Periodic Reporting for period 2 - InnCoCells (Innovative high-value cosmetic products from plants and plant cells)
Reporting period: 2022-11-01 to 2024-04-30
The aim of the InnCoCells project is to develop innovative and sustainable plant-based production processes for the exploitation of scientifically validated cosmetic ingredients based on underutilised plant resources. We will optimise these resources for profitable and sustainable production using cell and hairy root cultures, aeroponics and greenhouse/field cultivation. We will also apply systematic approaches such as metabolic engineering to optimise growth conditions and the yields of valuable bioactive compounds. The optimised processes will be demonstrated by pilot-scale production and product purification. We aim to bring at least 10 cosmetic ingredients to the pre-commercial stage or market. InnCoCells includes a cascade biorefinery concept in which by-products and biowaste are utilised to extract further bioactive molecules. The most promising processes will be characterised by techno-economic analysis and life cycle assessment to ensure economic feasibility and a smaller environmental footprint. The ingredients and extracts will be evaluated using a unique panel of innovative enzymatic, metabolomic and cell-based assays to ensure safety and validate claimed activities based on robust scientific data without animal testing. We will implement a unique stakeholder engagement strategy, including a Stakeholder Group to guide our research program based on the needs of industry, academia, farmers, policymakers and consumers. This industry-driven and interdisciplinary project will ultimately increase the strength of the European bioeconomy by supporting the development of innovative biobased cosmetics.
We selected a portfolio of plant species early in the project, some already available to the partners as cell suspension cultures, hairy roots or whole plants growing on aeroponic surfaces or in greenhouse/field plots. We initiated a bioprospecting program to mine these existing resources and to identify additional target species. Protocols for in vitro cultivation, as well as elicitors and genetic engineering tools, developed for model species have been adapted for our InnCoCells species portfolio to optimise biomass production and metabolite yields.
The cell suspension and hairy root cultures have been optimised at the laboratory scale (up to 20 L) in darkness or with illumination in two different bioreactor systems. Growth and compound accumulation were monitored, and four cell lines were scaled up to 300 L and one to 1000 L. Having established a robust seed train, one hairy root line was scaled up to 300 L. Two of four plants initially grown on 20-m2 aeroponic surfaces have been scaled up to 100 m2, and three plant species have been grown in large-scale field plots.
For the extraction of ingredients from plant biomass, we have investigated different drying and milling methods, pre-treatment with enzymes and pulsed electric fields, and a range of different hydrophilic, lipophilic and supercritical CO2 solvents to improve extraction efficiency and extract stability. A spiral filter press was suitable for the extraction of pilot-scale cell cultures but optimal process parameters and yields are species-dependent. Standard operating procedures for sample extraction, stability testing and bioactivity assays have been compiled, as well as targeted and untargeted metabolomics approaches to determine the chemical composition of bioactive extracts. More than 100 extracts have been tested for anti-inflammatory, anti-aging, antioxidant and antimicrobial properties, and the most promising have also been assessed for cytotoxicity and phototoxicity.
These assays were selected to match consumer demands for ingredients that are safe and efficacious, with a well-characterised mode of action against acne, inflammation, ageing, wrinkles, hyperpigmentation, sun damage, and skin dehydration. To meet these requirements, we have compiled the safety and toxicology data for our most promising ingredients as technical data sheets in line with EU cosmetic regulations and international standards such as EcoCert, COSMOS and Nature. Scientific dossiers based on bioactivity data are being prepared to substantiate product efficacy and safety claims, and to develop a market position. Techno-economic evaluation has been completed for ingredients derived from pilot-scale cell suspension cultures, and life cycle assessment is underway.
We have set out the project’s strategy for dissemination and communication as well as knowledge and IP management. Activities include a project website, social media channels, project brochures, promotional videos, a podcast series, press releases, newsletters and articles in industry periodicals, and collaboration with the three other projects funded under the same H2020 topic. We have published 10 scientific articles so far and have presented talks and posters at more than 50 conferences. We have also raised awareness of the project at high-profile cosmetic industry exhibitions. We have presented seven InnCoCells Academy public webinars as well as a training workshop on plant cell cultures and two invited webinars. Our Stakeholder Group has attended three project meetings and has helped with the selection of promising cosmetic ingredients as well as amplifying our dissemination and communication activities.
