- Acronym: SmartACT
- Project Code: PN-III-P1-1.1-TE-2021-1342
- Contract Number: 96TE
- Start: 15/05/2022
End: 13/09/2024 - Duration: 28 months
- Budget: 450.000,00 Lei
- Contracting Agency:: Executive Agency for Higher Education, Research, Development and Innovation Funding
- Coordinating Institution: : National Universuty for Science and Technology POLITEHNICA Bucharest – National Research Center for Micro and Nanomaterials
Address: Spl. Independentei 313, Sector 6, Bucharest, Romania
Phone: +4021.402.9465
Coordinator (CO) – UNIVERSITATEA POLITEHNICA DIN BUCURESTI – Project Manager – Conf. Univ. Ficai Denisa
Coordinator (CO) – UNIVERSITATEA POLITEHNICA DIN BUCURESTI – CS III Vasile Bogdan
Coordinator (CO) – UNIVERSITATEA POLITEHNICA DIN BUCURESTI – Lect. Univ. Neacsu Andreea-Ionela
Coordinator (CO) – UNIVERSITATEA POLITEHNICA DIN BUCURESTI – CS Angela Spoiala
Coordinator (CO) – UNIVERSITATEA POLITEHNICA DIN BUCURESTI – PhD Student Ludmila Motelica
Coordinator (CO) – UNIVERSITATEA POLITEHNICA DIN BUCURESTI – PhD Student Cornelia-Ioana Ilie
Coordinator (CO) – UNIVERSITATEA POLITEHNICA DIN BUCURESTI – PhD Student Alexandra Catalina Birca
The project scope is to improve medical strategies regarding cancer treatment, by the development of new nanostructured systems for targeted cancer therapy, based on natural catabolism products (dicarboxylic amino acids, hydroxyacids and keto acids as coating agents and natural or synthetic antitumor compounds, with induced cellular internalization. In order to achieve this main objective and minimize the negative side effects of current approaches, 4 specific objectives (SO) were taken into account, as follows:
SO 1. Fabrication and characterization of Fe3O4@ multifunctional compounds nanostructures, using dicarboxylic amino acids (e.g. glutamic acid, aspartic acid or their derivatives), hydroxyacids (e.g. tartaric acid, gluconic acid, vitamin C/ascorbic acid, folic acids) and keto acids (e.g. alpha-ketoglutaric acid), which, to the extent of our knowledge, were not yet reported as coating agents, representing a novelty in the field. The presence of these biological active agents, will allow an improved internalization because the tumoral cells are more active comparing to the normal cells and need to satisfy their higher need of food and oxygen. If required, these structures will be further stabilized to assure a better stability by PEG-ylation.
SO 2. Fabrication and characterization of Fe3O4@MC–Hydrophilic Cytostatic, as targeted cancer therapy systems. There are numerous hydrophilic antitumoral agents which can be used for cancer treatment. Their delivery usually occurs faster than the degradation of the matrix, the delivery mechanism being characteristic to a non-erodible matrix. The delivery can be easily triggered by an external alternative electromagnetic field.
SO 3. Fabrication and characterization of Fe3O4@MC – Hydrophobic Cytostatic, as targeted cancer therapy systems. At the expense of classical drugs used in cancer treatment, the project proposes fabrication of systems based on natural, non-toxic and commonly found in the diet, antitumor compounds, such as Bisabolol and Lycopene, which can be extracted from plants and vegetables. Their antitumor activity was demonstrated but only few reports were published so far. The hydrophobic cytostatics are especially important because their delivery is slow in aqueous media if no stimuli is applied.
