Abstract
BackgroundStem 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).MethodsColony-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.ResultsMSMSCs 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.ConclusionRAPA can promote osteogenic differentiation of MSMSCs, indicating a possible relationship between osteogenic differentiation and autophagy.
Highlights
Maxillary sinus membrane elevation is a frequent option when bone height in the posterior maxilla is less than four mm
maxillary sinus membrane stem cells (MSMSCs) possessed the active ability of proliferation and reached confluence after three days of culture in complete medium. 1 ×103 single cells were seeded at low density, forming 49 ± 4.0 colonies (1CFU-F per 204 cells) (Fig. 1A)
After osteogenic induction for 28 days, we observed calcium granules stained by Alizarin Red in MSMSCs
Summary
Maxillary sinus membrane elevation is a frequent option when bone height in the posterior maxilla is less than four mm. Maxillary sinus membrane elevation consists of two situations, bone augmentation with or without bone substitutes. Promoting effect of rapamycin on osteogenic differentiation of maxillary sinus membrane stem cells. 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). Real-time quantitative PCR (qPCR) evaluated subsequent autophagic levels and osteogenic differentiation. MSMSCs possessed an active proliferation and multi-differentiation capacity, showing a phenotype of mesenchymal stem cells. ALP activity and calcium nodules forming in four RAPA-treated groups on three-time points (7, 14, 21 d) showed significant differences. 100 nM and 10 nM RAPA-treated groups showed more bone formation in vivo. RAPA can promote osteogenic differentiation of MSMSCs, indicating a possible relationship between osteogenic differentiation and autophagy
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