VITROBIO

VITROBIO, based in France, is a self-financed R&D institute conducting research on new generation of cosmetic, nutritional products & medical devices since 1999. The aim of VITROBIO (and its sister companies Naturveda & Vedascience) was to conceive result oriented nutritional & cosmetic programs, developed like a drug, which are not “me too” and which cannot be copied, if successful. In this highly competitive market, we prefer to adapt a scientific strategy explaining the customer the problem – scientific solution – the story of our research & the product we propose. In the absence of any marketing or sales structure at Vitrobio, only 3 products were sold through 2 national TV shopping channels (TF1 & M6) in France & 1 in Greece between 2001-2010. Presentation was not “before” & “after” but scientific. More than 10 million programs were sold & these products still constitute sales record since 1990’s. This proves that an excellent product coupled with scientific proofs & a good presentation can still be highly successful in this competitive market. We continued research but stopped marketing the products on TV shopping in 2010 as we wanted to concentrate all the efforts to conduct research on Medical Devices. These new generation of MDs for the treatment of multiple skin & ENT diseases are now sold by known Pharma companies such as BMS -UPSA (US), URGO (France), Bouchara (Italy), Siemens (Germany), while discussions with GSK are in progress.

Taking into consideration the future EU regulations on the use of Essential Oils (Eos) in cosmetics & increasing public interest in Eos, we developed (and patented) a new form of long-acting & slow release filmogen Eos.

This research associates the most modern research technology with the knowledge of traditional Ayurvedic medicine.

WHAT IS AYURVEDA?

“Ayu” means life & “veda” means science. Ayurveda, over 3000 years old, is Indian traditional medicine, based on the use of a synergistic and specific association of some Earth (soil) & Natural ingredients (plants & essential oils) to treat various human pathologies. This science was practiced widely & regularly all over India with extraordinary results but always remained a scientific mystery. Modern science is now exploring Ayurvedic preparations because what was considered illogical before, starts looking very scientific today. Vedas were written 2000 years ago, but what surprised scientists is that they already mentioned excess weight and “low mood” (depression) in those times, since we thought that these were modern life diseases. According to Ayurveda, excess weight and “low mood” are not always related to excessive food intake and to our modern difficulties of life but to the poor functioning of our cells. The body is made up of cells, and disturbed cellular functions are manifested as a pathology.

This disturbs “doshas” (the energy) which may manifest externally or internally, in the body and the mind, as a cause or as a symptom. Therefore, normalizing cellular functions and removing the cause of the disturbance should cure the disease.

 

Ayurvedic theory on excess weight: Excess weight may develop not only due to excess food intake but also if the cells are not able to eliminate excessive reserves. Energy enters the cells, a part of it is used and the remainder is automatically eliminated when cellular functions are normal. If cellular functions are disturbed, they cannot remove all the waste, which starts accumulating into the cells and progressively manifests as excess weight.

A lack of some essential Earth elements justifies these poor cellular functions. Therefore, to reduce excess weight, Ayurvedic medicine suggests improving cellular functions of elimination by administering “Earth elements” & plant ingredients to accelerate waste removal.

Modern R&D proof: We also observe in our daily life that many health problems are not linked to our mode of life otherwise two persons, having identical mode of life should have the same problem! For example, some people eat a lot but don’t gain weight while some others accumulate whatever they eat! Why? Does it relate to cellular capacity of elimination?

To check this hypothesis, some scientists in our Institute created an in vitro model of disturbed cellular functions by modifying nutrients in the cells (in vitro). A high fat serum was prepared in rabbits and the cells in culture were exposed to this LDL-containing serum (see photo). The amount of fat inside the cells was quantified by staining the cultures with Oil-Red-O (a lipid-specific stain” which colors fat in red). The red color indicates fat deposits. We observed that poorly functioning cells use a part of this lipid, but the remainder is deposited inside the cells (see photo).

NUTRITIONAL SUPPLEMENTS

 

Discovery of “CELL COMPLEX” nutritional supplement:

We thought that the Ayurvedic “Earth elements” should be minerals, vitamins, and trace elements. Therefore, we exposed disturbed cell cultures accumulating lipids (see red colour inside the cells) to non-toxic, daily required amounts (DRA) of a mixture of these elements, but there was no change in the amount of red color inside the cells as normally functioning cells should remove the excess lipids. The Ayurvedic preparation was not effective, or there was something wrong with our technique!

