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Article ; Online: Screening strategies for surface modification of lipid-polymer hybrid nanoparticles.

Rouco, Helena / García-García, Patricia / Évora, Carmen / Díaz-Rodríguez, Patricia / Delgado, Araceli

2022 Volume 624, Page(s) 121973

Abstract: Lipid-polymer hybrid nanoparticles are promising platforms in the field of targeted drug delivery, integrating the positive features of polymeric and lipid nanocarriers. However, the use of bulk procedures in lipid-polymer hybrid nanoparticles ... ...

Abstract	Lipid-polymer hybrid nanoparticles are promising platforms in the field of targeted drug delivery, integrating the positive features of polymeric and lipid nanocarriers. However, the use of bulk procedures in lipid-polymer hybrid nanoparticles formulation is hindering their large-scale manufacturing. Therefore, the aim of this study is to explore the suitability of alternative formulation methods, such as microfluidics, to obtain surface-tunable nanoparticles displaying suitable characteristics. Formulations were prepared by single-step nanoprecipitation or using a micromixer chip. The nanocarriers were then surface-modified with an aptamer and an antibody, two common nanoparticle vectorization strategies, developing an optimized functionalization protocol. Both naked and surface-modified nanoparticles were characterized in terms of size, polydispersity, zeta potential and morphology. Moreover, the aptamer/antibody association efficiency was also determined. Nano-sized monodisperse nanoparticles, exhibiting a spherical core-shell structure, were obtained through both procedures. Furthermore, all the nanocarriers were successfully functionalized, showing association efficiency values above 70%. Interestingly, microfluidic-based nanoparticles displayed a smaller size and a more positive zeta potential than those prepared by single-step nanoprecipitation. Outcomes suggest both techniques led to lipid-polymer hybrid nanoparticles displaying a similar functionalization efficiency. Conversely, the microfluidic approach provided an improved control over critical parameters, as particle size or charge, constituting an interesting alternative to traditional formulation procedures.
MeSH term(s)	Drug Carriers/chemistry ; Drug Delivery Systems/methods ; Lipids/chemistry ; Nanoparticles/chemistry ; Particle Size ; Polymers/chemistry
Chemical Substances	Drug Carriers ; Lipids ; Polymers
Language	English
Publishing date	2022-07-08
Publishing country	Netherlands
Document type	Journal Article
ZDB-ID	428962-6
ISSN	1873-3476 ; 0378-5173
ISSN (online)	1873-3476
ISSN	0378-5173
DOI	10.1016/j.ijpharm.2022.121973
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Article ; Online: Osteoprotective effect of the marine alkaloid norzoanthamine on an osteoporosis model in ovariectomized rat.

García-García, Patricia / Reyes, Ricardo / Évora, Carmen / Delgado, Araceli / Fernández, José J / Daranas, Antonio Hernández

Biomedicine & pharmacotherapy = Biomedecine & pharmacotherapie

2022 Volume 147, Page(s) 112631

Abstract: Norzoanthamine (NZ), an alkaloid that has been isolated from the marine cnidiaria Zoanthus sp., has been shown an interesting anti-osteoporotic activity. Although its mechanism of action is not yet clear, it seems that it is different from those of ... ...

