Development and characterisation of biomimetic nanoparticles targeted to mesenchymal stem cells and osteoclasts
Authorship
A.A.P.
Master in Drug Research and Development
A.A.P.
Master in Drug Research and Development
Defense date
09.22.2025 10:00
09.22.2025 10:00
Summary
Osteoporosis is the most prevalent bone disease worldwide. Its treatment, primarily based on antiresorptive drugs, carries the risk of adverse side effects, highlighting the need for the development of new therapeutic strategies. Gene therapy represents a promising approach for the treatment of chronic diseases such as osteoporosis, as it enables the precise regulation of genes involved in bone remodelling. However, its effective application requires the development of delivery vectors capable of ensuring efficient and specific transfection of target cells with minimal adverse effects. In this study, biomimetic nanoparticles (BMNPs) were developed by coating polymeric cores with extracellular vesicle membranes isolated from mesenchymal stem cells (MSCs) and osteoclasts (OLS), for their potential use as gene therapy vectors. The resulting BMNPs were characterized in terms of physicochemical properties. Additionally, the composition of the isolated membranes was analyzed in terms of lipids, proteins, and DNA content. The cytocompatibility of the formulations was assessed, and their cellular association with MSCs and OLS was evaluated in the presence and absence of different endocytic pathway inhibitors. The obtained BMNPs exhibited size and zeta potential values consistent with those commonly reported in the literature for similar systems and showed no cytotoxicity. It was observed that MSCs internalized BMNPs predominantly via caveolin-mediated endocytosis, and to a lesser extent via FEME, while in OLS, the data suggested the fusion of the BMNP coating with the plasma membrane. These findings support the feasibility of producing BMNPs with suitable physicochemical properties and indicate that their cellular interaction is not only dependent on the intrinsic characteristics of the nanoparticles but also on the specific target cell type.
Osteoporosis is the most prevalent bone disease worldwide. Its treatment, primarily based on antiresorptive drugs, carries the risk of adverse side effects, highlighting the need for the development of new therapeutic strategies. Gene therapy represents a promising approach for the treatment of chronic diseases such as osteoporosis, as it enables the precise regulation of genes involved in bone remodelling. However, its effective application requires the development of delivery vectors capable of ensuring efficient and specific transfection of target cells with minimal adverse effects. In this study, biomimetic nanoparticles (BMNPs) were developed by coating polymeric cores with extracellular vesicle membranes isolated from mesenchymal stem cells (MSCs) and osteoclasts (OLS), for their potential use as gene therapy vectors. The resulting BMNPs were characterized in terms of physicochemical properties. Additionally, the composition of the isolated membranes was analyzed in terms of lipids, proteins, and DNA content. The cytocompatibility of the formulations was assessed, and their cellular association with MSCs and OLS was evaluated in the presence and absence of different endocytic pathway inhibitors. The obtained BMNPs exhibited size and zeta potential values consistent with those commonly reported in the literature for similar systems and showed no cytotoxicity. It was observed that MSCs internalized BMNPs predominantly via caveolin-mediated endocytosis, and to a lesser extent via FEME, while in OLS, the data suggested the fusion of the BMNP coating with the plasma membrane. These findings support the feasibility of producing BMNPs with suitable physicochemical properties and indicate that their cellular interaction is not only dependent on the intrinsic characteristics of the nanoparticles but also on the specific target cell type.
Direction
DIAZ RODRIGUEZ, PATRICIA (Tutorships)
ROUCO TABOADA, HELENA (Co-tutorships)
DIAZ RODRIGUEZ, PATRICIA (Tutorships)
ROUCO TABOADA, HELENA (Co-tutorships)
Court
LOZA GARCIA, MARIA ISABEL (Chairman)
GARCIA GONZALEZ, CARLOS ALBERTO (Secretary)
CSABA , NOEMI STEFANIA (Member)
LOZA GARCIA, MARIA ISABEL (Chairman)
GARCIA GONZALEZ, CARLOS ALBERTO (Secretary)
CSABA , NOEMI STEFANIA (Member)
Study of the phosphorylation of the CX3CR1 chemokine receptor and that of its natural genetic variant CX3CR1-A55T in cell lines
Authorship
P.F.R.
Master in Drug Research and Development
P.F.R.
Master in Drug Research and Development
Defense date
09.22.2025 10:00
09.22.2025 10:00
Summary
Fractalkine (CX3CL1, FKN) is a member of the chemokine family and is constitutively and abundantly expressed in neurons. The reciprocal interaction between the chemokine receptor CX3CR1, highly expressed in microglia cells, and its neuronal ligand CX3CL1, enables precise and effective communication between neurons and microglial cells, playing a key role in coordinating many aspects of brain function. In this context, the CX3CL1/CX3CR1 axis is of particular interest due to its potential involvement in psychiatric disorders such as schizophrenia. Alterations in the amino acid sequence of human CX3CR1 have been found in natural genetic variants of the receptor, among them the rare naturally occurring genetic variant CX3CR1-A55T. These changes in the amino acid sequence may modify the phosphorylation pattern of the receptor, altering its response to fractalkine and affecting microglial activation. Therefore, a comparative study of the phosphorylation pattern between the wild-type CX3CR1 isoform and its A55T genetic variant is essential to understand the functional impact of this mutation under both physiological and pathological conditions, such as schizophrenia.