Ayurvedic preparations to treat excess weight suggest using the Earth elements but they should be mixed in a very precise manner (red, green, blue) and in specific proportions. The association of these elements must be well dosed! In the absence of precise knowledge regarding the compositions of these Earth elements, we tested different associations of our Earth elements over 3 years: vitamins, minerals, and trace elements at different concentrations, and surprisingly we found a specific association of vitamins, minerals & trace elements which, when mixed in very specific proportions, was finally capable of stimulating cellular functions and allow the cells to remove all intracellularly deposited lipids within a few days (pharmacological studies & patents available).

This association of elements was called “Cell Complex” and was patented worldwide.

Upon request, the composition was also sent to the scientific committee of EU.

Ayurvedic theory on depression: According to Ayurvedic texts, “poor mood” may develop if the nerve cells are not working correctly due to slow development or poor functioning of “brain constituents” (neurons). Ayurveda suggests using some specific vegetable oils, rich in Omega-3 fatty acids, in association with specific “Earth elements” to get rid of “poor mood”.

Verifying the theory with Cell Complex: Testing Ayurvedic remedy for neuronal function deficiency problems (e.g. anxiety, excitability, depression): This work was performed at NEURONAX, Neuronal disfunction research Institute, CHU, Clermont Fd by Dr. Stephen GABRON.

These problems are probably related to the difficulties of neural transmission: if the length of the neurites is insufficient, electrical current may not pass and may cause irritability or depression, and if neurons are not functioning well, they may produce neurological disturbances. Increasing the length of the neurites & simultaneously improving neuronal functions should help to normalize these functions.

Omega-3 fatty acids are a key component of cellular membranes, but they are deficient in >7/10 persons (except in Japan where people regularly eat raw fish). Experiments were thus conducted by exposing disturbed neuronal cultures to cell function-stimulating “Cell Complex” in association with an EPA-DHA rich source of Omega-3.

Extraordinary results on neurite growth & neuronal functioning were obtained (For full details see: Shrivastava et al. J. Neuronal Neuroscience).

These results and further studies conducted in our laboratory prove that Ayurvedic natural remedies, suggested 1000s of years ago, are highly effective provided that the scientists understand the logic behind these safe recipes. All the nutritional supplements are based on Ayurvedic recipies and contains “earth elements” and/or synergistic association of plant extracts.

 

These “Earth elements” are presented either alone or in association with synergistic plant preparations as NUTRITIONAL SUPPLEMENTS.

DISCOVERY OF LONG ACTING – SLOW RELEASE EOs

 

Use of ESSENTIAL OILS (EOs) in Ayurveda

Introduction: Aromatherapy is now becoming one of the major complementary therapies which use essential oils, internally & externally as major therapeutic agents to treat several diseases. The global essential oils market size was valued at US$ 6.63 billion in 2016, growing at a CAGR of 9.7%. A market research study by Grand View Research estimates that the global essential oils market is expected to reach $11.67 billion by 2022.

The use of EOs was very common since Ayurvedic period and remains common today.

Ayurveda considered Eos as “Agni” or “Fire” (as they are highly concentrated pure fire elements of a plant) and suggested using only in very small quantities as aromatherapy, perfumes or massage but never “internally”.  Modern medicine is using EOs internally & externally, usually in high concentrations without any proofs of their safety. Only since last few years, the concern over the safety of EOs is becoming a major concern in all the developed countries.

These aromatic molecules are very potent organic volatile chemicals that make even the surroundings free from parasites, bacteria, viruses and fungi. Their efficacy as antibacterial, antiviral, anti-inflammatory; force, muscle, mood, memory, circulation, & immunity-booster, is documented scientifically.

Regulatory concerns over the use of Essential Oils!

EU is now concerned with EO vapour toxicity & side effects! It started with the allergens, in 2002, which was the reason for the foundation of EFEO – the European Federation of Essential Oils. Because of this regulation, several end users decided to change their formulas and minimized using substances containing allergens. Then came the REACH Cosmetic Regulation EC N° 1223/2009, with many more restrictions for marketing EOs in cosmetics.

