Interpenetrating Polymer Networks for Ocular Drug Delivery
Authorship
H.A.
Master's Degree in Drug Research and Development
H.A.
Master's Degree in Drug Research and Development
Defense date
07.14.2026 16:00
07.14.2026 16:00
Summary
The increasing prevalence of ocular diseases, combined with the limited bioavailability of conventional ophthalmic formulations, has highlighted the need for more effective drug delivery strategies. In recent years, interpenetrating polymer network (IPN)-based hydrogels have attracted considerable attention as an ocular drug delivery strategy because of their ability to combine the advantages of multiple polymeric networks. Compared with conventional single-network hydrogels, IPN systems provide improved mechanical strength, enhanced stability, higher drug loading, and more controlled drug release. These characteristics make them attractive for overcoming physiological barriers and prolonging drug residence time in ocular tissues. This master's thesis critically reviews recent advances in the development of IPN-based hydrogels for ocular drug delivery. It presents the fundamental concepts of IPN hydrogels, including their classification, structural design, fabrication strategies, and characterization techniques. In addition, it examines the influence of formulation parameters, such as polymer composition, crosslinking strategies, and network architecture, on the performance of these systems. Finally, recent experimental studies investigating the application of IPN-based hydrogels for the treatment of ocular diseases are critically analyzed, highlighting their therapeutic potential and limitations. Overall, IPN-based hydrogels represent a versatile and emerging approach for improving ocular therapeutics and addressing challenges associated with the treatment of eye diseases
The increasing prevalence of ocular diseases, combined with the limited bioavailability of conventional ophthalmic formulations, has highlighted the need for more effective drug delivery strategies. In recent years, interpenetrating polymer network (IPN)-based hydrogels have attracted considerable attention as an ocular drug delivery strategy because of their ability to combine the advantages of multiple polymeric networks. Compared with conventional single-network hydrogels, IPN systems provide improved mechanical strength, enhanced stability, higher drug loading, and more controlled drug release. These characteristics make them attractive for overcoming physiological barriers and prolonging drug residence time in ocular tissues. This master's thesis critically reviews recent advances in the development of IPN-based hydrogels for ocular drug delivery. It presents the fundamental concepts of IPN hydrogels, including their classification, structural design, fabrication strategies, and characterization techniques. In addition, it examines the influence of formulation parameters, such as polymer composition, crosslinking strategies, and network architecture, on the performance of these systems. Finally, recent experimental studies investigating the application of IPN-based hydrogels for the treatment of ocular diseases are critically analyzed, highlighting their therapeutic potential and limitations. Overall, IPN-based hydrogels represent a versatile and emerging approach for improving ocular therapeutics and addressing challenges associated with the treatment of eye diseases
Direction
ABBADESSA , ANNA (Tutorships)
LANDIN PEREZ, MARIANA (Co-tutorships)
ABBADESSA , ANNA (Tutorships)
LANDIN PEREZ, MARIANA (Co-tutorships)
Court
FONTENLA GIL, JOSE ANGEL (Chairman)
GOYANES GOYANES, ALVARO (Secretary)
CASTRO PEREZ, MARIA DE LOS ANGELES (Member)
FONTENLA GIL, JOSE ANGEL (Chairman)
GOYANES GOYANES, ALVARO (Secretary)
CASTRO PEREZ, MARIA DE LOS ANGELES (Member)
Development of a controlled ketamine delivery system
Authorship
F.N.B.R.
Master's Degree in Drug Research and Development
F.N.B.R.
Master's Degree in Drug Research and Development
Defense date
07.14.2026 16:00
07.14.2026 16:00
Summary
The present study aims to develop and characterize a controlled-release system of ketamine hydrochloride for subcutaneous administration. For this purpose, a hybrid system was designed based on the incorporation of drug-loaded poly(lactic-co-glycolic acid) nanoparticles, prepared by double emulsion followed by solvent evaporation, into a thermosensitive hydrogel composed of poloxamer 407 (P407) and hyaluronic acid (HA). The hydrogel properties were characterized by rheological analysis, while the system’s ability to modulate drug release was evaluated through in vitro release studies using Franz diffusion cells.
The present study aims to develop and characterize a controlled-release system of ketamine hydrochloride for subcutaneous administration. For this purpose, a hybrid system was designed based on the incorporation of drug-loaded poly(lactic-co-glycolic acid) nanoparticles, prepared by double emulsion followed by solvent evaporation, into a thermosensitive hydrogel composed of poloxamer 407 (P407) and hyaluronic acid (HA). The hydrogel properties were characterized by rheological analysis, while the system’s ability to modulate drug release was evaluated through in vitro release studies using Franz diffusion cells.
Direction
DIAZ RODRIGUEZ, PATRICIA (Tutorships)
LANDIN PEREZ, MARIANA (Co-tutorships)
DIAZ RODRIGUEZ, PATRICIA (Tutorships)
LANDIN PEREZ, MARIANA (Co-tutorships)
Court
FONTENLA GIL, JOSE ANGEL (Chairman)
GOYANES GOYANES, ALVARO (Secretary)
CASTRO PEREZ, MARIA DE LOS ANGELES (Member)
FONTENLA GIL, JOSE ANGEL (Chairman)
GOYANES GOYANES, ALVARO (Secretary)
CASTRO PEREZ, MARIA DE LOS ANGELES (Member)
Study of hypothalamic nuclear receptors as a potential pharmacological strategy for the treatment of obesity.
Authorship
E.M.B.V.
