Osteogenic factors are often used in orthopedics to promote bone growth, improve fracture healing, and induce spine fusion. Osteogenic oxysterols are naturally occurring molecules that were shown to induce osteogenic differentiation in vitro and promote spine fusion in vivo. The purpose of this study was to identify an osteogenic oxysterol more suitable for clinical development than those previously reported, and evaluate its ability to promote osteogenesis in vitro and spine fusion in rats in vivo. Among more than 100 oxysterol analogues synthesized, Oxy133 induced significant expression of osteogenic markers Runx2, osterix (OSX), alkaline phosphatase (ALP), bone sialoprotein (BSP), and osteocalcin (OCN) in C3H10T1/2 mouse embryonic fibroblasts and in M2-10B4 mouse marrow stromal cells. Oxy133-induced activation of an 8×-Gli luciferase reporter, its direct binding to Smoothened, and the inhibition of Oxy133-induced osteogenic effects by the Hedgehog (Hh) pathway inhibitor, cyclopamine, demonstrated the role of Hh pathway in mediating osteogenic responses to Oxy133. Oxy133 did not stimulate osteogenesis via BMP or Wnt signaling. Oxy133 induced the expression of OSX, BSP, and OCN and stimulated robust mineralization in primary human mesenchymal stem cells. In vivo, bilateral spine fusion occurred through endochondral ossification and was observed in animals treated with Oxy133 at the fusion site on xray after 4 weeks and confirmed with manual assessment, micro CT (μCT), and histology after 8 weeks, with equal efficiency to recombinant human bone morphogenetic protein-2 (rhBMP-2). Unlike rhBMP-2, Oxy133 did not induce adipogenesis in the fusion mass and resulted in denser bone evidenced by greater BV/TV ratio and smaller trabecular separation. Findings here suggest that Oxy133 has significant potential as an osteogenic molecule with greater ease of synthesis and improved time to fusion compared to previously studied oxysterols. Small molecule osteogenic oxysterols may serve as the next generation of bone anabolic agents for therapeutic development.

The present study aimed to explore the underlying mechanisms of bone morphogenetic protein 2 (BMP2) in alleviating intervertebral disc degeneration (Idd). A rat puncture Idd model was constructed, and the rats were randomly divided into six groups: control; Idd (model); Idd+PBS [containing 10 10 adeno-associated virus serotype 2 (AAV)]; and Idd + AAV2-BMP2 (10 6 , 10 8 and 10 10 ). IL-1β was used to treat primary nucleus pulposus (NP) cells to mimic Idd in vitro. The effects of BMP2 in Idd were determined by magnetic resonance imaging (MRI), hematoxylin and eosin staining and Alcian Blue staining in vivo. The levels of collagen II, aggrecan, transcription factor SOX9 (SOX9) and matrix metalloproteinase 13 (MMP-13) were examined using western blot analysis and reverse transcription quantitative polymerase chain reaction (RT-qPcR) in NP tissues and cells. The expression of c-telopeptide of type II collagen (cTX-II) in the sera or cell supernatants was determined by ELISA. In addition, the levels of phosphorylation of phosphoinositide 3-kinase (PI3K) and protein kinase B (Akt), and the levels of apoptosis-associated proteins and apoptosis ratio of NP cells were also determined by western blot analysis and flow cytometry, respectively. LY29400, an inhibitor of PI3K, was used to additionally confirm the signal pathway mechanism of BMP2 treatment in IDD. BMP2 significantly extended the interval between discs and alleviated the fibrous ring rupture and the decrease in the levels of glycoproteins in Idd rats, as determined by MRI and histological staining. Additionally, BMP2 treatment significantly upregulated the levels of collagen II, aggrecan and SOX9, but downregulated the levels of MMP-13 and cTX-II in Idd rats and NP cells in a dose-dependent manner. concurrently, recombinant human (rh)BMP2 pretreatment also significantly decreased the apoptosis ratio of interleukin (IL)-1β-treated NP cells via downregulating the level of cleaved caspase-3 and upregulating the level of uncleaved poly (adenosine 5'-diphosphate-ribose) polymerase. It was demonstrated that rhBMP2 also significantly decreased the inflammatory response in NP tissues and cells, based on levels of IL-6, TNF-α and IL-10. In addition, rhBMP2 inhibited cell apoptosis via upregulating the phosphorylation levels of the PI3K/Akt signaling pathway, and LY29400 pretreatment inhibited the effects of BMP2 in IL-1β treated NP cells. BMP2 alleviated Idd via the PI3K/Akt signaling pathway by inhibiting NP cell apoptosis and decreasing the levels of matrix proteins.

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In most developing countries, it has become a severe challenge for the limited medical resources and outdated healthcare technology to meet the high demand of large population. From the perspective of social development, this unbalanced healthcare system in developing counties has also exacerbated the contradiction between physicians and patients, particularly those suffering from malignant diseases (such as prostate cancer). Rapid improvements in artificial intelligence, computing power, parallel operation, and data storage management have contributed significantly to a credible medical data decision-making on the detection, diagnosis, treatment, and prognosis of malignant diseases. Consequently, to address these existing problems in the current healthcare field of developing countries, this paper proposes a novel big medical data decision-making model exploiting fuzzy inference logic for prostate cancer in developing countries, constructing an intelligent medical system for disease detection, medical data analysis and fusion, treatment recommendations, and risk management. Based on 1 933 535 items of hospitalization information from over 8000 prostate cancer cases in China, the experimental results demonstrate that the intelligent medical system could be adopted to assist physicians and medical specialists in coming up with a more dependable diagnosis scheme.INDEX TERMS Prostate cancer, fuzzy inference logic, intelligent medical system, big medical data decision-making model, fusion of multimodal medical data, machine-assisted diagnosis.