Tesi etd-02162023-150642
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Tipo di tesi
Dottorato
Autore
CAVALLO, AIDA
URN
etd-02162023-150642
Titolo
3D bioprinting of a fibrinogen-based bioink for a bi-layer skin substitute.
Settore scientifico disciplinare
ING-IND/34
Corso di studi
Istituto di Scienze della Vita - PHD IN MEDICINA TRASLAZIONALE
Commissione
relatore Prof. PASSINO, CLAUDIO
Parole chiave
- Nessuna parola chiave trovata
Data inizio appello
12/07/2023;
Disponibilità
parziale
Riassunto analitico
3D bioprinting has emerged as attractive technology due to the ability of mimicking native tissue architecture using different cell types and biomaterials. Engineered skin substitutes have received great attention due to the possibility to be employed in vivo as skin graft in patients with chronic wound and in vitro as an alternative to in vivo animal testing. Nowadays, skin tissue equivalents or cell-laden constructs development is still in an early stage.
In this study, a novel fibrinogen-alginate based bioink for the 3D bioprinting of a skin substitute is proposed.
The bioink formulation is composed of 30 mg/ml of fibrinogen, 6% of alginate and 25 mM of CaCl2. The sterile bioink was characterised in terms of homogeneity, rheological properties, printability, mechanical properties, degradation rate, water uptake and biocompatibility by the indirect method and by bioprinting using L929 mouse fibroblasts. The proposed bioink is a homogeneous blend with a shear thinning behaviour, excellent printability, adequate mechanical stiffness, porosity, biodegradability and water uptake and it is able to support L929 viability.
The proposed bioink was used for the biofabrication of skin substitutes composed by an epidermal layer with human keratinocytes (HaCaT) and by a dermal layer with human fibroblasts (NHDF). The construct has a round shape with a diameter of 12 mm and height of 2 mm with 3 layers of fibroblasts and 2 of keratinocytes. Skin substitute stained by live/dead and histological analysis (Haematoxylin and Eosin) immediately after printing and at days 7 and 14 of culture showed a tissue-like structure with two distinct layers characterised by the presence of viable and proliferating cells.
This bioprinted skin construct showed a similar native skin architecture with two distinct layers paving the way for its use as a skin substitute.
In this study, a novel fibrinogen-alginate based bioink for the 3D bioprinting of a skin substitute is proposed.
The bioink formulation is composed of 30 mg/ml of fibrinogen, 6% of alginate and 25 mM of CaCl2. The sterile bioink was characterised in terms of homogeneity, rheological properties, printability, mechanical properties, degradation rate, water uptake and biocompatibility by the indirect method and by bioprinting using L929 mouse fibroblasts. The proposed bioink is a homogeneous blend with a shear thinning behaviour, excellent printability, adequate mechanical stiffness, porosity, biodegradability and water uptake and it is able to support L929 viability.
The proposed bioink was used for the biofabrication of skin substitutes composed by an epidermal layer with human keratinocytes (HaCaT) and by a dermal layer with human fibroblasts (NHDF). The construct has a round shape with a diameter of 12 mm and height of 2 mm with 3 layers of fibroblasts and 2 of keratinocytes. Skin substitute stained by live/dead and histological analysis (Haematoxylin and Eosin) immediately after printing and at days 7 and 14 of culture showed a tissue-like structure with two distinct layers characterised by the presence of viable and proliferating cells.
This bioprinted skin construct showed a similar native skin architecture with two distinct layers paving the way for its use as a skin substitute.
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