Date: 2015-09-17 Session: Free Papers Research Cartilage & Bone Time: 08:30 - 10:00 Room: Dongguan Hall Abstract no.: 41405 EXPRESSION PROFILE OF LONG NONCODING RNAS IN HUMAN OSTEOARTHRITIC CARTILAGE Guangxin HUANG, Ming FU, Zhiqi ZHANG, Zhiyu HUANG, Baoxi YU, Hong SUN, Ziji ZHANG, Zibo YANG, Weiming LIAO Objectives: Long noncoding RNAs (lncRNAs) have emerged as a novel class of regulatory molecules involved in various biological processes, but their role in osteoarthritis (OA) remains unknown. Therefore, we aimed to reveal lncRNAs expression profile in human osteoarthritic cartilage and explore the potential functions of lncRNAs in OA. Methods: The expression profiles of lncRNAs and mRNAs in cartilage were obtained using microarray and verified by quantitative reverse transcription polymerase chain reaction (qRT-PCR). Bioinformatics analyses including lncRNA classification and subgroup analysis, gene ontology analysis, pathway analysis, network analysis and target prediction were performed. Results: There were 3007 upregulated lncRNAs and 1707 downregulated lncRNAs in OA cartilage compared with normal samples (Fold change>2.0). In addition, 2136 mRNAs were upregulated and 2,241 mRNAs were downregulated in OA cartilage (Fold change>2.0). The qRT-PCR results of six dysregulated lncRNAs were consistent with the microarray data. 106 lncRNAs and 291 mRNAs composed the coding-non-coding gene co-expression network. In the 600 top differentially expressed lncRNAs, 48 lncRNAs were predicted to have more than five cis-regulated target genes and up to 530 lncRNAs might regulate their trans target genes through collaboration with transcriptional factor SP1. The positive correlation between lncRNA uc.343 and predicted target homeobox gene C8 (HOXC8) expression in SW1353 cells treating with interleukin-1 beta confirmed the target prediction to some extent. Conclusions: This study revealed the expression pattern of lncRNAs in OA cartilage and predicted the potential function and targets, which indicated that lncRNAs may be new biomarkers for diagnosis or novel therapeutic targets of OA. Date: 2015-09-17 Session: Free Papers Research Cartilage & Bone Time: 08:30 - 10:00 Room: Dongguan Hall Abstract no.: 39141 CHONDROCYTE-SEEDED ECM DERIVED MICROCARRIERS AS BUILDING BLOCKS FOR ARTICULAR CARTILAGE REPAIR IN A RAT MODEL Heyong YIN, Zhen SUN, Jiang PENG, Xun SUN, Shibi LU We have developed innovative microcarriers derived from cartilage extracellular matrix (ECM). Articular cartilage slices from the porcine joint were physically shattered, filtrated by steel sieves with a sieve pore of 250µm and 125µm in diameter successively, and then decellularized using 1% SDS. After these processes, ECM derived microcarriers (CEMMs) with diameters of approximately 125 µm to 250 µm were prepared. On histology, CEMMs showed most of the ECM components after cell extraction, and scanning electron microscopy revealed a nearly-circular appearance and with pompons on the surface. Under Microgravity culture conditions in a rotary cell culture system (RCCS) bioreactor, chondrocytes can rapidly proliferate on the surface of CEMMs. Further, the CEMMs- adhered culture under dynamic culture condition displayed increased levels of the differentiation marker Collagen-II, SOX-9 , and ACAN and reduced levels of the dedifferentiation marker COL1A1 compared with static culture. In vivo study, building blocks of chondrocyte-seeded CEMMs were used to repair trochlear cartilage defect in a rat model (CEMMs group). Histological results at 6 and 12 weeks showed that the CEMMs group led to repair tissue that consisted of more hyaline-like cartilage tissue. While the control groups generally led to the formation of fibrocartilage repair tissue. Microcomputed tomography revealed that CEMMs group had more mineralized bone formation than that of control groups. To our knowledge, this is the first article that prepare innovative microcarrier derived from cartilage ECM and the derict use of chondrocyte-seeded CEMMs as building blocks for in vivo articular cartilage repair successfully. Date: 2015-09-17 Session: Free Papers Research Cartilage & Bone Time: 08:30 - 10:00 Room: Dongguan Hall Abstract no.: 39738 LONG NONCODING RNAS EXPRESSION SIGNATURES IN CHONDROGENIC DIFFERENTIATION HUMAN BONE MARROW MESENCHYMAL STEM CELLS Liang WANG, Yipeng WANG, Bin YU, Zhengyao LI, Ziquan LI Long non-coding RNAs (lncRNAs) have been established to participate in various biological processes that are crucial for development and differentiation. However, the roles of lncRNAs in the mechanisms of human bone marrow mesenchymal stem cells (MSCs) differentiation are not completely understood. The purpose of the study was to investigate the expression profiles of lncRNAs during the chondrogenic differentiation of human bone marrow MSCs, with a view to studying the biological function of lncRNAs and their involvement in the mechanism of differentiation. We compared the lncRNAs expression profiles of undifferentiated and differentiated cells during chondrogenic differentiation by microarray. 