Cancer is one of the most widespread group of diseases causing about 14.6% of all human deaths, representing a major public health issue worldwide. Standard treatment is usually causing side effects which can be reduced by targeting delivery with various nanostructures, including magnetic nanoparticles. Due to the fact that bare Fe3O4 nanoparticles are highly susceptible to degradation/dissolution in acidic and oxidative conditions as well as to the in vivo conditions, coating an outer protective layer to it is very important for maintaining the stability of the magnetic component until cellular internalization. Also, the proper choice of the shell will be essential in assisting the internalization of these nanostructures inside the tumor cells and further delivering the antitumoral agents directly inside the cells. The aim of the project is to improve medical strategies regarding the treatment of cancer, by developing new nanostructured systems for its targeted therapy, such as Fe3O4_dicarboxylic amino acids, hydroxy acids and keto acids, (as shell) and natural or synthetic antitumor compounds and thus to get improved cellular internalization and release of the antitumoral agents. The results obtained are promising, the Fe3O4@shell/cytostatic magnetic systems being potential solutions for the treatment of cancer, through the development of new nanostructured systems for its targeted therapy. The shell, natural products of catabolism (dicarboxylic amino acids, hydroxyacids and ketoacids) and polymers were used to assure stabilisation and/or internalization and natural or synthetic antitumor compounds were evaluated with the main aim to improve internalization into the tumoral cells and thus to reduce the harmful side effects (including systemic toxicity). The increase in internalization and targeted delivery of therapeutic agents was achieved by using amino acids, polyethylene glycol (PEG) and albumin as functionalization agent, with the role of increasing the stability of multifunctional magnetic systems and to control the release of embedded active substances in a triggered and targeted way. Polyethylene glycol (PEG) is frequently used as an agent to increase the stability of multifunctional magnetic systems in vitro and in vivo, due to its increased hydrophilicity in aqueous media, it can considerably reduce the resorption of magnetic nanoparticles by the reticuloendothelial system and, as a consequence, the circulation time compared to uncovered nanoparticles increases, nanoparticle aggregation decreases as a result of stabilization, and the association with non-targeted tissues and cells is reduced along with the systemic toxicity.
Also the “remote driving” of magnetic nanoparticles, under the influence of an external magnetic field, combined with the intrinsic penetrability of the magnetic field through the human body, magnetic nanoparticles are intended for targeted transport and triggered release of biologically active substances (transport agent in chemotherapy, antibiotics, analgesics, polyphenols, etc.) to a targeted region in the body. The advantage of the Fe3O4_CM type magnetic system using PEG, glutamic acid and aspartic acid as stabilizing and internalizing agents comes from the system’s multifunctionality, namely its role in treating cancer through the hyperthermia effect of magnetite as well as anti-inflammatory, antipyretic, antithermic, antiseptic and anti-infective thanks to the biologically active agents. The presence of these active biological agents will allow improved internalization and better stability at the cellular level because tumoral cells are more active compared to normal cells and must satisfy their greater need for specific compounds (vitamins, oligo-elements, …), nutrients and oxygen.
Based on these considerations, in stage I of the project, a series of multifunctional Fe3O4@amino acids, hydroxy acids and keto glutaric acid systems were synthesized and characterized. The magnetic systems were characterized from a structural and morphological point of view with the help of the physico-chemical methods characteristic for this type of materials. Depending on the nature of the CM, the change in the morphology of the mesoporous magnetic nanoparticles can be observed. Following the biological evaluation of the obtained Fe3O4@MC systems, most of the magnetic systems stabilized with the biologically active substances presented a better antimicrobial activity compared to the controls, and a synergistic effect was also observed. Among them, Fe3O4@ L-tryptophan determined the highest sensitivity of E. coli, P. aeruginosa and C. albicans, with MIC values of 0.001 mg/mL. Fe3O4@L-cysteine inhibited the adhesion of all strains studied. Cell viability, regardless of the nature of the stabilizer used (serine, tryptophan, cysteine, proline, glutaric, a-keto-glutaric, tartaric, usnic) is greater than 60%, which confirms good compatibility.
