PLLA scaffolds produced by thermally induced phase separation (TIPS) allow human chondrocyte growth and extracellular matrix formation dependent on pore size

Valerio Maria Bartolo Brucato, Vincenzo La Carrubba, Francesco Carfi' Pavia, Gioacchino Conoscenti, Gundula Schulze-Tanzil, Katharina Stoelzel, Tobias Schneider, Clemens Goegele

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    Abstract

    Damage of hyaline cartilage species such as nasoseptal or joint cartilage requires proper reconstruction, which remains challenging due to the low intrinsic repair capacity of this tissue. Implantation of autologous chondrocytes in combination with a biomimetic biomaterial represents a promising strategy to support cartilage repair. The aim of this work was to assess the viability, attachment, morphology, extracellular matrix (ECM) production of human articular and nasoseptal chondrocytes cultured in vitro in porous poly(L-lactic) (PLLA) scaffolds of two selected pore sizes (100 and 200 μm). The PLLA scaffolds with 100 and 200 μm pore sizes were prepared via ternary thermally induced phase separation (TIPS) technique and analyzed using scanning electron microscopy (SEM). Articular and nasoseptal chondrocytes were seeded on the scaffold and cultures maintained for 7 and 14 days. Live/dead staining, (immuno-)histology and gene expression analysis of type II, type I collagen, aggrecan and SOX9 were performed to assess scaffold cytocompatibility and chondrocyte phenotype. The majority of both chondrocyte types survived on both scaffolds for the whole culture period. Hematoxylin-eosin (HE), alcian blue (visualizing glycosaminoglycans) stainings, immunoreactivity and gene expression of ECM proteins and cartilage marker (type II, I collagen, aggrecan, SOX9) of the chondrocyte scaffold constructs indicated that the smaller pore dimensions promoted the differentiation of the chondrocytes compared with the larger pore size. The present work revealed that the scaffold pore size is an important factor influencing chondrocyte differentiation and indicated that the scaffolds with 100 μm pores serve as a cytocompatible basis for further future modifications.
    Lingua originaleEnglish
    pagine (da-a)449-459
    Numero di pagine11
    RivistaMATERIALS SCIENCE AND ENGINEERING. C, BIOMIMETIC MATERIALS, SENSORS AND SYSTEMS
    Volume80
    Stato di pubblicazionePublished - 2017

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    Scaffolds
    Phase separation
    Pore size
    cartilage
    Cartilage
    porosity
    matrices
    gene expression
    staining
    collagens
    Scaffolds (biology)
    Aggrecans
    Collagen
    Gene expression
    Repair
    phenotype
    Histology
    histology
    biomimetics
    Biomimetics

    All Science Journal Classification (ASJC) codes

    • Materials Science(all)
    • Mechanical Engineering
    • Mechanics of Materials
    • Condensed Matter Physics

    Cita questo

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    title = "PLLA scaffolds produced by thermally induced phase separation (TIPS) allow human chondrocyte growth and extracellular matrix formation dependent on pore size",
    abstract = "Damage of hyaline cartilage species such as nasoseptal or joint cartilage requires proper reconstruction, which remains challenging due to the low intrinsic repair capacity of this tissue. Implantation of autologous chondrocytes in combination with a biomimetic biomaterial represents a promising strategy to support cartilage repair. The aim of this work was to assess the viability, attachment, morphology, extracellular matrix (ECM) production of human articular and nasoseptal chondrocytes cultured in vitro in porous poly(L-lactic) (PLLA) scaffolds of two selected pore sizes (100 and 200{\^A} {\^I}¼m). The PLLA scaffolds with 100 and 200{\^A} {\^I}¼m pore sizes were prepared via ternary thermally induced phase separation (TIPS) technique and analyzed using scanning electron microscopy (SEM). Articular and nasoseptal chondrocytes were seeded on the scaffold and cultures maintained for 7 and 14{\^A} days. Live/dead staining, (immuno-)histology and gene expression analysis of type II, type I collagen, aggrecan and SOX9 were performed to assess scaffold cytocompatibility and chondrocyte phenotype. The majority of both chondrocyte types survived on both scaffolds for the whole culture period. Hematoxylin-eosin (HE), alcian blue (visualizing glycosaminoglycans) stainings, immunoreactivity and gene expression of ECM proteins and cartilage marker (type II, I collagen, aggrecan, SOX9) of the chondrocyte scaffold constructs indicated that the smaller pore dimensions promoted the differentiation of the chondrocytes compared with the larger pore size. The present work revealed that the scaffold pore size is an important factor influencing chondrocyte differentiation and indicated that the scaffolds with 100{\^A} {\^I}¼m pores serve as a cytocompatible basis for further future modifications.",
    author = "Brucato, {Valerio Maria Bartolo} and {La Carrubba}, Vincenzo and {Carfi' Pavia}, Francesco and Gioacchino Conoscenti and Gundula Schulze-Tanzil and Katharina Stoelzel and Tobias Schneider and Clemens Goegele",
    year = "2017",
    language = "English",
    volume = "80",
    pages = "449--459",
    journal = "Materials Science and Engineering C",
    issn = "0928-4931",
    publisher = "Elsevier BV",

