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Transcriptomic study on the impact of Temporomandibular joint Internal derangement in the Condylar Cartilage of Rabbits

ABSTRACT: Internal derangement (ID) in the temporomandibular joint (TMJ) compromises a group of clinical problems, and holds a relative high prevalence in populations. However, the temporal change in gene expression of condylar cartilage during continous ID remains unclear. The aim of current study is investigating the differential expressed gene pattern in condylar cartilage of rabbits with ID from 1 to 8 weeks. Immunohistochemical results indicated that abnormal collagen fiber arrangements, fragmentation of fibrils, inflammatory cells infiltration were detected from 1 week to 4 week in joint disc specimens, while newly formed vessels, mucoid degeneration, meniscal tears, osteoclasts and osteoblasts were observed later. The microarray analysis showed 6478 genes have more than two-fold changes among all the tested transcripts. The inflammation gene group including ACE and IL1βincreased rapidly in the early stage while decrease later. On the contrary, the bone construction related genes showed a low level at first and increased at later period in the ID progression. Besides, the current study found some genes such as HLA2G, which had never been reported, might be relevant with ID. Our findings might provide useful insights into the pathological mechanism of ID in TMJ.

ORGANISM(S): Oryctolagus cuniculus

PROVIDER: GSE66864 | GEO | 2015/03/14

SECONDARY ACCESSION(S): PRJNA278127

REPOSITORIES: GEO

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Project description:Background Extracellular matrix (ECM) protein malfunction or defect may lead to temporomandibular joint osteoarthritis (TMJ OA). Dentin sialophophoprotein (DSPP) is a mandibular condylar cartilage ECM protein, and its deletion impacted cell proliferation and other extracellular matrix alterations of postnatal condylar cartilage. However, it remains unclear if long-term loss of function of DSPP leads to TMJ OA. The study aimed to test the hypothesis that long-term haploinsufficiency of DSPP causes TMJ OA. Materials and Methods To determine whether Dspp+/- mice exhibit TMJ OA but no severe tooth defects, mandibles of wild-type (WT), Dspp+/-, and Dspp homozygous (Dspp-/-) mice were analyzed by Micro-computed tomography (micro-CT). To characterize the progression and possible mechanisms of osteoarthritic degeneration over time in Dspp+/- mice over time, condyles of Dspp+/- and WT mice were analyzed radiologically, histologically, and immunohistochemically. Results Micro-CT and histomorphometric analyses revealed that Dspp+/- and Dspp-/- mice had significantly lower subchondral bone mass, bone volume fraction, bone mineral density, and trabecular thickness compared to WT mice at 12 months. Interestingly, in contrast to Dspp-/- mice which exhibited tooth loss, Dspp+/- mice had minor tooth defects. RNA sequencing data showed that haplodeficency of DSPP affects the biological process of ossification and osteoclast differentiation. Additionally, histological analysis showed that Dspp+/- mice had condylar cartilage fissures, reduced cartilage thickness, decreased articular cell numbers and severe subchondral bone cavities, and with signs that were exaggerated with age. Radiographic data showed an increase in subchondral osteoporosis up to 18 months and osteophyte formation at 21 months. Moreover, Dspp+/- mice showed increased distribution of osteoclast in the subchondal bone and increased expression of MMP2, IL-6, FN-1, and TLR4 in the mandibular condylar cartilage. Conclusions Dspp+/- mice exhibit TMJ OA in a time-dependent manner, with lesions in the mandibular condyle attributed to hypomineralization of subchondral bone and breakdown of the mandibular condylar cartilage, accompanied by upregulation of inflammatory markers.

Project description:Objective The purpose of this study was to find the key regulatory genes and proteins in condylar ossification of temporomandibular joint (TMJ) by transcriptomics and proteomics in porcine embryos. Method A total of 12 miniature pig embryos (6 in each group) at E45 and E85 days were used in this study. Six embryos for tissue sections (3 in each group) . The remaining six embryos were cut down the condyle along the parallel line between the lowest point of the sigmoid notch and the highest point of the condyle, and each condylar tissue sample was divided into two parts averagely. After RNA extraction, library construction and sequencing, one part tissue was analyzed by mapping to compare the differential expression of mRNA before and after ossification of embryonic condylar tissue. Another tissue was analyzed by protein extraction, enzymolysis and mass spectrometry to compare the differential expression of protein before and after ossification of embryonic condylar tissue. Finally, the differential genes and proteins were analyzed by GO and KEGG enrichment analysis. The differentially expressed genes in transcriptome and proteomic analysis were verified by QPCR. Result A total of 25322 genes were detected by transcriptome analysis, and there were 19584 differential genes between E85 days and E45 days. There are 1592 differential genes were screened to meet the Fold change≥2 or ≤0.5 of which 1086 genes were up-regulated and 506 genes were down-regulated in E85 days compared with E45 days. A total of 4613 proteins were detected by proteomic analysis, there were 419 differential proteins between E85 and E45 days, including 313 up-regulated proteins and 106 down-regulated proteins. A total of 37 differentially expressed genes/proteins were found by cross transcriptome and proteomic analysis. QPCR analysis showed that 13 of 15 genes were consistent with transcriptome analysis. Conclusion Condylar transcriptome and proteomic analysis during the development of temporomandibular joint in miniature pigs revealed the regulatory genes/proteins of condylar ossification. These regulatory genes/proteins can provide basic data for human TMJ development research and disease treatment.

Project description:Menisci play a vital role in load transmission, shock absorption and joint stability. The current dogma is that the menisci simply protects the cartilage and play no role in osteoarthritis (OA) unless they are injured. However, there is increasing evidence suggesting that OA menisci may not merely be bystanders in the disease process of OA. This study sought: 1) to determine the prevalence of meniscal degeneration in OA patients, 2) to examine gene expression in OA meniscal cells compared to normal control meniscal cells, and 3) to test the hypothesis that OA meniscal cells are different from normal meniscal cells. The grades of meniscal degeneration correlated in a positive fashion with the grades of articular cartilage degeneration (r = 0.672; P < 0.0001). Many genes classified in the biological processes of immune response, inflammatory response, biomineral formation and cell proliferation, including major histocompatibility complex, class II, DP alpha 1 (HLA-DPA1), integrin, beta 2 (ITGB2), ectonucleotide pyrophosphatase/phosphodiesterase 1 (ENPP1), ankylosis, progressive homolog (ANKH) and fibroblast growth factor 7 (FGF7), were expressed at higher levels compared to normal control meniscal cells. In addition, many genes that were previously implicated in OA were also expressed at higher levels in OA meniscal cells, including ADAM metallopeptidase with thrombospondin type 1 motif, 5 (ADAMTS5). Our findings suggest that OA is a whole joint disease. Meniscal cells may play an active role in the development of OA. Investigation of the gene expression profiles of OA meniscal cells may reveal new therapeutic targets for OA therapy and also may uncover novel disease markers for early diagnosis of OA. Studies were approved by our human subjects Institutional Review Board. Menisci and articular cartilage were collected during joint replacement surgery for OA patients (OA specimens) and lower limb amputation surgery for osteosarcoma patients (normal control specimens), and graded. Meniscal cells were prepared from the meniscal tissues and expanded in monolayer culture. Differential gene expression in OA meniscal cells and normal control meniscal cells was examined using Affymetrix microarray and real time RT-PCR.

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Transcriptomic study on the impact of Temporomandibular joint Internal derangement in the Condylar Cartilage of Rabbits

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