In the second stage of the project, series of multifunctional Fe3O4@PEG_CM magnetic systems was obtained using PEG, glutamic acid and aspartic acid as stabilizing and internalizing agents. The presence of these active biological agents will allow improved internalization and better stability at the cellular level because tumor cells are more active compared to normal cells and must satisfy their greater need for nutrients and oxygen. These nanoparticle systems were compositionally and morphologically characterized by Fourier transform infrared spectroscopy (FTIR), X-ray diffraction (XRD), scanning electron microscopy (SEM), transmission electron microscopy (TEM-HRTEM), as well as chemical stability in specific environments. Detailed structural characterization using the specified techniques confirmed the successful formation of Fe3O4/amino acid (AA) and Fe3O4/PEG/amino acid (AA) core/shell hybrid structure. The X-ray diffractograms obtained on the magnetic nanopowders coated with PEG and/or amino acids indicate the presence of the magnetite core and thus the specific peaks of crystalline magnetite can be observed. TEM and DLS analyses showed good stability and polydispersity of these systems with an almost spherical shape.
These Fe3O4@MC/PEG nanostructures are able to adsorb cytostatic molecules and target them to the diseased organ/tissue followed by targeted drug release. This innovative and targeted drug delivery strategy involves coupling the drug to magnetic nanoparticles (NPs) that can be guided to the target using external magnetic fields. Once they reach the target organ/tissue, the nanoparticles release the drugs under the influence of the alternating magnetic field that can generate hypothermia and implicitly increase the release rate of cytostatics. At the present stage, the internalization capacity of the proposed and realized magnetic structures has been proven, and the studies will continue with a view to their optimization and access to higher levels of technological development
Also in the second stage of the project, a series of drug delivery systems based on citrate-stabilized MNPs, which act as nanocarriers for polyphenolic compounds (PCs) from extracts (BBE and BPE) with 5-fluorouracil. This study presents the development of drug delivery nanocarriers based on magnetite and trisodium citrate to improve the antitumor activity of 5-FU and mitigate the adverse effects on gut microbiota. MNPs were synthesized by a spray assisted method and loaded with bee pollen extracts (BPE) or bee bread extracts (BBE) and 5-FU. BPEs and BBEs have been characterized in previous studies. First, the loaded MNPs were morphologically and structurally characterized by physical analysis. The release behaviour of the bioactive agents was evaluated for 5-FU and PC, and their antioxidant activity was evaluated by three assays. The purpose of this research is based on the use of low concentrations of extracts and 5-FU to establish the influence of each bioactive compound and the effects on a selected bacteria with probiotic potential. In addition, the novelty of this study involves the increase of the inhibitory activity of the bacterial adhesion capacity to the inert substrate induced by the synthesized nanocarriers.
The results demonstrated that MNPs with narrow size distributions and an average NP size of ~10 nm were obtained. Moreover, it was shown that the antitumoral drug and PCs had fast release rates in the first 24 h from MNPs. Furthermore, the PCs exhibited prolonged release behaviour (up to 96 h), and loading MNPs with extracts enhanced their antioxidant and antibacterial properties. Additionally, the non-toxic properties of magnetite and the prebiotic potential of the developed DDS on L. rhamnosus MF9 were demonstrated, underline the role of the antioxidants (PCs) in supporting gut microbiota which is essential in the cancer treatment, microbiota being able to control the healing capacity of the body, in general via different mechanisms. The moderate antiproliferative activity on a colorectal tumoral cell line at low concentrations of MNPs and 5-FU suggests the potential of DDS to alleviate the adverse effects of anticancer drugs and improve gut balance.