    }

    TY - JOUR

    T1 - PLLA scaffolds produced by thermally induced phase separation (TIPS) allow human chondrocyte growth and extracellular matrix formation dependent on pore size

    AU - Brucato, Valerio Maria Bartolo

    AU - La Carrubba, Vincenzo

    AU - Carfi' Pavia, Francesco

    AU - Conoscenti, Gioacchino

    AU - Schulze-Tanzil, Gundula

    AU - Stoelzel, Katharina

    AU - Schneider, Tobias

    AU - Goegele, Clemens

    PY - 2017

    Y1 - 2017

    N2 - Damage of hyaline cartilage species such as nasoseptal or joint cartilage requires proper reconstruction, which remains challenging due to the low intrinsic repair capacity of this tissue. Implantation of autologous chondrocytes in combination with a biomimetic biomaterial represents a promising strategy to support cartilage repair. The aim of this work was to assess the viability, attachment, morphology, extracellular matrix (ECM) production of human articular and nasoseptal chondrocytes cultured in vitro in porous poly(L-lactic) (PLLA) scaffolds of two selected pore sizes (100 and 200 μm). The PLLA scaffolds with 100 and 200 μm pore sizes were prepared via ternary thermally induced phase separation (TIPS) technique and analyzed using scanning electron microscopy (SEM). Articular and nasoseptal chondrocytes were seeded on the scaffold and cultures maintained for 7 and 14 days. Live/dead staining, (immuno-)histology and gene expression analysis of type II, type I collagen, aggrecan and SOX9 were performed to assess scaffold cytocompatibility and chondrocyte phenotype. The majority of both chondrocyte types survived on both scaffolds for the whole culture period. Hematoxylin-eosin (HE), alcian blue (visualizing glycosaminoglycans) stainings, immunoreactivity and gene expression of ECM proteins and cartilage marker (type II, I collagen, aggrecan, SOX9) of the chondrocyte scaffold constructs indicated that the smaller pore dimensions promoted the differentiation of the chondrocytes compared with the larger pore size. The present work revealed that the scaffold pore size is an important factor influencing chondrocyte differentiation and indicated that the scaffolds with 100 μm pores serve as a cytocompatible basis for further future modifications.

    AB - Damage of hyaline cartilage species such as nasoseptal or joint cartilage requires proper reconstruction, which remains challenging due to the low intrinsic repair capacity of this tissue. Implantation of autologous chondrocytes in combination with a biomimetic biomaterial represents a promising strategy to support cartilage repair. The aim of this work was to assess the viability, attachment, morphology, extracellular matrix (ECM) production of human articular and nasoseptal chondrocytes cultured in vitro in porous poly(L-lactic) (PLLA) scaffolds of two selected pore sizes (100 and 200 μm). The PLLA scaffolds with 100 and 200 μm pore sizes were prepared via ternary thermally induced phase separation (TIPS) technique and analyzed using scanning electron microscopy (SEM). Articular and nasoseptal chondrocytes were seeded on the scaffold and cultures maintained for 7 and 14 days. Live/dead staining, (immuno-)histology and gene expression analysis of type II, type I collagen, aggrecan and SOX9 were performed to assess scaffold cytocompatibility and chondrocyte phenotype. The majority of both chondrocyte types survived on both scaffolds for the whole culture period. Hematoxylin-eosin (HE), alcian blue (visualizing glycosaminoglycans) stainings, immunoreactivity and gene expression of ECM proteins and cartilage marker (type II, I collagen, aggrecan, SOX9) of the chondrocyte scaffold constructs indicated that the smaller pore dimensions promoted the differentiation of the chondrocytes compared with the larger pore size. The present work revealed that the scaffold pore size is an important factor influencing chondrocyte differentiation and indicated that the scaffolds with 100 μm pores serve as a cytocompatible basis for further future modifications.

    UR - http://hdl.handle.net/10447/244820

    M3 - Article

    VL - 80

    SP - 449

    EP - 459

    JO - Materials Science and Engineering C

    JF - Materials Science and Engineering C

    SN - 0928-4931

    ER -