Regenerative Potential of Platelet Concentrate Lysate in Mechanically Injured Cartilage and Matrix-Associated Chondrocyte Implantation In Vitro
Abstract
:1. Introduction
2. Results
2.1. Apoptosis in Mechanically Injured Cartilage
2.2. Alterations of the Chondrogenic Phenotype in Mechanically Injured Chondrocytes
2.3. Post-Injurious Changes of Extracellular Matrix Compounds in Human Cartilage Explants
2.4. Chondrogenic Potential of IL-10 and PC Treatment in Cellularized ACI Grafts
3. Discussion
4. Materials and Methods
4.1. Preparation of PC
4.2. Isolation of Articular Cartilage Explants and Preparation of Human Chondrocyte-Laden Collagen Scaffolds
4.3. Injurious Compression and Treatment with PC and IL-10
4.4. DNA Quantification and Glycosaminoglycan Synthesis
4.5. Relative Quantification PCR (qPCR)
4.6. Histology—Detection of Cell Death, Toluidine Blue Staining and Immunohistochemistry
4.7. Statistics
5. Conclusions
Author Contributions
Funding
Institutional Review Board Statement
Informed Consent Statement
Data Availability Statement
Acknowledgments
Conflicts of Interest
References
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COL2A1 | Mean | ±SD # | p† | |
a | +IL-10 | 1.24 | 0.65 | ** e, ** f |
b | +PC | 2.1 | 1.4 | ** e, * f |
c | injury | 1.53 | 0.9 | ** e, * f |
d | injury + IL-10 | 3.74 | 2.19 | ns |
e | injury + PC | 7.56 | 5.5 | ns |
f | injury + IL-10 + PC | 7.44 | 3.91 | ns |
ACAN | Mean | ±SD | p | |
a | +IL-10 | 1.14 | 0.64 | ** e, * f |
b | +PC | 2.74 | 2 | * e |
c | injury | 2.13 | 1.34 | ** e |
d | injury + IL-10 | 4.44 | 2.68 | ns |
e | injury + PC | 7.22 | 5 | ns |
f | injury + IL-10 + PC | 5.88 | 2.87 | ns |
SOX9 | Mean | ±SD | p | |
a | +IL-10 | 2.21 | 0.81 | ns |
b | +PC | 5.88 | 5.79 | ** c |
c | injury | 0.63 | 0.37 | ns |
d | injury + IL-10 | 1.94 | 1.74 | ns |
e | injury + PC | 4.24 | 2.8 | ns |
f | injury + IL-10 + PC | 2.33 | 0.97 | ns |
COL1A1 | Mean | ±SD | p | |
a | +IL-10 | 1.02 | 1.3 | * c |
b | +PC | 1.88 | 1.59 | ns |
c | injury | 8.89 | 9.07 | ns |
d | injury + IL-10 | 4.19 | 4.42 | ns |
e | injury + PC | 4.53 | 5.53 | ns |
f | injury + IL-10 + PC | 2.03 | 2.01 | ns |
COL10A1 | Mean | ±SD | p | |
a | +IL-10 | 0.61 | 0.51 | **** c |
b | +PC | 0.9 | 0.58 | **** c |
c | injury | 2.84 | 1.38 | ns |
d | injury + IL-10 | 0.59 | 0.43 | **** c |
e | injury + PC | 0.67 | 0.62 | **** c |
f | injury + IL-10 + PC | 0.25 | 0.36 | **** c |
COL2/1 ratio | Mean | ±SD | p | |
a | +IL-10 | 1.98 | 2.27 | ns |
b | +PC | 1.87 | 1.48 | ns |
c | injury | 0.78 | 0.82 | ns |
d | injury + IL-10 | 3.9 | 4.05 | ns |
e | injury + PC | 2.35 | 2.71 | ns |
f | injury + IL-10 + PC | 2.95 | 2.34 | ns |
COL2A1 | Mean | ±SD # | p † | |
a | +IL-10 | 0.82 | 0.24 | ** c |
b | +PC | 1.09 | 0.16 | * c |
c | +IL-10 + PC | 1.66 | 0.29 | ns |
ACAN | Mean | ±SD | p | |
a | +IL-10 | 2.8 | 0.66 | ns |
b | +PC | 2.78 | 1.49 | ns |
c | +IL-10 + PC | 2.41 | 1.34 | ns |
COL1A1 | Mean | ±SD | p | |
a | +IL-10 | 0.81 | 0.21 | ns |
b | +PC | 1.04 | 0.53 | ** c |
c | +IL-10 + PC | 0.31 | 0.1 | ns |
COL10A1 | Mean | ±SD | p | |
a | +IL-10 | 1.6 | 0.66 | ns |
b | +PC | 2.12 | 1.49 | ns |
c | +IL-10 + PC | 1.04 | 1.34 | ns |
COL2/1 ratio | Mean | ±SD | p | |
a | +IL-10 | 0.94 | 0.12 | ** c |
b | +PC | 1.3 | 0.8 | ** c |
c | +IL-10 + PC | 5.91 | 1.77 | ns |
Human Target | Sequence (5′–3′) Sense | Sequence (5′–3′) Antisense |
---|---|---|
ACAN | GAGGCCAGCAGAGAAGATTCTG | GACGCCTCGCCTTCTTGAA |
COL2A1 | CAACACTGCCAACGTCCAGAT | CTGCTTCGTCCAGATAGGCAAT |
SOX9 | CTCGGAGACTTCTGAACGAGAG | CGTTCTTCACCGACTTCCTCC |
COL1A1 COL10A1 | AATTCCAAGGCCAAGAAGCATG CCCTTTTTGCTGCTAGTATCCTTGA | GGTAGCCATTTCCTTGGTGGTT AACTGTGTCTTGGTGTTGGGTAGTG |
GAPDH | GCCTCAAGATCATCAGCAATGC | TGGTCATGAGTCCTTCCACGAT |
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Weitkamp, J.-T.; Rolauffs, B.; Feldheim, M.; Bayer, A.; Lippross, S.; Weuster, M.; Smeets, R.; Naujokat, H.; Grodzinsky, A.J.; Kurz, B.; et al. Regenerative Potential of Platelet Concentrate Lysate in Mechanically Injured Cartilage and Matrix-Associated Chondrocyte Implantation In Vitro. Int. J. Mol. Sci. 2021, 22, 13179. https://doi.org/10.3390/ijms222413179
Weitkamp J-T, Rolauffs B, Feldheim M, Bayer A, Lippross S, Weuster M, Smeets R, Naujokat H, Grodzinsky AJ, Kurz B, et al. Regenerative Potential of Platelet Concentrate Lysate in Mechanically Injured Cartilage and Matrix-Associated Chondrocyte Implantation In Vitro. International Journal of Molecular Sciences. 2021; 22(24):13179. https://doi.org/10.3390/ijms222413179
Chicago/Turabian StyleWeitkamp, Jan-Tobias, Bernd Rolauffs, Moritz Feldheim, Andreas Bayer, Sebastian Lippross, Matthias Weuster, Ralf Smeets, Hendrik Naujokat, Alan Jay Grodzinsky, Bodo Kurz, and et al. 2021. "Regenerative Potential of Platelet Concentrate Lysate in Mechanically Injured Cartilage and Matrix-Associated Chondrocyte Implantation In Vitro" International Journal of Molecular Sciences 22, no. 24: 13179. https://doi.org/10.3390/ijms222413179