Periodic Reporting for period 1 - MAGNET-CELLPATCH (Multimodal magnetic cellular-patches with synergistic effects for high performance theranostics)
Período documentado: 2019-09-01 hasta 2021-08-31
In order to advance in the design of the multifunctional agents with enhanced theranostic efficency, it has been proposed the fabrication of different hybrid platforms that have the capacity to integrate different active modules. As it was highlighted in the MAGNET-CELLPATCH proposal, it is of utmost importance to minimize the chaotic aggregation effects of magnetic nanoparticles (MNPs) to achieve both a remarkable magnetothermal efficiency and a proper synergy among the different modules of the system.
Globally, one of the most common type of cancer is the colorectal cancer (CRC), after lung and breast cancers. In average more than 1 million people get colorectal cancer every year, which develops into hepatic metastasis in 70% of the cases and it is usually not curable. Consequently, there is an urgent need in this field to develop new materials with enhanced theranostic efficacy.
The transfer of the gained scientific knowledge to the clinical practice can promote progress and open new lines of work aimed at combating different types of tumors and diseases. Additionally, the theranostic platforms that have been developed during the project are completely adjustable to address the demands of the incipient “personalized medicine” that consists of adapting treatment to the genomic characteristics of each patient. In addition, the use of these kind of novel theranostic agents would save on expenses to treat collateral damage provoked by the conventional therapies.
Overall, the objectives of this project is to design, fabricate and study novel multifunctional theranostic platforms with high biomedical potential. For that purpose several properties of the platforms have to be improved, such as:
-Magnetothermal actuation
-Endocytosis resistance
-Dual-imaging
-Specific Targeting
-Biocompatibility
-Etc.
Two type of theranostic platforms have been proposed to accomplished the goals of the project: Platform I (MNPs individually coated by PEGylation technology) and Platform II (two-dimensional microdiscs with a monolayer of MNPs).
Globally, one of the most common type of cancer is the colorectal cancer (CRC), after lung and breast cancers. In average more than 1 million people get colorectal cancer every year, which develops into hepatic metastasis in 70% of the cases and it is usually not curable. Consequently, there is an urgent need in this field to develop new materials with enhanced theranostic efficacy.
The transfer of the gained scientific knowledge to the clinical practice can promote progress and open new lines of work aimed at combating different types of tumors and diseases. Additionally, the theranostic platforms that have been developed during the project are completely adjustable to address the demands of the incipient “personalized medicine” that consists of adapting treatment to the genomic characteristics of each patient. In addition, the use of these kind of novel theranostic agents would save on expenses to treat collateral damage provoked by the conventional therapies.
Overall, the objectives of this project is to design, fabricate and study novel multifunctional theranostic platforms with high biomedical potential. For that purpose several properties of the platforms have to be improved, such as:
-Magnetothermal actuation
-Endocytosis resistance
-Dual-imaging
-Specific Targeting
-Biocompatibility
-Etc.
Two type of theranostic platforms have been proposed to accomplished the goals of the project: Platform I (MNPs individually coated by PEGylation technology) and Platform II (two-dimensional microdiscs with a monolayer of MNPs).
Two kind of multifunctional theranostic platforms that prevent agglomeration among MNPs have been successfully fabricated by 2 different methods:
1) Wet-chemical synthesis: A surface modification of MNPs with Poly(maleic anhydride-alt-1-octadecene)-grafted Poly(ethylene glycol) copolymer (PMAO-PEG) in order to achieve single-particle polymer coatings. In this configuration, MNPs present minimal dipolar interactions. (Platform I)
2) Microfabrication: A homogeneous and controlled assembly of MNPs within polymeric-inorganic microdisks fabricated by photolithography. The monolayer of MNPs are immobilized within a polymeric multilayer, so clustering effects are significantly reduced. (Platform II)
In the following the most relevant achievements will be listed:
-Fine control over several physicochemical properties has been achieved in both platforms (size, morphology, composition, crystallinity, etc.).
-Highly reproducible and efficient magnetothermal performance have been achieved in physiological media and in-vitro.
-By controlling several conjugation parameters (the amount of reagents, reaction time, inert atmosphere, etc.), it has been possible to tailor the MNP-Ab system between 5-15 Ab/NP.
-It has been possible to tailor the arrangement of MNPs within platform II in different configurations:
i) When MNPs are sprayed in specific conditions and in absence of an external magnetic field, 2D assemblies with minimal agglomeration degree and with the magnetic moments randomly oriented are achieved
ii) Otherwise, when the MNPs are sprayed in the same conditions but under a permanent magnetic field of 245 mT, the MNPs are assembled in oriented patches.
The coercivity (Hc) at RT of the randomly oriented assembly does not change when the measurement is performed at 0º or 90º with respect to the magnetic field. On the contrary, the Hc of oriented assembly is significantly higher when measured at 0º. This indicates an increase of the shape anisotropy in this kind of 2D arrays, which very likely will lead to larger SLP values.
-The design and refinement of a magneto-optic system has been performed based on platform I. This platform is not only an efficient magneto-fluorescent marker, but also a highly suitable system to track the magnetic hyperthermia treatment locally.
-Toxicity assays in-vitro have revealed that platform I and II are completely biocompatible. In addition, it has been demonstrated the great endocytosis resistance of these systems.
