No abstract

Osteoarthritis (OA) is condition which poses a main concern to the aging population and its severity is expected to increase with the increasing life expectancy. In the future, several possible targets for OA treatment need to be defined. dickkopf-related protein 3 (dKK3) is an atypical member of the Wnt-antagonistic dickkopf-related protein (dKK) family. The availability of research into the role of dKK3 in the abnormal remodeling of subchondral bone in human knee joints is currently limited. Thus, the aim of the present study was the evaluation of dKK3 expression in the abnormal bone remodeling of subchondral bone in human knee OA in order to clarify the role of dKK3 in subchondral bone remodeling and to acknowledge its potential relevance to β-catenin. In total, 38 specimens were collected from osteotomies of the medial tibial plateau of the human knee. The patient samples were then divided into the normal, mild, moderate and severe symptom groups, according to the Osteoarthritis Research Society International (OARSI) score. Following hematoxylin and eosin (H&E) and Safranin O-fast green staining for alkaline phosphatase (AZO method), changes in the distribution and number of osteocytes in the subchondral bone and the degree of sclerosis of the subchondral bone were observed. Immunohistochemical staining, immunofluorescence, western blot analysis and reverse-transcription quantitative PcR (RT-qPcR) were used for the detection of dKK3 and β-catenin expression level changes in osteoblasts in the subchondral bone of the medial tibial plateau. H&E and alkaline phosphatase staining revealed that the total number of osteocytes in the subchondral bone increased with the severity of the disease. The samples were also evaluated using Safranin O-Fast Green staining and were attributed a score according to the OARSI scoring system: The scoring number and cartilage damage increased along with OA severity. Immunohistochemistry and immunofluorescence assays demonstrated that β-catenin expression in osteocytes increased from mild to moderate, whereas dKK3 expression decreased with the development of arthritis from normal, mild to moderate. According to the results of western blot analysis, β-catenin expression was higher in moderate OA and then decreased in severe OA. On the other hand, the dKK3 levels decreased along with the progression from normal, mild to moderate OA. The results of RT-qPcR demonstrated that β-catenin and dKK3 gene expression differed with the degree of OA. On the whole, the present study demonstrates that dKK3 and β-catenin may play opposite roles in OA subchondral bone remodeling.

An important causative factor in osteoarthritis (OA) is the abnormal mechanical stress-induced bone remodeling of the subchondral bone. β2-adrenergic receptor (Adrb2) plays a major role in mechanical stresses that induce bone remodeling. The medial tibial plateau (MTP) and lateral tibial plateau (LTP) of patients with varus Knee osteoarthritis (KO) bear different mechanical stresses. The present study aimed to investigate the expression of Adrb2 in medial tibial plateau subchondral bone (MTPSB) and lateral tibial plateau subchondral bone (LTPSB) in patients with varus KO. A total of 30 tibial plateau samples from patients undergoing total knee arthroplasty for varus KO and MTPSB and LTPSB were studied. Statistical analysis was performed using paired sample t-tests. Safranin O-Fast Green staining and Micro-computed tomography showed significant differences in the bone structure between MTPSB and LTPSB. Tartrate-resistant acid phosphatase (TRAP)-positive cell density in MTPSB was higher than that in LTPSB. Immunohistochemistry, reverse transcription-quantitative polymerase chain reaction, and Western blot analysis revealed that compared to LTPSB, the levels of Adrb2, tyrosine hydroxylase (TH), and osteocalcin increased significantly in MTPSB. Double-labeling immunofluorescence showed Adrb2 was present in the majority of TRAP-positive multinuclear cells of the MTPSB. The expression of Adrb2 and TH was significantly higher in MTPSB than in LTPSB, confirming the involvement of these molecules in the development of OA.

Hydroxyapatite/silk fibroin (HAp/SF) composite was prepared and applied to the posterolateral spinal fusion model in rats to observe the effect of bone fusion. Method: Calcium chloride, diammonium phosphate, SF, and polyvinyl alcohol were used as raw materials, HAp/SF composites were prepared by chemical precipitation. The microstructure of the composite, crystal phase composition, and chemical structure were analyzed by the scanning electron microscope (SEM) and X-ray diffraction (XRD), and fourier transform infrared spectrometer (FTIR Spectrometer). Through the cultivation of osteoblasts MC3T3-E1 in vitro, the adhesion and proliferation (A&P) of cells on the face of materials were investigated. Thereby, the biocompatibility of the material was characterized. HAp/SF material was applied to the rat posterolateral spinal fusion model. The osteogenesis and spinal fusion were evaluated by the imaging observation, histological observation and manual palpation. The results showed that the rod-shaped HAp with uniform size and high purity was obtained, with a diameter of 20∼40 nm and a length of 200∼500 nm, similar to the apatite crystal in natural bone tissue (BT). In composite materials, a spatial network structure was formed by the interweaving of the SF fibers, and HAp was deposited on the face of the SF or in the middle of its network structure. In the obtained HAp/SF materials, the calcium ions of HAp and the carbonyl groups of SF were used to form thermally stable complexes through strong chemical bonds. Besides, SF was a template for the directional induction of HAp crystal growth, and the growth of HAp crystal along the C axis was regulated by SF. The growth direction was parallel to the long axis of SF fibers, and was consistent with the structure of apatite crystals deposited on the face of collagen fibers in natural BT. The results of cell culture in vitro showed that: after comparison with the control group (CG) with pure Hap, the adhesion ability of cells to HAp/SF material was significantly improved. The proliferation capacity of bone artificial bone (BAM) material and HAp/SF material was also significantly improved. The nuclear and skeletal staining results of MC3T3-E1 cells on the face of three groups of materials (HAp, BAM and HAp/SF) were combined, and the results also indicated that BAM and HAp/SF materials had good ability to promote cell A&P. The results of posterolateral spinal fusion in rats showed that HAp/SF materials group palpated the posterolateral spine for fusion. The formation of new BT on the posterolateral side of the spine was revealed by the Micro-computed tomography (Micro-CT) examination. In conclusion, HAp/SF composite had good osteoblastic compatibility and can achieve good spinal fusion effect.