Abstract:
The blood-brain barrier (BBB) is a considerable obstacle to drug delivery into the central nervous system and explains the failure of many experimental therapies. The project proposal's basic concept is to use the nutrient transport proteins overexpressed in glioblastoma to uptake drugs-loaded human serum albumin (HAS) nanoparticles (NPs) and target them in the tumor. The project aims to design an NPs system based on HSA functionalized with low molecular weight protamine (LMWP), cross-linked with oxidized polysaccharides having co-encapsulated curcumin and temozolomide. LMWP will be used to improve the NPs diffusion through BBB. Through ultrasonication/precipitation and self-assembling techniques will be prepared, two types of drugs-loaded NPs will be characterized by FT-IR, SEM, TEM, DLS, and swelling degree. The polymer matrix's protective role and the antioxidant activity of the encapsulated polyphenol will be evaluated. The in vitro release kinetics studies of the drugs loaded in NPs will be performed at pH=7.4 and pH=5. The cytotoxicity and apoptosis induced by the drug-loaded NPs will be tested using two cell lines, U87 and C6. A BBB cell model will be used to determine the drug-loaded HSA NPs' ability to overcome it. The drug-loaded NPs obtained will determine the drug's protection, increased bioavailability, and side effects elimination, ensuring a controlled and sustained release at therapeutic doses.
The main project objective is to achieve a delivery system capable of exceeding BBB to treat human glioblastoma, consisting of HSA-based nanoparticles modified with and low molecular weight protamine and cross-linked with carbonyl groups from oxidized gellan and oxidized pectin having co-immobilized two drugs - temozolomide and curcumin..
Specific objectives and activities:
WP1. Obtaining and physicochemical characterization of precursors and functionalized and cross-linked nanoparticles containing or not encapsulated drugs.
Act 1.1 - Obtaining and spectroscopic characterization of human serum albumin (HSA) modified with low molecular mass protamine (LMWP).
Act 1.2 - Obtaining and physicochemical characterization of oxidized gellan and oxidized pectin.
Act 1.3 - Obtaining nanoparticles with and without encapsulated drugs using a method based on ultrasonication and precipitation.
Act 1.4 - Obtaining nanoparticles with and without encapsulated drugs by the self-assembly method.
Act 1.5 - Study of factors that can influence the physicochemical characteristics, morphology, and encapsulation capacity of drugs in biopolymer-based nanoparticles.
Act 1.6 - Determination of drug encapsulation efficiency.
Act 1.7 - Preliminary tests on the physicochemical characteristics of the drug-loaded nanoparticles.
WP.2 Kinetic studies of in vitro drug release from nanoparticles, ex vivo testing of nanoparticulate systems, evaluation of biomaterial characteristics of nanoparticles (cytotoxicity, hemocompatibility, microbiological activity), and the stability evaluation of the drugs encapsulated in nanoparticles at temperature, pH and light.
Act 2.1 - In vitro kinetic study of drug release from nanoparticles based on human serum albumin.
Act 2.2 - Determination of cell apoptosis induced by drug-loaded nanoparticles.
Act 2.3 - Testing in vitro the functionalized nanoparticles that contain co-encapsulated drugs on blood-brain barrier models and the permeability determination.
Act 2.4 - Testing the antioxidant properties of the drugs encapsulated in nanoparticles.
Act 2.5 - Evaluation of the cytotoxicity of the drugs encapsulated in nanoparticles using the MTT assay
Act 2.6 - Evaluation of microbiological activitye
Act 2.7 - Hemocompatibility analysis of drugs-loaded nanoparticles
Act 2.8 - Study of structural and thermal stability of drug-loaded nanoparticles
Act 2.9 - pH stability study of drugs immobilized in nanoparticles
Act 2.10 - Light stability study of drugs encapsulated in nanoparticles
WP3. The highlight of the polymer matrix's protective character for incorporated drugs at complexation with metal ions.
Act 3.1 - Study of the stability of drugs encapsulated in nanoparticles when complexed with metal ions.