Abstract	Norzoanthamine (NZ), an alkaloid that has been isolated from the marine cnidiaria Zoanthus sp., has been shown an interesting anti-osteoporotic activity. Although its mechanism of action is not yet clear, it seems that it is different from those of currently used drugs making it particularly interesting. Previous studies have been carried out mostly in vitro. Herein, we present an in vivo study that allows to check the real potential of NZ as a protector substance by direct application into ovariectomized rat bone using a sustained delivery system. Histological and histomorphometric results in ovariectomized rats showed higher bone quality as a result of greater number of trabeculae and osteogenic activity in the group implanted with NZ, compared to controls. In contrast with the untreated controls, NZ-treated groups showed a balanced osteoblast/osteoclast number ratio, similar to that found in the normal bone. These results suggest that NZ could be useful as adjunct to other osteoporosis treatments, but probably its main therapeutic role would be as preventive therapy against bone deterioration.
MeSH term(s)	Alkaloids/pharmacology ; Animals ; Azepines/pharmacology ; Biological Products/pharmacology ; Bone Density/drug effects ; Disease Models, Animal ; Drug Liberation ; Female ; Heterocyclic Compounds, 4 or More Rings/pharmacology ; Microspheres ; Osteoblasts/drug effects ; Osteoclasts/drug effects ; Osteoporosis/pathology ; Ovariectomy ; Polylactic Acid-Polyglycolic Acid Copolymer/chemistry ; Quinolines/pharmacology ; Rats ; Rats, Sprague-Dawley
Chemical Substances	Alkaloids ; Azepines ; Biological Products ; Heterocyclic Compounds, 4 or More Rings ; Quinolines ; norzoanthamine (164991-65-5) ; Polylactic Acid-Polyglycolic Acid Copolymer (1SIA8062RS)
Language	English
Publishing date	2022-01-13
Publishing country	France
Document type	Journal Article
ZDB-ID	392415-4
ISSN	1950-6007 ; 0753-3322 ; 0300-0893
ISSN (online)	1950-6007
ISSN	0753-3322 ; 0300-0893
DOI	10.1016/j.biopha.2022.112631
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Article ; Online: Tailor-made oligonucleotide-loaded lipid-polymer nanosystems designed for bone gene therapy.

García-García, Patricia / Briffault, Erik / Landin, Mariana / Evora, Carmen / Diaz-Rodriguez, Patricia / Delgado, Araceli

Drug delivery and translational research

2021 Volume 11, Issue 2, Page(s) 598–607

Abstract: Gene therapy has emerged as a tool for the treatment of systemic metabolic disorders as osteoporosis (OP). However, the design of a suitable vehicle able to efficiently load and release the genetic material on the target cells is still a challenge. ... ...

Abstract	Gene therapy has emerged as a tool for the treatment of systemic metabolic disorders as osteoporosis (OP). However, the design of a suitable vehicle able to efficiently load and release the genetic material on the target cells is still a challenge. Moreover, the internalization pathway of nanosystems has been described to be dependent on their surface characteristics and the cell type evaluated. In this study, we aim at obtaining PEGylated lipid-PLGA nanoparticles (NPs) with variable surface charge able to incorporate GapmeRs (single-strand antisense oligonucleotides) for OP treatment. Nanoparticles showing negative, positive, and neutral surface charge were obtained by modulating the lipid composition. All formulations showed a remarkably low polydispersity index with adequate size. NPs were loaded with GapmeRs showing a high encapsulation efficiency and a surface charge-independent oligonucleotide loading. All the formulations were adequately internalized by MSCs. Future experiments will be devoted to use the developed formulations to clarify if the intracellular distribution of hybrid NPs on mesenchymal stem cells (MSCs) is dependent on surface charge. This portfolio of NPs will serve as a tool to analyze the effect of NP surface charge on gene therapy efficiency.
MeSH term(s)	Drug Carriers ; Genetic Therapy ; Lipids ; Nanoparticles ; Oligonucleotides ; Particle Size ; Polymers
Chemical Substances	Drug Carriers ; Lipids ; Oligonucleotides ; Polymers
Language	English
Publishing date	2021-02-24
Publishing country	United States
Document type	Journal Article ; Research Support, Non-U.S. Gov't
ZDB-ID	2590155-2
ISSN	2190-3948 ; 2190-393X
ISSN (online)	2190-3948
ISSN	2190-393X
DOI	10.1007/s13346-021-00926-5
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Article: Stem Cell Growth and Differentiation in Organ Culture: New Insights for Uterine Fibroid Treatment.

Salas, Ana / Beltrán-Flores, Silvia / Évora, Carmen / Reyes, Ricardo / Montes de Oca, Francisco / Delgado, Araceli / Almeida, Teresa A

Biomedicines

2022 Volume 10, Issue 7

Abstract: Organ culture allows for the understanding of normal and tumor cell biology, and tissues generally remain viable for 5-7 days. Strikingly, we determined that myometrial ... ...

