Biofabrication of HepG2 Cells-Laden 3D Structures Using Nanocellulose-Reinforced Gelatin-Based Hydrogel Bioinks: Materials Characterization, Cell Viability Assessment, and Metabolomic Analysis.
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| Format: | Artículo científico |
| Langue: | en |
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ACS biomaterials science & engineering
2025
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| author | Lameirinhas, Nicole S Teixeira, Maria C Carvalho, João P F Valente, Bruno F A Luís, Jorge L Duarte, Iola F Pinto, Ricardo J B Oliveira, Helena Oliveira, José M Silvestre, Armando J D Vilela, Carla Freire, Carmen S R |
| author_facet | Lameirinhas, Nicole S Teixeira, Maria C Carvalho, João P F Valente, Bruno F A Luís, Jorge L Duarte, Iola F Pinto, Ricardo J B Oliveira, Helena Oliveira, José M Silvestre, Armando J D Vilela, Carla Freire, Carmen S R Lameirinhas, Nicole S Teixeira, Maria C Carvalho, João P F Valente, Bruno F A Luís, Jorge L Duarte, Iola F Pinto, Ricardo J B Oliveira, Helena Oliveira, José M Silvestre, Armando J D Vilela, Carla Freire, Carmen S R |
| collection | PubMed - marine biology |
| contents | Biofabrication of HepG2 Cells-Laden 3D Structures Using Nanocellulose-Reinforced Gelatin-Based Hydrogel Bioinks: Materials Characterization, Cell Viability Assessment, and Metabolomic Analysis. Lameirinhas, Nicole S Teixeira, Maria C Carvalho, João P F Valente, Bruno F A Luís, Jorge L Duarte, Iola F Pinto, Ricardo J B Oliveira, Helena Oliveira, José M Silvestre, Armando J D Vilela, Carla Freire, Carmen S R Humans Hep G2 Cells Gelatin Cellulose Cell Survival Hydrogels Bioprinting Ink Printing, Three-Dimensional Iridoids Metabolomics Rheology The successful replication of the intricate architecture of human tissues remains a major challenge in the biomedical area. Three-dimensional (3D) bioprinting has emerged as a promising approach for the biofabrication of living tissue analogues, taking advantage of the use of adequate bioinks and printing methodologies. Here, a hydrogel bioink based on gelatin (Gel) and nanofibrillated cellulose (NFC), cross-linked with genipin, was developed for the 3D extrusion-based bioprinting of hepatocarcinoma cells (HepG2). This formulation combines the biological characteristics of Gel with the exceptional mechanical and rheological attributes of NFC. Gel/NFC ink formulations with different Gel/NFC mass compositions, viz., 90:10, 80:20, 70:30, and 60:40, were prepared and characterized. The corresponding cross-linked hydrogels were obtained using 1.5% (w/w) genipin as the cross-linking agent. The rheological and mechanical performances of the inks were enhanced by the addition of NFC, as evidenced by the rise in the yield stress from 70.9 ± 28.6 to 627.9 ± 74.8 Pa, compressive stress at 80% strain from 0.5 ± 0.1 to 1.5 ± 0.2 MPa, and Young's modulus from 4.7 ± 0.9 to 12.1 ± 1.1 MPa, for 90:10 and 60:40 inks, respectively. Moreover, higher NFC contents translated into 3D structures with better shape fidelity and the possibility of printing more intricate structures. These hydrogels were noncytotoxic toward HepG2 cells for up to 48 h, with cell viabilities consistently above 80%. The ink 70:30 was loaded with HepG2 cells (2 × 10 cells mL) and bioprinted. Cell viability remained elevated (90 ± 4%) until day 14 postbioprinting, with cells maintaining their metabolic activity shown by H NMR metabolomics, proving the enormous potential of Gel/NFC-based bioinks for bioprinting HepG2 cells without jeopardizing their viability and metabolism. |
