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  1. Article ; Online: Matrix metalloproteinase-3 joins a growing list of proteases that regulate vascular calcification.

    Giachelli, Cecilia Maria / Donato, Maristella / Scatena, Marta

    Cardiovascular research

    2024  

    Language English
    Publishing date 2024-04-17
    Publishing country England
    Document type Journal Article
    ZDB-ID 80340-6
    ISSN 1755-3245 ; 0008-6363
    ISSN (online) 1755-3245
    ISSN 0008-6363
    DOI 10.1093/cvr/cvae064
    Database MEDical Literature Analysis and Retrieval System OnLINE

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  2. Article ; Online: Differentiation and Characterization of Osteoclasts from Human Induced Pluripotent Stem Cells.

    Blümke, Alexander / Simon, Jessica / Leber, Elizabeth / Scatena, Marta / Giachelli, Cecilia M

    Journal of visualized experiments : JoVE

    2024  , Issue 205

    Abstract: This protocol details the propagation and passaging of human iPSCs and their differentiation into osteoclasts. First, iPSCs are dissociated into a single-cell suspension for further use in embryoid body induction. Following mesodermal induction, embryoid ...

    Abstract This protocol details the propagation and passaging of human iPSCs and their differentiation into osteoclasts. First, iPSCs are dissociated into a single-cell suspension for further use in embryoid body induction. Following mesodermal induction, embryoid bodies undergo hematopoietic differentiation, producing a floating hematopoietic cell population. Subsequently, the harvested hematopoietic cells undergo a macrophage colony-stimulating factor maturation step and, finally, osteoclast differentiation. After osteoclast differentiation, osteoclasts are characterized by staining for TRAP in conjunction with a methyl green nuclear stain. Osteoclasts are observed as multinucleated, TRAP+ polykaryons. Their identification can be further supported by Cathepsin K staining. Bone and mineral resorption assays allow for functional characterization, confirming the identity of bona fide osteoclasts. This protocol demonstrates a robust and versatile method to differentiate human osteoclasts from iPSCs and allows for easy adoption in applications requiring large quantities of functional human osteoclasts. Applications in the areas of bone research, cancer research, tissue engineering, and endoprosthesis research could be envisioned.
    MeSH term(s) Humans ; Osteoclasts ; Induced Pluripotent Stem Cells ; Cell Differentiation ; Macrophage Colony-Stimulating Factor/pharmacology ; Bone and Bones ; Membrane Glycoproteins ; Bone Resorption ; RANK Ligand
    Chemical Substances Macrophage Colony-Stimulating Factor (81627-83-0) ; Membrane Glycoproteins ; RANK Ligand
    Language English
    Publishing date 2024-03-22
    Publishing country United States
    Document type Journal Article ; Video-Audio Media
    ZDB-ID 2259946-0
    ISSN 1940-087X ; 1940-087X
    ISSN (online) 1940-087X
    ISSN 1940-087X
    DOI 10.3791/66527
    Database MEDical Literature Analysis and Retrieval System OnLINE

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  3. Article ; Online: Dissecting specific Wnt components governing osteogenic differentiation potential by human periodontal ligament stem cells through interleukin-6.

    Purwaningrum, Medania / Giachelli, Cecilia M / Osathanon, Thanaphum / Rattanapuchpong, Sirirat / Sawangmake, Chenphop

    Scientific reports

    2023  Volume 13, Issue 1, Page(s) 9055

    Abstract: Periodontal ligament stem cells (PDLSCs) play a significant role on periodontal tissue and alveolar bone homeostasis. During inflammation, interleukin (IL)-6 serves as one of key cytokine players controlling tissue reaction as well as alveolar bone ... ...