Abstract	Organ culture allows for the understanding of normal and tumor cell biology, and tissues generally remain viable for 5-7 days. Strikingly, we determined that myometrial and
Language	English
Publishing date	2022-06-29
Publishing country	Switzerland
Document type	Journal Article
ZDB-ID	2720867-9
ISSN	2227-9059
ISSN	2227-9059
DOI	10.3390/biomedicines10071542
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Article ; Online: New local ganirelix sustained release therapy for uterine leiomyoma. Evaluation in a preclinical organ model.

Salas, Ana / García-García, Patricia / Díaz-Rodríguez, Patricia / Évora, Carmen / Almeida, Teresa A / Delgado, Araceli

Biomedicine & pharmacotherapy = Biomedecine & pharmacotherapie

2022 Volume 156, Page(s) 113909

Abstract: Currently, there is a limited number of treatment options available for patients with symptomatic leiomyomas, and surgical removal is by far the most frequent procedure. Previous studies found that GnRH agonists and antagonists acting through GnRH ... ...

Abstract	Currently, there is a limited number of treatment options available for patients with symptomatic leiomyomas, and surgical removal is by far the most frequent procedure. Previous studies found that GnRH agonists and antagonists acting through GnRH receptors led to cell death and decreased extracellular synthesis in cultured leiomyoma cells. In this study, we encapsulated the GnRH antagonist ganirelix in PLGA microspheres contained in an alginate scaffold that also supports a leiomyoma ex vivo tissue explant. Microspheres maintained ganirelix concentration stably during six days of culture, inducing significant cell death in 50-55% of tumor cells. Although no changes were observed in the expression of extracellular matrix genes, a decreased expression of the Nuclear Factor of Activated T cells 5, a transcription factor involved in osmotic stress and tumor size. Interestingly, all tumors analyzed experienced apoptosis independently of the original driver mutation. These data indicate that local therapy of ganirelix would induce tumor reduction in a wide range of uterine leiomyomas.
MeSH term(s)	Humans ; Female ; Delayed-Action Preparations ; Leiomyoma/metabolism ; Gonadotropin-Releasing Hormone/pharmacology ; Hormone Antagonists/pharmacology ; Hormone Antagonists/therapeutic use ; Uterine Neoplasms/pathology
Chemical Substances	ganirelix (IX503L9WN0) ; Delayed-Action Preparations ; Gonadotropin-Releasing Hormone (33515-09-2) ; Hormone Antagonists
Language	English
Publishing date	2022-10-21
Publishing country	France
Document type	Journal Article
ZDB-ID	392415-4
ISSN	1950-6007 ; 0753-3322 ; 0300-0893
ISSN (online)	1950-6007
ISSN	0753-3322 ; 0300-0893
DOI	10.1016/j.biopha.2022.113909
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Article: The Bone Regeneration Capacity of BMP-2 + MMP-10 Loaded Scaffolds Depends on the Tissue Status.

Garcia-Garcia, Patricia / Reyes, Ricardo / Rodriguez, José Antonio / Martín, Tomas / Evora, Carmen / Díaz-Rodríguez, Patricia / Delgado, Araceli

Pharmaceutics

2021 Volume 13, Issue 7

Abstract: Biomaterials-mediated bone formation in osteoporosis (OP) is challenging as it requires tissue growth promotion and adequate mineralization. Based on our previous findings, the development of scaffolds combining bone morphogenetic protein 2 (BMP-2) and ... ...