| format | Artículo científico |
| id | pubmed_40241282 |
| institution | PubMed |
| language | en |
| publishDate | 2025 |
| publisher | ACS biomaterials science & engineering |
| record_format | pubmed |
| spellingShingle | Biofabrication of HepG2 Cells-Laden 3D Structures Using Nanocellulose-Reinforced Gelatin-Based Hydrogel Bioinks: Materials Characterization, Cell Viability Assessment, and Metabolomic Analysis. Lameirinhas, Nicole S Teixeira, Maria C Carvalho, João P F Valente, Bruno F A Luís, Jorge L Duarte, Iola F Pinto, Ricardo J B Oliveira, Helena Oliveira, José M Silvestre, Armando J D Vilela, Carla Freire, Carmen S R Humans Hep G2 Cells Gelatin Cellulose Cell Survival Hydrogels Bioprinting Ink Printing, Three-Dimensional Iridoids Metabolomics Rheology Biofabrication of HepG2 Cells-Laden 3D Structures Using Nanocellulose-Reinforced Gelatin-Based Hydrogel Bioinks: Materials Characterization, Cell Viability Assessment, and Metabolomic Analysis. Lameirinhas, Nicole S Teixeira, Maria C Carvalho, João P F Valente, Bruno F A Luís, Jorge L Duarte, Iola F Pinto, Ricardo J B Oliveira, Helena Oliveira, José M Silvestre, Armando J D Vilela, Carla Freire, Carmen S R Humans Hep G2 Cells Gelatin Cellulose Cell Survival Hydrogels Bioprinting Ink Printing, Three-Dimensional Iridoids Metabolomics Rheology The successful replication of the intricate architecture of human tissues remains a major challenge in the biomedical area. Three-dimensional (3D) bioprinting has emerged as a promising approach for the biofabrication of living tissue analogues, taking advantage of the use of adequate bioinks and printing methodologies. Here, a hydrogel bioink based on gelatin (Gel) and nanofibrillated cellulose (NFC), cross-linked with genipin, was developed for the 3D extrusion-based bioprinting of hepatocarcinoma cells (HepG2). This formulation combines the biological characteristics of Gel with the exceptional mechanical and rheological attributes of NFC. Gel/NFC ink formulations with different Gel/NFC mass compositions, viz., 90:10, 80:20, 70:30, and 60:40, were prepared and characterized. The corresponding cross-linked hydrogels were obtained using 1.5% (w/w) genipin as the cross-linking agent. The rheological and mechanical performances of the inks were enhanced by the addition of NFC, as evidenced by the rise in the yield stress from 70.9 ± 28.6 to 627.9 ± 74.8 Pa, compressive stress at 80% strain from 0.5 ± 0.1 to 1.5 ± 0.2 MPa, and Young's modulus from 4.7 ± 0.9 to 12.1 ± 1.1 MPa, for 90:10 and 60:40 inks, respectively. Moreover, higher NFC contents translated into 3D structures with better shape fidelity and the possibility of printing more intricate structures. These hydrogels were noncytotoxic toward HepG2 cells for up to 48 h, with cell viabilities consistently above 80%. The ink 70:30 was loaded with HepG2 cells (2 × 10 cells mL) and bioprinted. Cell viability remained elevated (90 ± 4%) until day 14 postbioprinting, with cells maintaining their metabolic activity shown by H NMR metabolomics, proving the enormous potential of Gel/NFC-based bioinks for bioprinting HepG2 cells without jeopardizing their viability and metabolism. |
| title | Biofabrication of HepG2 Cells-Laden 3D Structures Using Nanocellulose-Reinforced Gelatin-Based Hydrogel Bioinks: Materials Characterization, Cell Viability Assessment, and Metabolomic Analysis. |
| topic | Humans Hep G2 Cells Gelatin Cellulose Cell Survival Hydrogels Bioprinting Ink Printing, Three-Dimensional Iridoids Metabolomics Rheology |
| url | https://pubmed.ncbi.nlm.nih.gov/40241282/ |