    Abstract Periodontal ligament stem cells (PDLSCs) play a significant role on periodontal tissue and alveolar bone homeostasis. During inflammation, interleukin (IL)-6 serves as one of key cytokine players controlling tissue reaction as well as alveolar bone tissue remodeling. It is believed that periodontal tissue inflammation causes periodontium degradation, especially alveolar bone. However, in this study, we show that an inflammatory mediator, IL-6, may serve another direction on alveolar bone homeostasis during inflammatory condition. We found that, IL-6 at 10 and 20 ng/mL was not cytotoxic and dose-dependently exerted beneficial effects on osteogenic differentiation of human PDLSCs (hPDLSCs), as demonstrated by increased alkaline phosphatase activity, mRNA expression of osteogenic markers, and matrix mineralization. The presence of physiological and inflammatory level of IL-6, the osteogenic differentiation potential by hPDLSCs was enhanced by several possible mechanisms including transforming growth factor (TGF), Wnt, and Notch pathways. After in-depth and thorough exploration, we found that Wnt pathway serves as key regulator controlling osteogenic differentiation by hPDLSCs amid the IL-6 presentation. Surprisingly, apart from other mesenchymal stem cells, distinct Wnt components are employed by hPDLSCs, and both canonical and non-canonical Wnt pathways are triggered by different mechanisms. Further validation by gene silencing, treatment with recombinant Wnt ligands, and β-catenin stabilization/translocation confirmed that IL-6 governed the canonical Wnt/β-catenin pathway via either WNT2B or WNT10B and employed WNT5A to activate the non-canonical Wnt pathway. These findings fulfill the homeostasis pathway governing periodontal tissue and alveolar bone regeneration and may serve for further therapeutic regimen design for restoring the tissues.
    MeSH term(s) Humans ; Periodontal Ligament ; Osteogenesis ; Interleukin-6/metabolism ; beta Catenin/metabolism ; Stem Cells/metabolism ; Wnt Signaling Pathway/physiology ; Inflammation/metabolism ; Immunologic Factors/metabolism ; Cell Differentiation ; Cells, Cultured
    Chemical Substances Interleukin-6 ; beta Catenin ; Immunologic Factors
    Language English
    Publishing date 2023-06-03
    Publishing country England
    Document type Journal Article ; Research Support, Non-U.S. Gov't ; Research Support, N.I.H., Extramural
    ZDB-ID 2615211-3
    ISSN 2045-2322 ; 2045-2322
    ISSN (online) 2045-2322
    ISSN 2045-2322
    DOI 10.1038/s41598-023-35569-8
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  4. Article ; Online: Dissecting specific Wnt components governing osteogenic differentiation potential by human periodontal ligament stem cells through interleukin-6

    Medania Purwaningrum / Cecilia M. Giachelli / Thanaphum Osathanon / Sirirat Rattanapuchpong / Chenphop Sawangmake

    Scientific Reports, Vol 13, Iss 1, Pp 1-

    2023  Volume 19

    Abstract: Abstract Periodontal ligament stem cells (PDLSCs) play a significant role on periodontal tissue and alveolar bone homeostasis. During inflammation, interleukin (IL)-6 serves as one of key cytokine players controlling tissue reaction as well as alveolar ... ...

    Abstract Abstract Periodontal ligament stem cells (PDLSCs) play a significant role on periodontal tissue and alveolar bone homeostasis. During inflammation, interleukin (IL)-6 serves as one of key cytokine players controlling tissue reaction as well as alveolar bone tissue remodeling. It is believed that periodontal tissue inflammation causes periodontium degradation, especially alveolar bone. However, in this study, we show that an inflammatory mediator, IL-6, may serve another direction on alveolar bone homeostasis during inflammatory condition. We found that, IL-6 at 10 and 20 ng/mL was not cytotoxic and dose-dependently exerted beneficial effects on osteogenic differentiation of human PDLSCs (hPDLSCs), as demonstrated by increased alkaline phosphatase activity, mRNA expression of osteogenic markers, and matrix mineralization. The presence of physiological and inflammatory level of IL-6, the osteogenic differentiation potential by hPDLSCs was enhanced by several possible mechanisms including transforming growth factor (TGF), Wnt, and Notch pathways. After in-depth and thorough exploration, we found that Wnt pathway serves as key regulator controlling osteogenic differentiation by hPDLSCs amid the IL-6 presentation. Surprisingly, apart from other mesenchymal stem cells, distinct Wnt components are employed by hPDLSCs, and both canonical and non-canonical Wnt pathways are triggered by different mechanisms. Further validation by gene silencing, treatment with recombinant Wnt ligands, and β-catenin stabilization/translocation confirmed that IL-6 governed the canonical Wnt/β-catenin pathway via either WNT2B or WNT10B and employed WNT5A to activate the non-canonical Wnt pathway. These findings fulfill the homeostasis pathway governing periodontal tissue and alveolar bone regeneration and may serve for further therapeutic regimen design for restoring the tissues.
    Keywords Medicine ; R ; Science ; Q
    Subject code 571
    Language English
    Publishing date 2023-06-01T00:00:00Z
    Publisher Nature Portfolio
    Document type Article ; Online
    Database BASE - Bielefeld Academic Search Engine (life sciences selection)

