Anti aggrecan

The samples were washed with PBS and fixed with 4% paraformaldehyde at 4°C for 30 minutes. The scaffolds were permeated with 0.5% Triton X‐100 for 30 minutes, sealed with 5% BSA (Invitrogen) for 1 hour, and then incubated with the primary antibodies anti‐aggrecan (Affinity Biosciences, USA) and anti‐Col II (Affinity Biosciences, USA) at 4°C overnight. Next, the primary antibodies were collected and the scaffolds were washed with PBS. Next, the scaffolds were incubated with Alexa Fluor 488‐conjugated Goat anti‐rabbit (Proteintech, USA) and Cy3‐conjugated Goat anti‐rabbit (Proteintech, USA) for 1 hour. Finally, the scaffolds were stained with DAPI for 15 minutes and were observed and photographed using LSCM.

Gibco-BRL (Waltham, MA, USA) provided Dulbecco's Modified Eagle Medium Nutrient Mixture F-12 (#8120482). GibcoBRL (Waltham, MA, USA) provided the fetal bovine serum (#16000044). We used many corresponding antibodies: anti-COX-2 (#ab283574), anti-ADAMTs-5 (#ab231595) from Abcam (Cambridge, UK); anti-NOS-2 (#AF0199), anti-GAPDH (#AF7012), anti-Aggrecan (#AF7561), anti-MMP-13 (#AF5355) and anti-Col-2 (#AF0135) from Affinity Biosciences (Cincinnati, OH, USA); anti-lamin B1 (#17416), anti-Bcl-2 (##3498), anti-Bax (#2772), anti-Cleaved-Caspase-3 (#9661), anti-P65 (#8242), anti-p-P65 (#3033), anti-p-IκBα (#2859), anti-IκBα (#9242) from Cell Signaling Technology (Danvers, MA, USA).
Maintained at Animal Center of Shandong University, all SD rats used in this experiment were 8-12 weeks male rats. The Shandong University Ethics Committee approved all animal studies conducted according to institutional guidelines. All animal experiments were performed according to the guidelines and through the approval of present University.

Dulbecco’s Modified Eagle Medium Nutrient Mixture F-12 (DMEM/F-12) (catalog#8120482) was obtained from Gibco-BRL (Waltham, MA, United States). The following specific antibodies were used: anti-NOS-2 (Catalog.#AF0199), anti-MMP-13 (Catalog.#AF5355), anti-TNF Receptor I (Catalog.#AF0282), anti-Aggrecan (Catalog.#DF7561), and anti-GAPDH (Catalog.# AF7021) from Affinity Biosciences (OH, United States); anti-cPLA2 (catalog#YP0868), anti-p-cPLA2 (catalog#YT1084) from ImmunoWay Biotechnology (TX, United States); lamin B1 (catalog#13435), anti-P38 (catalog#8690), anti-p-P38 (catalog#4511), anti-ERK1/2 (catalog#9194), anti-p-ERK1/2 (catalog#4370), anti-p-P65 (catalog#3033), and anti-P65 (catalog#8242) from Cell Signaling Technology (Danvers, MA, United States); anti-COX-2 (catalog#ab179800) from Abcam (Cambridge, United Kingdom).
The mice and SD rats used in the experiment were purchased from the Animal Center of Shandong University. All of the animals were 8–12 weeks old male mice and rats, living in the same specific pathogen-free (SPF) standard environmental conditions (25°C, light/dark cycle for 12 h) with free access to food, and allowed to move freely. All experiments involving animals were carried out in accordance with the guidelines of the Professional Committee of Animal Care and Use of Shandong University (Shandong, China), which approved this study.