Act 3.2 - Statistical analysis of data
Project impact and estimated results
The NPs based on HSA functionalized with LMWP having TMZ and curcumin immobilized determine drug protection, increased bioavailability, elimination of side effects, reduce or eliminate the tumor cell resistance to the drugs, ensuring a controlled and sustained release at therapeutic doses. Including both drugs in the delivery systems increases their ability to overcome BBB because the physicochemical characteristics of the nano-carrier become essential now, not the ones of the drug. The treatment efficiency and the possibility of an efficient dosage of the drugs are improved. The project will increase the visibility of research by publishing 3 articles in journals with a high impact factor (Q1) and 4 participation at international conferences. This research is in a multidisciplinary field and will also be able to continue because although these nanoparticles based on biopolymers are biocompatible and will overcome the BBB, more in vivo studies and their interaction with the immune system are needed to translate them into clinical research and produce them at industrial level. Significant contributions will be made to developing nanoparticles based on biopolymers as systems for the delivery and transport of active principles through the BBB. There is also the possibility of industrial production of the delivery systems with the highest therapeutic potential. We consider that the project will make contributions from the following perspectives: (1) Scientific (through fundamental studies that will contribute to the development of the addressed field; 2) Applicative (through the development and testing of new formulations with therapeutic potential; 3) Economic (production at industrial level with the possibility of using in the treatment of smaller quantities of the drug).
Dissemination:
- • 3 articles published in ISI-ranked Q1 journals;
- • Presentations at 4 international conferences;
- • 3 scientific reports;
- • 1 website.
Results obtained from Stage I of the Project in 2022
- 1. Participation with an oral communication of the paper "Biocompatible hydrogels films with the inclusion complex of
β-cyclodextrin/curcumin immobilized for biomedical applications" at an international conference:
Camelia Elena Iurciuc (Tincu), Christine Jérôme, Marcel Popa, Carmen Gafițanu, Eliza Grațiela Popa, Lăcrămioara Ochiuz, Biocompatible hydrogels films with the inclusion complex of β-cyclodextrin/curcumin immobilized for biomedical applications, International Conference on Natural Products in Drug Discovery and Development – Advances and Perspectives, PSE Meeting 2022, September 19 – 22, 2022, Iași, Romania. - 2. Submission for publication of the article with the title, "Immobilization of α-amylase in gellan-based supports and assay of enzyme activity in different hydrolysis cycles", authors: Camelia Elena Tincu (Iurciuc), Brahim Bouhadiba, Leonard Ionut Atanase, Corneliu Sergiu Stan, Marcel Popa, Lăcrămioara Ochiuz, at Polymers Journal
- 3. Submission for publication of the review article entitled "Overcoming the blood-brain barrier using albumin-based drug delivery nanosystems for the treatment of brain cancer" at the International Journal of Molecular Science. Authors: Camelia Elena Tincu (Iurciuc), Marcel Popa, Lăcrămioara Ochiuz.
- 4. Development of the project web page.
- 5. Writing and submission of the Scientific Report for Stage I/2022 entitled "Obtaining and physicochemical characterization of precursors and functionalized and cross-linked nanoparticles containing or not encapsulated drugs."
