Immunohistochemical Analysis of Knee Chondral Defect Repair after Autologous Particulated Cartilage and Platelet-Rich Plasma Treatment in Sheep
Abstract
:1. Introduction
2. Results
3. Discussion
4. Materials and Methods
4.1. Ethical Statement
4.2. Study Design and Surgical Procedure
4.3. Preparation and Use of Autologous PRP
4.4. Immunohistochemical Evaluation
4.5. Development of a Semiquantitative Scoring System
4.6. Statistical Analyses
5. Conclusions
Author Contributions
Funding
Institutional Review Board Statement
Informed Consent Statement
Data Availability Statement
Conflicts of Interest
Abbreviations
ECM | Extracellular matrix |
HA | Hyaluronic acid |
MCI | Minced cartilage implantation |
PACI | Particulated autograft cartilage implantation |
PACI + PRP | Therapeutic technique based on an autologous matrix composed of healthy hyaline cartilage chips included and mixed in a PRP clot and intraarticular infiltration of PRP |
PBS | Phosphate-buffered saline |
PRP | Platelet-rich plasma |
RLS | Ringer’s lactate solution |
RT | Room temperature |
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Month | Variable | Group | ||||
---|---|---|---|---|---|---|
HC | RLS | PACI + PRP (RLS) | HA | PACI + PRP (HA) | ||
9 | Col I | 6 ± 0 | 6 ± 2.04 | 6 ± 0.41 | 6 ± 1.33 | 6 ± 0 |
Col II | 6 ± 0 b | 5 ± 1.22 | 6 ± 1.22 | 4 ± 1.17 b | 5.50 ± 0.82 | |
Col III | 6 ± 0 | 4.50 ± 1.86 | 6 ± 1.67 | 6 ± 0.82 | 6 ± 2.04 | |
Col V | 6 ± 0 a | 2.50 ± 0.98 a | 4 ± 1.03 | 2.50 ± 1.79 | 4 ± 1.03 | |
Col VI | 6 ± 0 a,b | 3 ± 0.75 a | 4.50 ± 1.05 | 3.50 ± 0.82 b | 4.50 ± 1.21 | |
Col IX | 6 ± 0 | 3 ± 1.86 | 4 ± 1.22 | 3.50 ± 2.06 | 6 ± 1.03 | |
Col X | 6 ± 0 | 4.50 ± 1.37 | 6 ± 0.82 | 5 ± 1.17 | 6 ± 1.03 | |
Col XI | 6 ± 0 | 4 ± 1.17 | 4.50 ± 1.05 | 4.50 ± 1.67 | 4.50 ± 1.83 | |
Aggrecan | 6 ± 0 | 3.50 ± 1.87 | 3 ± 1.76 | 3.50 ± 1.26 | 4.50 ± 1.21 | |
18 | Col I | 6 ± 0 | 6 ± 2.04 | 6 ± 0 | 6 ± 2.27 | 6 ± 0 |
Col II | 6 ± 0 | 5 ± 1.03 | 6 ± 0.41 | 6 ± 1.25 | 6 ± 0.38 | |
Col III | 6 ± 0 | 3.50 ± 1.67 | 5.50 ± 1.27 | 3 ± 2.34 | 6 ± 1.53 | |
Col V | 6 ± 0 | 4.50 ± 0.82 | 6 ± 0.52 | 4 ± 1.90 | 6 ± 0.97 | |
Col VI | 6 ± 0 a,b | 4 ± 0.41 a | 6 ± 1.33 | 3 ± 1.21 b | 5 ± 0.75 | |
Col IX | 6 ± 0 b | 2 ± 2.34 | 4 ± 1.60 | 4 ± 1.99 b | 6 ± 0.75 | |
Col X | 6 ± 0 a | 3 ± 1.03 a | 4.5 ± 1.21 | 5 ± 1.60 | 6 ± 0.53 | |
Col XI | 6 ± 0 | 4.5 ± 1.05 | 5± 0.89 | 5 ± 0.90 | 5 ± 0.69 | |
Aggrecan | 6 ± 0 | 4.5 ± 1.47 | 5 ± 0.75 | 5 ± 1.38 | 5 ± 0.82 |
Target | Source | Reference | Host/Clonality | Dilution | Host Species |
---|---|---|---|---|---|
Collagen I | Abcam φ | ab34710 | Rabbit polyclonal | 1/200 | Rabbit |
Collagen II | Abcam φ | ab34712 | Rabbit polyclonal | 1/100 | Rabbit |
Collagen III | Abcam φ | ab7778 | Rabbit polyclonal | 1/200 | Rabbit |
Collagen V | Abcam φ | ab7046 | Rabbit polyclonal | 1/200 | Rabbit |
Collagen VI | Abcam φ | ab6588 | Rabbit polyclonal | 1/1000 | Rabbit |
Collagen IX | Abcam φ | ab134568 | Rabbit polyclonal | 1/200 | Rabbit |
Collagen X | Abcam φ | ab49945 | Mouse monoclonal | 1/500 | Mouse |
Collagen XI alpha 2 | Abcam φ | ab196613 | Rabbit polyclonal | 1/50 | Rabbit |
Aggrecan | Abcam φ | ab3778 | Mouse monoclonal | 1/100 | Mouse |
Parameters | Points | Final Score | |
---|---|---|---|
Immunostaining Pattern | Severely abnormal | 0 | 6 |
Abnormal | 1 | ||
Nearly normal | 2 | ||
Normal | 3 | ||
Immunostaining Intensity | Severely abnormal | 0 | |
Abnormal | 1 | ||
Nearly normal | 2 | ||
Normal | 3 |
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Alcaide-Ruggiero, L.; Molina-Hernández, V.; Morgaz, J.; Fernández-Sarmiento, J.A.; Granados, M.M.; Navarrete-Calvo, R.; Pérez, J.; Quirós-Carmona, S.; Carrillo, J.M.; Cugat, R.; et al. Immunohistochemical Analysis of Knee Chondral Defect Repair after Autologous Particulated Cartilage and Platelet-Rich Plasma Treatment in Sheep. Int. J. Mol. Sci. 2023, 24, 15157. https://doi.org/10.3390/ijms242015157
Alcaide-Ruggiero L, Molina-Hernández V, Morgaz J, Fernández-Sarmiento JA, Granados MM, Navarrete-Calvo R, Pérez J, Quirós-Carmona S, Carrillo JM, Cugat R, et al. Immunohistochemical Analysis of Knee Chondral Defect Repair after Autologous Particulated Cartilage and Platelet-Rich Plasma Treatment in Sheep. International Journal of Molecular Sciences. 2023; 24(20):15157. https://doi.org/10.3390/ijms242015157
Chicago/Turabian StyleAlcaide-Ruggiero, Lourdes, Verónica Molina-Hernández, Juan Morgaz, J. Andrés Fernández-Sarmiento, María M. Granados, Rocío Navarrete-Calvo, José Pérez, Setefilla Quirós-Carmona, José M. Carrillo, Ramón Cugat, and et al. 2023. "Immunohistochemical Analysis of Knee Chondral Defect Repair after Autologous Particulated Cartilage and Platelet-Rich Plasma Treatment in Sheep" International Journal of Molecular Sciences 24, no. 20: 15157. https://doi.org/10.3390/ijms242015157