Promoting effect of rapamycin on osteogenic differentiation of maxillary sinus membrane stem cells

Yanjun Lin; Min Zhang; Lin Zhou; Xuxi Chen; Jiang Chen; Dong Wu

doi:10.7717/peerj.11513

Promoting effect of rapamycin on osteogenic differentiation of maxillary sinus membrane stem cells

PeerJ. 2021 Jun 1:9:e11513. doi: 10.7717/peerj.11513. eCollection 2021.

Authors

Yanjun Lin^{1

2}, Min Zhang^{3

4}, Lin Zhou², Xuxi Chen², Jiang Chen², Dong Wu²

Affiliations

¹ Fujian Key Laboratory of Oral Diseases, Fujian Medical University, Fuzhou, Fujian, China.
² Research Center of Dental and Craniofacial Implants, Fujian Medical University, Fuzhou, Fujian, China.
³ Fujian Provincial Engineering Research Center of Oral Biomaterial, Fujian Medical University, Fuzhou, Fujian, China.
⁴ General Department of Hubin Campus, Hangzhou Stomatology Hospital, Hangzhou, Zhejiang, China.

Abstract

Background: Stem cells located in the maxillary sinus membrane can differentiate into osteocytes. Our study aimed to evaluate the effect of rapamycin (RAPA) on the osteogenic differentiation of maxillary sinus membrane stem cells (MSMSCs).

Methods: Colony-forming unit assay, immunophenotype identification assay, and multi-differentiation assay confirmed characteristics of MSMSCs obtained from SD rats. Transmission electron microscopy (TEM) and flow cytometry (FCM) identified the initial autophagic level of MSMSCs induced by RAPA. Real-time quantitative PCR (qPCR) evaluated subsequent autophagic levels and osteogenic differentiation. Alkaline phosphatase (ALP) activity assay and alizarin red staining (ARS) evaluated subsequent osteogenic differentiation. We performed a histological examination to clarify in vivo osteogenesis with ectopic bone mass from BALB/c nude mice.

Results: MSMSCs possessed an active proliferation and multi-differentiation capacity, showing a phenotype of mesenchymal stem cells. The autophagic level increased with increasing RAPA (0, 10, 100, 1,000 nM) and decreased over time. ALP activity and calcium nodules forming in four RAPA-treated groups on three-time points (7, 14, 21 d) showed significant differences. Col1a1, Runx2, and Spp1 expressed most in 100 nM RAPA group on 7 and 14 d. Osteogenesis-related genes except for Ibsp expression between four groups tended to be consistent on 21 d. 100 nM and 10 nM RAPA-treated groups showed more bone formation in vivo.

Conclusion: RAPA can promote osteogenic differentiation of MSMSCs, indicating a possible relationship between osteogenic differentiation and autophagy.

Keywords: Autophagy; Osteogenic differentiation; Rapamycin; Maxillary sinus membrane stem cells.

Grants and funding

The work was supported by the Natural Science Foundation of Fujian Province (2018J01819). The funders had no role in study design, data collection and analysis, decision to publish, or preparation of the manuscript.