Results obtained from Stage II of the Project in 2023
- Conference participation:
Oral communication:
1. Camelia Elena Tincu (Iurciuc), Oana-Maria Darabă, Leonard Ionuț Atanase, Marcel Popa, Lăcrămioara Ochiuz, β-cyclodextrin/curcumin complex immobilized in the albumin-based hydrogel films cross-linked with oxidized gellan for biomedical applications, International Congress of "Apollonia" University in Iași, Preparing the future by promoting excellence, XXXIIIth Edition, March 2 - 5, 2023, IAȘI, ROMANIA - Poster presentation:
2. Camelia Elena Tincu (Iurciuc), Silvia Vasiliu, Ștefania Racoviță, Leonard Ionuț Atanase, Marcel Popa, Lăcrămioara Ochiuz, New drug release system based on functionalized albumin cross- linked with oxidized polysaccharides for the potential treatment of brain tumors, International Congress of "Apollonia" University in Iași, Preparing the future by promoting excellence, XXXIIIth Edition, March 2 - 5, 2023, IAȘI, ROMANIA
3. Camelia Elena Tincu (Iurciuc), Silvia Vasiliu, Ștefania Racoviță, Marcel Popa, Lăcrămioara Ochiuz, Nanoparticles based on Functionalized Albumin Cross-linked with Oxidized Polysaccharides Used for Drug Delivery in Brain Tumors Therapy, 3rd International Conference on Bioengineering and Polymer Science, Bucharest, ROMANIA - June 7-10, 2023 - Published articles:
1. Tincu (Iurciuc) Camelia Elena, Brahim Bouhadiba, Leonard Ionut Atanase, Corneliu Sergiu Stan, Marcel Popa and Lăcrămioara Ochiuz. 2023. "An Accessible Method to Improve the Stability and Reusability of Porcine Pancreatic α-Amylase via Immobilization in Gellan-Based Hydrogel Particles Obtained by Ionic Cross-Linking with Mg2+ Ions" Molecules 28, no. 12:4695. https://doi.org/10.3390/molecules28124695
2. Tincu (Iurciuc) Camelia-Elena, Călin Vasile Andrițoiu, Marcel Popa, and Lăcrămioara Ochiuz. 2023. "Recent Advancements and Strategies for Overcoming the Blood–Brain Barrier Using Albumin-Based Drug Delivery Systems to Treat Brain Cancer, with a Focus on Glioblastoma" Polymers 15, no. 19: 3969. https://doi.org/10.3390/polym15193969
3. By December 31, 2023, an article will be submitted to Gels Journal. The article is titled "Albumin-based Hydrogel Films Covalently Cross-linked with Oxidized Gellan with Encapsulated Curcumin for Biomedical Applications." Authors: Tincu (Iurciuc) Camelia Elena, Oana-Maria Darabă, Christine Jerome, Marcel Popa, Lăcrămioara Ochiuz
4. Writing and completion of the Scientific Report for Stage II/2023 entitled "Kinetic studies of in vitro drug release from nanoparticles, ex vivo testing of nanoparticulate systems, evaluation of biomaterial characteristics of nanoparticles (cytotoxicity, hemocompatibility, microbiological activity) and evaluation of the drug encapsulated in nanoparticles at temperature, pH, and light."
Results obtained from Stage III of the Project in 2024
- Dissemination of results:
Participation in conferences:
Camelia Elena Tincu (Iurciuc), Silvia Vasiliu, Ștefania Racoviță, Gabriela Vochița, Daniela Gherghel, Leonard Ionuț Atanase, Marcel Popa, Lăcrămioara Ochiuz, Development of a drug delivery nanosystem composed of functionalized human serum albumin cross-linked with oxidized polysaccharides for treating brain tumors, International Congress of "Apollonia" University of Iași, Preparing the future by promoting excellence, XXXIV Edition, February 29 - March 3, 2024, IASI, ROMANIA - Published articles:
Submission for publication of the article entitled "Albumin-Based Hydrogel Films Covalently Cross-Linked With Oxidized Gellan With Encapsulated Curcumin For Biomedical Applications," Authors: Camelia Elena Tincu (Iurciuc), Oana-Maria Darabă, Christine Jerome, Marcel Popa, Lăcrămioara Ochiuz at Polymers. (Manuscript ID: polymers-2956947)
Results obtained
- Indicators to achieve:
