The specific objectives of the project BIOGELFARM are the following:

The synthesis and characterization of new structures of composite micro/nanogels able to transport and controllably release the embedded bioactive natural substances. The aim of obtaining these new composite nanogel structures is to use them as devices for subcutaneous or intravenous injection, thus achieving the release of bioactive substances in hard-to-reach areas, or directly to the affected area. The results of the project are aimed at achieving the following objectives:

• O1. The first specific objective consists of the synthesis of new structures of composite micro/nanogels and of the swelling of hydrogels of polyethylene glycol and natural nanozeolites by polymerization in inverse micro/nano-emulsion method. It is desirable that they exhibit enhanced properties in terms of slow and controlled release of incorporated active substances and also improved viscoelastic properties.
• O2. A new objective consists of the physico-chemical characterization of new structures of micro/nanogels. They will be characterized in terms of morphology, thermal stability, and mechanical properties and also of their capacity to controllably release the embedded natural bioactive substances.
• O3. Another important specific objective is represented by the reproducibility of the results in order to establish the laboratory technology for the obtaining micro/nanogels used in controlled release, which will allow for a patent application in this direction.
• O4. Another important objective of the project, will consist of the dissemination of the obtained scientific results, namely: (i). obtaining prestigious scientific results reflected by an increase in the number of publications of high international impact; (ii). protection of intellectual property rights of authors by filing a patent application; (iii). participation in national and international conferences (at least three international conferences).

Novelty and originality of the proposal consist of the obtaining of covalently crosslinked micro/nanogels based on polyethylene glycol and aliphatic polyamines that will contain nanoparticles of natural zeolite and natural bioactive substances from phytoextracts in their polymer structure. Natural zeolites exhibit various biological activities, being successfully used as compounds which activate the action of a drug (anti-cancer therapy, vaccine, etc.). The role of natural nanozeolites incorporation is to allow for a much slower and controlled release of the embedded active substances and improving the viscoelastic properties of hydrogels. It is also well known that zeolites play also a surfactant role. By placing them in the inverse emulsion polymerization, zeolites can act as a surfactant or co-surfactant making it possible to obtain nanoparticles. Bioactive substances from medicinal herbs have a significant role in the treatment of acute and chronic diseases, such as cancer, AIDS, hepatitis and acute kidney diseases. Nowadays bioactive substances play an important role in the development of controlled release systems in medicinal chemistry for the production of new drugs with fewer side effects. Limitations of this project consist of the fact that these controlled release systems will be tested only in simulated environments. In vivo tests will be carried out within forthcoming innovation type projects.

The overall objective of the project consists of obtaining micro/nanoparticles of covalently crosslinked hydrogels based on polyethylene glycol and aliphatic polyamines (injectable micro/nanogels) having embedded both natural nanozeolites but also of natural bioactive substances in their polymer structure, with potential pharmaceutical applications. These new micro/nanogel composite structures are of great interest as they are aimed at being used as devices for subcutaneous or intravenous injection, ensuring the release of bioactive substances in hard-to-reach areas (by macroscopic hydrogels). The role of natural nanozeolites incorporation is to allow for a much slower and controlled release of the embedded active substances and to improve the viscoelastic features of hydrogels. Nowadays bioactive substances play an important role in the development of controlled release systems in medicinal chemistry for the production of new drugs with fewer side effects. The use of phytoextracts as bioactive substances is a big challenge of the project proposal. These new micro/nanodevices with controlled release will have significant influence, both nationally and internationally on the scientific, social and economic field. The results will be disseminated as follows: three articles published in ISI journals; three scientific communications at international conferences, and patenting of any original aspect.

Getting new structures of micro/nanogels composite containing bioactive natural substances have significant potential in terms of research and development in chemistry and biomedicine. It is well known that the use of synthetic drugs has many side effects, thus the new systems of micro/nanogels for controlled release of bioactive natural substances; significantly reduce harmful effects on the human body. The main advantages of the controlled release of the active ingredient, compared to the conventional administration, consist of a sustained release of active ingredient over a longer period of time and in a concentration in the desired range so as to be used efficiently. This leads also to savings in active principle. Therefore, scientific impact, along with finally getting new controlled release of nanodevices will have significant influence, both nationally and internationally on the scientific, social and economic domain. Social effects consist of attracting PhD students and doctors in research alongside with experienced researchers in the field who will be part of this project team. Developing innovative technology for obtaining and implementing of new nanosystems for controlled release of bioactive natural substances will provide: (i). new devices with improved controlled release of the active substance; (ii). new devices that may have a strong social impact (health, economy and quality of life). Therefore, by financing this project, it is desired to establish synergy between research, biomedicine and medicinal plant processing companies.

Phases I / 2015

Title: Synthesis and characterization of simple, injectable micro/nanogels based on polyethylene glycol and aliphatic amines.