Abstract	Biomaterials-mediated bone formation in osteoporosis (OP) is challenging as it requires tissue growth promotion and adequate mineralization. Based on our previous findings, the development of scaffolds combining bone morphogenetic protein 2 (BMP-2) and matrix metalloproteinase 10 (MMP-10) shows promise for OP management. To test our hypothesis, scaffolds containing BMP-2 + MMP-10 at variable ratios or BMP-2 + Alendronate (ALD) were prepared. Systems were characterized and tested in vitro on healthy and OP mesenchymal stem cells and in vivo bone formation was studied on healthy and OP animals. Therapeutic molecules were efficiently encapsulated into PLGA microspheres and embedded into chitosan foams. The use of PLGA (poly(lactic-co-glycolic acid)) microspheres as therapeutic molecule reservoirs allowed them to achieve an in vitro and in vivo controlled release. A beneficial effect on the alkaline phosphatase activity of non-OP cells was observed for both combinations when compared with BMP-2 alone. This effect was not detected on OP cells where all treatments promoted a similar increase in ALP activity compared with control. The in vivo results indicated a positive effect of the BMP-2 + MMP-10 combination at both of the doses tested on tissue repair for OP mice while it had the opposite effect on non-OP animals. This fact can be explained by the scaffold's slow-release rate and degradation that could be beneficial for delayed bone regeneration conditions but had the reverse effect on healthy animals. Therefore, the development of adequate scaffolds for bone regeneration requires consideration of the tissue catabolic/anabolic balance to obtain biomaterials with degradation/release behaviors suited for the existing tissue status.
Language	English
Publishing date	2021-06-29
Publishing country	Switzerland
Document type	Journal Article
ZDB-ID	2527217-2
ISSN	1999-4923
ISSN	1999-4923
DOI	10.3390/pharmaceutics13070979
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Article ; Online: Injectable Scaffold for Bone Marrow Stem Cells and Bone Morphogenetic Protein-2 to Repair Cartilage.

Vayas, Raquel / Reyes, Ricardo / Arnau, María Rosa / Évora, Carmen / Delgado, Araceli

Cartilage

2019 Volume 12, Issue 3, Page(s) 293–306

Abstract: Objective: The limits of the microfracture (MFX) treatment in terms of lesion size and long-term tissue functionality makes it necessary to investigate different alternatives to repair focal cartilage lesions. The present study aims at evaluating the ... ...

Abstract	Objective: The limits of the microfracture (MFX) treatment in terms of lesion size and long-term tissue functionality makes it necessary to investigate different alternatives to repair focal cartilage lesions. The present study aims at evaluating the efficacy of a minimally invasive approach against the conventional MFX to repair a chondral defect in rabbits. An injectable scaffold of BMP-2 pre-encapsulated in PLGA microspheres dispersed in a Pluronic F-127 solution is proposed as support of cells and controlled delivery system for the growth factor. Design: MFX was compared versus the injectable system seeded with mesenchymal stem cells (MSCs), both without BMP-2 and under controlled release of BMP-2 at 2 different doses (3 and 12 µg/scaffold). The different treatments were evaluated on a 4-mm diameter chondral defect model using 9 experimental groups of 4 rabbits (8 knees) each, throughout 24 weeks. Results: Histologically, all the treated groups, except MFX treated, responded significantly better than the control group (nontreated defect). Although no significant differences were found between the treated groups, only BMP(12), MSC-BMP(12), and MFX-BMP(3) groups showed nonsignificant differences when compared with the normal cartilage. Conclusions: The hydrogel system proposed to control the release rate of the BMP-2 was safe, easily injectable, and also provided good support for cells. Treatments with MSCs or BMP-2 repaired efficiently the chondral lesion created in rabbits, being less invasive than MFX treatment.
MeSH term(s)	Animals ; Bone Marrow Cells ; Cartilage Diseases/therapy ; Cartilage, Articular ; Hydrogels ; Mesenchymal Stem Cells ; Rabbits
Chemical Substances	Hydrogels
Language	English
Publishing date	2019-04-11
Publishing country	United States
Document type	Journal Article ; Research Support, Non-U.S. Gov't
ZDB-ID	2515870-3
ISSN	1947-6043 ; 1947-6035
ISSN (online)	1947-6043
ISSN	1947-6035
DOI	10.1177/1947603519841682
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Article ; Online: Nanoparticle-mediated selective Sfrp-1 silencing enhances bone density in osteoporotic mice.

