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  1. AU="Pérez-Garrastachu, Miguel"
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  1. Artikel ; Online: Assessing Bioprinted Functionalized Grafts for Biological Tendon Augmentation In Vitro.

    Del Amo, Cristina / Perez-Garrastachu, Miguel / Jauregui, Ines / Llama-Pino, Xabier / Andia, Isabel

    International journal of molecular sciences

    2024  Band 25, Heft 9

    Abstract: Tendinopathy, characterized by inflammatory and degenerative changes, presents challenges in sports and medicine. In addressing the limitations of conservative management, this study focuses on developing tendon grafts using extrusion bioprinting with ... ...

    Abstract Tendinopathy, characterized by inflammatory and degenerative changes, presents challenges in sports and medicine. In addressing the limitations of conservative management, this study focuses on developing tendon grafts using extrusion bioprinting with platelet-rich plasma (PRP)-infused hydrogels loaded with tendon cells. The objective is to understand paracrine interactions initiated by bioprinted tendon grafts in either inflamed or non-inflamed host tissues. PRP was utilized to functionalize methacrylate gelatin (GelMA), incorporating tendon cells for graft bioprinting. Bioinformatic analyses of overexpressed proteins, predictive of functional enrichment, revealed insights into PRP graft behavior in both non-inflamed and inflamed environments. PRP grafts activated inflammatory pathways, including Interleukin 17 (IL-17), neuroinflammation, Interleukin 33 (IL-33), and chemokine signaling. Interleukin 1 beta (IL-1b) in the graft environment triggered p38 mitogen-activated protein kinase (MAPK) signaling, nuclear factor kappa light chain enhancer of activated B cells (NF-kB) canonical pathway, and Vascular Endothelial Growth Factor (VEGF) signaling. Biological enrichment attributed to PRP grafts included cell chemotaxis, collagen turnover, cell migration, and angiogenesis. Acellular PRP grafts differed from nude grafts in promoting vessel length, vessel area, and junction density. Angiogenesis in cellular grafts was enhanced with newly synthesized Interleukin 8 (IL-8) in cooperation with IL-1b. In conclusion, paracrine signaling from PRP grafts, mediated by chemokine activities, influences cell migration, inflammation, and angiogenic status in host tissues. Under inflammatory conditions, newly synthesized IL-8 regulates vascularization in collaboration with PRP.
    Mesh-Begriff(e) Tendons/metabolism ; Bioprinting/methods ; Animals ; Platelet-Rich Plasma/metabolism ; Humans ; Tissue Engineering/methods ; Hydrogels/chemistry ; Tissue Scaffolds/chemistry ; Tendinopathy/metabolism ; Tendinopathy/therapy ; Tendinopathy/pathology
    Chemische Substanzen Hydrogels
    Sprache Englisch
    Erscheinungsdatum 2024-04-26
    Erscheinungsland Switzerland
    Dokumenttyp Journal Article
    ZDB-ID 2019364-6
    ISSN 1422-0067 ; 1422-0067 ; 1661-6596
    ISSN (online) 1422-0067
    ISSN 1422-0067 ; 1661-6596
    DOI 10.3390/ijms25094752
    Datenquelle MEDical Literature Analysis and Retrieval System OnLINE

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  2. Artikel ; Online: Wnt-3a Induces Epigenetic Remodeling in Human Dental Pulp Stem Cells.

    Uribe-Etxebarria, Verónica / García-Gallastegui, Patricia / Pérez-Garrastachu, Miguel / Casado-Andrés, María / Irastorza, Igor / Unda, Fernando / Ibarretxe, Gaskon / Subirán, Nerea

    Cells

    2020  Band 9, Heft 3

    Abstract: Dental pulp stem cells (DPSCs) from adult teeth show the expression of a very complete repertoire of stem pluripotency core factors and a high plasticity for cell reprogramming. Canonical Wnt and Notch signaling pathways regulate stemness and the ... ...