Period: 01/10/2015 - 30/12/2015

On the basis of the literature data presented in the research report related to the stage I / 2015 it was proposed that the encapsulation of the bioactive substances to be done into the networks of micro/nano particles of simple hydrogels. This goal was achieved by the crosslinking reaction of Poly(ethylene glycol) diacrylate (PEGDA) in the presence of tetramethylethylenediamine (TMEDA) and ammonium persulfate (APS) by crosslinking in inverse emulsion technique, by dispersing the aqueous solution of reactants in an organic liquid immiscible with water, in the presence of an emulsifier system composed of sorbitan monooleate (SPAN80) and Poly(ethylene glycol) sorbitan monostearate (TWEEN60).

The singular stage of 2015 met the following objectives:

• Studies regarding the synthesis conditions of micro/nano simple gels by conducting experiments in various conditions;
• The physico-chemical characteristics of the new micro/nanostructures of simple hydrogels.

Thus, the synthesized macromers and the new micro/nanostructured gels were characterized through both physical and chemical methods by means of appropriate methods and devices, such as: Nuclear Magnetic Resonance (NMR) spectroscopy, Fourier Transform Infrared Spectroscopy (FTIR), and by Scanning Electron Microscopy (SEM). The micro/nanogels were characterized in terms of particle size, size distribution and nano-associations by means of Dynamic Light Scattering (DLS) technique.

Phases II/ 2016

Title: Synthesis and characterization of injectable composite micro/nanogels based on polyethylene glycol, aliphatic amines and encapsulated nanozeolites.

Period: 31/12/2015 - 30/12/2016

The main goal of the research study was the preparation of micro/nanocomposite hydrogels, based on Poly(ethylene glycol), aliphatic amines and encapsulated nanozeolites, and finally the elaboration of the technology for hydrogel production.

Thus, for achieving the main goal of this experimental study, the following steps have been completed:

• Synthesis, purification and characterization of the Poly(ethylene glycol) diacrylate (PEGDA) macromer by employing the reaction between PEG and acryloil chloride, which was confirmed in the previous stage as the most adequate method.
• Optimisation of the synthesis conditions of micro/nanogels based on Poly(ethylene glycol) and aliphatic amines and characterization of these simple gels.
• Synthesis and characterization of micro/nano composite gels based on Poly(ethylene glycol), aliphatic amines and encapsulated nanozeolites after previously tested from the point of view of particle size, which is an important parameter in encapsulation.
• The elaboration of the laboratory technology for the synthesis of the controlled release systems made from micro/nanocomposite gels based on Poly(ethylene glycol), aliphatic amines and nanozeolites.

Thus, in the second stage was studied the synthesis and characterization of the micro/nanocomposite gels used as controlled release systems of a bioactive substance in the form of micro/nanocomposite particles of crosslinked Poly(ethylene glycol) diacrylate and micro/nanocomposite particles based on composite hydrogels and natural nanozeolites. Was also considered the synthesis of micro/nanocomposite particles based on the composite hydrogels obtained through the crosslinking reaction of Poly(ethylene glycol) diacrylate in the presence of tetramethylethylenediamine (TMEDA) and ammonium persulfate (APS) by means of crosslinking in inverse emulsion technique by dispersing the aqueous solution of reactants in an organic liquid immiscible with water, in the presence of an emulsifier system composed of sorbitan monooleate (SPAN80) and Poly(ethylene glycol) sorbitan monostearate (TWEEN60). Besides, was studied the synthesis of micro/nanocomposite hydrogel particles, following the same procedure, succeeded by the addition of the inorganic phase (nanozeolite) in the aqueous phase. The synthesized macromer, the nanozeolite and also the new micro/nano(composite) gels were characterized through both physical and chemical methods by means of some appropriate methods and devices, such as: Gel Permeation Chromatography (GPC) with a view to determining the molecular weight distribution of the macromers, Nuclear Magnetic Resonance (NMR) spectroscopy in order to determine the functionality and purity of the synthesized PEGDA macromer, the BET method for determination of porosity and particle size distribution, Dynamic Light Scattering technique for particle size, nanoassociates and Polydispersity Index, Transmission Electron Microscopy for particle morphology and size and for the evidence of encapsulation of zeolite into the hydrogel network, Fourier Transform Infrared spectroscopy (FTIR) for both the chemical composition and identification of some hypothetical interactions between different compounds.

Phases III / 2017

Title: Study on the encapsulation of bioactive natural substances (SBA) in micro/nanogel composite structures with potential properties of controlled release of the encapsulated active substances and with potential applications in the pharmaceutical field.

Period: 31/12/2016 - 30/09/2017

The main objective of this research was to demonstrate the functionality and to optimize the production technology of the new micro/nanocomposite gels designed for controlled release of natural bioactive substances.

