Viscoelastic Behavior of Embroidered Scaffolds for ACL Tissue Engineering Made of PLA and P(LA-CL) After In Vitro Degradation
Abstract
:1. Introduction
2. Results
2.1. Relaxation Behavior during Hydrolytic Degradation
2.2. Comparison of the Relaxation Behavior between Scaffolds and Native Lapine ACL Tissue
2.3. Ultimate Tensile Properties of Degraded Scaffolds after Preconditioning
2.4. Lapine ACL Cell Survival on Embroidered Scaffolds
2.5. Protein Expression of Lapine ACL Cells on the Scaffolds in Comparison to Native ACLs
2.6. Mechanical Properties of Cell-Seeded Scaffolds
3. Discussion
4. Materials and Methods
4.1. Scaffold Fabrication
4.2. Lapine Anterior Cruciate Ligament Cell Isolation by Explant Culture
4.3. Scaffold Seeding: Dynamical Culture
4.4. Scaffold Seeding: Spheroid Culture
4.5. Live-Death Assay
4.6. Immunofluorescence Analysis of Marker Expression
4.7. Mechanical Testing
5. Conclusions
Author Contributions
Funding
Acknowledgments
Conflicts of Interest
Abbreviations
2D | Two-dimensional |
3D | Three-dimensional |
ACL | Anterior cruciate ligament |
bico | Bi-component scaffold made of PLA as upper thread and P(LA-CL) as lower thread |
DAPI | 4′,6-diamidino-2-phenylindole |
DMEM | Dulbecco’s Modified Eagle’s |
ECM | Extracellular matrix |
EO | Ethylene-oxide gas sterilization |
FCS | Fetal calf serum |
FDA | Fluorescein diacetate |
moco | Mono-component scaffold made of PLA threads |
NZW | New Zealand White rabbit |
PBS | Phosphate buffered saline |
PFA | Paraformaldehyde |
PI | Propidium iodide |
PLA | Polylactic acid |
P(LA-CL) | Poly(lactic-co-ε-caprolactone) |
PT | Patellar tendon |
RT | Room temperature |
S | Stiffness in N/mm |
SD | Standard deviation |
TBS | TRIS buffered saline |
TE | Tissue engineering |
UTE | Ultimate tensile elongation in % |
UTL | Ultimate tensile load in N |
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Relaxation Time (s) | Force (N) at Different Degradation Measuring Points (Days) (Mean ± SD) | ||||||
---|---|---|---|---|---|---|---|
0 | 7 | 14 | 28 | 84 | 168 | ||
moco | 10 | 63.8 ± 14.2 | 67.8 ± 6.6 | 66.6 ± 5.4 | 72.4 ± 7.7 | 84.4 ± 7.9 | 79.3 ± 6.6 |
180 | 53.0 ± 11.5 | 58.0 ± 5.4 | 56.8 ± 4.7 | 61.4 ± 6.9 | 71.1 ± 6.4 | 66.5 ± 5.8 | |
300 | 51.2 ± 11.1 | 56.2 ± 5.2 | 55.2 ± 4.5 | 59.6 ± 6.6 | 68.8 ± 6.1 | 64.8 ± 5.4 | |
600 | 48.6 ± 10.7 | 53.9 ± 5.0 | 53.0 ± 4.4 | 57.1 ± 6.4 | 65.8 ± 5.9 | 61.7 ± 5.6 | |
bico | 10 | 67.8 ± 5.0 | 71.6 ± 5.8 | 73.6 ± 7.6 | 76.2 ± 6.8 | 88.1 ± 11.3 | 35.4 ± 8.7 |
180 | 54.5 ± 4.2 | 59.4 ± 5.0 | 61.1 ± 7.1 | 63.7 ± 6.2 | 75.5 ± 10.4 | 28.5 ± 7.5 | |