Master's Degree in Drug Research and Development
E.M.B.V.
Master's Degree in Drug Research and Development
Defense date
07.14.2026 16:00
07.14.2026 16:00
Summary
Currently, several hypothalamic neurocircuits are known to be involved in the regulation of hunger, satiety, and energy expenditure, leading to the conclusion that the hypothalamus is essential in the integration of signals that affect energy status and body weight regulation. In the study of obesity, this knowledge takes on special relevance, since these circuits are positioned as a point of interest for the development of treatments for this disease, which increasingly affects the world population. For the integration and proper functioning of these circuits, nuclear receptors are essential, although not all of them have such a studied function, as is the case of retinoid X-receptors, present in this area of the brain and which have the ability to interact molecularly with other known nuclear receptors. With this in mind, the following work proposes a study of hypothalamic retinoid X-receptors and their potential function as a pharmacological target for the treatment of obesity. Various techniques will be performed in vivo, with the use of animal models treated with agonists of this receptor; likewise, molecular analyses by qPCR and histological analysis techniques will be performed.
Currently, several hypothalamic neurocircuits are known to be involved in the regulation of hunger, satiety, and energy expenditure, leading to the conclusion that the hypothalamus is essential in the integration of signals that affect energy status and body weight regulation. In the study of obesity, this knowledge takes on special relevance, since these circuits are positioned as a point of interest for the development of treatments for this disease, which increasingly affects the world population. For the integration and proper functioning of these circuits, nuclear receptors are essential, although not all of them have such a studied function, as is the case of retinoid X-receptors, present in this area of the brain and which have the ability to interact molecularly with other known nuclear receptors. With this in mind, the following work proposes a study of hypothalamic retinoid X-receptors and their potential function as a pharmacological target for the treatment of obesity. Various techniques will be performed in vivo, with the use of animal models treated with agonists of this receptor; likewise, molecular analyses by qPCR and histological analysis techniques will be performed.
Direction
LOZA GARCIA, MARIA ISABEL (Tutorships)
GONZALEZ GARCIA, ISMAEL (Co-tutorships)
LOZA GARCIA, MARIA ISABEL (Tutorships)
GONZALEZ GARCIA, ISMAEL (Co-tutorships)
Court
FONTENLA GIL, JOSE ANGEL (Chairman)
GOYANES GOYANES, ALVARO (Secretary)
CASTRO PEREZ, MARIA DE LOS ANGELES (Member)
FONTENLA GIL, JOSE ANGEL (Chairman)
GOYANES GOYANES, ALVARO (Secretary)
CASTRO PEREZ, MARIA DE LOS ANGELES (Member)
Optimization and comparison of human endothelial cell cultures for the development of an in vitro blood-brain bar model
Authorship
E.G.C.
Master's Degree in Drug Research and Development
E.G.C.
Master's Degree in Drug Research and Development
Defense date
07.14.2026 09:00
07.14.2026 09:00
Summary
The blood-brain barrier (BBB) is a highly selective barrier mainly composed of brain endothelial cells, which are the most important for maintaining barrier integrity due to the presence of tight junctions (TJs). BBB dysfunction has been associated with several central nervous system (CNS) disorders, and in vitro BBB models constitute valuable tools for studying BBB pathophysiology and evaluating novel therapeutic candidates. The aim of this work was to optimize an in vitro BBB model by comparing different human endothelial cell lines and evaluating the effect of coculture with different components of the neurovascular unit (NVU) on barrier properties. To this end, BBB models were developed using HUVEC, hCMEC/D3 and HBMEC endothelial cells in monoculture and in coculture with pericytes, astrocytes, microglia and neurons using Transwell systems. Barrier integrity was evaluated by measuring transendothelial electrical resistance (TEER) and a permeability assay using sodium fluorescein (NaFl) as a paracellular marker. In addition, the expression of the TJ proteins claudin-5 and occludin was analyzed by Western blot. HBMECs showed the highest TEER values and the lowest permeability, particularly in coculture with pericytes, suggesting they are the most suitable model for reproducing the properties of the BBB.
The blood-brain barrier (BBB) is a highly selective barrier mainly composed of brain endothelial cells, which are the most important for maintaining barrier integrity due to the presence of tight junctions (TJs). BBB dysfunction has been associated with several central nervous system (CNS) disorders, and in vitro BBB models constitute valuable tools for studying BBB pathophysiology and evaluating novel therapeutic candidates. The aim of this work was to optimize an in vitro BBB model by comparing different human endothelial cell lines and evaluating the effect of coculture with different components of the neurovascular unit (NVU) on barrier properties. To this end, BBB models were developed using HUVEC, hCMEC/D3 and HBMEC endothelial cells in monoculture and in coculture with pericytes, astrocytes, microglia and neurons using Transwell systems. Barrier integrity was evaluated by measuring transendothelial electrical resistance (TEER) and a permeability assay using sodium fluorescein (NaFl) as a paracellular marker. In addition, the expression of the TJ proteins claudin-5 and occludin was analyzed by Western blot. HBMECs showed the highest TEER values and the lowest permeability, particularly in coculture with pericytes, suggesting they are the most suitable model for reproducing the properties of the BBB.
