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  1. Article ; Online: Novel strategy for multi-material 3D bioprinting of human stem cell based corneal stroma with heterogenous design.

    Puistola, Paula / Miettinen, Susanna / Skottman, Heli / Mörö, Anni

    Materials today. Bio

    2023  Volume 24, Page(s) 100924

    Abstract: Three-dimensional (3D) bioprinting offers an automated, customizable solution to manufacture highly detailed 3D tissue constructs and holds great promise for regenerative medicine to solve the severe global shortage of donor tissues and organs. However, ... ...

    Abstract Three-dimensional (3D) bioprinting offers an automated, customizable solution to manufacture highly detailed 3D tissue constructs and holds great promise for regenerative medicine to solve the severe global shortage of donor tissues and organs. However, uni-material 3D bioprinting is not sufficient for manufacturing heterogenous 3D constructs with native-like microstructures and thus, innovative multi-material solutions are required. Here, we developed a novel multi-material 3D bioprinting strategy for bioprinting human corneal stroma. The human cornea is the transparent outer layer of your eye, and vision loss due to corneal blindness has serious effects on the quality of life of individuals. One of the main reasons for corneal blindness is the damage in the detailed organization of the corneal stroma where collagen fibrils are arranged in layers perpendicular to each other and the corneal stromal cells grow along the fibrils. Donor corneas for treating corneal blindness are scarce, and the current tissue engineering (TE) technologies cannot produce artificial corneas with the complex microstructure of native corneal stroma. To address this, we developed a novel multi-material 3D bioprinting strategy to mimic detailed organization of corneal stroma. These multi-material 3D structures with heterogenous design were bioprinted by using human adipose tissue -derived stem cells (hASCs) and hyaluronic acid (HA) -based bioinks with varying stiffnesses. In our novel design of 3D models, acellular stiffer HA-bioink and cell-laden softer HA-bioink were printed in alternating filaments, and the filaments were printed perpendicularly in alternating layers. The multi-material bioprinting strategy was applied for the first time in corneal stroma 3D bioprinting to mimic the native microstructure. As a result, the soft bioink promoted cellular growth and tissue formation of hASCs in the multi-material 3D bioprinted composites, whereas the stiff bioink provided mechanical support as well as guidance of cellular organization upon culture. Interestingly, cellular growth and tissue formation altered the mechanical properties of the bioprinted composite constructs significantly. Importantly, the bioprinted composite structures showed good integration to the host tissue in
    Language English
    Publishing date 2023-12-22
    Publishing country England
    Document type Journal Article
    ISSN 2590-0064
    ISSN (online) 2590-0064
    DOI 10.1016/j.mtbio.2023.100924
    Database MEDical Literature Analysis and Retrieval System OnLINE

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  2. Article ; Online: Bioprinting of human pluripotent stem cell derived corneal endothelial cells with hydrazone crosslinked hyaluronic acid bioink.

    Grönroos, Pyry / Mörö, Anni / Puistola, Paula / Hopia, Karoliina / Huuskonen, Maija / Viheriälä, Taina / Ilmarinen, Tanja / Skottman, Heli

    Stem cell research & therapy

    2024  Volume 15, Issue 1, Page(s) 81

    Abstract: Background: Human corneal endothelial cells lack regenerative capacity through cell division in vivo. Consequently, in the case of trauma or dystrophy, the only available treatment modality is corneal tissue or primary corneal endothelial cell ... ...

    Abstract Background: Human corneal endothelial cells lack regenerative capacity through cell division in vivo. Consequently, in the case of trauma or dystrophy, the only available treatment modality is corneal tissue or primary corneal endothelial cell transplantation from cadaveric donor which faces a high global shortage. Our ultimate goal is to use the state-of-the-art 3D-bioprint technology for automated production of human partial and full-thickness corneal tissues using human stem cells and functional bioinks. In this study, we explore the feasibility of bioprinting the corneal endothelium using human pluripotent stem cell derived corneal endothelial cells and hydrazone crosslinked hyaluronic acid bioink.
    Methods: Corneal endothelial cells differentiated from human pluripotent stem cells were bioprinted using optimized hydrazone crosslinked hyaluronic acid based bioink. Before the bioprinting process, the biocompatibility of the bioink with cells was first analyzed with transplantation on ex vivo denuded rat and porcine corneas as well as on denuded human Descemet membrane. Subsequently, the bioprinting was proceeded and the viability of human pluripotent stem cell derived corneal endothelial cells were verified with live/dead stainings. Histological and immunofluorescence stainings involving ZO1, Na
    Results: The bioink, modified for human pluripotent stem cell derived corneal endothelial cells successfully supported both the viability and printability of the cells. Following up to 10 days of ex vivo transplantations, STEM121 positive cells were confirmed on the Descemet membrane of rat and porcine cornea demonstrating the biocompatibility of the bioink. Furthermore, biocompatibility was validated on denuded human Descemet membrane showing corneal endothelial -like characteristics. Seven days post bioprinting, the corneal endothelial -like cells were viable and showed polygonal morphology with expression and native-like localization of ZO-1, Na
    Conclusions: Our results demonstrate the successful printing of human pluripotent stem cell derived corneal endothelial cells using covalently crosslinked hyaluronic acid bioink. This approach not only holds promise for a corneal endothelium transplants but also presents potential applications in the broader mission of bioprinting the full-thickness human cornea.
    MeSH term(s) Humans ; Rats ; Animals ; Swine ; Tissue Engineering/methods ; Endothelial Cells ; Bioprinting/methods ; Hyaluronic Acid/pharmacology ; Pluripotent Stem Cells ; Adenosine Triphosphatases
    Chemical Substances Hyaluronic Acid (9004-61-9) ; Adenosine Triphosphatases (EC 3.6.1.-)
    Language English
    Publishing date 2024-03-14
    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/s13287-024-03672-w
    Database MEDical Literature Analysis and Retrieval System OnLINE

