G1105 500ml

After completion
of the CT scan, the femurs were placed in a decalcification fluid
(Servicebio, G1105-500ML, Wuhan, China) for three consecutive weeks,
with the decalcification fluid being changed every 3 days. The decalcified
femur was embedded with paraffin and sectioned in the direction of
the long axis of the femoral stem with a thickness of 5 μm.
The sections were fixed on the slides and stained with H&E and
Alcian blue. The sections were imaged with an Olympus BX51 microscope
and a DP73 CCD Olympus Imaging System (Olympus Corporation, Tokyo).
Caseviewer 2.4 (3DHISTECH Ltd, Hungary) was used to observe and analyze
the morphology of the specimens at 0.8×, 5×, and 20×
magnifications. ImageJ was used to calculate the area of bone and
cartilage of the callus.

Human samples of medial tibia plateau were collected during total knee arthroplasty operations. Subchondral bone and articular cartilage samples were cut into 1- to 2-cm pieces, fixed in 4% paraformaldehyde (PFA) solution (G1101-500ML Servicebio, Wuhan, China) for 2 days, and decalcified in 10% EDTA (G1105-500ML, Servicebio, Wuhan, China) for 6 months. Samples were embedded in paraffins or optimal cutting temperature (OCT) compound and cut into 5-μm sections. The experiments were approved by the ethics committee of Shanghai University, no. ECSHU 2021-146. We informed the patient before the surgery that the medial tibia plateau would be used as a sample for scientific research. The patients expressed their willingness to participate and signed the informed consent form.