3638 differentially expressed lncRNAs were identified (fold- change >2.0 or <-2.0, P<0.05), consisting of 2166 up-regulated and 1472 down-regulated. Microarray data were validated using quantitative reverse transcription-polymerase chain reaction (qRT-PCR). Bioinformatic analyses were applied for further study of these differentially expressed lncRNAs. Among these lncRNAs, ZBED3-AS1 and CTA-941F9.9 were further analyzed with co-expression network and target prediction analysis. The results showed that the two up-regulated lncRNAs are likely to play important roles in chondrogenic differentiation process. In conclusion, the expression profile of lncRNAs was significantly altered during differentiation process. It provided a new insight on complicated regulation mechanisms of human bone marrow MSCs chondrogenic differentiation. Date: 2015-09-17 Session: Free Papers Research Cartilage & Bone Time: 08:30 - 10:00 Room: Dongguan Hall Abstract no.: 41851 ARE THE LEADING DRUGS TO FIGHT AGAINST STAPHYLOCOCCUS AUREUS REALLY TOXIC TO CARTILAGE? Mehmet ISYAR, Mustafa DOGAN, Ibrahim YILMAZ, Bulent BILIR, Duygu YASAR SIRIN, Selami CAKMAK, Mahir MAHIROGULLARI Objective: The aim of the present study was to investigate the effects of antibiotics, which are used against staphylococcus aureus, on human chondrocytes in vitro. Methods: Primary cell cultures obtained from gonarthrosis patients were divided into two main groups. One of these groups were designated as control chondrocyte culture. Other group was divided into three groups and each group was exposed to antibiotics vancomycin, teicoplanin, and linezolid respectively. Cell culture samples were characterized by immunophenotyping following incubation of three different antibiotics. Before and after administration of agents, cultures were subjected to inverted and environmental scanning electron microscopy. The number of live cells and the proliferation rate were monitored by MTT-assay. Results: It was showed that there is not a chondrotoxic effect of Vancomycin, teicoplanin, and linezolid. Conclusion: This present study indicates that vancomycin, teicoplanin, and linezolid can safely be used in orthopedic surgery, especially against to methicillin-resistant Staphylococcus aureus agents. Date: 2015-09-17 Session: Free Papers Research Cartilage & Bone Time: 08:30 - 10:00 Room: Dongguan Hall Abstract no.: 41203 THE EFFECT OF WHARTON'S JELLY ON MENISCUS FIBROCHONDROCYTES IN VITRO AND CELL-BASED SCAFFOLD FOR MENISCUS TISSUE ENGINEERING Wei NIU Clinical treatment for meniscus injury is still a hang-up because of weak self-repairing capability of inner avascular zone. Tissue-engineered meniscus is considered as a promising substitution for meniscus injury in future. This experiment was conducted to assess feasibility of Wharton’s jelly as a kind of material for meniscus tissue engineering. Collecting Wharton’s jelly from kibbling umbilical cord in differential centrifugation method, and produce a polyporous bioscaffold in freeze drying and cross-linking process. Fibrochondrocytes were isolated from rabbit’s meniscus by enzymatic digestion and were cultured to passage 3. Then the cells were seeded on the scaffold and six-well plate which was coated with Wharton’s jelly, and perform correlation detection after culturing for 7 and 14 days. DNA testing results showed that the material could accelerate cell proliferation, Histology and molecular biology results indicated that the material could promote secretion of collagen and glycosaminoglycan in extracellular matrix, slow down cell degeneration and maintain cell phenotype. Tissue-section result for cell-based structure showed that cells have good histocompatibility with the material, and this construct performed fibrocartilaginous characteristics. Compared to acellular scaffold, cell seeding can obviously enhance mechanical property of the scaffold. The results demonstrated that Wharton’s jelly may be select as a new resource for meniscus tissue engineering. Date: 2015-09-17 Session: Free Papers Research Cartilage & Bone Time: 08:30 - 10:00 Room: Dongguan Hall Abstract no.: 41312 IDENTIFICATION FOR LONG NONCODING RNA ASSOCIATED WITH OSTEOARTHRITIS IN HUMAN Dan XING, Yang CHEN, Yang CHEN Objective: Long noncoding RNAs (lncRNAs) are an important class of genes involved in various biological functions, but knowledge of lncRNAs in osteoarthritis (OA) is limited. The present study was to identify differential lncRNAs which expression in OA cartilage.