In the third stage of the project the antimicrobial, antitumoral and internalization capacity of the obtained systems in the first and second stage was evaluated. These magnetic nanoparticles were developed, these being able to function as Trojan Horses and to carry cytostatic drugs into the tumoral cells. This is important because the antitumoral activity is correlated with the cytostatic level from the cell and this is why the internalization is so important and many research teams are looking for solutions to improve the internalization. In the literature, most of the internalization studies are done using folic acid and here we proved that Aspartic acid could also improve the internalization capacity and thus improved antitumoral activity is obtained. Based on the high resolution images, it can see that PEG-ylated nanoparticles are quite stable and can reach and enter the mitochondria and organize around lipid vesicles in quite a high concentration. Best results, according to this study are obtained for Fe3O4_PEG_Asp_CisPt and Fe3O4-Asp-CisPt at both 100 and 500 µg/ml. Further research is needed to study the complex mechanism occurring inside the cells, which involves internalization and resorption and to correlate the ratio between Fe3O4, PEG and Aspartic acid, with the synthesis procedure and the type of the cancer. It is important to mention that a proper carrier have to be stable enough to assure internalization and, once these are inside, they can be destroyed and release the cargo (cytostatic drugs). Certainly, magnetic triggering could be also expected to be a solution to achieve a personalized therapy with external control of the delivery rate.
International conferences |
1. Denisa Ficai, Ludmila Motelica, Dan Adrian Vasile, Adrian Vasile Surdu, Roxana Doina Trusca, Ionela Andreea Neacsu, Bogdan Stefan Vasile, Ovidiu Cristian Oprea, Anton Ficai, Ecaterina Andronescu Design and synthesis of magnetic nanoparticles for applications in theranostics 22nd Romanian International Conference on Chemistry and Chemical Engineering (RICCCE), Septembrie 7-9, Sinaia 2. Motelica Ludmila, Ficai Denisa, Oprea Ovidiu, Ficai Anton, Andronescu Ecaterina, Truşcă Roxana, Surdu Adrian Synthesis and characterization of magnetite nanoparticles stabilized with amino acids, 22nd Romanian International Conference on Chemistry and Chemical Engineering (RICCCE), Septembrie 7-9, Sinaia 3. Angela SPOIALĂ, Cornelia-Ioana ILIE, Cristina UNCU, Ludmila MOTELICA, Roxana-Doina TRUȘCĂ, Denisa FICAI, Adrian SURDU, Ovidiu OPREA, and Anton FiCAI, New functionalized magnetic nanoparticles for targeted drug delivery systems, 6th EuChemS Inorganic Chemistry Conference, Viena, Austria, 02.09.2023 – 08.09.2023. 4. Denisa Ficai, Ludmila Motelica , Angela Spoiala, Cornelia Ioana Ilie, Cristina Chircov, Roxana Doina Trusca, Adrian Vasile Surdu, Bogdan Stefan Vasile, Ovidiu Cristian Oprea, Anton Ficai, Ecaterina Andronescu, Magnetic smart systems for theranostic , 8TH INTERNATIONAL CONFERENCE ON NANOTECHNOLOGY: FUNDAMENTALS AND APPLICATIONS (ICNFA’23), 08-12.08.2023, Londra, Marea Britanie. Best paper Award 5. Denisa Ficai, Ludmila Motelica, Cristina Chircov, Roxana Doina Trusca, Adrian Vasile Surdu, Bogdan Stefan Vasile, Ovidiu Cristian Oprea, Geanina Voicu, Manuela Calin, Victor-Eduard Peteu, Anton Ficai, PEG and aspartic acid-functionalized Fe3O4 nanoparticles as efficient theranostic vectors in cancer, 15th International Conference on Nanotechnology: Fundamentals and Applications (ICNFA 2024), Barcelona, Spain – August 25-27, 2024. 6. Denisa Ficai, Ludmila Motelica, Cristina Chircov, Roxana Doina Trusca, Adrian Vasile Surdu, Bogdan Stefan Vasile, Ovidiu Cristian Oprea, Geanina Voicu, Victor-Eduard Peteu, Anton Ficai, Manuela Calin, Magnetic Nanoparticles for Targeted Antitumoral Therapy, 23rd Romanian International Conference on Chemistry and Chemical Engineering, Constanța – Mamaia, ROMANIA – September 4 – 7, 2024 |
