Combining Innovative Bioink and Low Cell Density for the Production of 3D-Bioprinted Cartilage Substitutes: A Pilot Study
3D bioprinting offers interesting opportunities for 3D tissue printing by providing living cells with appropriate scaffolds with a dedicated structure. Biological advances in bioinks are currently promising for cell encapsulation, particularly that of mesenchymal stem cells (MSCs). We present herein...
Saved in:
| Main Authors: | , , , , , , , , |
|---|---|
| Format: | Article |
| Language: | English |
| Published: |
Wiley
2020-01-01
|
| Series: | Stem Cells International |
| Online Access: | http://dx.doi.org/10.1155/2020/2487072 |
| Tags: |
Add Tag
No Tags, Be the first to tag this record!
|
| _version_ | 1832567937588789248 |
|---|---|
| author | Christel Henrionnet Léa Pourchet Paul Neybecker Océane Messaoudi Pierre Gillet Damien Loeuille Didier Mainard Christophe Marquette Astrid Pinzano |
| author_facet | Christel Henrionnet Léa Pourchet Paul Neybecker Océane Messaoudi Pierre Gillet Damien Loeuille Didier Mainard Christophe Marquette Astrid Pinzano |
| author_sort | Christel Henrionnet |
| collection | DOAJ |
| description | 3D bioprinting offers interesting opportunities for 3D tissue printing by providing living cells with appropriate scaffolds with a dedicated structure. Biological advances in bioinks are currently promising for cell encapsulation, particularly that of mesenchymal stem cells (MSCs). We present herein the development of cartilage implants by 3D bioprinting that deliver MSCs encapsulated in an original bioink at low concentration. 3D-bioprinted constructs (10×10×4 mm) were printed using alginate/gelatin/fibrinogen bioink mixed with human bone marrow MSCs. The influence of the bioprinting process and chondrogenic differentiation on MSC metabolism, gene profiles, and extracellular matrix (ECM) production at two different MSC concentrations (1 million or 2 million cells/mL) was assessed on day 28 (D28) by using MTT tests, real-time RT-PCR, and histology and immunohistochemistry, respectively. Then, the effect of the environment (growth factors such as TGF-β1/3 and/or BMP2 and oxygen tension) on chondrogenicity was evaluated at a 1 M cell/mL concentration on D28 and D56 by measuring mitochondrial activity, chondrogenic gene expression, and the quality of cartilaginous matrix synthesis. We confirmed the safety of bioextrusion and gelation at concentrations of 1 million and 2 million MSC/mL in terms of cellular metabolism. The chondrogenic effect of TGF-β1 was verified within the substitute on D28 by measuring chondrogenic gene expression and ECM synthesis (glycosaminoglycans and type II collagen) on D28. The 1 M concentration represented the best compromise. We then evaluated the influence of various environmental factors on the substitutes on D28 (differentiation) and D56 (synthesis). Chondrogenic gene expression was maximal on D28 under the influence of TGF-β1 or TGF-β3 either alone or in combination with BMP-2. Hypoxia suppressed the expression of hypertrophic and osteogenic genes. ECM synthesis was maximal on D56 for both glycosaminoglycans and type II collagen, particularly in the presence of a combination of TGF-β1 and BMP-2. Continuous hypoxia did not influence matrix synthesis but significantly reduced the appearance of microcalcifications within the extracellular matrix. The described strategy is very promising for 3D bioprinting by the bioextrusion of an original bioink containing a low concentration of MSCs followed by the culture of the substitutes in hypoxic conditions under the combined influence of TGF-β1 and BMP-2. |
| format | Article |
| id | doaj-art-5f35686706ee4f65b86ee0b6544e873f |
| institution | Kabale University |
| issn | 1687-966X 1687-9678 |
| language | English |
| publishDate | 2020-01-01 |
| publisher | Wiley |
| record_format | Article |
| series | Stem Cells International |
| spelling | doaj-art-5f35686706ee4f65b86ee0b6544e873f2025-02-03T01:00:05ZengWileyStem Cells International1687-966X1687-96782020-01-01202010.1155/2020/24870722487072Combining Innovative Bioink and Low Cell Density for the Production of 3D-Bioprinted Cartilage Substitutes: A Pilot StudyChristel Henrionnet0Léa Pourchet1Paul Neybecker2Océane Messaoudi3Pierre Gillet4Damien Loeuille5Didier Mainard6Christophe Marquette7Astrid Pinzano8UMR 7365 CNRS-Université de Lorraine IMoPA, Biopôle de l’Université de Lorraine, Campus Brabois-Santé, 9, Avenue de la Forêt de Haye, BP20199, 54505 Vandœuvre-Lès-Nancy, FrancePlatform 3D Fab, University of Lyon, CNRS, INSA, CPE-Lyon, ICBMS, UMR 5246, 43, Bd du 11 Novembre 1918, F69622 Villeurbanne Cedex, FranceUMR 7365 CNRS-Université de Lorraine IMoPA, Biopôle de l’Université de Lorraine, Campus Brabois-Santé, 9, Avenue de la Forêt de Haye, BP20199, 54505 Vandœuvre-Lès-Nancy, FranceUMR 7365 CNRS-Université de Lorraine IMoPA, Biopôle de l’Université de Lorraine, Campus Brabois-Santé, 9, Avenue de la Forêt de Haye, BP20199, 54505 Vandœuvre-Lès-Nancy, FranceUMR 7365 CNRS-Université de Lorraine IMoPA, Biopôle de l’Université de Lorraine, Campus Brabois-Santé, 9, Avenue de la Forêt de Haye, BP20199, 54505 Vandœuvre-Lès-Nancy, FranceUMR 7365 CNRS-Université de Lorraine IMoPA, Biopôle de l’Université de Lorraine, Campus Brabois-Santé, 9, Avenue de la Forêt de Haye, BP20199, 54505 