Thus, in order to achieve the main goal of this experimental study, the following steps have been completed:

• Optimization of the synthesis conditions and of the ultimate properties for the micro/nanocomposite gels based on Poly(ethylene glycol) diacrylate, aliphatic amines and encapsulated nanozeolites. Furthermore, the controlled release ability of some natural bioactive substances from these hydrogels was also investigated.
• Elaboration of the laboratory technology for synthesis of micro/nanocomposite hydrogels based on Poly(ethylene glycol) diacrylate (PEGDA), aliphatic amines and nanozeolites used as controlled release system.
• Encapsulation of some natural bioactive substances into the nanostructured composite hydrogels network and investigation of their outcome on the ultimate properties.
• Demonstration of the effectiveness of products through encapsulation studies, release and cytotoxicity tests of the encapsulated natural bioactive substances.

The new micro/nanocomposite gel structures have been characterized through both physical and chemical methods by means of appropriate methods and devices, such as: the BET method for determination of porosity and particle size distribution, Dynamic Light Scattering for particle size, nanoassociates and Polydispersity Index, Transmission Electron Microscopy for particle morphology and size, and for revealing the evidence of encapsulation of zeolite into the hydrogel structure, Fourier Transform Infrared Spectroscopy (FTIR) and TGA / DTG analysis for both the chemical composition and revealing the existence of some hypothetical interactions between different compounds, and UV-VIS spectroscopy for the control release of the bioactive substances from the PEGDA hydrogels.

Moreover, some cytotoxicity tests were performed by using of the L929 murine fibroblast cell line. 

In conclusion, the formulated nanocomposite hydrogels are promising systems for controlled delivery of natural bioactive substances.

ü  Two patents application was filed:

1. “Nanohydrogels for the slow release of drugs and their synthesis procedure”, Authors: Anamaria Zaharia, Anita-Laura Radu, Andrei Sarbu, Mircea Teodorescu, Bogdan Cursaru, Paula Catalina Spatarelu, Tanta-Verona Iordache, Sandu Teodor, Ana Mihaela Florea, DOSAR OSIM:  A 2016 -00620/07.09.2016

2. “Inorganic-organic hydrides based on nanozeolite and nanogels for the controlled release of drugs and the process for their preparation”, Authors: Anita-Laura Radu, Anamaria Zaharia, Andrei Sarbu, Mircea Teodorescu, Bogdan Cursaru, Paula Catalina Spatarelu, Sandu Teodor, Tanta- Verona Iordache, DOSAR OSIM: A 2017 -00105/23.02.2017.

ü  The results of project were presented by team members within the framework of several scientific manifestations (participation at different national and international conferences), such as:

1. Hydrogels for controlled delivery of biologically active substances”, Catalina-Paula Spatarelu, Anamaria Zaharia, Anita-Laura Radu, Tanta-Verona Iordache, Ana-Mihaela Florea, Teodor Sandu, Andrei Sarbu, Mircea Teodorescu, Conferinta 5th Portuguese Young Chemists Meeting (5th PYCheM) and 1st European Young Chemists Meeting (1st EYCheM), 26-29 Aprilie 2016, Guimaraes, Portugalia, comunicare tip poster

2. “Synthesis paths for micro-hydrogels containing polyethylene glycol and aliphatic amines”, Anita-Laura Radu, Anamaria Zaharia, Bogdan Cursaru, Catalina-Paula Spatarelu, Tanta-Verona Iordache, Ana-Mihaela Florea, Raluca Ianchis, Teodor Sandu, Andrei Sarbu, Mircea Teodorescu, Conferinta 5th Portuguese Young Chemists Meeting (5th PYCheM) and 1st European Young Chemists Meeting (1st EYCheM), 26-29 Aprilie 2016, Guimaraes, Portugalia, comunicare tip poster

3. "L'obtention des hydrogels pour des systèmes a libération contrôlée", Catalina Paula Spatarelu, Anita-Laura Radu, Anamaria Zaharia, Bogdan Cursaru, Tanta-Verona Iordache, Ana Mihaela Florea, Raluca Ianchis, Teodor Sandu, Andrei Sarbu, Mircea Teodorescu, Le Neuvieme Colloque Franco-Roumain de Chimie Appliquee, 29 Iunie-2 Iulie 2016, Clermond-Ferrand, Franta, comunicare tip poster

4. "Hydrogels composites innovatrices pour la liberation controlee des engrais chimiques", Anita-Laura Radu, Anamaria Zaharia, Catalina Paula Spatarelu, Andrei Sarbu, Tanta-Verona Iordache, Ana Mihaela Florea, Teodor Sandu, Stela Iancu, Anicuta Stoica, Marta Stroescu, Le Neuvieme Colloque Franco-Roumain de Chimie Appliquee, 29 Iunie-2 Iulie 2016, Clermond-Ferrand, Franta, comunicare tip poster

