Alterbio

Leader of consortium:

Státní zdravotní ústav

Consortium members:

SYNPO, akciová společnost, Biomedica, spol. s r. o., Centrum organické chemie s.r.o., INOTEX spol. s r.o., Univerzita Palackého v Olomouci

Responsible person:

RNDr. Hana Bendová, Ph.D.

Duration of the work package:

01/2014 – 12/2019

Description:

Aim of this package is to evaluate and select the most efficient antimicrobial agents able of covalent or ionic bond within polymeric system in
comparison to antimicrobial effects of monomers with antimicrobial agents alone. The selected agents with efficient and stable antimicrobial
effects will be subjected to a battery of toxicological tests, focused on local toxicity, namely skin and eye irritation, phototoxicity, skin penetration.
The toxicological methods will comprise exclusively in vitro methods based on humans cells and reconstructed tissues, and QSAR applications
(respecting animal testing restrictions regarding safety evaluation of ingredients intended for consumer products, e.g. cosmetics) in compliance
with current legislation on cosmetics banning use of experimental animals (Regulation (EC) No. 1223/2009). Existing toxicological data on
systemic toxicity will be collected and examined for validity.

Based on the results of biological test results available, the most efficient and non-toxic forms of the antimicrobial agents will be incorporated
into the following types of consumer products: cosmetics, textiles, plastic foils (e.g. food contact materials), and varnishes for surface
antimicrobial protection of plastic materials.

The antimicrobial efficacy of antimicrobial agents as a part of polymeric system incorporated into the finished products will be determined and
health safety of finished products assessed in compliance with binding legislative measures in respective areas according to the type of product.

Task 8.1 Microbiological evaluation of antimicrobial agents

The task will comprise microbiological evaluation of efficiency of monomers and polymers with antimicrobial effects, containing chosen
antimicrobial agents, e.g. silver and photoactive photosensitisers.
The methods used will include antimicrobial tests for determining the biocidal/biostatic efficacy of preservatives against bacterial and fungal
growth. The methods will be modified on the basis of international standards for determination of antibacterial and antifungal activities with
respect to the type of finishing material surface (hydrophobic, hydrophilic), the temperature used for the employment of product etc. Strains of
bacteria and fungi used in tests will be obtained from the Collection of Microorganisms as well as from the environment.

Task 8.2 Toxicological evaluation of antimicrobial agents

Based on the microbiological tests, the most promising antimicrobial agents will be tested in order to obtain a basic toxicological profile (i.e. acute
toxicity and phototoxicity in vitro, skin and eye corrosion/irritation tested by means of reconstructed human models, skin penetration ex vivo using
porcine skin and reconstructed human models), according to EU, OECD, EN/ISO standard methods. Negative results for polymeric system will
be confirmed in clinical studies on human volunteers.

Task 8.3 Cosmetic products

The most efficient antimicrobial materials proven as safe on the basis of toxicological data available will be incorporated into final cosmetic
formulations.

The benefit of high-tech innovative antimicrobial additives incorporated into the formulation of cosmetic products will be determined using
microbiological challenge tests. In challenge testing, i.e. testing of antimicrobial efficacy of newly developed polymers contained in cosmetic
products a battery of microorganisms relevant for safety assessment of cosmetic products will be used. The tests will be designed so as to
enable to evaluate the speed, extent and stability of antimicrobial action. Namely the duration will be examined for at least 4-6 weeks period.
Specific attention will be focused on both types of emulsions used in cosmetic products (o/w, w/o) and on their pH values due to the nature of an
antimicrobial agent.

The safety assessment of finished cosmetic products according to the foreseen exposure scenario depending of the type of the product
will be performed, using in vitro methods and confirmatory clinical studies on human volunteers according to COLIPA guidelines (issued by
COSMETICS EUROPE – The Personal Care Association).

Task 8.4 Textiles

Multifunctional textiles with selected functional finishing agents will be subjected to microbiological testing with the aim to confirm the extent and
durability of the antimicrobial effect.

The tests will comprise methods for the Resistance to dry microbial penetration (clothing for protection against infection agents) EN ISO 22612,
the Determination of antibacterial activity of antibacterial finished products EN ISO 20743, the Testing method for antibacterial textiles JIS L 1902,
and Testing of textiles – Evaluation of the action of microfungi EN 14119.

Textiles with proven antimicrobial effect foreseen to be used for personal protection, health care or as household products will be assessed
regarding their health safety in a battery of in vitro tests and confirmatory clinical studies.

Task 8.5 Plastics

Plastic products with antimicrobial agents integrated into the polymeric matrix or with antimicrobial coating, intended e.g. as food contact
materials, will be tested by means of the challenge tests for antimicrobial efficiency.

Measurement of antibacterial activity on plastic surfaces will be performed using methods according to International standard ISO 22196 Plastics
– Measurement of antibacterial activity on plastics surfaces.

As all food contact materials (FCM) must comply with the general requirements set by the EU Framework Regulation No. 1935/2004 on
materials and articles intended to come into contact with foods, the final articles/products under normal and foreseeable conditions of use shall
not transfer their constituents into foodstuffs in quantities, which could endanger human health, bring about an unacceptable change in the
composition of the foodstuffs or bring about deterioration in the organoleptic characteristics thereof. Plastic products with antimicrobial agents
integrated into polymeric matrix or with antimicrobial coating are included in the scope of the specific EU Regulation No. 450/2009 on active and
intelligent materials and articles. Active packaging is a type of food packaging with an extra function, in addition to that of providing a protective
barrier against external influence. Materials and articles with antimicrobial agents are intended to influence the packed food. The individual active
substance or, if relevant, the combination of substances which constitute the antimicrobial components should be evaluated to guarantee that
they are safe and comply with the requirements laid down in the Framework Regulation and in the relevant specific measures.

The safety evaluation will be carried out in the National Reference Laboratory for Food Contact Materials, using migration tests, and will include
determination of antimicrobial agents level released into food simulants and foodstuffs under real conditions of use according to EU rules set out
in the Regulation No. 10/2011 on plastics and plastic articles intended for food contact. Sensory analysis also will be performed to verify that the
basic requirements under the FCM legislation are fulfilled.

Task 8.6 Varnishes for surface antimicrobial protection of plastic materials

Different types of plastic materials and foils will undergo surface modification by special antimicrobial varnishes. Chosen plastic materials or
foils with applied varnishes will be tested for antimicrobial efficacy by means of methods according to European standard EN15457 (Paints
and warnishes – Laboratory method for testing the efficacy of film preservatives in coating against fungi) and International standard ISO 22196
(Measurement of antibacterial activity on plastic surfaces – Plastics – Measurement of antibacterial activity on plastics surfaces).
Varnishes with proven antimicrobial effect foreseen to be used for indoor or which may come into direct contact with human body will be
assessed regarding their health safety using a battery of in vitro tests and confirmatory clinical studies will be considered.