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  3. Article ; Online: Cornea-Specific Human Adipose Stem Cell-Derived Extracellular Matrix for Corneal Stroma Tissue Engineering.

    Puistola, Paula / Kethiri, Abhinav / Nurminen, Antti / Turkki, Johannes / Hopia, Karoliina / Miettinen, Susanna / Mörö, Anni / Skottman, Heli

    ACS applied materials & interfaces

    2024  Volume 16, Issue 13, Page(s) 15761–15772

    Abstract: Utilizing tissue-specific extracellular matrices (ECMs) is vital for replicating the composition of native tissues and developing biologically relevant biomaterials. Human- or animal-derived donor tissues and organs are the current gold standard for the ... ...

    Abstract Utilizing tissue-specific extracellular matrices (ECMs) is vital for replicating the composition of native tissues and developing biologically relevant biomaterials. Human- or animal-derived donor tissues and organs are the current gold standard for the source of these ECMs. To overcome the several limitations related to these ECM sources, including the highly limited availability of donor tissues, cell-derived ECM offers an alternative approach for engineering tissue-specific biomaterials, such as bioinks for three-dimensional (3D) bioprinting. 3D bioprinting is a state-of-the-art biofabrication technology that addresses the global need for donor tissues and organs. In fact, there is a vast global demand for human donor corneas that are used for treating corneal blindness, often resulting from damage in the corneal stromal microstructure. Human adipose tissue is one of the most abundant tissues and easy to access, and adipose tissue-derived stem cells (hASCs) are a highly advantageous cell type for tissue engineering. Furthermore, hASCs have already been studied in clinical trials for treating corneal stromal pathologies. In this study, a corneal stroma-specific ECM was engineered without the need for donor corneas by differentiating hASCs toward corneal stromal keratocytes (hASC-CSKs). Furthermore, this ECM was utilized as a component for corneal stroma-specific bioink where hASC-CSKs were printed to produce corneal stroma structures. This cost-effective approach combined with a clinically relevant cell type provides valuable information on developing more sustainable tissue-specific solutions and advances the field of corneal tissue engineering.
    MeSH term(s) Animals ; Humans ; Tissue Engineering/methods ; Corneal Stroma/metabolism ; Cornea ; Extracellular Matrix/chemistry ; Biocompatible Materials/metabolism ; Adipose Tissue ; Stem Cells ; Tissue Scaffolds ; Bioprinting/methods
    Chemical Substances Biocompatible Materials
    Language English
    Publishing date 2024-03-21
    Publishing country United States
    Document type Journal Article
    ISSN 1944-8252
    ISSN (online) 1944-8252
    DOI 10.1021/acsami.3c17803
    Database MEDical Literature Analysis and Retrieval System OnLINE

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  4. Article: Inkjet Printable Polydimethylsiloxane for All-Inkjet-Printed Multilayered Soft Electrical Applications

    Mikkonen, Riikka / Puistola, Paula / Jönkkäri, Ilari / Mäntysalo, Matti

    ACS applied materials & interfaces. 2020 Feb. 13, v. 12, no. 10

    2020  

    Abstract: In recent years, additive manufacturing of polydimethylsiloxane (PDMS) has gained interest for the development of soft electronics. To build complex electrical devices, fabrication of multilayered structures is required. We propose here a straightforward ...

