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OBJECTIVES OF STAGE 3/2020

 

       Within this stage of the complex project, the following specific objectives were proposed (in part):

Objective 1. Development of new high-performance regenerative dermato-cosmetic and pharmaceutical products based on marine resources with synergistic action, physico-chemical and microbiological evaluation and therapeutic effect of dermato-cosmetic and pharmaceutical products obtained on optimal experimental models in cell cultures specific to dermal tissue;

Objective 2. Incorporation of polyphenolic extracts obtained from red pomace varieties and characterization of materials with encapsulated extracts by different techniques, thermal analysis, FTIR spectroscopy, in vitro determination of free radical scavenging activity, adsorption-desorption isotherms and determination of release profiles of polyphenols in functionalized mesoporous silica matrices;

Objective 3. Obtaining and characterization of mesoporous silica-ulvan composite materials, by preliminary studies on the coating of silica particles with ulvan, sulfonated polysaccharide extracted from green alga Ulva lactuca that has proven good biocompatibility with an anticancer agent, resveratrol, for which its release profiles were determined from different matrices of pure and functionalized silica.

 

STAGE SUMMARY

 

             In order to achieve the objectives proposed at this stage, in the framework of the component 1 project, 4 variants of new dermato-cosmetic products (P1, P2, P3 and P4) were obtained, conditioned as bioadhesive gels, by incorporating biologically active compounds, respectively gelatin (GEL) and glycosaminoglycans (GAG), isolated and purified from Rapana venosa as well as hydroalcoholic extracts of green seaweed (Cladophora vagabunda, Ulva lactuca and Enteromorpha intestinalis) in hydrophilic gel bases consisting of Carbopol Ultrez 10 NF and Carbopol 940, respectively NF, in different concentrations and combination ratios. The variants also contain a natural preservative, namely citromycin extract from grapefruit seeds as well as citric acid and triethanolamine for pH regulation. The stages of the technologies for obtaining gelatin powder from Rapana venosa and these dermato-cosmetic products were established. The physico-chemical properties of the pharmaceutical preparations were determined in accordance with the provisions of the Romanian Pharmacopoeia, 10th Ed., These being: firmness, adhesiveness, consistency, stretching capacity, pH, degree of penetration into the skin. Optimized experimental methods and models were used to determine the antioxidant capacity by the photochemiluminescence method using the ACL technique. The antibacterial effect of the samples was tested according to the standard SR EN ISO6579 / 2002 / (E) on specific microbial strains. The therapeutic effect of the obtained products was tested on optimal experimental models in cell cultures specific to dermal tissue. In the present step, the non-irritating potential of the samples was evaluated using an in vitro experimental model by dosing the pro-inflammatory cytokines IL-1α and IL-6, secreted by the skin cells - keratinocytes HACaT. The effect of stimulating cell viability and proliferation by the tested bio-adhesive gels was also demonstrated in vitro, on an experimental model using NCTC L929 connective tissue fibroblasts, by MTT and LDH methods. The results of physico-chemical and biological tests showed that the four variants of bio-adhesive gels showed good preservation over time, had free radical scavenging activity, a high degree of biocompatibility and did not induce an irritating effect. All the variants of pharmaceutical products were selected as optimal, and in the next stage their antimicrobial activity, the effect on the tissue regeneration process, the photoprotective action on skin cells, etc. will be tested. We obtained 2 variants of biostimulator-regenerator based on marine and wine bio-resources, with fertilization potential. Following their testing on vine crop soils, the positive effect on the growth of vine shoots was demonstrated, as well as on the content of chlorophyll pigments and carotenoids in the leaves.

             In order to achieve the objectives proposed at this stage, in the component 2 project, experiments were performed to determine the in vitro release profiles of cytostatic agent, irinotecan, from mesoporous matrices of silica type SBA-15 and a natural compound with effect anticancer, trans-resveratrol. Studies have also been made to obtain mesoporous silica-ulvan composite materials. Coating the mesoporous silica matrix with ulvan will ensure the targeting of the carcinogenic tissue. By approaching the two directions of research, we will aim to determine both the optimal conditions for the release profile of the cytostatic agent (irinotecan) and trans-resveratrol (silica matrix, concentration of biologically active substance), as well as the method and content of solvent covers silica particles.