    Abstract Dental pulp stem cells (DPSCs) from adult teeth show the expression of a very complete repertoire of stem pluripotency core factors and a high plasticity for cell reprogramming. Canonical Wnt and Notch signaling pathways regulate stemness and the expression of pluripotency core factors in DPSCs, and even very short-term (48 h) activations of the Wnt pathway induce a profound remodeling of DPSCs at the physiologic and metabolic levels. In this work, DPSC cultures were exposed to treatments modulating Notch and Wnt signaling, and also induced to differentiate to osteo/adipocytes. DNA methylation, histone acetylation, histone methylation, and core factor expression levels where assessed by mass spectroscopy, Western blot, and qPCR. A short-term activation of Wnt signaling by WNT-3A induced a genomic DNA demethylation, and increased histone acetylation and histone methylation in DPSCs. The efficiency of cell reprogramming methods relies on the ability to surpass the epigenetic barrier, which determines cell lineage specificity. This study brings important information about the regulation of the epigenetic barrier by Wnt signaling in DPSCs, which could contribute to the development of safer and less aggressive reprogramming methodologies with a view to cell therapy.
    Mesh-Begriff(e) Cell Differentiation/physiology ; Cells, Cultured ; DNA Methylation/physiology ; Dental Pulp/cytology ; Epigenesis, Genetic/physiology ; Humans ; Stem Cells/cytology ; Wnt Signaling Pathway/physiology
    Sprache Englisch
    Erscheinungsdatum 2020-03-07
    Erscheinungsland Switzerland
    Dokumenttyp Journal Article ; Research Support, Non-U.S. Gov't
    ZDB-ID 2661518-6
    ISSN 2073-4409 ; 2073-4409
    ISSN (online) 2073-4409
    ISSN 2073-4409
    DOI 10.3390/cells9030652
    Datenquelle MEDical Literature Analysis and Retrieval System OnLINE

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  3. Artikel: Plasma-Based Bioinks for Extrusion Bioprinting of Advanced Dressings.

    Del Amo, Cristina / Perez-Valle, Arantza / Perez-Garrastachu, Miguel / Jauregui, Ines / Andollo, Noelia / Arluzea, Jon / Guerrero, Pedro / de la Caba, Koro / Andia, Isabel

    Biomedicines

    2021  Band 9, Heft 8

    Abstract: Extrusion bioprinting based on the development of novel bioinks offers the possibility of manufacturing clinically useful tools for wound management. In this study, we show the rheological properties and printability outcomes of two advanced dressings ... ...

    Abstract Extrusion bioprinting based on the development of novel bioinks offers the possibility of manufacturing clinically useful tools for wound management. In this study, we show the rheological properties and printability outcomes of two advanced dressings based on platelet-rich plasma (PRP) and platelet-poor plasma (PPP) blended with alginate and loaded with dermal fibroblasts. Measurements taken at 1 h, 4 days, and 18 days showed that both the PRP- and PPP-based dressings retain plasma and platelet proteins, which led to the upregulation of angiogenic and immunomodulatory proteins by embedded fibroblasts (e.g., an up to 69-fold increase in vascular endothelial growth factor (VEGF), an up to 188-fold increase in monocyte chemotactic protein 1 (MCP-1), and an up to 456-fold increase in hepatocyte growth factor (HGF) 18 days after printing). Conditioned media harvested from both PRP and PPP constructs stimulated the proliferation of human umbilical vein endothelial cells (HUVECs), whereas only those from PRP dressings stimulated HUVEC migration, which correlated with the VEGF/MCP-1 and VEGF/HGF ratios. Similarly, the advanced dressings increased the level of interleukin-8 and led to a four-fold change in the level of extracellular matrix protein 1. These findings suggest that careful selection of plasma formulations to fabricate wound dressings can enable regulation of the molecular composition of the microenvironment, as well as paracrine interactions, thereby improving the clinical potential of dressings and providing the possibility to tailor each composition to specific wound types and healing stages.
    Sprache Englisch
    Erscheinungsdatum 2021-08-16
    Erscheinungsland Switzerland
    Dokumenttyp Journal Article
    ZDB-ID 2720867-9
    ISSN 2227-9059
    ISSN 2227-9059
    DOI 10.3390/biomedicines9081023
    Datenquelle MEDical Literature Analysis and Retrieval System OnLINE

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  4. Artikel ; Online: Characterisation of corneas following different time and storage methods for their use as a source of stem-like limbal epithelial cells.

    Romo-Valera, Cristina / Pérez-Garrastachu, Miguel / Hernáez-Moya, Raquel / Rodriguez-Astigarraga, Maddalen / Romano-Ruiz, Paula / Etxebarria, Jaime / Arluzea, Jon / Andollo, Noelia

    Experimental eye research

    2021  Band 211, Seite(n) 108720

    Abstract: The transplantation of expansions of limbal epithelial stem cells (LESC) remains one of the most efficient therapies for the treatment of limbal stem cell deficiency (LSCD) to date. However, the available donor corneas are scarce, and the corneas ... ...

    Abstract The transplantation of expansions of limbal epithelial stem cells (LESC) remains one of the most efficient therapies for the treatment of limbal stem cell deficiency (LSCD) to date. However, the available donor corneas are scarce, and the corneas conserved for long time, under hypothermic conditions (after 7 days) or in culture (more than 28 days), are usually discarded due to poor viability of the endothelial cells. To establish an objective criterion for the utilisation or discarding of corneas as a source of LESC, we characterized, by immunohistochemistry analysis, donor corneas conserved in different conditions and for different periods of time. We also studied the potency of LESCs isolated from these corneas and maintained in culture up to 3 cell passages. We hoped that the study of markers of LESCs present in both the corneoscleral histological sections and the cell cultures would show the adequacy of the methods used for cell isolation and how fit the LESC enrichment of the obtained cell populations to be expanded was. Thus, the expressions of markers of the cells residing in the human limbal and corneal epithelium (cytokeratin CK15 and CK12, vimentin, Collagen VII, p63α, ABCG2, Ki67, Integrin β4, ZO1, and melan A) were analysed in sections of corneoscleral tissues conserved in hypothermic conditions for 2-9 days with post-mortem time (pmt) < 8 h or for 1 day with pmt > 16 h, and in sclerocorneal rims maintained in an organ culture medium for 29 days. Cell populations isolated from donor corneoscleral tissues were also assessed based on these markers to verify the adequacy of isolation methods and the potential of expanding LESCs from these tissues. Positivity for several putative stem cell markers such as CK15 and p63α was detected in all corneoscleral tissues, although a decrease was recorded in the ones conserved for longer times. The barrier function and the ability to adhere to the extracellular matrix were maintained in all the analysed tissues. In limbal epithelial cell cultures, a simultaneous decrease in the melan A melanocyte marker and the putative stem cell markers was detected, suggesting a close relationship between the melanocytes and the limbal stem cells of the niche. Holoclones stained with putative stem cell markers were obtained from long-term, hypothermic, stored sclerocorneal rims. The results showed that the remaining sclerocorneal rims after corneal transplantation, which were conserved under hypothermic conditions for up to 7 days and would have been discarded at a first glance, still maintained their potential as a source of LESC cultures.
    Mesh-Begriff(e) Adult ; Aged ; Aged, 80 and over ; Biomarkers/metabolism ; Cell Separation ; Cells, Cultured ; Collagen/metabolism ; Cornea/cytology ; Cornea/metabolism ; Epithelium, Corneal/cytology ; Epithelium, Corneal/metabolism ; Humans ; Keratins/metabolism ; Limbus Corneae/cytology ; Limbus Corneae/metabolism ; Middle Aged ; Organ Culture Techniques/methods ; Stem Cells/cytology ; Stem Cells/metabolism ; Time Factors ; Tissue Donors ; Tissue Preservation/methods ; Vimentin/metabolism
    Chemische Substanzen Biomarkers ; Vimentin ; Keratins (68238-35-7) ; Collagen (9007-34-5)
    Sprache Englisch
    Erscheinungsdatum 2021-08-11
    Erscheinungsland England
    Dokumenttyp Journal Article ; Research Support, Non-U.S. Gov't
    ZDB-ID 80122-7
    ISSN 1096-0007 ; 0014-4835
    ISSN (online) 1096-0007
    ISSN 0014-4835
    DOI 10.1016/j.exer.2021.108720
    Datenquelle MEDical Literature Analysis and Retrieval System OnLINE

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  5. Artikel: Nucleoporins redistribute inside the nucleus after cell cycle arrest induced by histone deacetylases inhibition

    Pérez-Garrastachu, Miguel / Arluzea, Jon / Andrade, Ricardo / Díez-Torre, Alejandro / Urtizberea, Marta / Silió, Margarita / Aréchaga, Juan

    Nucleus. 2017 Sept. 3, v. 8, no. 5

    2017  

    Abstract: Nucleoporins are the main components of the nuclear-pore complex (NPC) and were initially considered as mere structural elements embedded in the nuclear envelope, being responsible for nucleocytoplasmic transport. Nevertheless, several recent scientific ... ...

    Abstract Nucleoporins are the main components of the nuclear-pore complex (NPC) and were initially considered as mere structural elements embedded in the nuclear envelope, being responsible for nucleocytoplasmic transport. Nevertheless, several recent scientific reports have revealed that some nucleoporins participate in nuclear processes such as transcription, replication, DNA repair and chromosome segregation. Thus, the interaction of NPCs with chromatin could modulate the distribution of chromosome territories relying on the epigenetic state of DNA. In particular, the nuclear basket proteins Tpr and Nup153, and the FG-nucleoporin Nup98 seem to play key roles in all these novel functions. In this work, histone deacetylase inhibitors (HDACi) were used to induce a hyperacetylated state of chromatin and the behavior of the mentioned nucleoporins was studied. Our results show that, after HDACi treatment, Tpr, Nup153 and Nup98 are translocated from the nuclear pore toward the interior of the cell nucleus, accumulating as intranuclear nucleoporin clusters. These transitory structures are highly dynamic, and are mainly present in the population of cells arrested at the G0/G1 phase of the cell cycle. Our results indicate that the redistribution of these nucleoporins from the nuclear envelope to the nuclear interior may be implicated in the early events of cell cycle initialization, particularly during the G1 phase transition.
    Schlagwörter DNA ; DNA repair ; cell cycle checkpoints ; chromatin ; chromosome segregation ; epigenetics ; histone deacetylase ; interphase ; nuclear pore ; nucleocytoplasmic transport ; nucleoporins ; phase transition
    Sprache Englisch
    Erscheinungsverlauf 2017-0903
    Umfang p. 515-533.
    Erscheinungsort Taylor & Francis
    Dokumenttyp Artikel
    Anmerkung NAL-AP-2-clean
    ZDB-ID 2619626-8
    ISSN 1949-1042 ; 1949-1034
    ISSN (online) 1949-1042
    ISSN 1949-1034
    DOI 10.1080/19491034.2017.1320001
    Datenquelle NAL Katalog (AGRICOLA)

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  6. Artikel ; Online: Nucleoporins redistribute inside the nucleus after cell cycle arrest induced by histone deacetylases inhibition.

    Pérez-Garrastachu, Miguel / Arluzea, Jon / Andrade, Ricardo / Díez-Torre, Alejandro / Urtizberea, Marta / Silió, Margarita / Aréchaga, Juan

    Nucleus (Austin, Tex.)

    2017  Band 8, Heft 5, Seite(n) 515–533

    Abstract: Nucleoporins are the main components of the nuclear-pore complex (NPC) and were initially considered as mere structural elements embedded in the nuclear envelope, being responsible for nucleocytoplasmic transport. Nevertheless, several recent scientific ... ...

    Abstract Nucleoporins are the main components of the nuclear-pore complex (NPC) and were initially considered as mere structural elements embedded in the nuclear envelope, being responsible for nucleocytoplasmic transport. Nevertheless, several recent scientific reports have revealed that some nucleoporins participate in nuclear processes such as transcription, replication, DNA repair and chromosome segregation. Thus, the interaction of NPCs with chromatin could modulate the distribution of chromosome territories relying on the epigenetic state of DNA. In particular, the nuclear basket proteins Tpr and Nup153, and the FG-nucleoporin Nup98 seem to play key roles in all these novel functions. In this work, histone deacetylase inhibitors (HDACi) were used to induce a hyperacetylated state of chromatin and the behavior of the mentioned nucleoporins was studied. Our results show that, after HDACi treatment, Tpr, Nup153 and Nup98 are translocated from the nuclear pore toward the interior of the cell nucleus, accumulating as intranuclear nucleoporin clusters. These transitory structures are highly dynamic, and are mainly present in the population of cells arrested at the G0/G1 phase of the cell cycle. Our results indicate that the redistribution of these nucleoporins from the nuclear envelope to the nuclear interior may be implicated in the early events of cell cycle initialization, particularly during the G1 phase transition.
    Mesh-Begriff(e) Acetylation/drug effects ; Active Transport, Cell Nucleus/drug effects ; Cell Cycle Checkpoints/drug effects ; Cell Line, Tumor ; Cell Nucleus/drug effects ; Cell Nucleus/metabolism ; Chromatin/metabolism ; Histone Deacetylase Inhibitors/pharmacology ; Histone Deacetylases/metabolism ; Humans ; Hydroxamic Acids/pharmacology ; Nuclear Pore Complex Proteins/metabolism
    Chemische Substanzen Chromatin ; Histone Deacetylase Inhibitors ; Hydroxamic Acids ; Nuclear Pore Complex Proteins ; trichostatin A (3X2S926L3Z) ; Histone Deacetylases (EC 3.5.1.98)
    Sprache Englisch
    Erscheinungsdatum 2017-07-11
    Erscheinungsland United States
    Dokumenttyp Journal Article ; Research Support, Non-U.S. Gov't
    ZDB-ID 2619626-8
    ISSN 1949-1042 ; 1949-1034
    ISSN (online) 1949-1042
    ISSN 1949-1034
    DOI 10.1080/19491034.2017.1320001
    Datenquelle MEDical Literature Analysis and Retrieval System OnLINE

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