Vandœuvre-Lès-Nancy, FranceUMR 7365 CNRS-Université de Lorraine IMoPA, Biopôle de l’Université de Lorraine, Campus Brabois-Santé, 9, Avenue de la Forêt de Haye, BP20199, 54505 Vandœuvre-Lès-Nancy, FrancePlatform 3D Fab, University of Lyon, CNRS, INSA, CPE-Lyon, ICBMS, UMR 5246, 43, Bd du 11 Novembre 1918, F69622 Villeurbanne Cedex, FranceUMR 7365 CNRS-Université de Lorraine IMoPA, Biopôle de l’Université de Lorraine, Campus Brabois-Santé, 9, Avenue de la Forêt de Haye, BP20199, 54505 Vandœuvre-Lès-Nancy, France3D bioprinting offers interesting opportunities for 3D tissue printing by providing living cells with appropriate scaffolds with a dedicated structure. Biological advances in bioinks are currently promising for cell encapsulation, particularly that of mesenchymal stem cells (MSCs). We present herein the development of cartilage implants by 3D bioprinting that deliver MSCs encapsulated in an original bioink at low concentration. 3D-bioprinted constructs (10×10×4 mm) were printed using alginate/gelatin/fibrinogen bioink mixed with human bone marrow MSCs. The influence of the bioprinting process and chondrogenic differentiation on MSC metabolism, gene profiles, and extracellular matrix (ECM) production at two different MSC concentrations (1 million or 2 million cells/mL) was assessed on day 28 (D28) by using MTT tests, real-time RT-PCR, and histology and immunohistochemistry, respectively. Then, the effect of the environment (growth factors such as TGF-β1/3 and/or BMP2 and oxygen tension) on chondrogenicity was evaluated at a 1 M cell/mL concentration on D28 and D56 by measuring mitochondrial activity, chondrogenic gene expression, and the quality of cartilaginous matrix synthesis. We confirmed the safety of bioextrusion and gelation at concentrations of 1 million and 2 million MSC/mL in terms of cellular metabolism. The chondrogenic effect of TGF-β1 was verified within the substitute on D28 by measuring chondrogenic gene expression and ECM synthesis (glycosaminoglycans and type II collagen) on D28. The 1 M concentration represented the best compromise. We then evaluated the influence of various environmental factors on the substitutes on D28 (differentiation) and D56 (synthesis). Chondrogenic gene expression was maximal on D28 under the influence of TGF-β1 or TGF-β3 either alone or in combination with BMP-2. Hypoxia suppressed the expression of hypertrophic and osteogenic genes. ECM synthesis was maximal on D56 for both glycosaminoglycans and type II collagen, particularly in the presence of a combination of TGF-β1 and BMP-2. Continuous hypoxia did not influence matrix synthesis but significantly reduced the appearance of microcalcifications within the extracellular matrix. The described strategy is very promising for 3D bioprinting by the bioextrusion of an original bioink containing a low concentration of MSCs followed by the culture of the substitutes in hypoxic conditions under the combined influence of TGF-β1 and BMP-2.http://dx.doi.org/10.1155/2020/2487072 |
| spellingShingle | Christel Henrionnet Léa Pourchet Paul Neybecker Océane Messaoudi Pierre Gillet Damien Loeuille Didier Mainard Christophe Marquette Astrid Pinzano Combining Innovative Bioink and Low Cell Density for the Production of 3D-Bioprinted Cartilage Substitutes: A Pilot Study Stem Cells International |
| title | Combining Innovative Bioink and Low Cell Density for the Production of 3D-Bioprinted Cartilage Substitutes: A Pilot Study |
| title_full | Combining Innovative Bioink and Low Cell Density for the Production of 3D-Bioprinted Cartilage Substitutes: A Pilot Study |
| title_fullStr | Combining Innovative Bioink and Low Cell Density for the Production of 3D-Bioprinted Cartilage Substitutes: A Pilot Study |
| title_full_unstemmed | Combining Innovative Bioink and Low Cell Density for the Production of 3D-Bioprinted Cartilage Substitutes: A Pilot Study |
| title_short | Combining Innovative Bioink and Low Cell Density for the Production of 3D-Bioprinted Cartilage Substitutes: A Pilot Study |
| title_sort | combining innovative bioink and low cell density for the production of 3d bioprinted cartilage substitutes a pilot study |
| url | http://dx.doi.org/10.1155/2020/2487072 |
| work_keys_str_mv | AT christelhenrionnet combininginnovativebioinkandlowcelldensityfortheproductionof3dbioprintedcartilagesubstitutesapilotstudy AT leapourchet combininginnovativebioinkandlowcelldensityfortheproductionof3dbioprintedcartilagesubstitutesapilotstudy AT paulneybecker combininginnovativebioinkandlowcelldensityfortheproductionof3dbioprintedcartilagesubstitutesapilotstudy AT oceanemessaoudi combininginnovativebioinkandlowcelldensityfortheproductionof3dbioprintedcartilagesubstitutesapilotstudy AT pierregillet combininginnovativebioinkandlowcelldensityfortheproductionof3dbioprintedcartilagesubstitutesapilotstudy AT damienloeuille combininginnovativebioinkandlowcelldensityfortheproductionof3dbioprintedcartilagesubstitutesapilotstudy AT didiermainard combininginnovativebioinkandlowcelldensityfortheproductionof3dbioprintedcartilagesubstitutesapilotstudy AT christophemarquette combininginnovativebioinkandlowcelldensityfortheproductionof3dbioprintedcartilagesubstitutesapilotstudy AT astridpinzano combininginnovativebioinkandlowcelldensityfortheproductionof3dbioprintedcartilagesubstitutesapilotstudy |