Even if EOs are diluted before use, they are applied on live skin. Skin is keratinized and doesn’t allow substances to enter easily into the body. EOs contain high concentrations of 100s of chemicals such as a-pinene, canfene, β-pinene, Sabinene, β-myrcene, a-felandrene, limonene, β-felandrene, g-terpinene, β-ocimene, a-ocimene, linalool, caryophyllene, terpinen-4-ol, azulene derivative, a-humelene, a-terpineol, germacrene D, bicyclogermacrene, isoledene, g-cadinene, d-cadinene, copaene, elemene, caryophilene oxide, spathulenol, eudol, germacrone, unidentified sesquiterpene alcohol, epi-bicyclosesquiphelandrene, Α-cadinol, elemol, neril hexanoate, g-eudesmol, g-cadinol, unidentified sesquiterpenes, …

Traditional use suggests that they are probably not harmful to the health but their toxicity, mutagenicity, and carcinogenicity were never studied.

They are highly volatile and 50% oil evaporates within a few minutes, as shown below (see: https://link.springer.com/article/10.1007/BF02324825).

50% evaporation rate in minutes: Sandalwood: 46 min; Jasmin absolute: 46 min; Patchouli: 28.5 min; Cloves: 29 min; Anise: 24.5 min; Rose: 23.5 min; Geranium: 22.5 min; Ylang-Ylang: 22 min; Citronella: 19.5 min; Fennel seed: 19.5 min; Clary sage: 17 min; Coriander: 15.5 min; Peppermint: 15.5 min; Lavender: 13 min; Dillweed: 10.5 min; Bergamot: 9 min; Orange: 8.5 min; Mandarin: 6 min.

The key problems with the use of current diluted EOs are:

  1. Poor absorption / poor efficacy,
  2. Limited duration of activity but flash absorption (max within 10-50 min after skin application) instead of slow release to minimize side effects and to maximize efficacy,
  3. More oil (nearly 90%) is volatilized and enters the body through the lungs than is acting topically where it was applied,
  4. Nearly instant release of 100’s of chemicals in the air with unknown toxicity / side effects,
  5. High chances of toxicity due to sudden high blood/air concentration, if inhaled.
  6. Not being fatty, aromatic oils do not spread well on a large skin surface. Mixed with a carrier, they are poorly absorbed.

What the September 2018 EU guidelines may change?

Not known yet, but surely those guidelines shall:

  • further reduce the concentration limits for use, in order to minimize air evaporation and sudden peak of unknown chemicals absorbed from the air (using low concentrations, however, may lead to more frequent/multiple applications),
  • call for stopping the use in children & pregnant women altogether

(we will update this section in September 2018).

Development of specific low-concentration, slow release, long-acting, filmogen Essential Oil (EO) formulations

Not only due to the absence of novelty in this field, but also due to changing regulations and the probability of withdrawal of many currently best-selling EO formulations at the end of the year, we launched a R&D program with the objective to find new EO formulations which:

  • could be used in low concentrations, as per the legislation/regulations,
  • could remain over the biological surface (skin, mucosa) over a period of at least 4-6h, so as to deploy their therapeutic potential;
  • could be released slowly into the biological tissue;
  • are not volatilized within an hour;
  • are not irritant & have very few/slight side effects, due to the low concentrations used / absorbed.

R&D PROGRAM

  • Phase 1: Searching for an EO-storing, non-irritant film for topical application

The initial R&D phase was concentrated mainly on searching for an ingredient with hydrating and slow-fading filmogen properties, capable to trap EOs for slow release.

To achieve these objectives, it was not possible to use any ingredient which is a chemical, which does not trap but absorbs essential oils, which has a density lower than the density of EOs (ex: normal or salted water, saline, since EOs float in water), which may interact with EOs, and which may have any pharmacological, metabolic, receptor, immunological or biological interaction with the underlying live tissue.

The secondary objective was to render the film adherent & resistant to mechanical pressures.

After testing different filmogen substances for their cellular interaction and cellular toxicity using in vitro cell culture models, we identified some natural, cell-friendly, filmogen ingredients:

  • plant-based
  • sugar-based (ex. sucrose, glucose, fructose, saccharose)
  • sugar–alcohol-based solutions

Finally, we conceived as a viscous, colorless, odorless, solution with humectant properties. It had no interactions with any EOs and the film was capable of trapping EOs. This solution was patented worldwide.

