Transcriptome sequencing analysis of osteogenic rat bone marrow mesenchymal stem cells induced by osteomyelitis

doi:10.12307/2023.387

Abstract

Abstract: BACKGROUND: Recently, osteomyelitis environment has been found to significantly inhibit the osteogenic differentiation of bone marrow mesenchymal stem cells, thereby affecting bone formation and bone metabolism, but the specific mechanism remains unclear.
OBJECTIVE: Transcriptome analysis was conducted on rat bone marrow mesenchymal stem cells with osteogenic differentiation induced by staphylococcal protein A to search for potential differentially expressed lncRNAs and related regulatory networks.
METHODS: Bone marrow mesenchymal stem cells of rats were divided into experimental group and control group. The former group was induced osteogenic differentiation for 7 and 14 days in staphylococcal protein A (1 μg/mL) (to simulate osteomyelitis environment), while the control group was induced osteogenic differentiation for 7 and 14 days without any treatment. The content detection of alkaline phosphatase and alizarin red staining were used to observe the osteogenic induction of the cells. Two groups of cells were collected for transcriptome sequencing, gene ontology and KEGG bioinformatics analysis. Differential expression levels of lncRNAs were further verified by real-time fluorescence quantitative PCR.
RESULTS AND CONCLUSION: (1) After 7 and 14 days of osteogenic differentiation, the staining area of alizarin red in the control group was larger than that in the experimental group, and the alkaline phosphatase activity in the control group was significantly higher than that in the experimental group (P < 0.01). (2) Transcriptome sequencing results found 328 lncRNAs with significant differential expression level, with 184 significantly up-regulated and 144 significantly down-regulated. (3) Gene enrichment analysis showed that the function of differentially expressed genes mainly involved biological process, cell classification and molecular function. KEGG enrichment analysis found that the main signal pathways included tumor necrosis factor, MAPK and intestinal inflammatory disease signaling pathways, which were related to bone formation and bone metabolism. (4) Six differentially expressed lncRNAs related to osteogenesis differentiation were selected, and the expression trends of ENSRNOT00000092811, TCONS_00023877, and XR_001836916.1 were consistent with high-throughput sequencing after qPCR validation. (5) The results showed that multiple lncRNAs were involved in the regulation of osteogenic differentiation ability of rat bone marrow mesenchymal stem cells in osteomyelitis, and they may play a role through signal pathway, such as tumor necrosis factor and MAPK signaling pathways.

Key words: osteomyelitis, bone marrow mesenchymal stem cell, osteogenic differentiation, transcriptome sequencing, lncRNA

CLC Number:

Wen Hongjie, Chen Zhong, Yang Huagang, Xu Yongqing. Transcriptome sequencing analysis of osteogenic rat bone marrow mesenchymal stem cells induced by osteomyelitis[J]. Chinese Journal of Tissue Engineering Research, 2023, 27(15): 2333-2338.

Figures/Tables 7

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