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  5. Article ; Online: Noncanonical Wnts at the Cusp of Fibrocalcific Signaling Processes in Human Calcific Aortic Valve Disease.

    Giachelli, Cecilia M / Speer, Mei Y

    Arteriosclerosis, thrombosis, and vascular biology

    2017  Volume 37, Issue 3, Page(s) 387–388

    MeSH term(s) Aortic Valve ; Aortic Valve Stenosis ; Calcinosis ; Heart Valve Diseases ; Humans ; Wnt Signaling Pathway
    Language English
    Publishing date 2017-02-08
    Publishing country United States
    Document type Editorial ; Research Support, U.S. Gov't, Non-P.H.S. ; Comment
    ZDB-ID 1221433-4
    ISSN 1524-4636 ; 1079-5642
    ISSN (online) 1524-4636
    ISSN 1079-5642
    DOI 10.1161/ATVBAHA.116.308842
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  6. Article: Comparison of osteoclast differentiation protocols from human induced pluripotent stem cells of different tissue origins.

    Blümke, Alexander / Ijeoma, Erica / Simon, Jessica / Wellington, Rachel / Purwaningrum, Medania / Doulatov, Sergei / Leber, Elizabeth / Scatena, Marta / Giachelli, Cecilia M

    Research square

    2023  

    Abstract: Background: Ever since their discovery, induced pluripotent stem cells (iPSCs) have been extensively differentiated into a large variety of cell types. However, a limited amount of work has been dedicated to differentiating iPSCs into osteoclasts. While ...

    Abstract Background: Ever since their discovery, induced pluripotent stem cells (iPSCs) have been extensively differentiated into a large variety of cell types. However, a limited amount of work has been dedicated to differentiating iPSCs into osteoclasts. While several differentiation protocols have been published, it remains unclear which protocols or differentiation methods are preferrable regarding the differentiation of osteoclasts.
    Methods: In this study we compare the osteoclastogenesis capacity of a peripheral blood mononuclear cell (PBMC)-derived iPSC line to a fibroblast-derived iPSC line in conjunction with either embryoid body-based or monolayer-based differentiation strategies. Both cell lines and differentiation protocols were investigated regarding their ability to generate osteoclasts and their inherent robustness and ease of use. The ability of both cell lines to remain undifferentiated while propagating using a feeder-free system was assessed using alkaline phosphatase staining. This was followed by evaluating mesodermal differentiation and the characterization of hematopoietic progenitor cells using flow cytometry. Finally, osteoclast yield and functionality based on resorptive activity, Cathepsin K and tartrate-resistant acid phosphatase (TRAP) expression were assessed. Results were validated using qRT-PCR throughout the differentiation stages.
    Results: Embryoid-body based differentiation yielded CD45
    Conclusions: The outcome of this study demonstrates the successful differentiation of osteoclasts from iPSCs in conjunction with the embryoid-based differentiation method, while the monolayer-based method did not yield osteoclasts. No differences were observed regarding osteoclast differentiation between the PBMC and fibroblast-derived iPSC lines.
    Language English
    Publishing date 2023-07-07
    Publishing country United States
    Document type Preprint
    DOI 10.21203/rs.3.rs-3089289/v1
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  7. Article ; Online: Comparison of osteoclast differentiation protocols from human induced pluripotent stem cells of different tissue origins.