Fractalkine (CX3CL1, FKN) is a member of the chemokine family and is constitutively and abundantly expressed in neurons. The reciprocal interaction between the chemokine receptor CX3CR1, highly expressed in microglia cells, and its neuronal ligand CX3CL1, enables precise and effective communication between neurons and microglial cells, playing a key role in coordinating many aspects of brain function. In this context, the CX3CL1/CX3CR1 axis is of particular interest due to its potential involvement in psychiatric disorders such as schizophrenia. Alterations in the amino acid sequence of human CX3CR1 have been found in natural genetic variants of the receptor, among them the rare naturally occurring genetic variant CX3CR1-A55T. These changes in the amino acid sequence may modify the phosphorylation pattern of the receptor, altering its response to fractalkine and affecting microglial activation. Therefore, a comparative study of the phosphorylation pattern between the wild-type CX3CR1 isoform and its A55T genetic variant is essential to understand the functional impact of this mutation under both physiological and pathological conditions, such as schizophrenia.
Direction
CASTRO PEREZ, MARIA DE LOS ANGELES (Tutorships)
CASTRO PEREZ, MARIA DE LOS ANGELES (Tutorships)
Court
LOZA GARCIA, MARIA ISABEL (Chairman)
GARCIA GONZALEZ, CARLOS ALBERTO (Secretary)
CSABA , NOEMI STEFANIA (Member)
LOZA GARCIA, MARIA ISABEL (Chairman)
GARCIA GONZALEZ, CARLOS ALBERTO (Secretary)
CSABA , NOEMI STEFANIA (Member)
3D Printing for Filling Sustained-Release Capsules
Authorship
I.C.O.O.
Master in Drug Research and Development
I.C.O.O.
Master in Drug Research and Development
Defense date
09.22.2025 10:00
09.22.2025 10:00
Summary
The development of personalized oral dosage forms using 3D printing is advancing rapidly. In this study, gelatin capsules using the Semi-Solid Extrusion (SSE) method of 3D Printing were filled with formulations containing paracetamol as the model drug. Two doses were prepared; 20 mg and 50 mg, each composed of 25% paracetamol, 37.5% polyethylene glycol 4000 (PEG 4000), and 37.5% Witepsol PMF 166 or PMF 123 as excipients. One hundred capsules of each dose were manufactured, and content uniformity was assessed. The results showed that SSE 3D printing can fill gelatin capsules with precise and reproducible drug content, offering potential for on-demand manufacturing of customized oral formulations with flexible dosing. The formulation with Witepsol PMF 123 could not be used due to a clogged tip, so only the formulation with Witepsol PMF 166 was validated. For quality control, three capsules were randomly selected from each dose and analyzed by High-Performance Liquid Chromatography (HPLC). The mean recovery of paracetamol was 98.27 +/- 1.35% (RSD = 1.37%) for 20 mg capsules and 97.73 +/- 3.64% (RSD = 3.72%) for 50 mg capsules, in accordance with the target doses and acceptable variability according to the pharmacopoeia. The sustained-release profile obtained supports the primary objective of the study: to demonstrate that SSE 3D printing can produce customized pharmaceutical capsules with controlled and sustained release.
The development of personalized oral dosage forms using 3D printing is advancing rapidly. In this study, gelatin capsules using the Semi-Solid Extrusion (SSE) method of 3D Printing were filled with formulations containing paracetamol as the model drug. Two doses were prepared; 20 mg and 50 mg, each composed of 25% paracetamol, 37.5% polyethylene glycol 4000 (PEG 4000), and 37.5% Witepsol PMF 166 or PMF 123 as excipients. One hundred capsules of each dose were manufactured, and content uniformity was assessed. The results showed that SSE 3D printing can fill gelatin capsules with precise and reproducible drug content, offering potential for on-demand manufacturing of customized oral formulations with flexible dosing. The formulation with Witepsol PMF 123 could not be used due to a clogged tip, so only the formulation with Witepsol PMF 166 was validated. For quality control, three capsules were randomly selected from each dose and analyzed by High-Performance Liquid Chromatography (HPLC). The mean recovery of paracetamol was 98.27 +/- 1.35% (RSD = 1.37%) for 20 mg capsules and 97.73 +/- 3.64% (RSD = 3.72%) for 50 mg capsules, in accordance with the target doses and acceptable variability according to the pharmacopoeia. The sustained-release profile obtained supports the primary objective of the study: to demonstrate that SSE 3D printing can produce customized pharmaceutical capsules with controlled and sustained release.
Direction
GOYANES GOYANES, ALVARO (Tutorships)
GOYANES GOYANES, ALVARO (Tutorships)
Court
LOZA GARCIA, MARIA ISABEL (Chairman)
GARCIA GONZALEZ, CARLOS ALBERTO (Secretary)
CSABA , NOEMI STEFANIA (Member)
LOZA GARCIA, MARIA ISABEL (Chairman)
GARCIA GONZALEZ, CARLOS ALBERTO (Secretary)
CSABA , NOEMI STEFANIA (Member)