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Constructing condylar organoid to explore its difference to the traditional two-dimensional (2D) culture condylar chondrocyte

ABSTRACT: constructing condylar organoid to explore its difference to the traditional two-dimensional (2D) culture condylar chondrocyte

PROVIDER: PRJNA962030 | ENA |

REPOSITORIES: ENA

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			Action	DRS
	SRR24313646_1.fastq.gz	Fastqsanger.gz
	SRR24313646_2.fastq.gz	Fastqsanger.gz
	SRR24313647_1.fastq.gz	Fastqsanger.gz
	SRR24313647_2.fastq.gz	Fastqsanger.gz
	SRR24313648_1.fastq.gz	Fastqsanger.gz

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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:Background: Functional orthoses are commonly used to treat skeletal Class II malocclusion, but the specific mechanism through which they do this has been a challenging topic in orthodontics. In the present study, we aimed to explore the effect of tensile stress on the osteogenic differentiation of condylar chondrocytes from an exosomal perspective. Methods: We cultured rat condylar chondrocytes under resting and tensile stress conditions and subsequently extracted cellular exosomes from them. We then screened miRNAs that were differentially expressed between the two exosome extracts by high-throughput sequencing and performed bioinformatics analysis and osteogenesis-related target gene prediction using the TargetScan and miRanda softwares. Exosomes cultured under resting and tensile stress conditions were co-cultured with condylar chondrocytes for 24 h to form the Control-Exo and Force-Exo exosome groups, respectively. Quantitative real time PCR(RT-qPCR) and western blotting were then used to determine the mRNA and protein expression levels of Runx2 and Sox9 in condylar chondrocytes. Results: The mRNA and protein expression levels of Runx2 and Sox9 in the Force-Exo group were significantly higher than those in the Control-Exo group (p < 0.05). The differential miRNA expression results were consistent with our sequencing results. Bioinformatics analysis and target gene prediction results showed that the main biological processes and molecular functions involved in differential miRNA expression in exosomes under tensile stress were biological processes and protein binding, respectively. Kyoto Gene and Genome Data Bank (KEGG) pathway enrichment analysis showed significant enrichment of differentially expressed miRNAs in the mTOR signaling pathway. The differentially expressed miRNAs were found to target osteogenesis-related genes. Conclusion: These results suggest that stimulation of rat condylar chondrocytes with tensile stress can alter the expression levels of certain miRNAs in their exosomes and promote their osteogenic differentiation. Exosomes under tensile stress culture conditions thus have potential applications in the treatment of Osteoarthritis (OA).

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Constructing condylar organoid to explore its difference to the traditional two-dimensional (2D) culture condylar chondrocyte

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