International salons of inventions and innovations |
Denisa FICAI, Ludmila MOTELICA, Cornelia Ioana ILIE, Angela SPOIALA, Ovidiu Cristian OPREA; Anton FICAI, Magnetic smart drug delivery systems for theranostic using a personalized approach; Project PN-III-P1-1.1-TE-2021-1342, no 96, 15th European Exhibition of Creativity and Innovation-EuroInvent 2023, 13 May 2023, Iaşi, Romania, 4 Premii obtinute- Carol Davila Award – UMFCD, Diploma de excelenta Asociatia Justin Capra, Diploma de excelenta cu mentiune speciala USMF, Bronze medal |
ISI Articles |
1. Spoială Angela, Ilie Cornelia-Ioana, Ludmila Motelica, Ficai Denisa, Augustin Semenescu, Oprea Ovidiu Cristian, Ficai Anton, Smart Magnetic Drug Delivery Systems for the Treatment of Cancer, Nanomaterials, Nanomaterials (Basel). 2023 Mar; 13(5): 876, Published online 2023 Feb 26. doi: 10.3390/nano13050876, 2. Denisa-Maria Radulescu, Vasile-Adrian Surdu , Anton Ficai , Denisa Ficai, Alexandru-Mihai Grumezescu and Ecaterina Andronescu, Green Synthesis of Metal and Metal Oxide Nanoparticles: A Review of the Principles and Biomedical Applications, Int. J. Mol. Sci. 2023, 24(20), 15397; https://doi.org/10.3390/ijms242015397 3. Spoială Angela, Motelica Ludmila, Ilie Cornelia-Ioana, Ficai Denisa, Chircov Cristina, Joanna Kisala, Surdu Adrian-Vasile, Trușcă Roxana, Vasile Bogdan, Oprea Ovidiu, Ficai Anton, Aminoacid functionalised magnetite nanoparticles Fe3O4@AA (AA = Ser, Cys, Pro, Trp) as Biocompatible Magnetite Nanoparticles with Potential Therapeutic Applications,, Scientific reports-Nature, trimis spre publicare, major revision. 4. Ludmila Motelica, Geanina Voicu, Cristina Uncu, Adrian Vasile Surdu, Roxana Doina Trusca, Bogdan Stefan Vasile, Denisa Ficai, Ovidiu Cristian Oprea, Daciana Silvia Marta, Victor-Eduard Peteu, Maria Anghelache, Anton Ficai and Manuela Calin, Aspartic Acid Functionalized Magnetic Nanoparticles for Enhanced Internalization in Tumoral Cell, Journal of the Australian Ceramic Society Editorial Office, 2024, acepted for publication. 5. Cornelia-Ioana Ilie, Angela Spoiala, Cristina Chircov, Georgiana Dolete, Ovidiu-Cristian Oprea, Bogdan-Stefan Vasile, Simona Adriana Crainiceanu, Adrian-Ionut Nicoara, Ioana Cristina Marinas, Miruna Silvia Stan , Lia-Mara Ditu, Anton Ficai and Eliza Oprea, Antioxidant, Antitumoral, Antimicrobial, and Prebiotic Activity of Magnetite Nanoparticles Loaded with Bee Pollen/Bee Bread Extracts and 5-Fluorouracil, Antioxidants 2024, 13, 895. https://doi.org/10.3390/antiox13080895 |
Patent application | Denisa FICAI, Ludmila MOTELICA, Angela SPOIALA, Cornelia Ioana ILIE, Ovidiu Cristian OPREA, Anton FICAI, Nanoparticule magnetice stabilizate cu aminoacizi pentru internalizare celulară îmbunătățită și tehnologia de obținere aferentă, Cerere depusa la OSIM, Nr.a/00791 |
PhD Thesis | Doctorand Cornelia Ioana ILIE, îndrumător Prof. Dr. Ing. Anton FICAI |
Obtained materials (magnetic systems) |
The following were obtained: 1 control sample NPFe3O4 18 materials of type NPFe3O4/ stabilization agent/ and or Internalization 18 materials of type NPFe3O4/ stabilizing agent/ and or Internalization/ Cytostatic |