The cell suspension and hairy root cultures have been optimised at the laboratory scale (up to 20 L) in darkness or with illumination in two different bioreactor systems. Growth and compound accumulation were monitored, and four cell lines were scaled up to 300 L and one to 1000 L. Having established a robust seed train, one hairy root line was scaled up to 300 L. Two of four plants initially grown on 20-m2 aeroponic surfaces have been scaled up to 100 m2, and three plant species have been grown in large-scale field plots.
For the extraction of ingredients from plant biomass, we have investigated different drying and milling methods, pre-treatment with enzymes and pulsed electric fields, and a range of different hydrophilic, lipophilic and supercritical CO2 solvents to improve extraction efficiency and extract stability. A spiral filter press was suitable for the extraction of pilot-scale cell cultures but optimal process parameters and yields are species-dependent. Standard operating procedures for sample extraction, stability testing and bioactivity assays have been compiled, as well as targeted and untargeted metabolomics approaches to determine the chemical composition of bioactive extracts. More than 100 extracts have been tested for anti-inflammatory, anti-aging, antioxidant and antimicrobial properties, and the most promising have also been assessed for cytotoxicity and phototoxicity.
These assays were selected to match consumer demands for ingredients that are safe and efficacious, with a well-characterised mode of action against acne, inflammation, ageing, wrinkles, hyperpigmentation, sun damage, and skin dehydration. To meet these requirements, we have compiled the safety and toxicology data for our most promising ingredients as technical data sheets in line with EU cosmetic regulations and international standards such as EcoCert, COSMOS and Nature. Scientific dossiers based on bioactivity data are being prepared to substantiate product efficacy and safety claims, and to develop a market position. Techno-economic evaluation has been completed for ingredients derived from pilot-scale cell suspension cultures, and life cycle assessment is underway.
We have set out the project’s strategy for dissemination and communication as well as knowledge and IP management. Activities include a project website, social media channels, project brochures, promotional videos, a podcast series, press releases, newsletters and articles in industry periodicals, and collaboration with the three other projects funded under the same H2020 topic. We have published 10 scientific articles so far and have presented talks and posters at more than 50 conferences. We have also raised awareness of the project at high-profile cosmetic industry exhibitions. We have presented seven InnCoCells Academy public webinars as well as a training workshop on plant cell cultures and two invited webinars. Our Stakeholder Group has attended three project meetings and has helped with the selection of promising cosmetic ingredients as well as amplifying our dissemination and communication activities.
We will move beyond the state of the art by addressing four major bottlenecks: (1) target compounds tend to accumulate in plants at low and variable concentrations; (2) compounds can only be isolated from extremely rare or endangered plants, which are poorly characterised and unsuitable for large-scale production; (3) bioactive compounds cannot be produced on a scale that meets demand and costs, and (4) the discovery of bioactive compounds in plants is not based on systematic analysis. These bottlenecks will be overcome by implementing innovative and systematic approaches to establish and scale up plant lines and cell/tissue cultures and corresponding extraction and bioactivity screening processes. The expected results are: (1) sustainable exploitation of at least 10 relevant metabolic pathways in plant species; (2) a multi-step evaluation pipeline yielding at least 50 scientifically verified active ingredients; (3) optimised production processes and technologies for at least 20 ingredients; (4) evaluation of at least 10 agri-food by-products/waste fractions to generate added value; and (5) pilot-scale production of at least 10 active ingredients, along with economic and sustainability evaluation. The major direct scientific and technological impacts will include novel, natural, sustainable and eco-friendly cultivation and processing methods leading to products with clear scientifically proven bioactive properties as well as the full exploitation of waste streams in a cascade biorefinery approach. The longer-term societal and socioeconomic impacts will include more public-private cooperation in the European biotechnology industry, greater public knowledge of biodiversity, ecosystems and the potential for sustainable exploitation as well as new market opportunities in the European cosmetics industry.