IN VITRO EFFICACY TESTS
The nutraceutical claim is the pact you make with your consumers: we back up your promises.
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Digestive system
Anti-inflammatory activity on AGS cells
To determine the effectiveness of the product on the digestive system, we can evaluate its anti-inflammatory properties on a specific cell line derived from human gastric tissue (AGS cell). An inflammatory state will be induced in the cell line using an appropriate inflammatory 'stimulus', then the cells will be treated with the product and the concentration of the COX2 enzyme will be determined as an indication of the level of residual inflammation. COX-2 is the inducible isoform of the enzyme cyclooxygenase and is activated following tissue damage and is responsible for the synthesis of pro-inflammatory prostaglandins.Activity against Helycobacter pylorii
If the product has an antimicrobial claim, we can carry out an activity test against Helycobacter pylorii. This bacterium, once established in the stomach, produces toxic metabolites within the gastric mucosa which, over time, can lead to gastritis and ulcers. With this test, we will demonstrate in vitro the efficacy of the compound and establish the inhibitory concentration against Helycobacter pylorii.Intestinal system
Anti-inflammatory activity on Caco cells
To determine the effectiveness of the product on the intestinal tract, its anti-inflammatory properties can be assessed on a human colon cell line (CACO2). An inflammatory state will be induced in the cell line and, following treatment with the product, the amount of specific inflammatory markers will be measured. In our tests we can detect different inflammatory markers such as cytokines and chemokines. Interleukin is usually present at a basal level in resting cells, and its production is significantly increased by the pro-inflammatory cytokine Tumor Necrosis Factor alpha (TNFα), whose expression is in turn triggered by harmful inflammatory stimuli. Inducible nitric oxide synthase (iNOS) is activated during inflammation with both signalling and bactericidal functions and is therefore another important marker of the inflammatory state.Activity on the intestinal mucosa
In order to assess whether a particular compound has activity on the intestinal mucosa, we can perform tests on reconstructed mucosa in vitro. Using the HT29-MTX cell line, a subpopulation of mucus-secreting intestinal cells, we reconstruct a mucosal layer on which the product is tested. We will then evaluate how the product is able to affect the mucous layer and its protective capacity against toxic agents that could compromise it.Prebiotic activity
Prebiotic activity reflects the ability of a given compound to support the growth of a specific probiotic microorganism in relation to pathogenic microorganisms. In this test, we will assess whether your product is able to promote the growth of beneficial microorganisms while discouraging potentially pathogenic microorganisms in the intestine.Focus Probiotics
We have developed specific tests to evaluate the effectiveness of your probiotic strains:Immune system
In our laboratories, we can investigate the effect of products in modulating the immune response in in vitro assays on cells of the immune system by determining the levels of certain signal molecules and thus understand their systemic effect.
In particular, we can perform:
Cytotoxicity tests on cell lines such as colon CACO-2, immune system Natural Killer and macrophages RAW 264.7.
Determination of the anti-inflammatory activity of compounds on the colon CACO-2 cell line by monitoring the level of the cytokine IL8 secreted in an inflammatory context induced by TNFalpha (tumour necrosis factor alpha).
On the Natural Killer cell line through a membrane-based antibody array, we can simultaneously assess several cytokines, chemokines and growth factors to determine the levels of extracellular signalling. These molecules are released by cells and play critical roles in many biological processes such as cell growth, differentiation, gene expression, migration, immunity and inflammation. In most biological processes, several cytokines operate in correlation, where the action of one cytokine is regulated by the presence or absence of other cytokines.
Determination of the anti-inflammatory activity of products on macrophages RAW 264.7 by monitoring the level of NO (nitric oxide) produced in an inflammatory context induced by LPS (bacterial lipopolysaccharide).
Neuropathic pain
For products acting at the nerve level, which are useful in the treatment of neuropathic pain, we can perform a specific test to support this claim.
On an isolated microglia cell line, we test for various markers such as Cannabinoid receptor type 2 (CB2) and Metabotropic glutamate receptor (mGlu2).
As microglia play a key role in neuroinflammation, such as brain and spinal cord injury, ischaemic stroke, infection and neurodegenerative diseases, detailed cell biological exploration of microglia is therefore of paramount importance.
Joints
Anti-inflammatory activity on SW1353 chondrocytes
To determine the efficacy of the product at the joint level, its anti-inflammatory properties can be assessed on a chondrocyte cell line (SW1353). On the cell line induced in an inflammatory state and treated with the test product, we evaluate the expression levels of Interleukin 1 beta and the inducible isoform of the cyclooxygenase enzyme COX-2. These markers are important indices of the joint inflammatory state.Diabetes
Glucose uptake assay
Glucose metabolism is a key process and a deficiency in insulin-stimulated uptake is associated with type 2 diabetes. Determination of glucose uptake is therefore useful for determining the effects of various compounds on diabetes. Through our tests, we can measure the level of glucose uptake through an in vitro assay on a liver cell line (HEP G2) and thus determine how a compound intervenes in this metabolic process.Weight control
Alpha amylase inhibition
In our laboratories, we can evaluate the alpha-amylase enzyme inhibiting activity of natural extracts for nutraceutical use. Alpha-amylase is an enzyme belonging to the class of hydrolases, which catalyses the endohydrolysis of 1,4-α-D-glucosidic bonds in oligosaccharides and polysaccharides containing three residues or more. Using a standard enzyme of known titre (of porcine origin, usually used as a standard for simulating digestion in the gastro-intestinal tract), tests are performed to quantify the inhibition of enzyme activity in the presence of different concentrations of the test compound. This test evaluates the change in specific enzyme activity in the presence of the test compound and thus determines its inhibitory effect.Probiotic focus
We have developed specific tests and activities to evaluate the effectiveness of your probiotic strains:
- Analysis of the specific prebiotic power of fibres and natural extracts towards probiotics
- Analysis of the degradation of toxic compounds by probiotics
- Analysis of metabolite and peptide production by probiotics
- Optimisation studies of metabolite production by probiotics
- Stability testing of probiotic formulations and additives/coformulants
- Testing of synergistic activity with natural extracts
In collaboration with the Bioindustry division, we can also offer you:
- Strain improvement
- Development of high-density fermentation methods
- Identification and formulation of probiotic mixtures for specific properties