Direction
CAMPOS TOIMIL, MANUEL (Tutorships)
VIÑA CASTELAO, MARÍA DOLORES (Co-tutorships)
CAMPOS TOIMIL, MANUEL (Tutorships)
VIÑA CASTELAO, MARÍA DOLORES (Co-tutorships)
Court
FONTENLA GIL, JOSE ANGEL (Chairman)
GOYANES GOYANES, ALVARO (Secretary)
CASTRO PEREZ, MARIA DE LOS ANGELES (Member)
FONTENLA GIL, JOSE ANGEL (Chairman)
GOYANES GOYANES, ALVARO (Secretary)
CASTRO PEREZ, MARIA DE LOS ANGELES (Member)
Master Dissertation
Authorship
U.G.L.
Master's Degree in Drug Research and Development
U.G.L.
Master's Degree in Drug Research and Development
Defense date
07.14.2026 09:00
07.14.2026 09:00
Summary
Nanostructured drug delivery systems allow for improvements in both bioavailability and release control of drugs in the eye. The present work aims to develop a nanosystem consisting in a polymer core and lipidic surface enabling a good encapsulation of hydroxocobalamin (OHCbl) with the purpose of treating CblC type diseases. Physicochemical properties, stability, morphology, encapsulation capacity and release of OHCbl have been studied. Toxicity of the formulation, using organotypical models, and both in-vitro and in-vivo assays have also been tested. To evaluate the possibility of freeze-drying the final formulation, some lyoprotectants were tested. Particle size and distribution was consistent for the double emulsion and solvent evaporation method. Final encapsulation efficiency was 48%. Lipid polymer hybrid nanoparticles (LPHNP) were deemed secure by the tests made on organotypical systems. In conclusion, LPHNPs could be a possible drug delivery system (DDS) for OHCbl administration in the eye. These results could provide a basis for the development of a suitable DDS in the treatment of ocular pathologies related to CblC type diseases
Nanostructured drug delivery systems allow for improvements in both bioavailability and release control of drugs in the eye. The present work aims to develop a nanosystem consisting in a polymer core and lipidic surface enabling a good encapsulation of hydroxocobalamin (OHCbl) with the purpose of treating CblC type diseases. Physicochemical properties, stability, morphology, encapsulation capacity and release of OHCbl have been studied. Toxicity of the formulation, using organotypical models, and both in-vitro and in-vivo assays have also been tested. To evaluate the possibility of freeze-drying the final formulation, some lyoprotectants were tested. Particle size and distribution was consistent for the double emulsion and solvent evaporation method. Final encapsulation efficiency was 48%. Lipid polymer hybrid nanoparticles (LPHNP) were deemed secure by the tests made on organotypical systems. In conclusion, LPHNPs could be a possible drug delivery system (DDS) for OHCbl administration in the eye. These results could provide a basis for the development of a suitable DDS in the treatment of ocular pathologies related to CblC type diseases
Direction
OTERO ESPINAR, FRANCISCO JAVIER (Tutorships)
Díaz Tomé, Victoria (Co-tutorships)
OTERO ESPINAR, FRANCISCO JAVIER (Tutorships)
Díaz Tomé, Victoria (Co-tutorships)
Court
FONTENLA GIL, JOSE ANGEL (Chairman)
GOYANES GOYANES, ALVARO (Secretary)
CASTRO PEREZ, MARIA DE LOS ANGELES (Member)
FONTENLA GIL, JOSE ANGEL (Chairman)
GOYANES GOYANES, ALVARO (Secretary)
CASTRO PEREZ, MARIA DE LOS ANGELES (Member)
Lithium modulates endotoxemia associated inflammatory responses independently of canonical NFkB pathway in ATDC5 chondrocytes
Authorship
E.A.A.H.
Master's Degree in Drug Research and Development
E.A.A.H.
Master's Degree in Drug Research and Development
Defense date
07.14.2026 09:00
07.14.2026 09:00
Summary
Osteoarthritis (OA) is a chronic joint disease characterized by cartilage degradation and low grade inflammation. Inflammatory responses in chondrocytes play an important role in OA progression. Recent studies have linked gut dysbiosis and endotoxemia to OA development through increased intestinal permeability, which allows bacterial lipopolysaccharide (LPS) to enter the circulation and activate inflammatory pathways in chondrocytes. One of the main pathways involved is nuclear factor kappa B (NFkB), which regulates the expression of proinflammatory genes through p65 nuclear translocation. LPS activates toll like receptor 4 (TLR4) and is commonly used to induce inflammation in chondrocytes. This study investigated the potential antii nflammatory effect of lithium in LPS stimulated chondrocytes and whether this effect is mediated by inhibition of NFkB pathway through blocking p65 nuclear translocation. p65 nuclear translocation in mouse ATDC5 chondrocytes stimulated with LPS for 1 h was evaluated by immunofluorescence. Nitric oxide (NO) production was indirectly assessed by measuring nitrite accumulation using the Griess reaction in cell supernatants collected after 48 h of stimulation with LPS. LPS stimulation increased p65 nuclear translocation in ATDC5 cells. However, neither lithium nor indomethacin, the anti inflammatory control, reduced this effect. In contrast, lithium significantly decreased LPS induced NO accumulation, whereas indomethacin did not. These findings suggest that lithium exerts anti inflammatory effects in chondrocytes, but these effects are not associated with inhibition of NF kB p65 nuclear translocation. Lithium may, therefore, regulate TLR4 mediated inflammatory responses through alternative signaling pathways.