- Publication of 3 articles in ISI journals with a high impact factor, in open access regime
- Participation with scientific papers at 4 international conferences
- Writing the annual research reports and the final research report
- 1 web page of the project
- Realized indicators:
- Publication of 3 articles in ISI journals with a high impact factor, in open access mode
- Participation with scientific papers at 5 International Conferences
- Drafting of annual research reports and the final research report
- 1 web page of the project
- Participation in conferences:
Oral communication:
1. Camelia Elena Iurciuc (Tincu), Christine Jérôme, Marcel Popa, Carmen Gafițanu, Eliza Grațiela Popa, Lăcrămioara Ochiuz, Biocompatible hydrogels films with the inclusion complex of β-cyclodextrin/curcumin immobilized for biomedical applications, International Conference on Natural Products in Drug Discovery and Development – Advances and Perspectives, PSE Meeting 2022, September 19 – 22, 2022, Iași, Romania
2. Camelia Elena Tincu (Iurciuc), Oana-Maria Darabă, Leonard Ionuț Atanase, Marcel Popa, Lăcrămioara Ochiuz, β-cyclodextrin/curcumin complex immobilized in the albumin-based hydrogel films cross-linked with oxidized gellan for biomedical applications, International Congress of "Apollonia" University in Iași, Preparing the future by promoting excellence, XXXIIIth Edition, March 2 - 5, 2023, IASI, ROMANIA -
Poster presentations:
1. Camelia Elena Tincu (Iurciuc), Silvia Vasiliu, Ștefania Racoviță, Leonard Ionuț Atanase, Marcel Popa, Lăcrămioara Ochiuz, New drug release system based on functionalized albumin cross-linked with oxidized polysaccharides for the potential treatment of brain tumors, International Congress of "Apollonia" University in Iași, Preparing the future by promoting excellence, XXXIIIth Edition, March 2 - 5, 2023, IASI, ROMANIA
2. Camelia Elena Tincu (Iurciuc), Silvia Vasiliu, Ștefania Racoviță, Marcel Popa, Lăcrămioara Ochiuz, Nanoparticles based on Functionalized Albumin Cross-linked with Oxidized Polysaccharides Used for Drug Delivery in Brain Tumors Therapy, 3rd International Conference on Bioengineering and Polymer Science, Bucharest, ROMANIA - June 7-10, 2023
3. Camelia Elena Tincu (Iurciuc), Silvia Vasiliu, Ștefania Racoviță, Gabriela Vochița, Daniela Gherghel, Leonard Ionuț Atanase, Marcel Popa, Lăcrămioara Ochiuz, Development of a drug delivery nanosystem composed of functionalized human serum albumin cross-linked with oxidized polysaccharides for treating brain tumors, International Congress of "Apollonia" University of Iași, Preparing the future by promoting excellence, XXXIV Edition, February 29 - March 3, 2024, IAȘI, ROMANIA -
Published articles:
1. Tincu (Iurciuc) Camelia Elena, Brahim Bouhadiba, Leonard Ionut Atanase, Corneliu Sergiu Stan, Marcel Popa, and Lăcrămioara Ochiuz. 2023. “An Accessible Method to Improve the Stability and Reusability of Porcine Pancreatic α-Amylase via Immobilization in Gellan-Based Hydrogel Particles Obtained by Ionic Cross-Linking with Mg2+ Ions” Molecules 28, no. 12: 4695. https://doi.org/10.3390/polym15193969
2. Tincu (Iurciuc), Camelia-Elena, Călin Vasile Andrițoiu, Marcel Popa, and Lăcrămioara Ochiuz. 2023. “Recent Advancements and Strategies for Overcoming the Blood–Brain Barrier Using Albumin-Based Drug Delivery Systems to Treat Brain Cancer, with a Focus on Glioblastoma” Polymers 15, no. 19: 3969. https://doi.org/10.3390/polym15193969
3. Submission for publication of the article entitled "Albumin-Based Hydrogel Films Covalently Cross-Linked With Oxidized Gellan With Encapsulated Curcumin For Biomedical Applications", Authors: Camelia Elena Tincu (Iurciuc), Oana-Maria Darabă, Christine Jerome, Marcel Popa, Lăcrămioara Ochiuz at Polymers magazine. (Manuscript ID: polymers-2956947)