Synergy between Nature and Science: the world of food supplements serving intestinal health
At the Root of Intestinal Inflammation
Inflammatory bowel diseases are among the major medical challenges of our time. Chronic intestinal inflammatory diseases, collectively referred to as IBD (Inflammatory Bowel Disease), mainly include two widespread types: Crohn’s Disease (CD – Crohn’s Disease) and Ulcerative Colitis (UC – Ulcerative Colitis). Understanding these conditions can lead to more effective therapies and a better quality of life for patients. Crohn’s disease and ulcerative colitis are two of the most common, characterized by an alternating pattern of relapses and remissions. These conditions are often the result of an inappropriate immune response to intestinal dysmicrobisms in genetically predisposed individuals.
In fact, over 150 genetic susceptibility loci have been identified, of which 70% are shared between both pathologies.
Common symptoms of IBD include abdominal pain, bloody diarrhea, weight loss, fatigue, fever, intestinal obstruction, and, in children, developmental delay. Crohn’s disease, in particular, can affect any part of the gastrointestinal tract, from the oral cavity to the anus, with a greater incidence in the ileocolic region. It is characterized by discontinuous extension, with granulomas, masses of inflamed tissue identifiable in almost every layer of the intestinal wall, from the tunica mucosa to the serosa. Furthermore, granulomas are considered a hallmark of established CD, although they are not present in all patients with this disease. CD can manifest itself in three types: inflammatory, obstructive and fistulous.
Food supplements and European legislation
Food supplements are widely used in the daily diet. Their regulation is crucial to ensure consumer safety. In Europe, Directive No. 46 of 10 June 2002 laid the foundations for the harmonization of the supplements sector in individual countries.
- Main Objective: Guarantee the protection of public health and allow free circulation of these products.
- Definition: Products intended to supplement the normal diet, representing a concentrated source of nutrients or other substances with a nutritional or physiological effect.
- Forms and Regulations: Supplements can come in different pre-measured forms and are considered foods, therefore subject to the same rules that concern the treatment and safety of food products.
Italy has implemented this European legislation with Legislative Decree n°169/2004. Food supplements are defined as food products intended to supplement the common diet and constitute a concentrated source of nutrients or other substances with a nutritional or physiological effect. These substances include, but are not limited to, amino acids, essential fatty acids, fibers and plant extracts.
Supplements come in various pre-measured, typically pharmaceutical forms, such as capsules, lozenges, tablets, chewing gum, powders in sachets, liquids in vials or dropper bottles, and other similar forms. These products in Italy are considered foods and are therefore regulated by the same rules that concern the treatment and safety of food products (EU Reg. 852/2004). They have the task of integrating the normal diet without replacing any food and have an exclusively health-related role.
Food supplements are subject to vertical regulations to ensure the safety and effectiveness of the products. Some key considerations regarding the regulation of dietary supplements are:
- Supplements must not have curative or preventive properties for human diseases. However, they must specify the vitamins, minerals and their respective maximum daily quantities allowed.
- Extracts and preparations of plant origin, also known as “botanicals”, in food supplements are currently regulated by the ministerial decree of 10 August 2018, with subsequent amendments defined by the executive decree of 26 July 2019 and subsequent Rev. 01 August 2022 attached 1.
- The term “Novel Food” refers to those foods not used in the EU before 15 May 1997. This includes foods derived from new sources, new production processes, or foods not traditionally consumed for their nutritional value.
- The field of probiotics has undergone changes over the years. The first probiotic supplements were authorized in Italy about 30 years ago, and generally contained Saccharomyces cerevisiae, Streptococcus thermophilus and Lactobacillus bulgaricus. With the introduction of Directive 2002/46/EC, the field of application of food supplements has been extended to also include probiotic-based products. Today, reference is made to EFSA regarding the safety of strains of microorganisms considered GRAS (Generally Recognize As Safe)
Andrographis Paniculata: A Bridge between Tradition and Modern Science
Traditional medicinal plants have been used for millennia and now, with modern science, we are discovering the rationale for their effectiveness. Andrographis paniculata is an example of this. Traditionally used for the treatment of infections and inflammatory conditions, it belongs to the Acanthaceae family and appears as an annual herb with dark green lanceolate leaves and quadrangular stems. Its flowers are bilabial, with petals that vary from white to dark purple. Flowering occurs from December to April.
This plant is also known by several popular names that reflect its different characteristics and properties. Among the most common in Europe are: “King of the Bitter” or “Bile of the Earth”, due to its extremely bitter taste, and “Chiretta Verde”, because it shares the same bitterness as the Gentianaceae Swertia chirayita.
Andrographis paniculata was not known in the Greco-Roman world, but was already widespread in Asia as early as 2000-3000 BC. Furthermore, it has been identified as a potential source of antioxidant and anti-inflammatory compounds, including andrographolide. This compound has shown anti-inflammatory properties through modulation of several cellular pathways.
Extensive studies have outlined details about the pharmacodynamics (biochemistry) of andrographolide, its interactions with various enzymes, and its potential therapeutic applications.
