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. 2022 Feb 23:10:e12897.
doi: 10.7717/peerj.12897. eCollection 2022.

Synovial fluid proteome profile of surgical versus chemical induced osteoarthritis in rabbits

Sharifah Zakiah Syed Sulaiman  1 , Wei Miao Tan  1 , Rozanaliza Radzi  1 , Intan Nur Fatiha Shafie  1 , Mokrish Ajat  2 , Rozaihan Mansor  3 , Suhaila Mohamed  4 , Norasfaliza Rahmad  5 , Angela Min Hwei Ng  6 , Seng Fong Lau  1   4
Affiliations

Synovial fluid proteome profile of surgical versus chemical induced osteoarthritis in rabbits

Sharifah Zakiah Syed Sulaiman et al. PeerJ. .

Abstract

Background: Animal models are significant for understanding human osteoarthritis (OA). This study compared the synovial fluid proteomics changes in surgical and chemical induced OA models.

Methods: Thirty rabbits either had anterior cruciate ligament transection (ACLT) procedure or injected intra-articularly with monosodium iodoacetate (MIA, 8 mg) into the right knee. The joints were anatomically assessed, and the synovial fluid proteins analyzed using two-dimensional polyacrylamide gel electrophoresis (2DGE) and MALDI TOF/TOF mass spectrometry analysis at 4, 8 and 12 weeks. The proteins' upregulation and downregulation were compared with control healthy knees.

Results: Seven proteins (histidine-rich glycoprotein, beta-actin-like protein 2 isoform X1, retinol-binding protein-4, alpha-1-antiproteinase, gelsolin isoform, serotransferrin, immunoglobulin kappa-b4 chain-C-region) were significantly expressed by the surgical induction. They characterized cellular process (27%), organization of cellular components or biogenesis (27%), localization (27%) and biological regulation (18%), which related to synovitis, increased cellularity, and subsequently cartilage damage. Three proteins (apolipoprotein I-IV precursor, serpin peptidase inhibitor and haptoglobin precursor) were significantly modified by the chemical induction. They characterized stimulus responses (23%), immune responses (15%), biological regulations (15%), metabolism (15%), organization of cellular components or biogenesis (8%), cellular process (8%), biological adhesions (8%) and localization (8%), which related to chondrocytes glycolysis/death, neovascularization, subchondral bone necrosis/collapse and inflammation.

Conclusions: The surgical induced OA model showed a wider range of protein changes, which were most upregulated at week 12. The biological process proteins expressions showed the chemical induced joints had slower OA progression compared to surgical induced joints. The chemical induced OA joints showed early inflammatory changes, which later decreased.

Keywords: Chemical; MALDI TOF/TOF mass spectrometry; Osteoarthritis; Proteomics; Rabbits; Surgical; Synovial fluid; Two-dimensional polyacrylamide gel electrophoresis.

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Conflict of interest statement

The authors declare there are no competing interests.

Figures

Figure 1
Figure 1. Definition of tissue compartment in subchondral bone.
White square dotted line, cancellous bone (Cn. B), yellow round dotted line, bone marrow (BM). Calculation of BV/TV: Bone volume (BV/TV, %) = (Cn.B region –BM regions)*/Cn.B region ×100, * Cn.B region –BM regions = Tb.B regions.
Figure 2
Figure 2. OARSI histopathological scores for (A) surgically induced femoral cartilage (B) chemically induced femoral cartilage (C) surgically induced tibial cartilage (D) chemically induced tibial cartilage Values are presented in median with 95% confidence interval (n = 5).
Significant difference (Dunn’s multiple comparisons, p < 0.05) between column with the same letter within the groups.
Figure 3
Figure 3. Representative histology images of femoral articular cartilage.
(A–D) Surgically induced group. (E–H) Chemically induced group. Safranin-O/fast green stained with 20×magnification. Surgically induced group showed apoptosis indicated by empty chondrons (red arrow) during week 4 and superficial fibrillation (red asterisk) during week 8. Chemically induced group showed surface erosion as early as week 8 (red arrowhead).
Figure 4
Figure 4. Representative histology images of tibial articular cartilage (A–D) surgically induced group. (E–H) chemically induced group. Safranin-O/fast green stained with 20x magnification.
During week 4, surgically induced group showed chondrocyte apoptosis (red arrow) and fibrillation (red asterisk) during week 8. Cartilage erosion (red arrowhead) were present during week 12. For chemically induced group, erosion can be observed during week 8 and week 12.
Figure 5
Figure 5. Subchondral bone BV/TV values for (A) femur and (B) tibia in week 4, week 8 and week 12 for the control, surgically (ACLT) and chemically (MIA) induced group.
No significant difference between groups were reported. The results are represented as mean (±SD).
Figure 6
Figure 6. Representative 2DGE (pI 3-10) image for rabbit synovial fluid in (A) surgically induced group and (B) chemically induced group after staining with colloidal Coomassie stain.
Gels from respective groups were scanned and analyzed with Progenesis SameSpot software and protein spots (circled in red) were selected to be picked.
See this image and copyright information in PMC

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