Osteoarthritis (OA) is a chronic joint disease characterized by cartilage degradation and low grade inflammation. Inflammatory responses in chondrocytes play an important role in OA progression. Recent studies have linked gut dysbiosis and endotoxemia to OA development through increased intestinal permeability, which allows bacterial lipopolysaccharide (LPS) to enter the circulation and activate inflammatory pathways in chondrocytes. One of the main pathways involved is nuclear factor kappa B (NFkB), which regulates the expression of proinflammatory genes through p65 nuclear translocation. LPS activates toll like receptor 4 (TLR4) and is commonly used to induce inflammation in chondrocytes. This study investigated the potential antii nflammatory effect of lithium in LPS stimulated chondrocytes and whether this effect is mediated by inhibition of NFkB pathway through blocking p65 nuclear translocation. p65 nuclear translocation in mouse ATDC5 chondrocytes stimulated with LPS for 1 h was evaluated by immunofluorescence. Nitric oxide (NO) production was indirectly assessed by measuring nitrite accumulation using the Griess reaction in cell supernatants collected after 48 h of stimulation with LPS. LPS stimulation increased p65 nuclear translocation in ATDC5 cells. However, neither lithium nor indomethacin, the anti inflammatory control, reduced this effect. In contrast, lithium significantly decreased LPS induced NO accumulation, whereas indomethacin did not. These findings suggest that lithium exerts anti inflammatory effects in chondrocytes, but these effects are not associated with inhibition of NF kB p65 nuclear translocation. Lithium may, therefore, regulate TLR4 mediated inflammatory responses through alternative signaling pathways.
Direction
DIAZ RODRIGUEZ, PATRICIA (Tutorships)
Gómez Vaamonde, Rodolfo (Co-tutorships)
Alonso Pérez, Ana (Co-tutorships)
DIAZ RODRIGUEZ, PATRICIA (Tutorships)
Gómez Vaamonde, Rodolfo (Co-tutorships)
Alonso Pérez, Ana (Co-tutorships)
Court
FONTENLA GIL, JOSE ANGEL (Chairman)
GOYANES GOYANES, ALVARO (Secretary)
CASTRO PEREZ, MARIA DE LOS ANGELES (Member)
FONTENLA GIL, JOSE ANGEL (Chairman)
GOYANES GOYANES, ALVARO (Secretary)
CASTRO PEREZ, MARIA DE LOS ANGELES (Member)
3D-printed calcium phosphate-based scaffolds with osteogenic capacity and their application in regenerative medicine
Authorship
J.S.L.M.
Master's Degree in Drug Research and Development
J.S.L.M.
Master's Degree in Drug Research and Development
Defense date
07.14.2026 09:00
07.14.2026 09:00
Summary
The regeneration of critical-sized bone defects remains a major clinical challenge due to the limitations of conventional treatments, including autologous and allogeneic grafts, as well as metallic implants. In this context, tissue engineering and additive manufacturing have emerged as promising alternatives for the development of personalized bone substitutes capable of enhancing bone repair and regeneration. The aim of this study was to develop and characterize osteogenic 3D-printed scaffolds based on a formulation containing alpha-tricalcium phosphate (alpha-TCP), hydroxypropyl methylcellulose (HPMC) and disodium phosphate (Na2HPO4). A ceramic ink suitable for extrusion-based 3D printing was optimized, resulting in excellent printability, structural stability, and geometric fidelity. The fabricated scaffolds exhibited a woodpile macroporous architecture with interconnected pores and dimensions compatible with bone tissue engineering applications. A controlled setting process performed at 37 graos Celsius and 100% relative humidity promoted the partial conversion of alpha-TCP into calcium-deficient hydroxyapatite (CDHA), as confirmed by X-ray diffraction and scanning electron microscopy analyses. SEM micrographs revealed adequate particle cohesion and the formation of apatite-like precipitates. Furthermore, dissolution studies showed a slight mass increase associated with apatite precipitation, highlighting the bioactive nature of the material. Biological evaluation demonstrated the absence of cytotoxic effects and the ability of the scaffolds to support cell proliferation. In addition, the scaffolds exhibited suitable mechanical properties and positive osteoinductive behavior, evidenced by the expression of early osteogenic markers. Overall, the results demonstrate that the developed scaffolds represent a promising platform for future applications in bone regenerative medicine and localized therapeutic agent delivery
The regeneration of critical-sized bone defects remains a major clinical challenge due to the limitations of conventional treatments, including autologous and allogeneic grafts, as well as metallic implants. In this context, tissue engineering and additive manufacturing have emerged as promising alternatives for the development of personalized bone substitutes capable of enhancing bone repair and regeneration. The aim of this study was to develop and characterize osteogenic 3D-printed scaffolds based on a formulation containing alpha-tricalcium phosphate (alpha-TCP), hydroxypropyl methylcellulose (HPMC) and disodium phosphate (Na2HPO4). A ceramic ink suitable for extrusion-based 3D printing was optimized, resulting in excellent printability, structural stability, and geometric fidelity. The fabricated scaffolds exhibited a woodpile macroporous architecture with interconnected pores and dimensions compatible with bone tissue engineering applications. A controlled setting process performed at 37 graos Celsius and 100% relative humidity promoted the partial conversion of alpha-TCP into calcium-deficient hydroxyapatite (CDHA), as confirmed by X-ray diffraction and scanning electron microscopy analyses. SEM micrographs revealed adequate particle cohesion and the formation of apatite-like precipitates. Furthermore, dissolution studies showed a slight mass increase associated with apatite precipitation, highlighting the bioactive nature of the material. Biological evaluation demonstrated the absence of cytotoxic effects and the ability of the scaffolds to support cell proliferation. In addition, the scaffolds exhibited suitable mechanical properties and positive osteoinductive behavior, evidenced by the expression of early osteogenic markers. Overall, the results demonstrate that the developed scaffolds represent a promising platform for future applications in bone regenerative medicine and localized therapeutic agent delivery
Direction
DIAZ GOMEZ, LUIS ANTONIO (Tutorships)
DIAZ GOMEZ, LUIS ANTONIO (Tutorships)
Court
FONTENLA GIL, JOSE ANGEL (Chairman)
GOYANES GOYANES, ALVARO (Secretary)
CASTRO PEREZ, MARIA DE LOS ANGELES (Member)
FONTENLA GIL, JOSE ANGEL (Chairman)
GOYANES GOYANES, ALVARO (Secretary)
CASTRO PEREZ, MARIA DE LOS ANGELES (Member)
Optimization of Colloidal Systems for the Delivery of Products with Metabolic Activity
Authorship
M.A.L.P.
