A Comparison of 3 Bioinks for 3D Bioprinting of Articular Cartilage

2021 ◽  
Author(s):  
Christel Henrionnet ◽  
Océane Messaoudi ◽  
Léa Pourchet ◽  
Pierre Gillet ◽  
Damien Loeuille ◽  
...  
Keyword(s):  
Collagen Synthesis ◽  
Type Ii Collagen ◽  
Type Ii ◽  
3D Bioprinting ◽  
Matrix Synthesis ◽  
Cartilage Engineering ◽  
Low Viscosity ◽  
Promising Tool ◽  
Marrow Mesenchymal Stem Cells ◽  
Hematoxylin Eosin

Background: 3D printing has become a promising tool for cartilage engineering, combining 3D deposition of cells seeded in supporting biomaterials. Objective: Our goal was to evaluate the chondrogenic properties of three different bioinks, seeded with human bone marrow mesenchymal stem cells (bMSCs). Methods: The three different tested bioinks are seeded with 2 × 106 cells/mL bMSCs. The bioink#1 is composed of gelatin, fibrinogen, and very low viscosity alginate. The bioink#2 has the same composition, excepted for the alginate that is a low viscosity one. The bioink#3 is manufactured by CELLINK®. The cartilaginous substitutes were cultivated for 28 days in the presence of ITS vs TGF-ß1. The extracellular matrix synthesis is evaluated at D28 by histology (Hematoxylin-Eosin-Saffron & Alcian Blue) and immunostaining (type II collagen). Results: The bioink#1 better promoted type II collagen synthesis, although the three bioink were equipotent in terms of proteoglycan content. Despite its universal characteristics, the bioink#3 failed to encourage the hyaline-like matrix synthesis. Conclusion: The bioink#1 containing gelatin, fibrinogen, and very low viscosity seems to be the fittest of the three bio-inks to obtain a cartilaginous substitute presenting a remarkable matrix synthesis. This study confirms the importance of the choice of bioink for cartilage engineering.

scholarly journals Combining Innovative Bioink and Low Cell Density for the Production of 3D-Bioprinted Cartilage Substitutes: A Pilot Study

2020 ◽  
Vol 2020 ◽  
pp. 1-16 ◽  
Author(s):  
Christel Henrionnet ◽  
Léa Pourchet ◽  
Paul Neybecker ◽  
Océane Messaoudi ◽  
Pierre Gillet ◽  
...  
Keyword(s):  
Gene Expression ◽  
Extracellular Matrix ◽  
Cell Encapsulation ◽  
Type Ii Collagen ◽  
Mitochondrial Activity ◽  
Type Ii ◽  
3D Bioprinting ◽  
Matrix Synthesis ◽  
M Cell ◽  
Low Concentration

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.

scholarly journals Levels of matrix metalloproteinase-2 in committed differentiation of bone marrow mesenchymal stem cells induced by kartogenin

2019 ◽  
Vol 47 (7) ◽  
pp. 3261-3270
Author(s):  
Cheng Wang ◽  
Qiaohui Liu ◽  
Xiaoyuan Ma ◽  
Guofeng Dai
Keyword(s):  
Stem Cells ◽  
Bone Marrow ◽  
Mesenchymal Stem Cells ◽  
Matrix Metalloproteinase ◽  
Type Ii Collagen ◽  
Matrix Metalloproteinase 2 ◽  
Type Ii ◽  
Proliferation And Differentiation ◽  
Marrow Mesenchymal Stem Cells

Objective To measure the inductive effect of kartogenin on matrix metalloproteinase-2 levels during the differentiation of human bone marrow mesenchymal stem cells (hMSCs) into chondrocytes in vitro. Methods In vitro cultured bone marrow hMSCs were grown to the logarithmic phase and then divided into three groups: control group (0 µM kartogenin), 1 µM kartogenin group and 10 µM kartogenin group. After 72 h of culture, cell proliferation and differentiation were observed microscopically. Matrix metalloproteinase-2 (MMP-2) in the cell supernatant and type II collagen levels in the cells were detected by enzyme linked immunosorbent assay and immunofluorescence staining, respectively. Results Kartogenin induced the proliferation and differentiation of hMSCs. With the increase of kartogenin concentration, the level of type II collagen was increased, while the level of MMP-2 decreased. Conclusion These findings indicate that kartogenin can induce hMSCs to differentiate into chondrocytes, and with the increase of kartogenin concentration, degeneration of the cartilage extracellular matrix may be inhibited.

