Accelerated osteogenic differentiation of human bone-derived cells in ankylosing spondylitis

Ankylosing spondylitis (AS) is characterized by excessive bone formation with syndesmophytes, leading to bony ankylosis. The contribution of osteoblasts to the pathogenesis of ankylosis is poorly understood. The aim of this study was to determine molecular differences between disease controls (Ct) and AS bone-derived cells (BdCs) during osteogenic differentiation with or without inflammation using AS patient serum. We confirmed osteoblastic differentiation of Ct and AS BdCs under osteogenic medium by observing morphological changes and measuring osteoblastic differentiation markers. Osteoblast differentiation was detected by alkaline phosphatase (ALP) staining and activity, and alizarin red and hydroxyapatite staining. Osteoblast-specific markers were analyzed by quantitative reverse-transcriptase-polymerase chain reaction, immunoblotting, and immunostaining. To examine the effects of inflammation, we added AS and healthy control serum to Ct and AS BdCs, and then analyzed osteoblast-specific markers. AS BdCs showed elevated basal intercellular and extracellular ALP activity compared to Ct. When osteoblast differentiation was induced, AS BdCs exhibited higher expression of osteoblast-specific marker genes and faster mineralization than Ct, indicating that these cells differentiated more rapidly into osteoblasts. ALP activity and mineralization accelerated when serum from AS patients was added to Ct and AS BdCs. Our results revealed that AS BdCs showed significantly increased osteoblastic activity and differentiation capacity by regulating osteoblast-specific transcription factors and proteins compared to Ct BdCs. Active inflammation of AS serum accelerated osteoblastic activity. Our study could provide useful basic data for understanding the molecular mechanism of ankylosis in AS.This work was supported by Basic Science Research Program through the National Research Foundation of Korea (NRF) funded by the Ministry of Science, ICT & Future (NRF-2016R1A2B4008606)

A novel role for bone-derived cells in ankylosing spondylitis: Focus on IL-23

The main aim of this study are to explore the role of bone-derived cells (BdCs) in anIcylosing spondylitis (AS) and determine the underlying molecular mechanisms of IL-23 production. Primary BdCs were isolated from diced bone of facet joints obtained during surgery from seven AS patients and seven disease control (Ct) patients. Osteoblastic activity of BdCs was assessed by measuring their alkaline phosphatase activity and by alizarin red staining. Osteoblast and endoplasmic reticulum (ER) stress related genes were assessed by quantitative PCR, immunoblotting, immunofluorescence, and immunohistochemistry. In addition, expression of IL-23 in response to BIX (selective BIP inducer X)-induced ER stress was evaluated by qPCR and ELISA. Protein interaction and binding to IL-23 promoter were confirmed by Immunoprecipitation and Chromatin immunoprecipitation, respectively. Transcript levels of genes involved in osteoblast function, as well as of the ER stress marker were higher in the AS group than the Ct group, and elevated RUNX2, BiP and IL-23 expression were observed in the BdCs, serum, and bone biopsies from the AS group. BIX-induced ER stress stimulated osteoblastic activity and IL-23 secretion by upregulating RUNX2 expression. Furthermore, in AS BdCs, RUNX2 interacted with C/EBP beta to bind to IL-23 promoter and RUNX2 knockdown suppressed IL-23 secretion. These finding may provide a molecular mechanism involved in sustained ER stress in AS BdCs stimulates the activation of RUNX2 and C/EBP beta genes, leading to IL-23 production. (C) 2017 Elsevier Inc. All rights reserved.We thank Prof. Young Wha Koh (Department of Pathology, Ajou University School of Medicine, Suwon, Republic of Korea) for help in evaluating the histologic. This work was supported by Basic Science Research Program through the National Research Foundation of Korea (NRF) funded by the Ministry of Science, ICT& Future (NRF-2016R1A2B4008606) and the Ministry of Education, Science and Technology (NRF-2013R1A1A2009617)

Poly-γ-glutamic acid suppresses osteoclastogenesis in human osteoclast precursor and prevents joint damage in a collagen-induced murin arthritis model