Master's Degree in Drug Research and Development
M.A.L.P.
Master's Degree in Drug Research and Development
Defense date
07.14.2026 16:00
07.14.2026 16:00
Summary
Colloidal systems (CSs) containing active pharmaceutical ingredients offer advantages in terms of bioavailability and absorption. The aim of this study was to design, develop, and physicochemically characterize colloidal systems loaded with Molecule A (CS-A) and Molecule B (CS-B) as an alternative approach for the treatment of obesity.
Colloidal systems (CSs) containing active pharmaceutical ingredients offer advantages in terms of bioavailability and absorption. The aim of this study was to design, develop, and physicochemically characterize colloidal systems loaded with Molecule A (CS-A) and Molecule B (CS-B) as an alternative approach for the treatment of obesity.
Direction
OTERO ESPINAR, FRANCISCO JAVIER (Tutorships)
Díaz Tomé, Victoria (Co-tutorships)
OTERO ESPINAR, FRANCISCO JAVIER (Tutorships)
Díaz Tomé, Victoria (Co-tutorships)
Court
FONTENLA GIL, JOSE ANGEL (Chairman)
GOYANES GOYANES, ALVARO (Secretary)
CASTRO PEREZ, MARIA DE LOS ANGELES (Member)
FONTENLA GIL, JOSE ANGEL (Chairman)
GOYANES GOYANES, ALVARO (Secretary)
CASTRO PEREZ, MARIA DE LOS ANGELES (Member)
Supramolecular nanomaterials for antimicrobial applications
Authorship
M.M.D.
Master's Degree in Drug Research and Development
M.M.D.
Master's Degree in Drug Research and Development
Defense date
07.14.2026 09:00
07.14.2026 09:00
Summary
Bacterial resistance to conventional antibiotics has been the key to the development of new antimicrobial strategies, with nanofibres standing out for their ability to act via non-specific mechanisms, against which it is more difficult to develop resistance. However, their potential toxicity to healthy cells limits their application. In this Master’s thesis, molecular precursors capable of self-assembling only within the pathogen are designed. These precursors consist of a self-assembling peptide linked to a sugar via a disulfide bond. The presence of the carbohydrate blocks the formation of nanofibres outside the cell and facilitates their entry into the bacteria. Once internalised, the reducing environment of the bacterial cytosol breaks the disulphide bond, releasing the peptide and promoting the formation of nanofibres in situ. This strategy represents an innovative approach to the development of antimicrobial materials generated directly within pathogens.
Bacterial resistance to conventional antibiotics has been the key to the development of new antimicrobial strategies, with nanofibres standing out for their ability to act via non-specific mechanisms, against which it is more difficult to develop resistance. However, their potential toxicity to healthy cells limits their application. In this Master’s thesis, molecular precursors capable of self-assembling only within the pathogen are designed. These precursors consist of a self-assembling peptide linked to a sugar via a disulfide bond. The presence of the carbohydrate blocks the formation of nanofibres outside the cell and facilitates their entry into the bacteria. Once internalised, the reducing environment of the bacterial cytosol breaks the disulphide bond, releasing the peptide and promoting the formation of nanofibres in situ. This strategy represents an innovative approach to the development of antimicrobial materials generated directly within pathogens.
Direction
INSUA LOPEZ, IGNACIO (Tutorships)
INSUA LOPEZ, IGNACIO (Tutorships)
Court
FONTENLA GIL, JOSE ANGEL (Chairman)
GOYANES GOYANES, ALVARO (Secretary)
CASTRO PEREZ, MARIA DE LOS ANGELES (Member)
FONTENLA GIL, JOSE ANGEL (Chairman)
GOYANES GOYANES, ALVARO (Secretary)
CASTRO PEREZ, MARIA DE LOS ANGELES (Member)
Analysis of the Quality of Pharmaceutical Tablets Using X-ray Computed Microtomography
Authorship
P.M.P.
Master's Degree in Drug Research and Development
P.M.P.
Master's Degree in Drug Research and Development
Defense date
07.14.2026 09:00
07.14.2026 09:00
Summary
Quality assurance in pharmaceutical tablets requires microstructural characterization methods that are accurate and comprehensive. The usual characterization approaches have been performed using techniques such as mercury porosimetry or helium pycnometry, two useful methods that still present limitations today, including being destructive, not detecting closed porosity, and not providing information on the spatial distribution of pores. Given these limitations, X-ray microcomputed tomography (Micro-CT) is presented as a non-destructive alternative. This work proposes Micro-CT as a non-destructive technique for evaluating the microstructural quality of tablets. Based on the Beer-Lambert attenuation law, Micro-CT makes it possible to scan and reconstruct three-dimensional models from 2D projections, distinguishing components and porosity. First, paracetamol- and starch-formulated tablets were analyzed to evaluate the workflows at the level of porosity and component distribution. Then, sertraline tablets, both branded Pfizer and generic, were analyzed and the methods were evaluated again. After the analyses, it was observed that through these Micro-CT analyses, the complete distribution of the components of a tablet can be visualized, as well as the pores, both open and closed. This suggests that it is a technique that, without destroying tablet batches, allows their quality to be characterized.