             Also at this stage, studies were continued on the determination of polyphenol release profiles from extracts obtained from tescovine encapsulated in mesoporous matrices of pure MCM 41 type and functionalized with acid groups, which favor a better stability over time of polyphenolic extracts and which have proven better bioavailability by increasing their solubility in simulated biological fluids. The encapsulation of the polyphenolic extract from the Mamaia pomace variety was the most efficient in the silica support functionalized with propionic acid groups. In this stage, 4 variants of pharmaceutical preparations such as bio-adhesive gels were obtained, by incorporating biologically active compounds, respectively gelatin powder isolated and purified from marine mollusks Rapana venosa, as well as hydro alcoholic extracts from by-products of the vinification-pomace process. red varieties and hydro alcoholic extracts of green seaweed of the genus Chlorophyta incorporated in hydrophilic gel bases consisting of the carbomers Carbopol Ultrez 10 NF and Carbopol 940 NF, in well-specified concentrations and combination ratios, associated with the preservatives citromycin and citric acid, respectively triethanolamine pH regulator. The pharmaceutical preparations obtained have biological safety and have been evaluated for their antimicrobial activity against 2 bacterial strains and a fungal strain specific to dermal tissue and have shown good antibacterial activity, no cytotoxicity, which recommends them as potential products with antibacterial properties and with stimulating-biodegenerating effect.

 

             Therefore, by carrying out the activities of this stage, all the proposed objectives were achieved, all achievement indicators were met and exceeded, and the value of the results obtained is supported by the dissemination activity, being presented in 4 international conferences and by publishing 4 ISI articles with impact factor (of which 1 article in a top ISI journal, Antioxidants, FI = 5.014, area Q1), 2 articles in an ISI journal are being published, 2 articles in journals have been published indexed BDI and 2 patent applications were filed with OSIM, which consist in obtaining dermato-cosmetic preparations such as bioadhesive gels based on extracts from marine organisms and respectively obtaining a biostimulator-regenerator as a biomass-based biofertilizer residues from marine organisms and by-products from viticulture. Also, by implementing the third stage of the complex project, the infrastructure of all partners was improved and their service offer was diversified. All 6 new employed researchers, with a very good professional training (doctoral students and master students), were actively involved in carrying out the activities of the complex project.

 

The human resource involved in carrying out the activities from Stage 3/2020.

Jobs supported by the program, including newly hired human resources:

 

  • UOC Coordinator: 11 experienced researchers prof. (3), conf. (5), head of works (2), CSIII (1). No new hires made.

  • P1-INCDM partner: 9 experienced researchers with CSI (4), CSIII (2) positions, of which 2 post-doctoral students, 3 doctors, 1 doctoral student, 3 technicians. No new hires made.

  • P2-UPB partner: 2 experienced researchers and 2 research assistants (PhD students) employed full time.

  • P3-INCDSB partner: 14 specialist researchers with CS, CS III, CS II, CS I positions, of which 7 doctors, 3 post-doctoral students, 2 doctoral students as well as 2 technicians. The new employees (1 CS and 1 ACS) held the 2 positions in the field of chemistry-biochemistry.

  • P4-ICF partner: 6 experienced researchers and 1 young full-time doctoral student at UPB.

  • P5-SCDVV partner: 3 experienced researchers and 1 full-time research assistant. The young new employee enrolled for a doctorate at UPB, Scientific Coordinator Prof. Dr. Daniela Berger, Project Manager component 2, the topic of the doctoral thesis being in the field of the project. P5 did not purchase equipment at this stage.