Disadvantages of VD-binder film:

  1. The product was poorly filmogen as it disintegrated within 90-120 minutes because of its humectant properties.
  2. The EO release was prolonged up to 1.5-2h, but it was not satisfactory enough.

 

  • Phase 2: Improving the duration of retention of the film

 

To improve the duration of film retention, so as to avoid frequent EO applications and to allow slow EO release, multiple experiments were conducted, adding different substances into the standard VD-binder solution.

We observed that some natural and synthetic polymers, which are very big, branched and inert molecules, have the properties to bind specifically with VD-binder macromolecules without interacting with EOs.

This binding improved the film resistance and flexibility to mechanical pressures. The duration of retention over a live biological surface was improved up to 4-6h.  The EO-incorporating film was designated “F-VD-binder” film.

The desired quantities of selected Ayurvedic EOs were incorporated in the F-VD-binder film to conceive a new, slow-release device for EOs.

This research was conducted in collaboration with Inflamlab, University of Toulouse, France.

Phase 3: Verifying speed of EO release from F-VD-binder film

These experiments were conducted at LEXVA Laboratoire Analytique, St Beauzire in France.

Example:

Aim: To compare the release of Eucalyptus EO, the EO was mixed in water or in F-VD-binder solution at a concentration of 0.20% and kept overnight. The concentration of volatile compounds (Alpha-Pinene, Limonene, and Eucalyptol) was quantified through aqueous phase GC / FID chromatography on a semi-polar column after 20h storage. The concentrations were determined by measuring the area covered compared to water control.

The following mean results were obtained:

Blue curve: Saline solution + EO; Orange curve: F-VD-binder  + EO

Component Alpha pinene Limonene Eucalyptol
Mean area M1 (EO mixed in water) 1032.288       298.375  1.209 e4
Mean area M2 (EO mixed in F-VD-binder)   242.527         69.077    160 e4
% decrease in EO release vs water solution –          76.55% –          76.84% –          86.76%

These results show that when incorporated in a filmogen F-VD-binder solution, the EO volatility decreases 4-5 times, compared to the same concentration of EO in another non-filmogen solution.

The filmogenicity of F-VD-binder was also determined in vitro by applying the film over live skin tissue (n=6 each) and washing the film with the culture medium (isotonic liquid) at 6h intervals. Depending upon the concentration of VD-binder in F-VD-binder film, the mechanical resistance of the film could be increased 5 to 6-fold (maximum effects between 40-50% VD-binder concentration in water).

Slow-release & long-acting essential oil formulations

Based on these findings, we conceived a range of topical EO preparations, containing 1 or more EOs for each specific pathology to be treated.

Precautions: In the absence of scientific data to demonstrate safety, the use of essential oils is not recommended in pregnant women & children below 8 of age.

Conclusion: To take full advantage of the beneficial properties of EO by topical application, the EOs were incorporated in a filmogen liquid. The EO evaporation and in consequence their concentration was reduced 4-5 times. The efficacy was improved due to long term adherence, environmental & pulmonary contamination was highly minimized. Long acting & slow release EO formulations should represent the 1st and one of the major improvement in EO therapy

NUTRITIONAL & EO PROGRAMS BASED ON THESE DISCOVERIES

Why a program containing a nutritional supplement & a cosmetic?

Because Ayurveda suggests acting:

  • Internally & Externally (to prepare a healthy ground 1st)
  • On the cause & the symptoms

These preparations usually contain a synergistic association of “Earth elements”, Plant Extracts and Essential Oils, which cannot be mixed easily. Therefore, the solids are presented as nutritional supplements & the Essential Oils as cosmetic preparations.

PRESENTATION:

Each program containing 2 packs:

  1. Slow-release Essential Oil spray (2-3 sprays twice /d)
  2. A container or strips with 60 or 90 vegetal capsules (2-4 caps/d)

Note: These programs are manufactured in France under GMP or ISO13485 norms.

Complete program is presented in a box which may contain capsules (variable presentation) & an essential oil spray for each pathology. Examples: