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  1. Article ; Online: The nuclei of human adult stem cells can move within the cell and generate cellular protrusions to contact other cells.

    Bueno, Carlos / García-Bernal, David / Martínez, Salvador / Blanquer, Miguel / Moraleda, José M

    Stem cell research & therapy

    2024  Volume 15, Issue 1, Page(s) 32

    Abstract: Background: The neuronal transdifferentiation of adult bone marrow cells (BMCs) is still considered an artifact based on an alternative explanation of experimental results supporting this phenomenon obtained over decades. However, recent studies have ... ...

    Abstract Background: The neuronal transdifferentiation of adult bone marrow cells (BMCs) is still considered an artifact based on an alternative explanation of experimental results supporting this phenomenon obtained over decades. However, recent studies have shown that following neural induction, BMCs enter an intermediate cellular state before adopting neural-like morphologies by active neurite extension and that binucleated BMCs can be formed independent of any cell fusion events. These findings provide evidence to reject the idea that BMC neural transdifferentiation is merely an experimental artifact. Therefore, understanding the intermediate states that cells pass through during transdifferentiation is crucial given their potential application in regenerative medicine and disease modelling.
    Methods: In this study, we examined the functional significance of the variety of morphologies and positioning that cell nuclei of human bone marrow-derived mesenchymal stem cells (hBM-MSCs) can adopt during neural-like differentiation using live-cell nuclear fluorescence labelling, time-lapse microscopy, and confocal microscopy analysis.
    Results: Here, we showed that after neural induction, hBM-MSCs enter an intermediate cellular state in which the nuclei are able to move within the cells, switching shapes and positioning and even generating cellular protrusions as they attempt to contact the cells around them. These findings suggest that changes in nuclear positioning occur because human cell nuclei somehow sense their environment. In addition, we showed the process of direct interactions between cell nuclei, which opens the possibility of a new level of intercellular interaction.
    Conclusions: The present study advances the understanding of the intermediate stage through which hBM-MSCs pass during neural transdifferentiation, which may be crucial to understanding the mechanisms of these cell conversion processes and eventually harness them for use in regenerative medicine. Importantly, our study provides for the first time evidence that the nuclei of hBM-MSC-derived intermediate cells somehow sense their environment, generating cellular protrusions to contact other cells. In summary, human mesenchymal stromal cells could not only help to increase our understanding of the mechanisms underlying cellular plasticity but also facilitate the exact significance of nuclear positioning in cellular function and in tissue physiology.
    MeSH term(s) Adult ; Humans ; Cell Differentiation/physiology ; Neurons ; Adult Stem Cells ; Cell Surface Extensions ; Cell Nucleus ; Bone Marrow Cells ; Cells, Cultured
    Language English
    Publishing date 2024-02-07
    Publishing country England
    Document type Journal Article
    ZDB-ID 2548671-8
    ISSN 1757-6512 ; 1757-6512
    ISSN (online) 1757-6512
    ISSN 1757-6512
    DOI 10.1186/s13287-024-03638-y
    Database MEDical Literature Analysis and Retrieval System OnLINE

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  2. Article ; Online: Comparative bioactivity and immunomodulatory potential of the new Bioroot Flow and AH Plus Bioceramic sealer: An in vitro study on hPDLSCs.

    Sanz, José Luis / López-García, Sergio / García-Bernal, David / Rodríguez-Lozano, Francisco Javier / Forner, Leopoldo / Lozano, Adrián / Murcia, Laura

    Clinical oral investigations

    2024  Volume 28, Issue 3, Page(s) 195

    Abstract: Objectives: To evaluate the cytocompatibility, bioactivity, and anti-inflammatory potential of the new pre-mixed calcium silicate cement-based sealers Bioroot Flow (BrF) and AH Plus Bioceramic Sealer (AHPbcs) on human periodontal ligament stem cells ( ... ...

    Abstract Objectives: To evaluate the cytocompatibility, bioactivity, and anti-inflammatory potential of the new pre-mixed calcium silicate cement-based sealers Bioroot Flow (BrF) and AH Plus Bioceramic Sealer (AHPbcs) on human periodontal ligament stem cells (hPDLSCs) compared to the epoxy resin-based sealer AH Plus (AHP).
    Materials and methods: Standardized discs and 1:1, 1:2, and 1:4 eluates of BrF, AHPbcs and AHP after setting were prepared. The following assays were performed: cell attachment and morphology via SEM, cell viability via a MTT assay, cell migration/proliferation via a wound-healing assay, cytoskeleton organization via immunofluorescence staining; cytokine release via ELISA; osteo/cemento/odontogenic marker expression via RT-qPCR, and cell mineralized nodule formation via Alizarin Red S staining. HPDLSCs were isolated from extracted third molars from healthy patients. Comparisons were made with hPDLSCs cultured in unconditioned (negative control) or osteogenic (positive control) culture media. Statistical significance was established at p < 0.05.
    Results: Both BrF and AHPbcs showed significantly positive results in the cytocompatibility assays (cell metabolic activity, migration, attachment, morphology, and cytoskeleton organization) compared with a negative control group, while AHP showed significant negative results. BrF exhibited an upregulation of at least one osteo/cementogenic marker compared to the negative and positive control groups. BrF showed a significantly higher calcified nodule formation than AHPbcs, the negative and positive control groups, while AHPbcs was higher than the negative control group. Both were also significantly higher than AHP group.
    Conclusion: BrF and AHPbcs exhibit adequate and comparable cytocompatibility on hPDLSCs. BrF also promoted the osteo/cementogenic differentiation of hPDLSCs. Both calcium silicate-based sealers favored the downregulation of the inflammatory cytokine IL-6 and the calcified nodule formation from hPDLSCs. BrF exerted a significantly higher influence on cell mineralization than AHPbcs.
    Clinical relevance: This is the first study to elucidate the biological properties and immunomodulatory potential of Bioroot Flow and AH Plus Bioceramic Sealer. The results act as supporting evidence for their use in root canal treatment.
    MeSH term(s) Humans ; Periodontal Ligament ; Epoxy Resins ; Cytokines ; Alopecia/congenital ; Root Canal Filling Materials ; Calcium Compounds ; Silicates
    Chemical Substances calcium silicate (S4255P4G5M) ; epoxy resin-based root canal sealer ; Epoxy Resins ; Cytokines ; Root Canal Filling Materials ; Calcium Compounds ; Silicates
    Language English
    Publishing date 2024-03-05
    Publishing country Germany
    Document type Journal Article
    ZDB-ID 1364490-7
    ISSN 1436-3771 ; 1432-6981
    ISSN (online) 1436-3771
    ISSN 1432-6981
    DOI 10.1007/s00784-024-05593-7
    Database MEDical Literature Analysis and Retrieval System OnLINE

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  3. Article: The Cytocompatibility of Silver Diamine Fluoride on Mesenchymal Stromal Cells from Human Exfoliated Deciduous Teeth: An In Vitro Study.

    García-Bernal, David / Pecci-Lloret, Maria Pilar / López-García, Sergio

    Materials (Basel, Switzerland)

    2022  Volume 15, Issue 6

    Abstract: Silver diamine fluoride (SDF) has been used for many years for the treatment of caries, and minimally invasive dentistry concepts have made it popular again. The fact that its application does not require the administration of anesthesia makes its use in ...

    Abstract Silver diamine fluoride (SDF) has been used for many years for the treatment of caries, and minimally invasive dentistry concepts have made it popular again. The fact that its application does not require the administration of anesthesia makes its use in children more desirable. The aim of this study was to determine the cytotoxicity of two new commercial SDF products: Riva Star (SDI Dental Limited) and e-SDF (Kids-e-Dental) on mesenchymal stromal cells from human exfoliated deciduous teeth (SHEDs). SHEDs were exposed to SDF products at different concentrations (0.1%, 0.01% and 0.005%). Then different assays were performed to evaluate their cytocompatibility on SHEDs: IC50, MTT, cell migration (wound healing), cell cytoskeleton staining, cell apoptosis, generation of intracellular reactive oxygen species (ROS), and ion chromatography. Statistical analyses were performed using one-way ANOVA and Tukey’s post hoc test (p < 0.05). Riva Star Step 2 showed the same cell metabolic activity when compared to the control condition at any time and concentration. Meanwhile, e-SDF displayed high cytotoxicity at any time and any concentration (*** p < 0.001), whereas Riva Star Step 1 displayed high cytotoxicity at any time at 0.1% and 0.01% (*** p < 0.001). Only e-SDF showed a statistically significant decreased cell migration rate (*** p < 0.001) at all times and in all concentrations. At 0.1%, e-SDF and Riva Star Step 1 only showed 4.37% and 4.47% of viable cells, respectively. These results suggest that Riva Star has better in vitro cytocompatibility on SHEDs than does e-SDF. Riva Star Step 1 was found to be as cytotoxic as e-SDF, but it had better biological properties when mixed with Riva Star Step 2. Our findings suggest that Riva Star is more suitable when used in deciduous teeth due to its lower cytotoxicity compared to e-SDF.
    Language English
    Publishing date 2022-03-12
    Publishing country Switzerland
    Document type Journal Article
    ZDB-ID 2487261-1
    ISSN 1996-1944
    ISSN 1996-1944
    DOI 10.3390/ma15062104
    Database MEDical Literature Analysis and Retrieval System OnLINE

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  4. Article: Interplay between mesenchymal stromal cells and the immune system after transplantation: implications for advanced cell therapy in the retina.

    Norte-Muñoz, María / García-Bernal, David / García-Ayuso, Diego / Vidal-Sanz, Manuel / Agudo-Barriuso, Marta

    Neural regeneration research

    2023  Volume 19, Issue 3, Page(s) 542–547

    Abstract: Advanced mesenchymal stromal cell-based therapies for neurodegenerative diseases are widely investigated in preclinical models. Mesenchymal stromal cells are well positioned as therapeutics because they address the underlying mechanisms of ... ...

    Abstract Advanced mesenchymal stromal cell-based therapies for neurodegenerative diseases are widely investigated in preclinical models. Mesenchymal stromal cells are well positioned as therapeutics because they address the underlying mechanisms of neurodegeneration, namely trophic factor deprivation and neuroinflammation. Most studies have focused on the beneficial effects of mesenchymal stromal cell transplantation on neuronal survival or functional improvement. However, little attention has been paid to the interaction between mesenchymal stromal cells and the host immune system due to the immunomodulatory properties of mesenchymal stromal cells and the long-held belief of the immunoprivileged status of the central nervous system. Here, we review the crosstalk between mesenchymal stromal cells and the immune system in general and in the context of the central nervous system, focusing on recent work in the retina and the importance of the type of transplantation.
    Language English
    Publishing date 2023-09-27
    Publishing country India
    Document type Journal Article ; Review
    ZDB-ID 2388460-5
    ISSN 1876-7958 ; 1673-5374
    ISSN (online) 1876-7958
    ISSN 1673-5374
    DOI 10.4103/1673-5374.380876
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  5. Article: Comparative Cytotoxicity of Menthol and Eucalyptol: An In Vitro Study on Human Gingival Fibroblasts.

    Puig-Herreros, Clara / Sanz, José Luis / García-Bernal, David / Rodríguez-Lozano, Francisco Javier / Murcia, Laura / Forner, Leopoldo / Ghilotti, James / Oñate-Sánchez, Ricardo E / López-García, Sergio

    Pharmaceutics

    2024  Volume 16, Issue 4

    Abstract: The aim of this study was to assess the influence of eucalyptol and menthol on the cell viability, migration, and reactive oxygen species production of human gingival fibroblasts (GFs) in vitro. Three different concentrations of eucalyptol and menthol ... ...

    Abstract The aim of this study was to assess the influence of eucalyptol and menthol on the cell viability, migration, and reactive oxygen species production of human gingival fibroblasts (GFs) in vitro. Three different concentrations of eucalyptol and menthol were prepared following ISO 10993-5 guidelines (1, 5, and 10 mM). GFs were isolated from extracted teeth from healthy donors. The following parameters were assessed: cell viability via MTT, Annexin-V-FITC and 7-AAD staining, and IC
    Language English
    Publishing date 2024-04-09
    Publishing country Switzerland
    Document type Journal Article
    ZDB-ID 2527217-2
    ISSN 1999-4923
    ISSN 1999-4923
    DOI 10.3390/pharmaceutics16040521
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  6. Article ; Online: Binucleated human bone marrow-derived mesenchymal cells can be formed during neural-like differentiation with independence of any cell fusion events.

    Bueno, Carlos / Blanquer, Miguel / García-Bernal, David / Martínez, Salvador / Moraleda, José M

    Scientific reports

    2022  Volume 12, Issue 1, Page(s) 20615

    Abstract: Although it has been reported that bone marrow-derived cells (BMDCs) can transdifferentiate into neural cells, the findings are considered unlikely. It has been argued that the rapid neural transdifferentiation of BMDCs reported in culture studies is ... ...

    Abstract Although it has been reported that bone marrow-derived cells (BMDCs) can transdifferentiate into neural cells, the findings are considered unlikely. It has been argued that the rapid neural transdifferentiation of BMDCs reported in culture studies is actually due to cytotoxic changes induced by the media. While transplantation studies indicated that BMDCs can form new neurons, it remains unclear whether the underlying mechanism is transdifferentiation or BMDCs-derived cell fusion with the existing neuronal cells. Cell fusion has been put forward to explain the presence of gene-marked binucleated neurons after gene-marked BMDCs transplantation. In the present study, we demostrated that human BMDCs can rapidly adopt a neural-like morphology through active neurite extension and binucleated human BMDCs can form with independence of any cell fusion events. We also showed that BMDCs neural-like differentiation involves the formation of intermediate cells which can then redifferentiate into neural-like cells, redifferentiate back to the mesenchymal fate or even repeatedly switch lineages without cell division. Furthermore, we have discovered that nuclei from intermediate cells rapidly move within the cell, adopting different morphologies and even forming binucleated cells. Therefore, our results provide a stronger basis for rejecting the idea that BMDCs neural transdifferentiation is merely an artefact.
    MeSH term(s) Humans ; Cell Fusion ; Bone Marrow ; Cell Differentiation ; Cell Communication ; Neurons
    Language English
    Publishing date 2022-11-30
    Publishing country England
    Document type Journal Article
    ZDB-ID 2615211-3
    ISSN 2045-2322 ; 2045-2322
    ISSN (online) 2045-2322
    ISSN 2045-2322
    DOI 10.1038/s41598-022-24996-8
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  7. Article ; Online: Premixed calcium silicate-based ceramic sealers promote osteogenic/cementogenic differentiation of human periodontal ligament stem cells: A microscopy study.

    López-García, Sergio / Sánchez-Bautista, Sonia / García-Bernal, David / Lozano, Adrián / Forner, Leopoldo / Sanz, José L / Murcia, Laura / Rodríguez-Lozano, Francisco J / Oñate-Sánchez, Ricardo E

    Microscopy research and technique

    2024  

    Abstract: To evaluate the effects of premixed calcium silicate based ceramic sealers on the viability and osteogenic/cementogenic differentiation of human periodontal ligament stem cells (hPDLSCs). The materials evaluated were TotalFill BC Sealer (TFbc), AH Plus ... ...

    Abstract To evaluate the effects of premixed calcium silicate based ceramic sealers on the viability and osteogenic/cementogenic differentiation of human periodontal ligament stem cells (hPDLSCs). The materials evaluated were TotalFill BC Sealer (TFbc), AH Plus Bioceramic Sealer (AHPbc), and Neosealer Flo (Neo). Standardized discs and 1:1, 1:2, and 1:4 eluates of the tested materials were prepared. The following in vitro experiments were carried out: ion release, cell metabolic activity 3-(4,5-dimethylthiazol-2-yl)-2,5-diphenyltetrazolium bromide (MTT) assay, cell migration, immunofluorescence experiment, cell attachment, gene expression, and mineralization assay. Statistical analyses were performed using one-way ANOVA followed by Tukey's post hoc test (p < .05). Increased Ca
    Language English
    Publishing date 2024-03-03
    Publishing country United States
    Document type Journal Article
    ZDB-ID 1099714-3
    ISSN 1097-0029 ; 1059-910X
    ISSN (online) 1097-0029
    ISSN 1059-910X
    DOI 10.1002/jemt.24545
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  8. Article ; Online: Assessment of the anti-inflammatory and biological properties of Bioroot Flow: A novel bioceramic sealer.

    López-García, Sergio / Sanz, José L / Murcia, Laura / García-Bernal, David / Lozano, Adrián / Forner, Leopoldo / Rodríguez-Lozano, Francisco J / Oñate-Sánchez, Ricardo E

    Tissue & cell

    2024  Volume 88, Page(s) 102391

    Abstract: Introduction: BioRoot Flow (BRF) is a novel premixed bioceramic sealer indicated for endodontic treatments, but the biological and immunomodulatory effects of this endodontic sealer on human periodontal ligament stem cells (hPDLSCs) have not been ... ...

    Abstract Introduction: BioRoot Flow (BRF) is a novel premixed bioceramic sealer indicated for endodontic treatments, but the biological and immunomodulatory effects of this endodontic sealer on human periodontal ligament stem cells (hPDLSCs) have not been elucidated.
    Methods: To ascertain the biological impact of BRF, TotalFill BC Sealer (TFbc), and AH Plus (AHP) on human Periodontal Ligament Stem Cells (hPDLSCs), assessments were conducted to evaluate the cytocompatibility, cellular proliferation, migratory capacity, osteo/cementogenic differentiation potential, the ability to form mineralized nodules, and the immunomodulatory characteristics of hPDLSCs following treatment with these endodontic sealers.
    Results: Biological assays showed adequate cell metabolic activity and cell migration in BRF, while SEM assay evidenced that TFbc and BRF groups demonstrated a superior cell adhesion process, including substrate adhesion, cytoskeleton development, and spreading on the niche-like structures of the cement as compared to the AHP group. TFbc and BRF-treated groups exhibited a significantly lower IL6 and IL8 production than AHP (* p <.05). The bioceramic sealers stimulated heightened expression of BSP, CEMP-1, and CAP genes within a 7-14 day period. Notably, BRF and TFbc demonstrated a significant enhancement in the mineralization of hPDLSCs when compared to the negative control. Among these, cells treated with BRF showed a more substantial accumulation of calcium (*** p < .001).
    Conclusions: Taken together, these findings indicate that BRF can potentially enhance cell differentiation by promoting the expression of essential genes related to bone and cement formation. In addition, BRF and TFbc displayed anti-inflammatory effects.
    Language English
    Publishing date 2024-04-17
    Publishing country Scotland
    Document type Journal Article
    ZDB-ID 204424-9
    ISSN 1532-3072 ; 0040-8166
    ISSN (online) 1532-3072
    ISSN 0040-8166
    DOI 10.1016/j.tice.2024.102391
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  9. Article ; Online: Optimizing cryopreservation conditions for use of fucosylated human mesenchymal stromal cells in anti-inflammatory/immunomodulatory therapeutics.

    Gil-Chinchilla, Jesús I / Bueno, Carlos / Martínez, Carlos M / Ferrández-Múrtula, Ana / García-Hernández, Ana M / Blanquer, Miguel / Molina-Molina, Mar / Zapata, Agustín G / Sackstein, Robert / Moraleda, Jose M / García-Bernal, David

    Frontiers in immunology

    2024  Volume 15, Page(s) 1385691

    Abstract: Mesenchymal stem/stromal cells (MSCs) are being increasingly used in cell-based therapies due to their broad anti-inflammatory and immunomodulatory properties. Intravascularly-administered MSCs do not efficiently migrate to sites of inflammation/ ... ...

    Abstract Mesenchymal stem/stromal cells (MSCs) are being increasingly used in cell-based therapies due to their broad anti-inflammatory and immunomodulatory properties. Intravascularly-administered MSCs do not efficiently migrate to sites of inflammation/immunopathology, but this shortfall has been overcome by cell surface enzymatic fucosylation to engender expression of the potent E-selectin ligand HCELL. In applications of cell-based therapies, cryopreservation enables stability in both storage and transport of the produced cells from the manufacturing facility to the point of care. However, it has been reported that cryopreservation and thawing dampens their immunomodulatory/anti-inflammatory activity even after a reactivation/reconditioning step. To address this issue, we employed a variety of methods to cryopreserve and thaw fucosylated human MSCs derived from either bone marrow or adipose tissue sources. We then evaluated their immunosuppressive properties, cell viability, morphology, proliferation kinetics, immunophenotype, senescence, and osteogenic and adipogenic differentiation. Our studies provide new insights into the immunobiology of cryopreserved and thawed MSCs and offer a readily applicable approach to optimize the use of fucosylated human allogeneic MSCs as immunomodulatory/anti-inflammatory therapeutics.
    MeSH term(s) Humans ; Glycosylation ; Immunomodulation ; Mesenchymal Stem Cells/metabolism ; Cryopreservation/methods ; Anti-Inflammatory Agents/metabolism
    Chemical Substances Anti-Inflammatory Agents
    Language English
    Publishing date 2024-03-28
    Publishing country Switzerland
    Document type Journal Article ; Research Support, Non-U.S. Gov't
    ZDB-ID 2606827-8
    ISSN 1664-3224 ; 1664-3224
    ISSN (online) 1664-3224
    ISSN 1664-3224
    DOI 10.3389/fimmu.2024.1385691
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  10. Article ; Online: In Vitro Biocompatibility of Several Children's Toothpastes on Human Gingival Fibroblasts.

    Pecci-Lloret, María Pilar / López-García, Sergio / Rodríguez-Lozano, Francisco Javier / Álvarez-Novoa, Pablo / García-Bernal, David

    International journal of environmental research and public health

    2022  Volume 19, Issue 5

    Abstract: The European Academy of Paediatric Dentistry has recommended fluoride toothpastes from the eruption of the first teeth in children. Toothpastes stay in the mouth in contact with human gingival fibroblasts (hGFs) for a long time. Thus, the objective of ... ...

    Abstract The European Academy of Paediatric Dentistry has recommended fluoride toothpastes from the eruption of the first teeth in children. Toothpastes stay in the mouth in contact with human gingival fibroblasts (hGFs) for a long time. Thus, the objective of this study was to compare the cytotoxicity of five different commonly used children’s toothpastes: Oral B Kids +3 (Procter & Gamble, Alicante, Spain), Fluor Kin Calcium (Kin, Madrid, Spain), PHB Junior (PHB, Barcelona, Spain), Colgate +3 (Colgate Palmolive, Madrid, Spain) and Vitis Kids (Dentaid, Valencia, Spain) on hGFs. The children’s toothpastes were exposed to hGFs at different concentrations (1:1, 1:2, 1:4). Afterwards, several tests were performed: MTT assays, cell cycle analyses, cell cytoskeleton staining assays, apoptosis/necrosis assays, and ICP-MS and ion chromatography. Oral B displayed the lowest cytotoxicity and was the toothpaste with the highest fluoride ion release; meanwhile, the other toothpastes were cytotoxic (*** p < 0.0001); Fluor Kin being the one with the lowest fluoride ion release. Among all the toothpastes analyzed, Oral B exhibited the best results in vitro in terms of biocompatibility. Future evaluations, both in vitro and in vivo, are required to confirm the biocompatibility of sodium lauryl sarcosinate and sodium lauryl sulfate containing toothpastes.
    MeSH term(s) Child ; Fibroblasts ; Fluorides ; Gingiva ; Humans ; Sodium Fluoride ; Toothpastes/toxicity
    Chemical Substances Toothpastes ; Sodium Fluoride (8ZYQ1474W7) ; Fluorides (Q80VPU408O)
    Language English
    Publishing date 2022-03-03
    Publishing country Switzerland
    Document type Journal Article ; Research Support, Non-U.S. Gov't
    ZDB-ID 2175195-X
    ISSN 1660-4601 ; 1661-7827
    ISSN (online) 1660-4601
    ISSN 1661-7827
    DOI 10.3390/ijerph19052954
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