- Highly efficient magnetic hyperthermia in-vitro (in colon cancer-derived cell line (HCT116) ) was achieved using low/modarate concentration of MNPs out at 17 kA/m and 650 kHz. Two different nanoparticle concentrations were used (C1=0.25 and C2=0.5 mg/ml) and the determined initial heating rate was 2 ºC/min and 4 ºC/min, respectively. The magnetic field was applied for one hour and the cells with NPs reached 48 and 44 ºC, while the control stayed at the T of the background. And 48 h post-hyperthermia a complete cell death was achieved when using C2. Although the decrease in cell number for C1 was not statistically significant, the expression of apoptosis genes (BCL10 and CASP8) was enhanced, meaning that the apoptotic process had been already activated and they would die within a short period of time.
1) Wet-chemical synthesis: A surface modification of MNPs with Poly(maleic anhydride-alt-1-octadecene)-grafted Poly(ethylene glycol) copolymer (PMAO-PEG) in order to achieve single-particle polymer coatings. In this configuration, MNPs present minimal dipolar interactions. (Platform I)
2) Microfabrication: A homogeneous and controlled assembly of MNPs within polymeric-inorganic microdisks fabricated by photolithography. The monolayer of MNPs are immobilized within a polymeric multilayer, so clustering effects are significantly reduced. (Platform II)
In the following the most relevant achievements will be listed:
-Fine control over several physicochemical properties has been achieved in both platforms (size, morphology, composition, crystallinity, etc.).
-Highly reproducible and efficient magnetothermal performance have been achieved in physiological media and in-vitro.
-By controlling several conjugation parameters (the amount of reagents, reaction time, inert atmosphere, etc.), it has been possible to tailor the MNP-Ab system between 5-15 Ab/NP.
-It has been possible to tailor the arrangement of MNPs within platform II in different configurations:
i) When MNPs are sprayed in specific conditions and in absence of an external magnetic field, 2D assemblies with minimal agglomeration degree and with the magnetic moments randomly oriented are achieved
ii) Otherwise, when the MNPs are sprayed in the same conditions but under a permanent magnetic field of 245 mT, the MNPs are assembled in oriented patches.
The coercivity (Hc) at RT of the randomly oriented assembly does not change when the measurement is performed at 0º or 90º with respect to the magnetic field. On the contrary, the Hc of oriented assembly is significantly higher when measured at 0º. This indicates an increase of the shape anisotropy in this kind of 2D arrays, which very likely will lead to larger SLP values.
-The design and refinement of a magneto-optic system has been performed based on platform I. This platform is not only an efficient magneto-fluorescent marker, but also a highly suitable system to track the magnetic hyperthermia treatment locally.
-Toxicity assays in-vitro have revealed that platform I and II are completely biocompatible. In addition, it has been demonstrated the great endocytosis resistance of these systems.
- Highly efficient magnetic hyperthermia in-vitro (in colon cancer-derived cell line (HCT116) ) was achieved using low/modarate concentration of MNPs out at 17 kA/m and 650 kHz. Two different nanoparticle concentrations were used (C1=0.25 and C2=0.5 mg/ml) and the determined initial heating rate was 2 ºC/min and 4 ºC/min, respectively. The magnetic field was applied for one hour and the cells with NPs reached 48 and 44 ºC, while the control stayed at the T of the background. And 48 h post-hyperthermia a complete cell death was achieved when using C2. Although the decrease in cell number for C1 was not statistically significant, the expression of apoptosis genes (BCL10 and CASP8) was enhanced, meaning that the apoptotic process had been already activated and they would die within a short period of time.
The development of nano-/micro-platforms that allow multiplexing images and creating multi-therapeutic action is an ambitious challenge that has been accomplished to a notable extent. Therefore, the goals achieved during the MAGNET-CELLPATCH project will likely have a great impact on a scientific level. The transfer of the gained scientific knowledge to the clinical practice can promote progress and open new lines of work aimed at combating different types of tumors and diseases. Additionally, the theranostic platforms that have been developed during the project are completely adjustable to address the demands of the incipient “personalized medicine” that consists of adapting treatment to the genomic characteristics of each patient.
In relation to social impact, MAGNET-CELLPATCH project has addressed an applied research in the design of Advanced Materials to cover important demands of the biomedical field. The project is completely aligned with the area of "Health, demographic change and well-being" (identified as task 1 of the Spanish strategy of Science, Technology and Innovation), with the European Strategy included in the Program Horizon Europe, as well as with the Smart Specialization Strategy of PCTI Euskadi 2030. Hence, the results obtained from this project will contribute to the scientific development and to a better quality of life and well-being of the society.
Regarding the economic impact, the need to fabricate multifunctional materials for novel biomedical procedures would lead to an increase in employment to respond to the scaling and production demands. In addition, the use of these kind of novel theranostic agents would save on expenses to treat collateral damage provoked by the conventional therapies.
In relation to social impact, MAGNET-CELLPATCH project has addressed an applied research in the design of Advanced Materials to cover important demands of the biomedical field. The project is completely aligned with the area of "Health, demographic change and well-being" (identified as task 1 of the Spanish strategy of Science, Technology and Innovation), with the European Strategy included in the Program Horizon Europe, as well as with the Smart Specialization Strategy of PCTI Euskadi 2030. Hence, the results obtained from this project will contribute to the scientific development and to a better quality of life and well-being of the society.
Regarding the economic impact, the need to fabricate multifunctional materials for novel biomedical procedures would lead to an increase in employment to respond to the scaling and production demands. In addition, the use of these kind of novel theranostic agents would save on expenses to treat collateral damage provoked by the conventional therapies.