1Department of Foot and Ankle Surgery, Honghui Hospital of Xi'an Jiaotong University, Xi'an 710054, China; 2The Second Department of Cardiovascular, Shanxi Provincial People's Hospital, Xi'an 710068, China; 3School of Public Health, Xi'an Jiaotong University, Xi'an 710049, China; 4Cancer Center, the First Affiliated Hospital of Xi'an Jiaotong University, Xi'an 710061, China; 5Spine Hospital, Honghui Hospital of Xi'an Jiaotong University, Xi'an 710054, China
Abstract：Objective To investigate the pathological characters and relative molecular pathogenesis of diabetic neuropathic osteoarthropathy(DNOAP).Methods The articular cartilages of tibial talus, subtalar joint and talo-scaphoid joint of 8 patients with DNOAP admitted to Honghui Hospital of Xi'an Jiaotong University from March 2017 to June 2018 were selected as DNOAP group, including 3 males and 5 females, aged 20-66(55.7±3.8) years. From April 2017 to July 2018, the articular cartilage of tibial talus joint, subtalar joint and talus-navicular joint of 8 patients with amputation due to traffic accident or serious trauma in Honghui Hospital of Xi'an Jiaotong University were normal control group, including 4 males and 4 females, aged 19-65(57.6±3.7) years. Cartilage samples from DNOAP group and normal control group were taken. Masson staining and safranine O/fixed green staining were used to observe the histopathological characteristics of cartilage, and the ultrastructural changes of chondrocytes were observed by electron microscopy. Chondrocyte culture was carried out in DNOAP group and normal control group. DCFH-DA probe was used to detect the level of reactive oxygen species in chondrocyte. Western blot was used to detect the expression of receptor activator of nuclear factor-κ B ligand(RANKL), osteoprotegerin(OPG), IL-1β, IL-6, TNF-α, aggrecan protein in chondrocyte. Flow cytometry(FCM) was used to detect the apoptotic rate of chondrocyte.Results In DNOAP group, chondrocytes decreased, subchondral bone proliferated, structure disordered and osteoclasts formed in a large number in subchondral bone area. Under transmission electron microscopy, mitochondria swelled, membrane structure was incomplete, arrangement disordered, endoplasmic reticulum was severely swollen and nucleus of DNOAP chondrocytes were observed. The chromatin was partially broken and concentrated at the edge of nuclear membrane. In the normal control group, chondrocyte was found in the cartilage lacuna, with homogeneous staining of cartilage matrix and neat arrangement of subchondral bone. Under transmission electron microscopy, chondrocyte showed abundant rough endoplasmic reticulum and normal concentration of chromatin in the nucleus. The ROS fluorescence intensity of chondrocyte in DNOAP group was 28.1±2.3, which was higher than that of normal control group 11.9±0.7. The difference was statistically significant (t=19.059, P＜0.01). The relative expression of RANKL, TNF-α, IL-1β and IL-6 protein in DNOAP group was higher than that in normal control group, while the relative expression of OPG and aggrecan protein was lower than that in normal control group (t=6.062, 9.780, 12.479, 11.696, 8.792, 7.726, all P values＜0.01). FCM showed that the apoptotic rate of chondrocyte in DNOAP group was 3.3% ± 0.2%, which was higher than that in normal control group 1.2%±0.1%. The difference was significant (t=26.563, P＜0.01).Conclusions The pathological characteristics of DNOAP are serious destruction of cartilage structure and organelles. Inflammation plays a role in promoting the pathogenesis of DNOAP.
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2020, Vol. 25 