    Abstract In recent years, additive manufacturing of polydimethylsiloxane (PDMS) has gained interest for the development of soft electronics. To build complex electrical devices, fabrication of multilayered structures is required. We propose here a straightforward digital printing fabrication process of silicone rubber-based, multilayered electronics. An inkjet-printable PDMS solution was developed for the digital patterning of elastomeric structures. The silicone ink was used together with a highly conductive silver nanoparticle (Ag NP) ink for the fabrication of all-inkjet-printed multilayered electrical devices. The application of the multilayered circuit board was successful. The sheet resistances were below 0.3 Ω/□, and the conductive layer thickness was less than 1 μm. The electrical insulation between the conductive layers was done by printing a 20–25 μm-thick dielectric PDMS layer selectively on top of the bottommost conductive layer.
    Keywords electronics ; insulating materials ; manufacturing ; nanosilver ; polydimethylsiloxane
    Language English
    Dates of publication 2020-0213
    Size p. 11990-11997.
    Publishing place American Chemical Society
    Document type Article
    ISSN 1944-8252
    DOI 10.1021/acsami.9b19632
    Database NAL-Catalogue (AGRICOLA)

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  5. Article ; Online: Inkjet Printable Polydimethylsiloxane for All-Inkjet-Printed Multilayered Soft Electrical Applications.

    Mikkonen, Riikka / Puistola, Paula / Jönkkäri, Ilari / Mäntysalo, Matti

    ACS applied materials & interfaces

    2020  Volume 12, Issue 10, Page(s) 11990–11997

    Abstract: In recent years, additive manufacturing of polydimethylsiloxane (PDMS) has gained interest for the development of soft electronics. To build complex electrical devices, fabrication of multilayered structures is required. We propose here a straightforward ...

    Abstract In recent years, additive manufacturing of polydimethylsiloxane (PDMS) has gained interest for the development of soft electronics. To build complex electrical devices, fabrication of multilayered structures is required. We propose here a straightforward digital printing fabrication process of silicone rubber-based, multilayered electronics. An inkjet-printable PDMS solution was developed for the digital patterning of elastomeric structures. The silicone ink was used together with a highly conductive silver nanoparticle (Ag NP) ink for the fabrication of all-inkjet-printed multilayered electrical devices. The application of the multilayered circuit board was successful. The sheet resistances were below 0.3 Ω/□, and the conductive layer thickness was less than 1 μm. The electrical insulation between the conductive layers was done by printing a 20-25 μm-thick dielectric PDMS layer selectively on top of the bottommost conductive layer.
    Language English
    Publishing date 2020-02-28
    Publishing country United States
    Document type Journal Article
    ISSN 1944-8252
    ISSN (online) 1944-8252
    DOI 10.1021/acsami.9b19632
    Database MEDical Literature Analysis and Retrieval System OnLINE

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  6. Article ; Online: Toward Corneal Limbus In Vitro Model: Regulation of hPSC-LSC Phenotype by Matrix Stiffness and Topography During Cell Differentiation Process.

    Kauppila, Maija / Mörö, Anni / Valle-Delgado, Juan José / Ihalainen, Teemu / Sukki, Lassi / Puistola, Paula / Kallio, Pasi / Ilmarinen, Tanja / Österberg, Monika / Skottman, Heli

    Advanced healthcare materials

    2023  Volume 12, Issue 29, Page(s) e2301396

    Abstract: A functional limbal epithelial stem cells (LSC) niche is a vital element in the regular renewal of the corneal epithelium by LSCs and maintenance of good vision. However, little is known about its unique structure and mechanical properties on LSC ... ...

    Abstract A functional limbal epithelial stem cells (LSC) niche is a vital element in the regular renewal of the corneal epithelium by LSCs and maintenance of good vision. However, little is known about its unique structure and mechanical properties on LSC regulation, creating a significant gap in development of LSC-based therapies. Herein, the effect of mechanical and architectural elements of the niche on human pluripotent derived LSCs (hPSC-LSC) phenotype and growth is investigated in vitro. Specifically, three formulations of polyacrylamide gels with different controlled stiffnesses are used for culture and characterization of hPSC-LSCs from different stages of differentiation. In addition, limbal mimicking topography in polydimethylsiloxane is utilized for culturing hPSC-LSCs at early time point of differentiation. For comparison, the expression of selected key proteins of the corneal cells is analyzed in their native environment through whole mount staining of human donor corneas. The results suggest that mechanical response and substrate preference of the cells is highly dependent on their developmental stage. In addition, data indicate that cells may carry possible mechanical memory from previous culture matrix, both highlighting the importance of mechanical design of a functional in vitro limbus model.
    MeSH term(s) Humans ; Stem Cells ; Limbus Corneae/metabolism ; Cornea ; Phenotype ; Cell Differentiation
    Language English
    Publishing date 2023-07-21
    Publishing country Germany
    Document type Journal Article ; Research Support, Non-U.S. Gov't
    ZDB-ID 2649576-4
    ISSN 2192-2659 ; 2192-2640
    ISSN (online) 2192-2659
    ISSN 2192-2640
    DOI 10.1002/adhm.202301396
    Database MEDical Literature Analysis and Retrieval System OnLINE

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