Quality assurance in pharmaceutical tablets requires microstructural characterization methods that are accurate and comprehensive. The usual characterization approaches have been performed using techniques such as mercury porosimetry or helium pycnometry, two useful methods that still present limitations today, including being destructive, not detecting closed porosity, and not providing information on the spatial distribution of pores. Given these limitations, X-ray microcomputed tomography (Micro-CT) is presented as a non-destructive alternative. This work proposes Micro-CT as a non-destructive technique for evaluating the microstructural quality of tablets. Based on the Beer-Lambert attenuation law, Micro-CT makes it possible to scan and reconstruct three-dimensional models from 2D projections, distinguishing components and porosity. First, paracetamol- and starch-formulated tablets were analyzed to evaluate the workflows at the level of porosity and component distribution. Then, sertraline tablets, both branded Pfizer and generic, were analyzed and the methods were evaluated again. After the analyses, it was observed that through these Micro-CT analyses, the complete distribution of the components of a tablet can be visualized, as well as the pores, both open and closed. This suggests that it is a technique that, without destroying tablet batches, allows their quality to be characterized.
Direction
DIAZ GOMEZ, LUIS ANTONIO (Tutorships)
ALVAREZ LORENZO, CARMEN ISABEL (Co-tutorships)
DIAZ GOMEZ, LUIS ANTONIO (Tutorships)
ALVAREZ LORENZO, CARMEN ISABEL (Co-tutorships)
Court
FONTENLA GIL, JOSE ANGEL (Chairman)
GOYANES GOYANES, ALVARO (Secretary)
CASTRO PEREZ, MARIA DE LOS ANGELES (Member)
FONTENLA GIL, JOSE ANGEL (Chairman)
GOYANES GOYANES, ALVARO (Secretary)
CASTRO PEREZ, MARIA DE LOS ANGELES (Member)
Pharmacological inhibition of exosome release as a therapeutic strategy in heterotopic ossification
Authorship
J.N.O.
Master's Degree in Drug Research and Development
J.N.O.
Master's Degree in Drug Research and Development
Defense date
07.14.2026 15:00
07.14.2026 15:00
Summary
Neurogenic heterotopic ossification (NHO) is characterized by the formation of mature lamellar bone in soft tissues following central nervous system (CNS) injuries. Recent research indicates that exosomes may act as key mediators in pathological inter-organ communication, transporting signals from the CNS to peripheral tissues. The main objective of this study is to evaluate the potential of the exosome signalling inhibitor GW4869 as a therapeutic strategy against NHO. To this end, an in vitro model was stablished using human reactive astrocytes (CCF-STTG1) and pre-osteoblastic cells (SaOS-2). The inhibitor GW4869 was used to block exosomal biogenesis in astrocytes, and it was evaluated whether this inhibition modified the osteoinductive potential of their secretome. Results revealed that the treatment with GW4869 was insufficient to reverse sclerostin (SOST) inhibition or reduce osteopontin (SPP1) overexpression. These findings suggest that the pro-osteoblastogenic effect of the astrocytic secretome might be independent of the GW4869-sensitive exosomal pathway. In conclusion, although the employed model constitutes an effective tool for studying inter-organ communication in NHO, the inhibition of exosome release via GW4869 is, under the tested conditions, insufficient to interrupt pathological signalling. Nonetheless, due to the variability of the results, additional studies are necessary to reach more robust conclusions.
Neurogenic heterotopic ossification (NHO) is characterized by the formation of mature lamellar bone in soft tissues following central nervous system (CNS) injuries. Recent research indicates that exosomes may act as key mediators in pathological inter-organ communication, transporting signals from the CNS to peripheral tissues. The main objective of this study is to evaluate the potential of the exosome signalling inhibitor GW4869 as a therapeutic strategy against NHO. To this end, an in vitro model was stablished using human reactive astrocytes (CCF-STTG1) and pre-osteoblastic cells (SaOS-2). The inhibitor GW4869 was used to block exosomal biogenesis in astrocytes, and it was evaluated whether this inhibition modified the osteoinductive potential of their secretome. Results revealed that the treatment with GW4869 was insufficient to reverse sclerostin (SOST) inhibition or reduce osteopontin (SPP1) overexpression. These findings suggest that the pro-osteoblastogenic effect of the astrocytic secretome might be independent of the GW4869-sensitive exosomal pathway. In conclusion, although the employed model constitutes an effective tool for studying inter-organ communication in NHO, the inhibition of exosome release via GW4869 is, under the tested conditions, insufficient to interrupt pathological signalling. Nonetheless, due to the variability of the results, additional studies are necessary to reach more robust conclusions.