Kefir: The Hidden Ferment of Wellbeing
Kefir, commonly known as a fermented drink, has gained popularity in Western countries due to its numerous health benefits. Native to the Caucasus, it is produced through the fermentation of milk and hosts a wide range of beneficial microorganisms. Its consistency is slightly viscous and has a sour taste, and is enriched by abundant fermentation products such as organic acids and volatile compounds.
Kefir granules, with a gelatinous consistency and a white-yellow colour, are a distinctive feature of this drink. These granules host a greater quantity of lactococci in the outer layers, while greater quantities of lactobacilli and yeasts are found inside. The major polysaccharide that gives structure to kefir granules is kefiran, produced mainly by Lactobacillus kefiranofaciens.
Research has shown that kefir has beneficial health properties, including cholesterol-lowering, antimicrobial, hypotensive and anti-tumor effects. Some lactobacilli isolated from kefir have been shown to have the ability to protect cells from toxin damage and to inhibit the adherence of certain pathogens to cells. Furthermore, some species of bifidobacteria show an increased growth rate when grown in kefir, with an overexpression of genes associated with flagella transmigration.
The Science Behind the Discovery
PNK Farmaceutici’s research is not just a set of results, but a journey through the rigorous scientific process. The scientific publication shows the tools and techniques that made these discoveries possible.
Science, in its essence, is based on accurate and detailed methods. The accuracy of the conclusions depends on how precisely the experiments are conducted. In the R&D laboratories of PNK Farmaceutici, the methodological framework behind the research is revealed. This publication is not just a list of tools and techniques; it is a journey through the scientific rigor, meticulousness and precision with which each experiment was designed and conducted. From unique bacterial strains to advanced sequencing techniques, every detail is critical to understanding the impact and importance of the study. Through this overview, you have the opportunity to appreciate not only the results, but also the process that led to these discoveries.
Andrographis paniculata:
Sample
Several samples of Andrographis paniculata extracts were found on the market and analyzed in parallel. These samples varied in the titration of the active ingredient andrographolide and highlighted distinct characteristics, such as the color and volatility of the powder: for example, very volatile brownish powders compared to more granular white or cream-colored ones. The samples were also heterogeneous in terms of the active content (from 40% to 98%).
Materials and methods
The reagents and tools for evaluating the suitability of the extract were:
- GC-MS for the verification of impurities contaminating the extract
- Solvents: High purity methanol (MeOH), ethanol (EtOH) and acetonitrile (ACN) were used.
- HPLC-DAD for the determination of the titer of andrographolide and neoandrographolide
- Andrographolide and Neoandrographolide standards, key compounds used as controls in analyses.
Bacterial strains:
Sample
Different bacterial strains were used as test samples in antimicrobial activity analyses. These include strains such as S. mutans, S. pyogenes, S. sanguinis, S. uberis, E. faecalis, E. cloacae, S. aureus, L. monocytogenes, L. innocua, E. faecium, E. coli and K. pneumoniae.
The Lentilactobacillus kefiri SGL 13 strain belongs to the company strain library and is registered with the identified number DSM 27331
Materials and methods
- Test microorganisms used were obtained from different collections, such as the ATCC and the DSMZ.
- The L. kefiri SGL 13 strain was isolated from Turkish kefir grains, which were purchased at a local market in Turkey. This product did not have a specific brand, but was a traditional and artisanal product. Its identification was made through whole genome sequencing and physiological stress survival studies have determined its suitability for probiotic use.
- Agar Well Diffusion techniques were conducted to test the inhibition of A. paniculata against various pathogens. Different concentrations of the dry plant extract titrated at 60% andrographolide were tested.
- The survival of SGL 13 in the presence of the titrated extract was also evaluated.
Results and Implications
In-depth investigation of Andrographis paniculata has revealed facets that go beyond the simple presence of andrographolide. The concentration of this active ingredient, its effect in combination with other compounds such as neoandrographolide, and the challenges in determining the actual concentration compared to that declared by suppliers were crucial issues addressed.
But, like any good story, there are also surprises. The discovery of traces of contaminants in some samples raises questions about the safety and integrity of extraction processes. And the presence of neoandrographolide in significant quantities opens a new chapter of investigation into the potential synergistic or antagonistic interactions between the various compounds.
From the analysis of the survival properties of Lentilactobacillus kefiriSGL13, the importance of evaluating each individual bacterial strain according to the context in which it will be used clearly emerges. Its tolerance to certain conditions, such as the presence of bile salts or pH variations, and its sensitivity to antibiotics are essential aspects to understand the potential health use of the strain.
Paniculin 13TM the brand derived from this study, which combines the anti-inflammatory properties of A. paniculata with the anti-inflammatory properties of L. kefiri SGL 13. The combination in a single formula of the probiotic with the plant extract of Andrographis paniculata, act synergistically to increase the effectiveness of their individual actions, proposing an alternative to current treatments that involve either the use of a single element or traditional pharmacological approaches .
But beyond the conclusions, this work highlights the dynamic nature of scientific research. Each finding leads to new questions, each observation challenges previous understandings, and each analysis opens new avenues to explore.