 

             Presentation of the structure of the research and technological services offer with the indication of the link from the Erris platform:

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DESCRIPTION OF STAGE No. 3

 

Name: Development of high-performance dermato-cosmetic and pharmaceutical products based on resources of marine origin with synergistic action and demonstration of their functionality - Part 1

(Reporting period: 01/01/2020 - 31/12/2020)

 

EXPECTED RESULTS:

• Optimal technological parameters

• Encapsulation methods

• Embedding methods

• Microbial loading parameters

• In vitro experimental methods and models for evaluating the regenerative, anti-inflammatory and anti-tumor effect

• Manufacturing formulas

• Technology for obtaining biostimulator-regenerator

• Technology for obtaining a bioregenerative pharmaceutical product

• Technology for obtaining a pharmaceutical product with cytostatic action

• Technology for obtaining a product with antimicrobial activity

• Controlled drug delivery systems with synergistic action and in vitro release profiles

• High performance pharmaceutical preparations

• Patenting of the obtained results

• Communication and publication of results

 

COMPONENT PROJECT NO. 1

• Activity 3.1.: Development of dermato-cosmetic products and their conditioning in different forms - Part 1

 

Achievement indicators:

• 2 biotechnologies

• 3 variants of conditioning dermato-cosmetic products

 

• Activity 3.2.: Physico-chemical and microbiological evaluation of dermato-cosmetic products - Part 1

 

Achievement indicators:

• 1 method of antioxidant activity analysis

• 1 bulletin for the analysis of physico-chemical parameters

• 1 analysis bulletin for antimicrobial activity

 

• Activity 3.3.: Demonstration of the therapeutic effect of the products obtained on optimal experimental models in cell cultures specific to dermal tissue - Part 1

 

Achievement indicators:

• 2 optimized methods for biocompatibility analysis

• 1 in vitro experimental model

 

• Activity. 3.4.: Selection of optimal variants and development of prototype products with superior dermato-cosmetic efficiency - Part 1

 

Achievement indicators:

• 2 dermato-cosmetic products

 

• Activity 3.5.: Processing of biological waste resulting from intermediate technological processes for conversion to obtain a biostimulator-regenerator for viticultural soils - Part 1

 

Achievement indicators:

• 1 biostimulator-regenerator - preliminary version

 

• Activity 3.11.: Dissemination of results - Part 1

 

Achievement indicators:

• 1 paper communicated at a scientific event

• 1 article published in the ISI / BDI magazine

 

• Activity 3.12.: Innovation activity and patent application - Part 1

 

Achievement indicators:

• 1 patent application

 

COMPONENT PROJECT NO. 2

 

• Activity 3.6.: Separation from natural extracts of components with synergistic action and their biochemical characterization - Part 1

 

Achievement indicators:

• extracted technological parameters - partially

 

• Activity 3.7.: Incorporation of a cytostatic agent into the mesopores of the functionalized silica-type matrix with compounds from natural extracts and characterization of the resulting composites - Part 1

 

Achievement indicators:

• 1 functionalized mesoporous matrix of silica type

 

• Activity 3.8.: Assessment of the influence of the natural extract on the in vitro release profiles of the cytostatic agent - Part 1

 

Achievement indicators:

• kinetic parameters - partially

 

• Activity 3.9.: Evaluation of the synergistic activity of the cytostatic agent and the component of the marine extract on cell cultures - Part 1

 

Achievement indicators:

• 1 analysis report for cell viability

• 1 cell morphology report

 

• Activity 3.10.: Formulation of pharmaceutical / bioregenerative products - Part 1

 

Achievement indicators:

• 1 pharmaceutical product

• 1 analysis bulletin for antimicrobial activity

 

• Activity 3.13.: Widespread dissemination through national or international communication and publication of results - Part 1

 

Achievement indicators:

• 2 papers communicated at scientific events

 

• Activity 3.14.: Innovation activity and patent application - Part 1

 

Achievement indicators:

• 1 patent application

Objective of stage 3/2020
Stage summary
Expected results
Component project 1
Component project 2
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