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CA1 Modulates the Osteogenic Differentiation of Dental Follicle Stem Cells by Activating the BMP Signaling Pathway In Vitro

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Tissue Engineering and Regenerative Medicine Aims and scope

Abstract

Background:

Carbonic anhydrase 1 (CA1) has been found to be involved in osteogenesis and osteoclast in various human diseases, but the molecular mechanisms are not completely understood. In this study, we aim to use siRNA and lentivirus to reduce or increase the expression of CA1 in Dental follicle stem cells (DFSCs), in order to further elucidate the role and mechanism of CA1 in osteogenesis, and provide better osteogenic growth factors and stem cell selection for the application of bone tissue engineering in alveolar bone fracture transplantation.

Methods:

The study used RNA interference and lentiviral vectors to manipulate the expression of the CA1 gene in DFSCs during in vitro osteogenic induction. The expression of osteogenic marker genes was evaluated and changes in CA1, alkaline phosphatase (ALP), Runt-related transcription factor 2 (RUNX2), and Bone morphogenetic proteins (BMP2) were measured using quantitative real-time polymerase chain reaction (qRT-PCR) and Western blotting (WB). The osteogenic effect was assessed through Alizarin Red staining.

Results:

The mRNA and protein expression levels of CA1, ALP, RUNX2, and BMP2 decreased distinctly in the si-CA1 group than other groups (p < 0.05). In the Lentivirus-CA1 (LV-CA1) group, the mRNA and protein expressions of CA1, ALP, RUNX2, and BMP2 were amplified to varying degrees than other groups (p < 0.05). Apart from CA1, BMP2 (43.01%) and ALP (36.69%) showed significant upregulation (p < 0.05). Alizarin red staining indicated that the LV-CA1 group produced more calcified nodules than other groups, with a higher optical density (p < 0.05), and the osteogenic effect was superior.

Conclusions:

CA1 can impact osteogenic differentiation via BMP related signaling pathways, positioning itself upstream in osteogenic signaling pathways, and closely linked to osteoblast calcification and ossification processes.

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Data availability

The data that support the findings of this study are available on request from the corresponding author. The data are not publicly available due to privacy or ethical restrictions.

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Acknowledgements

Research was funded by grants from the National Natural Science Foundation of China (81070817) and Natural Science Foundation of Shandong Province (ZR2010HM054; ZR2015HM022). The funding body played no role in the design of the study and collection, analysis, and interpretation of data and in writing the manuscript.

Funding

National Natural Science Foundation of China, 81070817, Wen-lin **ao,Natural Science Foundation of Shandong Province, ZR2010HM054, Wen-lin **ao, ZR2015HM022, Wen-lin **ao

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Authors and Affiliations

  1. Department of Oral and Maxillofacial Surgery, The Affiliated Hospital of Qingdao University, Qingdao, 266003, China

    **-ze Zhao, Ying-Ying Ge, Ling-fa Xue, Yao-xiang Xu, ** Yue, Cong Li & Wen-lin **ao

  2. School of Stomatology, Qingdao University, Qingdao, 266023, China

    **-ze Zhao, Ying-Ying Ge, Ling-fa Xue, Yao-xiang Xu, ** Yue, Cong Li & Wen-lin **ao

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  1. **-ze Zhao
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  6. Cong Li
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Contributions

**-ze Zhao and Ying-Ying Ge: conducted the experiments, performed data collection and/or assembly; data analysis and interpretation; and wrote the manuscript. Cong Li: data collection and analysis. Ling-fa Xue, Yao-xiang Xu, ** Yue and Wen-lin **ao: developed the study conception and design; performed data analysis and interpretation; administrative and manuscript proof.All authorsread and approvedthe manuscript.

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Correspondence to Wen-lin **ao.

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Ethical statement

This study was approved by the Ethics Committee of Qingdao University (Approval Number: QYFYWZLL27632), and all samples were obtained with informed consent from patients.

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Zhao, Jz., Ge, YY., Xue, Lf. et al. CA1 Modulates the Osteogenic Differentiation of Dental Follicle Stem Cells by Activating the BMP Signaling Pathway In Vitro. Tissue Eng Regen Med (2024). https://doi.org/10.1007/s13770-024-00642-4

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