Direction
DIAZ RODRIGUEZ, PATRICIA (Tutorships)
Gómez Vaamonde, Rodolfo (Co-tutorships)
Piñeiro Ramil, María (Co-tutorships)
DIAZ RODRIGUEZ, PATRICIA (Tutorships)
Gómez Vaamonde, Rodolfo (Co-tutorships)
Piñeiro Ramil, María (Co-tutorships)
Court
FONTENLA GIL, JOSE ANGEL (Chairman)
GOYANES GOYANES, ALVARO (Secretary)
CASTRO PEREZ, MARIA DE LOS ANGELES (Member)
FONTENLA GIL, JOSE ANGEL (Chairman)
GOYANES GOYANES, ALVARO (Secretary)
CASTRO PEREZ, MARIA DE LOS ANGELES (Member)
Studies of SEDN2 35 Mediated Platelet Inhibition
Authorship
R.C.P.G.
Master's Degree in Drug Research and Development
R.C.P.G.
Master's Degree in Drug Research and Development
Defense date
07.14.2026 15:00
07.14.2026 15:00
Summary
Cardiovascular diseases remain the leading cause of mortality worldwide and are largely associated with the formation of arterial thrombi. Although current antiplatelet therapies effectively reduce the risk of thrombotic events, their use is limited by an increased risk of bleeding. Consequently, the search for new therapeutic targets has focused on platelet glycoprotein VI (GPVI), a collagen receptor that plays a pivotal role in thrombosis while having a more limited role in physiological hemostasis. In this context, the aim of this study was to evaluate the antiplatelet potential of the small molecule named SEDN2 35 by investigating its effects on GPVI mediated signaling and thrombus formation. To this end, western blot analyses were performed to assess the phosphorylation of key proteins involved in the GPVI signaling pathway, and ex vivo microfluidic assays using whole blood from healthy donors were conducted to evaluate thrombus formation and platelet activation markers under physiological flow conditions. The results showed that SEDN2 35 reduced the phosphorylation of proteins involved in GPVI signaling and decreased thrombus formation and complexity, as well as integrin activation, phosphatidylserine exposure, and P selectin expression. Overall, these findings support the potential of SEDN2 35 as a promising GPVI targeted antiplatelet agent and provide further evidence for the development of safer antithrombotic therapies aimed at preventing atherothrombotic events while minimizing the bleeding risk associated with current treatments.
Cardiovascular diseases remain the leading cause of mortality worldwide and are largely associated with the formation of arterial thrombi. Although current antiplatelet therapies effectively reduce the risk of thrombotic events, their use is limited by an increased risk of bleeding. Consequently, the search for new therapeutic targets has focused on platelet glycoprotein VI (GPVI), a collagen receptor that plays a pivotal role in thrombosis while having a more limited role in physiological hemostasis. In this context, the aim of this study was to evaluate the antiplatelet potential of the small molecule named SEDN2 35 by investigating its effects on GPVI mediated signaling and thrombus formation. To this end, western blot analyses were performed to assess the phosphorylation of key proteins involved in the GPVI signaling pathway, and ex vivo microfluidic assays using whole blood from healthy donors were conducted to evaluate thrombus formation and platelet activation markers under physiological flow conditions. The results showed that SEDN2 35 reduced the phosphorylation of proteins involved in GPVI signaling and decreased thrombus formation and complexity, as well as integrin activation, phosphatidylserine exposure, and P selectin expression. Overall, these findings support the potential of SEDN2 35 as a promising GPVI targeted antiplatelet agent and provide further evidence for the development of safer antithrombotic therapies aimed at preventing atherothrombotic events while minimizing the bleeding risk associated with current treatments.
Direction
GARCIA ALONSO, ANGEL (Tutorships)
GARCIA ALONSO, ANGEL (Tutorships)
Court
FONTENLA GIL, JOSE ANGEL (Chairman)
GOYANES GOYANES, ALVARO (Secretary)
CASTRO PEREZ, MARIA DE LOS ANGELES (Member)
FONTENLA GIL, JOSE ANGEL (Chairman)
GOYANES GOYANES, ALVARO (Secretary)
CASTRO PEREZ, MARIA DE LOS ANGELES (Member)
Role of apoptosis in the generation of paracrine senescence signals
Authorship
A.R.R.
Master's Degree in Drug Research and Development
A.R.R.
Master's Degree in Drug Research and Development
Defense date
07.14.2026 09:00
07.14.2026 09:00
Summary
Apoptosis and cellular senescence represent opposing responses to damage, yet both cooperate in key physiological processes and jointly contribute to age-related deterioration. Previous evidence from our laboratory demonstrated that apoptotic cells can induce paracrine senescence in neighbouring cells through their secretome, and that this effect is reduced in the absence of mitochondrial involvement. However, the molecular pathways mediating this intercellular communication remain uncharacterised. In this study, the A549 cell line was used as a model of staurosporine-induced mitochondrial apoptosis to investigate the mechanisms underlying the propagation of pro-senescent signals to the cellular microenvironment. Two complementary strategies were designed: a pharmacological screening using inhibitors of the cGAS STING (H151) and NFkB (BAY117082 and IKK16) pathways in producer cells, whose conditioned media were subsequently transferred to target cells to assess paracrine senescence induction; and a constitutive genetic silencing of the NFkB p65 subunit via RNA interference (shRNA), to overcome the potential limitations of chemical inhibition. Results showed that pharmacological screening effectively reduced the inflammatory profile in producer cells, but this blockade was not sufficient to attenuate paracrine senescence in target cells. In contrast, p65 silencing reduced the senescent response in target cells, identifying NFkB as a relevant effector in this process. These findings open new avenues for the study of the crosstalk between apoptosis and senescence, with potential therapeutic implications in the investigation of ageing-associated pathologies.
