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. 2024 Feb 3;11(1):21.
doi: 10.1186/s40643-024-00731-1.

Collagen type II solution extracted from supercritical carbon dioxide decellularized porcine cartilage: regenerative efficacy on post-traumatic osteoarthritis model

Srinivasan Periasamy  1 , Yun-Ju Chen  1 , Dur-Zong Hsu  2 , Dar-Jen Hsieh  3
Affiliations

Collagen type II solution extracted from supercritical carbon dioxide decellularized porcine cartilage: regenerative efficacy on post-traumatic osteoarthritis model

Srinivasan Periasamy et al. Bioresour Bioprocess. .

Abstract

Osteoarthritis (OA) of the knee is a common degenerative articular disorder and is one of the main causes of pain and functional disability. Cartilage damage is frequently linked to elevated osteoarthritis incidence. Supercritical carbon dioxide (scCO2) decellularized cartilage graft produced from the porcine cartilage is an ideal candidate for cartilage tissue engineering. In the present study, we derived collagen type II (Col II) solution from the scCO2 decellularized porcine cartilage graft (dPCG) and compared its efficacy with hyaluronic acid (HA) in the surgical medial meniscectomy (MNX) induced post-traumatic osteoarthritis (PTOA) model. Dose-dependent attenuation of the OA (12.3 ± 0.8) progression was observed in the intra-articular administration of Col II solution (7.3 ± 1.2) which significantly decreased the MNX-induced OA symptoms similar to HA. The pain of the OA group (37.4 ± 2.7) was attenuated dose-dependently by Col II solution (45.9 ± 4.1) similar to HA (43.1 ± 3.5) as evaluated by a capacitance meter. Micro-CT depicted a dose-dependent attenuation of articular cartilage damage by the Col II solution similar to HA treatment. A significant (p < 0.001) dose-dependent elevation in the bone volume was also observed in Col II solution-treated OA animals. The protective competence of Col II solution on articular cartilage damage is due to its significant (p < 0.001) increase in the expression of type II collagen, aggrecan and SOX-9 similar to HA. To conclude, intra-articular administration of type II collagen solution and HA reestablished the injured cartilage and decreased osteoarthritis progression in the experimental PTOA model.

Keywords: Decellularized porcine cartilage graft (dPCG); Medial meniscectomy (MNX); Osteoarthritis (OA); Supercritical carbon dioxide (scCO2); Type II collagen solution.

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Conflict of interest statement

The authors declare that they have no competing interests.

Figures

Fig. 1
Fig. 1
Experimental protocol of the MNX‐induced OA and the treatment schedule of different groups. The body weight changes were recorded 6 weeks after MNX surgery in rats. Data are means ± standard deviation (SD) (n = 6)
Fig. 2
Fig. 2
Effects of the Col II solution on knee pain in MNX-induced osteoarthritis in rats. Rats were divided into eight groups (n = 6). N group, Normal control; OA group, MNX alone; CS1 group, MNX + type II collagen (1 mg/ml); CS5 group, MNX + type II collagen (5 mg/ml); CS10 group, MNX + type II collagen (10 mg/ml); CCS group, MNX + cartilage + type II collagen; and HA group, MNX + Synvice-One Hyaln G-F 20 (8 mg/ml). Data are expressed as mean ± SD (n = 6). *p < 0.05 compared with N group; #p < 0.05 compared with OA group; †p < 0.05 compared with HA group (n = 6)
Fig. 3
Fig. 3
Effects of the Col II solution on cartilage and bone damage in MNX-induced osteoarthritis in rats by micro-CT image. Micro-CT images and coronal sagittal and axial images (a), bone volume (b) of the hind knee joint of sham and MNX-treated legs were generated by micro-CT 6 weeks after treatments. Data are expressed as mean ± SD (n = 6). The differences between treatments with different letters are statistically significant (p < 0.05)
Fig. 4
Fig. 4
Effects of the Col II solution on cartilage and bone damage in MNX-induced osteoarthritis in rats by histological analysis and scoring for cartilage repair in rat knee joints. The pathological changes in rat knee joints were examined by H&E stain (a), scoring (b) 6 weeks after MNX treatment (magnification: × 40). The red colour arrow indicates cartilage damage. The blue colour arrow indicates mild cartilage damage. The green and black colour arrow indicates cartilage damage preservation. Data are means ± standard deviation (SD) (n = 6). *p < 0.05 compared with N Group; #p < 0.05 compared with OA group; ##p < 0.001 compared with OA group
Fig. 5
Fig. 5
Effects of the Col II solution on cartilage and bone damage in MNX-induced osteoarthritis in rats by safranin-O stain histological analysis for cartilage repair in rat knee joints. The histological and pathological changes in rat knee joints were examined by safranin-O stain 6 weeks after MNX treatment (magnification: × 40). The red colour arrow indicates cartilage damage. The blue colour arrow indicates mild cartilage damage. The green colour arrow indicates cartilage damage preservation
Fig. 6
Fig. 6
Effect of Col II solution on cartilage collagen II expressions in MNX-induced rat OA. Immunostaining of collagen II expression (a), the quantification optical density of collagen II expression (b). Data are expressed as mean ± SD. *p < 0.05 compared with N group; #p < 0.05 compared with OA group; †p < 0.05 compared with HA group (N = 6)
Fig. 7
Fig. 7
Effects of Col II solution on cartilage aggrecan expression in MNX-induced rat OA. The immunostaining of aggrecan expression (a), the quantification optical density of aggrecan expression (b). Data are expressed as mean ± SD. *p < 0.05 compared with N group; #p < 0.05 compared with OA group (N = 6)
Fig. 8
Fig. 8
Effects of Col II solution on cartilage SOX9 expression in MNX-induced rat OA. The immunostaining of SOX‐9 expression (a), the quantification optical density of SOX‐9 expression (b). Data are expressed as mean ± SD. *p < 0.05 compared with N group; #p < 0.05 compared with OA group; †p < 0.05 compared with HA group (n = 6)
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