García-García, Patricia / Reyes, Ricardo / García-Sánchez, Daniel / Pérez-Campo, Flor María / Rodríguez-Rey, José Carlos / Évora, Carmen / Díaz-Rodríguez, Patricia / Delgado, Araceli

Journal of nanobiotechnology

2022 Volume 20, Issue 1, Page(s) 462

Abstract: Osteoporosis (OP) is characterized by a loss in bone mass and mineral density. The stimulation of the canonical Wnt/β-catenin pathway has been reported to promote bone formation, this pathway is controlled by several regulators as secreted frizzled- ... ...

Abstract	Osteoporosis (OP) is characterized by a loss in bone mass and mineral density. The stimulation of the canonical Wnt/β-catenin pathway has been reported to promote bone formation, this pathway is controlled by several regulators as secreted frizzled-related protein-1 (Sfrp-1), antagonist of the pathway. Thus, Sfrp-1 silencing therapies could be suitable for enhancing bone growth. However, the systemic stimulation of Wnt/β-catenin has been correlated with side effects. This work hypothesizes the administration of lipid-polymer NPs (LPNPs) functionalized with a MSC specific aptamer (Apt) and carrying a SFRP1 silencing GapmeR, could favor bone formation in OP with minimal undesired effects. Suitable SFRP1 GapmeR-loaded Apt-LPNPs (Apt-LPNPs-SFRP1) were administered in osteoporotic mice and their biodistribution, toxicity and bone induction capacity were evaluated. The aptamer functionalization of the NPs modified their biodistribution profile showing a four-fold increase in the bone accumulation and a ten-fold decrease in the hepatic accumulation compared to naked LPNPs. Moreover, the histological evaluation revealed evident changes in bone structure observing a more compact trabecular bone and a cortical bone thickness increase in the Apt-LPNPs-SFRP1 treated mice with no toxic effects. Therefore, these LPNPs showed suitable properties and biodistribution profiles leading to an enhancement on the bone density of osteoporotic mice.
MeSH term(s)	Mice ; Animals ; beta Catenin/metabolism ; Bone Density/physiology ; Tissue Distribution ; Nanoparticles/chemistry ; Polymers/chemistry
Chemical Substances	WD repeat containing planar cell polarity effector ; beta Catenin ; APT (7320-57-2) ; Polymers
Language	English
Publishing date	2022-10-29
Publishing country	England
Document type	Journal Article
ZDB-ID	2100022-0
ISSN	1477-3155 ; 1477-3155
ISSN (online)	1477-3155
ISSN	1477-3155
DOI	10.1186/s12951-022-01674-5
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Article ; Online: Nanoclay-reinforced HA/alginate scaffolds as cell carriers and SDF-1 delivery-platforms for bone tissue engineering.

Erezuma, Itsasne / Lukin, Izeia / Pimenta-Lopes, Carolina / Ventura, Francesc / Garcia-Garcia, Patricia / Reyes, Ricardo / Arnau, Mª Rosa / Delgado, Araceli / Taebnia, Nayere / Kadumudi, Firoz Babu / Dolatshahi-Pirouz, Alireza / Orive, Gorka

International journal of pharmaceutics

2022 Volume 623, Page(s) 121895

Abstract: Bone tissue engineering has come on the scene to overcome the difficulties of the current treatment strategies. By combining biomaterials, active agents and growth factors, cells and nanomaterials, tissue engineering makes it possible to create new ... ...

Abstract	Bone tissue engineering has come on the scene to overcome the difficulties of the current treatment strategies. By combining biomaterials, active agents and growth factors, cells and nanomaterials, tissue engineering makes it possible to create new structures that enhance bone regeneration. Herein, hyaluronic acid and alginate were used to create biologically active hydrogels, and montmorillonite nanoclay was used to reinforce and stabilize them. The developed scaffolds were found to be biocompatible and osteogenic with mMSCs in vitro, especially those reinforced with the nanoclay, and allowed mineralization even in the absence of differentiation media. Moreover, an in vivo investigation was performed to establish the potential of the hydrogels to mend bone and act as cell-carriers and delivery platforms for SDF-1. Scaffolds embedded with SDF-1 exhibited the highest percentages of bone regeneration as well as of angiogenesis, which confirms the suitability of the scaffolds for bone. Although there are a number of obstacles to triumph over, these bioengineered structures showed potential as future bone regeneration treatments.
MeSH term(s)	Alginates/chemistry ; Biocompatible Materials/chemistry ; Bone Regeneration ; Bone and Bones ; Cell Differentiation ; Hydrogels/chemistry ; Osteogenesis ; Tissue Engineering ; Tissue Scaffolds/chemistry
Chemical Substances	Alginates ; Biocompatible Materials ; Hydrogels
Language	English
Publishing date	2022-06-09
Publishing country	Netherlands
Document type	Journal Article
ZDB-ID	428962-6
ISSN	1873-3476 ; 0378-5173
ISSN (online)	1873-3476
ISSN	0378-5173
DOI	10.1016/j.ijpharm.2022.121895
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Article ; Online: Organotypic culture as a research and preclinical model to study uterine leiomyomas.