Apoptosis and cellular senescence represent opposing responses to damage, yet both cooperate in key physiological processes and jointly contribute to age-related deterioration. Previous evidence from our laboratory demonstrated that apoptotic cells can induce paracrine senescence in neighbouring cells through their secretome, and that this effect is reduced in the absence of mitochondrial involvement. However, the molecular pathways mediating this intercellular communication remain uncharacterised. In this study, the A549 cell line was used as a model of staurosporine-induced mitochondrial apoptosis to investigate the mechanisms underlying the propagation of pro-senescent signals to the cellular microenvironment. Two complementary strategies were designed: a pharmacological screening using inhibitors of the cGAS STING (H151) and NFkB (BAY117082 and IKK16) pathways in producer cells, whose conditioned media were subsequently transferred to target cells to assess paracrine senescence induction; and a constitutive genetic silencing of the NFkB p65 subunit via RNA interference (shRNA), to overcome the potential limitations of chemical inhibition. Results showed that pharmacological screening effectively reduced the inflammatory profile in producer cells, but this blockade was not sufficient to attenuate paracrine senescence in target cells. In contrast, p65 silencing reduced the senescent response in target cells, identifying NFkB as a relevant effector in this process. These findings open new avenues for the study of the crosstalk between apoptosis and senescence, with potential therapeutic implications in the investigation of ageing-associated pathologies.
Direction
DIAZ RODRIGUEZ, PATRICIA (Tutorships)
Collado Rodríguez, Manuel (Co-tutorships)
DIAZ RODRIGUEZ, PATRICIA (Tutorships)
Collado Rodríguez, Manuel (Co-tutorships)
Court
FONTENLA GIL, JOSE ANGEL (Chairman)
GOYANES GOYANES, ALVARO (Secretary)
CASTRO PEREZ, MARIA DE LOS ANGELES (Member)
FONTENLA GIL, JOSE ANGEL (Chairman)
GOYANES GOYANES, ALVARO (Secretary)
CASTRO PEREZ, MARIA DE LOS ANGELES (Member)
In vivo diffusion assessment of mRNA nanocarriers
Authorship
A.S.T.
Master's Degree in Drug Research and Development
A.S.T.
Master's Degree in Drug Research and Development
Defense date
07.14.2026 15:00
07.14.2026 15:00
Summary
This study develops and evaluates a new mRNA-loaded nanocarrier (NC) for central nervous system (CNS) delivery and compares its in vivo brain diffusion behaviour with a canonical cholesterol-based lipid nanoparticle (LNP). Nanosystems studied (Cy5-mFLuc NC, and Cy5-mFLuc LNP) showed optimal physicochemical properties for their diffusion in the brain microenvironment, presenting hydrodynamic diameters less than 100 nm, polydispersity indices (less than 0.14), and a neutral zeta potential. Electrophoresis and RiboGreen assays confirmed that mRNA encapsulation efficiencies were optimal (more than 80% for the NC and more than 70% for the LNP). In vivo kinetic studies were performed using multiphoton microscopy in healthy mice, and it was observed that both formulations have a characteristic pattern that begins with an initial increase in apparent volume followed by signal attenuation due to the dilution effect. However, NC significantly outperformed the dispersion of the conventional LNP, with an average maximum diffusion volume achieved of 347% at 4 hours (considering the initial volume of the first timepoint 100%), while the LNP reached a significantly lower volume increase (131.67%) at the same timepoint. These findings indicate that subtle variations in lipid composition and their resulting changes in the physicochemical and biophysical properties of the NC, critically influence their diffusion in the brain. Overall, this work highlights the dependence of the lipid composition in the control of mRNA distribution within the brain as a key parameter for CNS nanomedicine design.
This study develops and evaluates a new mRNA-loaded nanocarrier (NC) for central nervous system (CNS) delivery and compares its in vivo brain diffusion behaviour with a canonical cholesterol-based lipid nanoparticle (LNP). Nanosystems studied (Cy5-mFLuc NC, and Cy5-mFLuc LNP) showed optimal physicochemical properties for their diffusion in the brain microenvironment, presenting hydrodynamic diameters less than 100 nm, polydispersity indices (less than 0.14), and a neutral zeta potential. Electrophoresis and RiboGreen assays confirmed that mRNA encapsulation efficiencies were optimal (more than 80% for the NC and more than 70% for the LNP). In vivo kinetic studies were performed using multiphoton microscopy in healthy mice, and it was observed that both formulations have a characteristic pattern that begins with an initial increase in apparent volume followed by signal attenuation due to the dilution effect. However, NC significantly outperformed the dispersion of the conventional LNP, with an average maximum diffusion volume achieved of 347% at 4 hours (considering the initial volume of the first timepoint 100%), while the LNP reached a significantly lower volume increase (131.67%) at the same timepoint. These findings indicate that subtle variations in lipid composition and their resulting changes in the physicochemical and biophysical properties of the NC, critically influence their diffusion in the brain. Overall, this work highlights the dependence of the lipid composition in the control of mRNA distribution within the brain as a key parameter for CNS nanomedicine design.
Direction
ALONSO FERNANDEZ, MARIA JOSEFA (Tutorships)
ALONSO FERNANDEZ, MARIA JOSEFA (Tutorships)
Court
FONTENLA GIL, JOSE ANGEL (Chairman)
GOYANES GOYANES, ALVARO (Secretary)
CASTRO PEREZ, MARIA DE LOS ANGELES (Member)
FONTENLA GIL, JOSE ANGEL (Chairman)
GOYANES GOYANES, ALVARO (Secretary)
CASTRO PEREZ, MARIA DE LOS ANGELES (Member)