    Blümke, Alexander / Ijeoma, Erica / Simon, Jessica / Wellington, Rachel / Purwaningrum, Medania / Doulatov, Sergei / Leber, Elizabeth / Scatena, Marta / Giachelli, Cecilia M

    Stem cell research & therapy

    2023  Volume 14, Issue 1, Page(s) 319

    Abstract: Background: Ever since their discovery, induced pluripotent stem cells (iPSCs) have been extensively differentiated into a large variety of cell types. However, a limited amount of work has been dedicated to differentiating iPSCs into osteoclasts. While ...

    Abstract Background: Ever since their discovery, induced pluripotent stem cells (iPSCs) have been extensively differentiated into a large variety of cell types. However, a limited amount of work has been dedicated to differentiating iPSCs into osteoclasts. While several differentiation protocols have been published, it remains unclear which protocols or differentiation methods are preferable regarding the differentiation of osteoclasts.
    Methods: In this study, we compared the osteoclastogenesis capacity of a peripheral blood mononuclear cell (PBMC)-derived iPSC line to a fibroblast-derived iPSC line in conjunction with either embryoid body-based or monolayer-based differentiation strategies. Both cell lines and differentiation protocols were investigated regarding their ability to generate osteoclasts and their inherent robustness and ease of use. The ability of both cell lines to remain undifferentiated while propagating using a feeder-free system was assessed using alkaline phosphatase staining. This was followed by evaluating mesodermal differentiation and the characterization of hematopoietic progenitor cells using flow cytometry. Finally, osteoclast yield and functionality based on resorptive activity, Cathepsin K and tartrate-resistant acid phosphatase (TRAP) expression were assessed. The results were validated using qRT-PCR throughout the differentiation stages.
    Results: Embryoid body-based differentiation yielded CD45
    Conclusions: The outcome of this study demonstrates the successful differentiation of osteoclasts from iPSCs in conjunction with the embryoid-based differentiation method, while the monolayer-based method did not yield osteoclasts. No differences were observed regarding osteoclast differentiation between the PBMC and fibroblast-derived iPSC lines.
    MeSH term(s) Humans ; Induced Pluripotent Stem Cells/metabolism ; Osteoclasts ; Leukocytes, Mononuclear ; Cathepsin K/metabolism ; Cell Differentiation
    Chemical Substances Cathepsin K (EC 3.4.22.38)
    Language English
    Publishing date 2023-11-07
    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/s13287-023-03547-6
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  8. Article: Monocyte Adhesion and Transmigration Through Endothelium Following Cardiopulmonary Bypass Shearing is Mediated by IL-8 Signaling.

    Zhou, Hao / Tu, Lan N / Giachelli, Cecilia / Nigam, Vishal / Scatena, Marta

    bioRxiv : the preprint server for biology

    2023  

    Language English
    Publishing date 2023-06-06
    Publishing country United States
    Document type Preprint
    DOI 10.1101/2023.06.05.543811
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  9. Article ; Online: Vascular calcification in CKD-MBD: Roles for phosphate, FGF23, and Klotho.

    Yamada, Shunsuke / Giachelli, Cecilia M

    Bone

    2016  Volume 100, Page(s) 87–93

    Abstract: Vascular calcification (VC) is highly prevalent in aging, diabetes mellitus, and chronic kidney disease (CKD). VC is a strong predictor of cardiovascular morbidity and mortality in the CKD population. Complex pathological mechanisms are involved in the ... ...