5. « Modelling the separation of algae extracts by a coupled ultrafiltration/pervaporation process », Bogdan Trica, Tanase Dobre, Gholamreza Djelveh, Raluca Somoghi,Le Neuvieme Colloque Franco-Roumain de Chimie Appliquee, 29 Iunie-2 Iulie 2016, Clermond-Ferrand, Franta, comunicare orala

6. "Influence of synthesis parameters on the obtaining of PEG covalently cross-linked nanogels", Catalina-Paula Spatarelu, Anita-Laura Radu, Anamaria Zaharia, Bogdan Cursaru, Andrei Sarbu, Tanta-Verona Iordache, Ana-Mihaela Florea, Bogdan Trica, Teodor Sandu, 14th Young Chemists’ Congress (YOUNGCHEM), 5-9 Octombrie 2016, Czestochowa, Polonia, comunicare orala

7. ”Hybrid hydrogels based on bio and synthetic polymers with applications in agriculture”, Anita-Laura Radu, Anamaria Zaharia, Catalina Paula Spatarelu, Andrei Sarbu, Tanta-Verona Iordache, Ana Mihaela Florea, Teodor Sandu, Stela Iancu, 14th Young Chemists’ Congress (YOUNGCHEM), 5-9 Octombrie 2016, Czestochowa, Polonia, comunicare tip poster

8. “Obtaining of new nanogels for controlled delivery of biologically active substances”, Bogdan Cursaru, Catalina-Paula Spatarelu, Anita-Laura Radu, Anamaria Zaharia, Andrei Sarbu, Tanta-Verona Iordache, Ana-Mihaela Florea, Bogdan Trica, Teodor Sandu, Mircea Teodorescu, XXXIX-th Romanian Chemistry Conference, 4-7 Octombrie 2016, Caciulata, Romania, comunicare tip poster

9. “Novel nanogel composites for controlled-drug delivery of chemotherapeutics”, A-L. Radu, A. Zaharia, B. Cursaru, C-P. Spatarelu, T-V. Iordache, A-M. Florea, R. Ianchis, T. Sandu, A. Sarbu, M. Teodorescu; Fifth International Symposium Frontiers in Polymer Science”; in Spania, Seville, in perioada 16.05.2017 - 20.05.2017.

10. „New PEGDA/nano-zeolite composite hydrogels for controlled delivery of biologically active substances”, B. Cursaru, A.L. Radu, A. Zaharia, C-P. Spatarelu, A. Sarbu, T-V. Iordache, A-M. Florea, B. Trica, T. Sandu, M. Teodorescu, “13th International Conference on Materials Chemistry” desfasurat in in Marea Britanie, Liverpool, perioada 09.07.2017 - 13.07.2017.

       4 ISI articles were sent for publication, of which one published and three were submitted for evaluation and possible publication, with cumulative IF of  7.189.

       1. “Novel poly(ethylene glycol) diacrylate-nanogels synthesized by mini-emulsion technique”,Anita-Laura Radu,Bogdan Cursaru, Catalina Paula Spatarelu, François-Xavier Perrin, Tanta-Verona Iordache,Ana-Mihaela Florea, Raluca Ianchis, Bogdan Trica, Teodor Sandu, Andrei Sarbu,Mircea Teodorescu, Anamaria Zaharia, trimis spre evaluare in Journal of Dispersion Science and Technology, IF : 1,591 (editura Taylor&Francis)

       2. “Straightforward preparation of naphtodianthrone-rich ethanolic extracts from wild St. John’s Wort”,Carmen Elena ?ebrencu, Tan?a-Verona Iordache, Elena Ionescu, Ruxandra Mihaela Cre?u, Ana-Mihaela Florea, Sandu Teodor, Steluta Apostol, Anamaria Zaharia, Anita-Laura Radu, Andrei Sarbu, trimis spre evaluare in Journal Of Dietary Supplements, IF : 1,05 (editura Taylor&Francis)

       3. “One-step preparation of molecularly imprinted hollow beads for pseudohypericin separation from Hypericum perforatum L. Extracts”, Ana-Mihaela Florea, Tan?a-Verona Iordache, Catherine Branger, Hugues Brisset, Anamaria Zaharia, Anita-Laura Radu, Gheorghe Hubca, Andrei Sarbu, trimis spre evaluare in Polymer, IF : 3.77 (editura Elsevier)

       4. "Evaluation of molecularly imprinted polymer pearls for selective isolation of hypericins", Ana-Mihaela Florea, Tan?a-Verona Iordache, Anamaria Zaharia, Bianca Georgescu, Andreea-Elena Voicu, Boiko Tsyntsarski, Gheorghe Hubca, Andrei Sarbu, MATERIALE PLASTICE, 54 (No. 3), 2017, (IF: 0,778).