Poly-gamma-glutamic acid (gamma-PGA), a natural polymer derived from Bacillus subtilis, shows anti-inflammatory activity. However, the effects of gamma-PGA on osteoclasts, which are important cells for joint destruction in inflammatory diseases such as rheumatoid arthritis (RA), have not yet been reported. In this study, we show that gamma-PGA markedly inhibits osteoclast differentiation in normal PBMC-derived osteoclast precursors and in synovial fluid macrophages of patients with RA. gamma-PGA also reduces RANK expression by down-regulating M-CSF receptors. Additionally, oral administration of gamma-PGA attenuated bone destruction in a collagen-induced arthritis (CIA) model, demonstrating decreases in inflammation, cartilage damage, and osteoclast formation in histological analyses. Taken together, these data suggest that gamma-PGA could be a good candidate for therapeutic prevention of joint destruction in RA.This research was supported by the Basic Science Research Program through the National Research Foundation of Korea (NRF) funded by the Ministry of Education, Science, and Technology (2013R1A1A2009617) and the Ministry of Science, ICT, & Future (2016R1A2B4008606), and a grant of the Korea Health Technology R&D project through the Korea Health Industry Development Institute (KHIDI), funded by the Ministry of Health & Welfare, Republic of Korea (HI17C0888 and HI15C1062). We thank Dr. Dong Soo Han (Department of Internal Medicine, Hanyang University Guri Hospital, Guri, Korea) for providing experimental materials

IL-17A induces osteoblast differentiation by activating JAK2/STAT3 in ankylosing spondylitis

Background: IL-17A has recently emerged as a potential target that regulates the extensive inflammation and abnormal bone formation observed in ankylosing spondylitis (AS). Blocking IL-17A is expected to inhibit bony ankylosis. Here, we investigated the effects of anti IL-17A agents in AS.Methods: TNF alpha, IL-17A, and IL-12/23 p40 levels in serum and synovial fluid from patients with ankylosing spondylitis (AS), rheumatoid arthritis (RA), osteoarthritis (OA), or healthy controls (HC) were measured by ELISA. Bone tissue samples were obtained at surgery from the facet joints of ten patients with AS and ten control (Ct) patients with noninflammatory spinal disease. The functional relevance of IL-17A, biological blockades, Janus kinase 2 (JAK2), and non-receptor tyrosine kinase was assessed in vitro with primary bone-derived cells (BdCs) and serum from patients with AS.Results: Basal levels of IL-17A and IL-12/ 23 p40 in body fluids were elevated in patients with AS. JAK2 was also highly expressed in bone tissue and primary BdCs from patients with AS. Furthermore, addition of exogenous IL-17A to primary Ct-BdCs promoted the osteogenic stimulus-induced increase in ALP activity and mineralization. Intriguingly, blocking IL-17A with serum from patients with AS attenuated ALP activity and mineralization in both Ct and AS-BdCs by inhibiting JAK2 phosphorylation and downregulating osteoblast-involved genes. Moreover, JAK2 inhibitors effectively reduced JAK2-driven ALP activity and JAK2-mediated events.Conclusions: Our findings indicate that IL-17A regulates osteoblast activity and differentiation via JAK2/STAT3 signaling. They shed light on AS pathogenesis and suggest new rational therapies for clinical AS ankylosis.This work was supported by the Basic Science Research Program through the National Research Foundation of Korea (NRF) funded by the Ministry of Science, ICT and Future (NRF-2016R1A2B4008606) and the Ministry of Education (2017R1A6A3A11034394). It was also supported by a Korea Health Technology R&D grant through the Korea Health Industry Development Institute (KHIDI), which is funded by the Ministry of Health and Welfare, Republic of Korea (HI17C0888)

영재학생과 일반학생의 학교생활만족도 비교

학위논문(석사)--아주대학교 교육대학원 :교육학과,2011. 2Ⅰ. 서론 1 1. 연구의 필요성과 목적 1 2. 연구문제 3 3. 용어의 정리 3 4. 연구의 제한점 4 Ⅱ. 이론적 배경 5 1. 학교생활만족도 5 2. 만족의 정의 8 3. 선행연구의 고찰 11 Ⅲ. 연구방법 15 1. 연구대상 15 2. 측정도구 16 3. 자료처리 18 Ⅳ. 연구의 결과 및 논의 20 1. 영재학생과 일반학생의 학교생활만족도 비교 20 2. 성별에 따른 학교생활만족도 분석 25 3. 학년에 따른 학교생활만족도 분석 29 4. 영재학생 중 소속학교와 영재교육기관의 학교생활만족도 분석 33 5. 영재교육 경력에 따른 학교생활만족도 분석 40 Ⅴ. 요약 및 결론 42 1. 요약 42 2. 결론 및 제언 45 참고문헌 47 ABSTRACT 50 부 록 52Maste