Salas, Ana / López, Judith / Reyes, Ricardo / Évora, Carmen / de Oca, Francisco Montes / Báez, Delia / Delgado, Araceli / Almeida, Teresa A

Scientific reports

2020 Volume 10, Issue 1, Page(s) 5212

Abstract: Organotypic cultures of tissue slices have been successfully established in lung, prostate, colon, gastric and breast cancer among other malignancies, but until now an ex vivo model based on tissue slices has not been established for uterine leiomyoma. ... ...

Abstract	Organotypic cultures of tissue slices have been successfully established in lung, prostate, colon, gastric and breast cancer among other malignancies, but until now an ex vivo model based on tissue slices has not been established for uterine leiomyoma. In the present study, we describe a method for culturing tumour slides onto an alginate scaffold. Morphological integrity of tissue slices was maintained for up to 7 days of culture, with cells expressing desmin, estrogen and progesterone receptors. Driver mutations were present in the ex vivo slices at all-time points analyzed. Cultivated tumour slices responded to ovarian hormones stimulation upregulating the expression of genes involved in leiomyoma pathogenesis. This tissue model preserves extracellular matrix, cellular diversity and genetic background simulating more in-vivo-like situations. As a novelty, this platform allows encapsulation of microspheres containing drugs that can be tested on the ex vivo tumour slices. After optimizing drug release rates, microspheres would then be directly tested in animal models through local injection.
MeSH term(s)	Alginates ; Animals ; Antineoplastic Agents/pharmacology ; DNA Mutational Analysis ; Drug Compounding ; Drug Screening Assays, Antitumor ; Estradiol/pharmacology ; Exons/genetics ; Extracellular Matrix ; Female ; Leiomyoma/drug therapy ; Leiomyoma/genetics ; Leiomyoma/metabolism ; Leiomyoma/pathology ; Mediator Complex/genetics ; Neoplasm Proteins/biosynthesis ; Neoplasm Proteins/genetics ; Neoplasms, Hormone-Dependent/drug therapy ; Neoplasms, Hormone-Dependent/genetics ; Neoplasms, Hormone-Dependent/metabolism ; Neoplasms, Hormone-Dependent/pathology ; Progesterone/pharmacology ; RNA, Messenger/biosynthesis ; RNA, Messenger/genetics ; RNA, Neoplasm/biosynthesis ; RNA, Neoplasm/genetics ; Tissue Culture Techniques ; Tissue Scaffolds ; Uterine Neoplasms/drug therapy ; Uterine Neoplasms/genetics ; Uterine Neoplasms/metabolism ; Uterine Neoplasms/pathology
Chemical Substances	Alginates ; Antineoplastic Agents ; MED12 protein, human ; Mediator Complex ; Neoplasm Proteins ; RNA, Messenger ; RNA, Neoplasm ; Progesterone (4G7DS2Q64Y) ; Estradiol (4TI98Z838E)
Language	English
Publishing date	2020-03-23
Publishing country	England
Document type	Journal Article ; Research Support, Non-U.S. Gov't
ZDB-ID	2615211-3
ISSN	2045-2322 ; 2045-2322
ISSN (online)	2045-2322
ISSN	2045-2322
DOI	10.1038/s41598-020-62158-w
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