300 | 52.2 ± 4.0 | 57.5 ± 4.9 | 59.3 ± 6.9 | 61.8 ± 5.9 | 73.5 ± 10.1 | 27.4 ± 7.3 | |
600 | 49.2 ± 4.0 | 54.8 ± 4.6 | 56.7 ± 6.7 | 59.2 ± 5.8 | 70.7 ± 9.8 | 26.1 ± 7.1 |
moco | bico | ||||||
---|---|---|---|---|---|---|---|
UTL (N) | UTE (%) | S (N/mm) | UTL (N) | UTE (%) | S (N/mm) | ||
Group A | 0 d | 405.8 ± 12.1 | 53.6 ± 5.4 | 151.0 ± 23.0 | 334.4 ± 14.8 | 40.2 ± 3.0 | 196.0 ± 19.2 |
7 d | 394.6 ± 16.0 | 62.4 ± 2.6 | 170.8 ± 22.4 | 338.1 ± 6.0 | 55.1 ± 1.1 | 255.4 ± 18.6 | |
14 d | 409.3 ± 17.9 | 62.2 ± 3.5 | 202.2 ± 29.8 | 323.4 ± 17.5 | 53.5 ± 2.6 | 239.1 ± 31.5 | |
28 d | 388.1 ± 13.4 | 58.6 ± 5.6 | 210.7 ± 36.0 | 326.5 ± 12.9 | 54.9 ± 2.4 | 281.2 ± 9.2 | |
84 d | 360.4 ± 5.0 | 41.8 ± 1.8 | 224.8 ± 14.3 | 290.6 ± 15.0 | 43.4 ± 2.3 | 206.9 ± 34.5 | |
168 d | 330.0 ± 9.6 | 44.0 ± 2.7 | 261.8 ± 27.9 | 232.6 ± 36.5 | 46.8 ± 5.5 | 259.8 ± 70.5 | |
Group B | 0 d | 401.2 ± 19.8 | 50.4 ± 7.2 | 160.2 ± 28.8 | 333.1 ± 7.7 | 39.4 ± 2.6 | 192.0 ± 8.6 |
7 d | 410.1 ± 15.3 | 61.6 ± 3.2 | 204.3 ± 17.1 | 329.4 ± 19.0 | 53.0 ± 3.9 | 240.7 ± 9.0 | |
14 d | 402.2 ± 13.6 | 61.2 ± 3.2 | 199.8 ± 19.6 | 319.2 ± 9.4 | 52.4 ± 2.3 | 246.8 ± 14.2 | |
28 d | 394.9 ± 8.7 | 58.4 ± 3.0 | 218.3 ± 21.9 | 323.0 ± 16.5 | 53.0 ± 2.6 | 254.7 ± 14.6 | |
84 d | 348.7 ± 6.8 | 43.8 ± 2.0 | 192.6 ± 11.8 | 295.9 ± 15.6 | 45.6 ± 2.1 | 231.3 ± 46.4 | |
168 d | 329.2 ± 29.8 | 44.8 ± 4.5 | 235.4 ± 26.7 | 210.9 ± 27.8 | 47.0 ± 5.8 | 114.7 ± 31.9 |
Test | Group A | Group B 1 |
---|---|---|
1 | | |
pause | 45 min | |
2 | | |
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Hahn, J.; Schulze-Tanzil, G.; Schröpfer, M.; Meyer, M.; Gögele, C.; Hoyer, M.; Spickenheuer, A.; Heinrich, G.; Breier, A. Viscoelastic Behavior of Embroidered Scaffolds for ACL Tissue Engineering Made of PLA and P(LA-CL) After In Vitro Degradation. Int. J. Mol. Sci. 2019, 20, 4655. https://doi.org/10.3390/ijms20184655
Hahn J, Schulze-Tanzil G, Schröpfer M, Meyer M, Gögele C, Hoyer M, Spickenheuer A, Heinrich G, Breier A. Viscoelastic Behavior of Embroidered Scaffolds for ACL Tissue Engineering Made of PLA and P(LA-CL) After In Vitro Degradation. International Journal of Molecular Sciences. 2019; 20(18):4655. https://doi.org/10.3390/ijms20184655
Chicago/Turabian StyleHahn, Judith, Gundula Schulze-Tanzil, Michaela Schröpfer, Michael Meyer, Clemens Gögele, Mariann Hoyer, Axel Spickenheuer, Gert Heinrich, and Annette Breier. 2019. "Viscoelastic Behavior of Embroidered Scaffolds for ACL Tissue Engineering Made of PLA and P(LA-CL) After In Vitro Degradation" International Journal of Molecular Sciences 20, no. 18: 4655. https://doi.org/10.3390/ijms20184655