Experimental Study on Construction of Tissue Engineered Bone by Autologous Oxygen Release Nano-Bionic Scaffold Combined with Bone Morphogenetic Protein-2 Induced Bone Marrow Mesenchymal Stem Cells

2019 ◽  
Vol 9 (9) ◽  
pp. 1254-1260
Author(s):  
Fei Wang ◽  
Hongfang Wei ◽  
Chengdong Hu ◽  
Dongfeng Li ◽  
Xiwei Huo ◽  
...  
Keyword(s):  
Bone Marrow ◽  
Type Ii Collagen ◽  
Negative Control Group ◽  
Negative Control ◽  
Control Group ◽  
Type Ii ◽  
Oxygen Release ◽  
Fracture Group ◽  
Bionic Scaffold ◽  
Marrow Mesenchymal Stem Cells

Bone marrow mesenchymal stem cells (BMSCs) are used for bone tissue engineering. BMP-2 and autologous oxygen-releasing nano-bionic scaffolds promote bone differentiation of BMSCs. Our study intends to evaluate the role of autologous oxygen-releasing nano-bionic scaffolds combined with BMP-2-induced BMSCs in the construction of tissue engineered bone. Rat BMSCs were isolated and transfected with NC (negative control group) and BMP-2 (BMP-2 plasmid group), respectively. Healthy male SD rats were randomly and equally divided into fracture group, negative control group and the BMP-2 group which was implanted with autologous oxygen-releasing nano-bionic scaffolds to synthesize BMSCs and transfected with BMP-2 plasmids respectively followed by analysis of osteophytes growth, ALP activity, expression of BMP-2, type II collagen, Runx2 and OC by real time PCR, TGF-β1 secretion by ELISA and BMP-2 protein expression by western blot. BMSCs induced by autologous oxygen release nano-bionic scaffold combined with BMP-2 can significantly promote the increase of bone mineral density, increase the expression of Runx2 and OC, promote ALP activity, upregulate type II collagen, BMP-2 mRNA and protein, and TGF-β1 secretion compared to fracture group (P < 0.05). The BMSCs induced by autologous oxygen-releasing nanobionic scaffolds transfected with BMP-2 had a more significant effect on bone repair. Autologous oxygen-releasing nano-bionic scaffolds combined with BMP-2-induced BMSCs can promote bone healing by regulating BMP-2 and increasing osteogenesis at the bone defect.

Mir-149 Promotes Survival and Differentiation of Bone Marrow Mesenchymal Stem Cells by Regulating Interleukin-6/Signal Transducer and Activator of Transcription 3 Signaling Pathway

2019 ◽  
Vol 9 (8) ◽  
pp. 1160-1166
Author(s):  
Guozhong Qin ◽  
Shaochuan Huo ◽  
Juehui Li ◽  
Yin Lian ◽  
Xiaoli Jin
Keyword(s):  
Bone Marrow ◽  
Signaling Pathway ◽  
Caspase 3 ◽  
Type Ii Collagen ◽  
Type Ii ◽  
Alp Activity ◽  
Stat3 Signaling ◽  
Stat3 Signaling Pathway ◽  
Marrow Mesenchymal Stem Cells ◽  
Sirna Group

Bone marrow mesenchymal stem cells (BMSCs) can self-renew with multi-directional differentiation. Mir-149 is involved in various diseases, but whether Mir-149 regulates the survival and differentiation of BMSCs and related mechanisms remains unclear. BMSCs were isolated and randomly divided into Si-NC group, Mir-149 siRNA group, and Mir-149 siRNA + STAT3 inhibitor WP1066 group followed by analysis of the expression of Mir-149, RUNX2 and OPN mRNA by real time PCR, BMSCs proliferation by MTT assay, Caspase 3 activity, ALP activity, formation of type II collagen and IL-6 level by ELISA, as well as STAT3 signaling pathway expression by Western blot. Mir-149 expression was reduced in BMSCs of Mir-149 siRNA group, with promoted survival of BMSCs, decreased Caspase 3 activity, increased expression of RUNX2 and OPN, type II collagen formation, ALP activity, IL-6 secretion, as well as elevated pSTAT3 phosphorylation. The differences were statistically significant compared to Si-NC group (P < 0.05). Mir-149 siRNA + WP1066 inhibited pSTAT3 phosphorylation, reduced BMSCs survival, increased Caspase 3 activity, decreased RUNX2 and OPN expression, type II collagen production, ALP activity, as well as reduced IL-6 secretion. Compared with Mir-149 siRNA group, there were significant differences (P < 0.05). Down-regulation of Mir-149 in BMSCs can promote BMSCs survival and osteogenic differentiation by regulating IL-6/STAT3 signaling pathway.

scholarly journals Uncoupling of type II collagen synthesis and degradation predicts progression of joint damage in patients with knee osteoarthritis

2002 ◽  
Vol 46 (10) ◽  
pp. 2613-2624 ◽  
Author(s):  
Patrick Garnero ◽  
Xavier Ayral ◽  
Jean-Charles Rousseau ◽  
S. Christgau ◽  
Linda J. Sandell ◽  
...  
Keyword(s):  
Knee Osteoarthritis ◽  
Collagen Synthesis ◽  
Joint Damage ◽  
Type Ii Collagen ◽  
Type Ii ◽  
Synthesis And Degradation
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