The Anti-Arthritic Activity of Diclofenac Lipid-Core Nanocapsules: Stereological Analysis Showing More Protection of Deep Joint Components
Abstract
:1. Introduction
2. Results
2.1. Preparation of DIC-LNC Dispersed in Water
2.2. DIC-LNC Reduces Edema Formation at the Hind Paws
2.3. DIC-LNC Reduces the Serum Levels of Proinflammatory Cytokines and CRP
2.4. Absence of Liver and Renal Toxicity with DIC-LNC
2.5. Cavalieri’s Volume of Metatarsophalangeal (MTP) Joints
2.6. Density of the Joint Components and Absolute Volume
2.7. Surface Area of MTP Joints
2.8. Quantification of Chondrocytes
3. Discussion
4. Materials and Methods
4.1. Materials
4.2. Preparation of the Lipid-Core Nanocapsules Formulations
4.3. Physicochemical Characterization of the Formulations
4.3.1. pH Measurements
4.3.2. Electrophoretic Mobility and Zeta Potential
4.3.3. Laser Diffraction
4.3.4. Dynamic Light Scattering
4.3.5. Nanoparticle Tracking Analysis
4.3.6. Drug Content and Encapsulation Efficiency
4.4. Animals
4.5. Induction of Arthritis by Freund’s Complete Adjuvant and Experimental Design
4.6. Randomization, Blinding, and Allocation Concealment
4.7. Edema Volume and Arthritis Score of the Hind Paw
4.8. Quantification of Cytokines and Biochemical Markers
4.9. Stereological Analysis
4.10. Determination of Cavalieri’s Volume
4.11. Determination of Relative Volume
4.12. Determination of Surface Area
4.13. Counting the Number of Cellular Profiles
4.14. Statistical Analysis
5. Conclusions
Author Contributions
Funding
Institutional Review Board Statement
Informed Consent Statement
Data Availability Statement
Conflicts of Interest
References
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LNC | DIC-LNC | |
---|---|---|
Potentiometry | ||
pH | 5.43 ± 0.24 | 5.39 ± 0.16 |
Laser diffraction | ||
D[4,3] (nm) | 153 ± 10 | 204 ± 46 |
Span | 1.4 ± 0.2 | 1.7 ± 0.1 |
Dynamic Light Scattering | ||
Dh (nm) | 170 ± 13 | 166 ± 13 |
PDI | 0.06 ± 0.02 | 0.08 ± 0.02 |
Electrophoretic mobility | ||
Zeta Potential (mV) | −13 ± 6 | −11 ± 2 |
Nanoparticle tracking analysis | ||
Dh (nm) | 182 ± 9 | 196 ± 14 |
D50 (nm) | 173 ± 13 | 186 ± 2 |
D90 (nm) | 257 ± 10 | 309 ± 9 |
PND (× 1012 particles mL−1) | 4.98 ± 0.25 | 4.76 ± 0.78 |
Volume (mm3) | ||||||
---|---|---|---|---|---|---|
Groups | Joint | Cartilage | Bone | Capsule | Synovial Space | Synovial Membrane |
Arthritis | 3.02 ± 0.08 | 0.50 ± 0.10 | 1.20 ± 0.05 | 0.97 ± 0.14 | 0.16 ± 0.01 | 0.08 ± 0.02 |
LNC | 2.80 ± 0.32 | 0.36 ± 0.08 | 1.08 ± 0.16 | 0.89 ± 0.15 | 0.17 ± 0.01 | 0.04 ± 0.01 |
Diclofenac | 2.46 ± 0.12 | 0.34 ± 0.03 | 1.02 ± 0.12 | 0.67 ± 0.02 | 0.19 ± 0.02 | 0.03 ± 0.00 |
Diclofenac-LNC | 2.10 ± 0.10 | 0.21 ± 0.01 | 0.75 ± 0.05 | 0.58 ± 0.02 | 0.26 ± 0.01 | 0.02 ± 0.00 |
No Arthritis | 1.95 ± 0.05 | 0.23 ± 0.03 | 0.61 ± 0.06 | 0.52 ± 0.03 | 0.41 ± 0.03 | 0.01 ± 0.05 |
Surface Area (mm2) | Cellular Count (Cell/mm2) | ||||
---|---|---|---|---|---|
Groups | Cartilage | Synovial Membrane | Chondrocytes | Isogenous Groups | Total Chondrocytes |
Arthritis | 38.25 ± 4.32 | 13.26 ± 2.78 | 1416 ± 126.3 | 80.55 ± 8.77 | 1517 ± 54.30 |
LNC | 29.09 ± 3.75 | 9.98 ± 1.49 | 1543 ± 49.92 | 111.5 ± 4.51 | 1701 ± 91.96 |
Diclofenac | 27.87 ± 3.31 | 9.92 ± 0.46 | 1627 ± 95.61 | 161.1 ± 11.84 | 1849 ± 16.73 |
Diclofenac-LNC | 20.45 ± 1.58 | 8.01 ± 0.19 | 1894 ± 47.00 | 226.8 ± 20.5 | 2095 ± 52.33 |
No Arthritis | 19.45 ± 0.28 | 7.43 ± 1.04 | 2208 ± 217.4 | 297.9 ± 19.5 | 2367 ± 225.8 |
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Ureña, N.M.; de Oliveira, C.P.; Guterres, S.S.; Pohlmann, A.R.; da Costa, O.T.F.; Boechat, A.L. The Anti-Arthritic Activity of Diclofenac Lipid-Core Nanocapsules: Stereological Analysis Showing More Protection of Deep Joint Components. Molecules 2023, 28, 5219. https://doi.org/10.3390/molecules28135219
Ureña NM, de Oliveira CP, Guterres SS, Pohlmann AR, da Costa OTF, Boechat AL. The Anti-Arthritic Activity of Diclofenac Lipid-Core Nanocapsules: Stereological Analysis Showing More Protection of Deep Joint Components. Molecules. 2023; 28(13):5219. https://doi.org/10.3390/molecules28135219
Chicago/Turabian StyleUreña, Nathalie Marte, Catiúscia Padilha de Oliveira, Silvia Stanisçuaski Guterres, Adriana Raffin Pohlmann, Oscar Tadeu Ferreira da Costa, and Antonio Luiz Boechat. 2023. "The Anti-Arthritic Activity of Diclofenac Lipid-Core Nanocapsules: Stereological Analysis Showing More Protection of Deep Joint Components" Molecules 28, no. 13: 5219. https://doi.org/10.3390/molecules28135219