    Abstract Vascular calcification (VC) is highly prevalent in aging, diabetes mellitus, and chronic kidney disease (CKD). VC is a strong predictor of cardiovascular morbidity and mortality in the CKD population. Complex pathological mechanisms are involved in the development of VC, including osteochondrogenic differentiation and apoptosis of vascular smooth muscle cells, instability and release of extracellular vesicles loaded calcium and phosphate, and elastin degradation. Elevated serum phosphate is a late manifestation of CKD, and has been shown to accelerate mineral deposition in both the vessel wall and heart valves. α-Klotho and fibroblast growth factor 23 (FGF23) are emerging factors in CKD-mineral and bone disorder (CKD-MBD) and are thought to be involved in the pathogenesis of uremic VC. There are discordant reports regarding the biomedical effects of FGF23 on VC. In contrast, mounting evidence supports a well-supported protective role for α-Klotho on VC. Further studies are warranted to elucidate potential roles of FGF23 and α-Klotho in VC and to determine where and how they are synthesized in normal and disease conditions. A thorough systemic evaluation of the biomedical interplay of phosphate, FGF23, and α-Klotho may potentially lead to new therapeutic options for patients with CKD-MBD.
    MeSH term(s) Chronic Kidney Disease-Mineral and Bone Disorder/metabolism ; Fibroblast Growth Factors/metabolism ; Glucuronidase/metabolism ; Phosphates/metabolism ; Vascular Calcification/metabolism
    Chemical Substances Phosphates ; Fibroblast Growth Factors (62031-54-3) ; fibroblast growth factor 23 (7Q7P4S7RRE) ; Glucuronidase (EC 3.2.1.31) ; klotho protein (EC 3.2.1.31)
    Language English
    Publishing date 2016-11-12
    Publishing country United States
    Document type Journal Article ; Review
    ZDB-ID 632515-4
    ISSN 1873-2763 ; 8756-3282
    ISSN (online) 1873-2763
    ISSN 8756-3282
    DOI 10.1016/j.bone.2016.11.012
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  10. Article ; Online: Comparison of osteoclast differentiation protocols from human induced pluripotent stem cells of different tissue origins

    Alexander Blümke / Erica Ijeoma / Jessica Simon / Rachel Wellington / Medania Purwaningrum / Sergei Doulatov / Elizabeth Leber / Marta Scatena / Cecilia M. Giachelli

    Stem Cell Research & Therapy, Vol 14, Iss 1, Pp 1-

    2023  Volume 20

    Abstract: Abstract Background Ever since their discovery, induced pluripotent stem cells (iPSCs) have been extensively differentiated into a large variety of cell types. However, a limited amount of work has been dedicated to differentiating iPSCs into osteoclasts. ...

    Abstract Abstract Background Ever since their discovery, induced pluripotent stem cells (iPSCs) have been extensively differentiated into a large variety of cell types. However, a limited amount of work has been dedicated to differentiating iPSCs into osteoclasts. While several differentiation protocols have been published, it remains unclear which protocols or differentiation methods are preferable regarding the differentiation of osteoclasts. Methods In this study, we compared the osteoclastogenesis capacity of a peripheral blood mononuclear cell (PBMC)-derived iPSC line to a fibroblast-derived iPSC line in conjunction with either embryoid body-based or monolayer-based differentiation strategies. Both cell lines and differentiation protocols were investigated regarding their ability to generate osteoclasts and their inherent robustness and ease of use. The ability of both cell lines to remain undifferentiated while propagating using a feeder-free system was assessed using alkaline phosphatase staining. This was followed by evaluating mesodermal differentiation and the characterization of hematopoietic progenitor cells using flow cytometry. Finally, osteoclast yield and functionality based on resorptive activity, Cathepsin K and tartrate-resistant acid phosphatase (TRAP) expression were assessed. The results were validated using qRT-PCR throughout the differentiation stages. Results Embryoid body-based differentiation yielded CD45+, CD14+, CD11b+ subpopulations which in turn differentiated into osteoclasts which demonstrated TRAP positivity, Cathepsin K expression and mineral resorptive capabilities. This was regardless of which iPSC line was used. Monolayer-based differentiation yielded lower quantities of hematopoietic cells that were mostly CD34+ and did not subsequently differentiate into osteoclasts. Conclusions The outcome of this study demonstrates the successful differentiation of osteoclasts from iPSCs in conjunction with the embryoid-based differentiation method, while the monolayer-based method did not yield ...
    Keywords Human induced pluripotent stem cells ; Osteoclasts ; Osteoclastogenesis ; Hematopoietic differentiation ; Mesodermal differentiation ; Mineral resorption ; Medicine (General) ; R5-920 ; Biochemistry ; QD415-436
    Subject code 571
    Language English
    Publishing date 2023-11-01T00:00:00Z
    Publisher BMC
    Document type Article ; Online
    Database BASE - Bielefeld Academic Search Engine (life sciences selection)

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