Carbon Dots from Lycium barbarum Attenuate Radiation-Induced Bone Injury by Inhibiting Senescence via METTL3/Clip3 in an m6A-Dependent Manner

Radiation-induced bone injury management remains a challenge in clinical practice, and there is no effective medicine. Recently, biomass-derived carbon dots (CDs) have attracted attention in biomedical engineering due to the advantages of abundant heteroatoms, low toxicity, and no need to drug loading. Here, we report that CDs, synthesized from Lycium barbarum via hydrothermal strategy, can effectively alleviate radiation-induced bone injury. CCK-8, apoptosis analysis, beta-galactosidase staining, quantitative polymerase chain reaction, and western blots demonstrate that CDs can mediate radiation-induced damage and senescence of bone marrow mesenchymal stem cells (BMSCs). CDs regulate osteogenic-and adipogenic-balance after irradiation, shown by alizarin red and oil red O staining. In vivo experiments reveal that CDs prevent the occurrence of osteoradionecrosis in rats, demonstrated by micro-CT and histology examination. The osseointegration of titanium implants installed in irradiated bone is promoted by CDs. Mechanistically, CDs increase the N6-methyladenosine (m6A) level of irradiated BMSCs via the increased methyltransferase-like 3 (METTL3). High-throughput sequencing facilitates detection of increased m6A levels located in the 3 '-untranslated regions (UTR) of the CAP-Gly domain containing linker protein 3 (Clip3) mRNA. The dual-luciferase reporter assay shows that 3 ' UTR is the direct target of METTL3. Subsequently, the increased m6A modification led to enhanced degradation of mRNA and downregulated CLIP3 expression, eventually resulting in the alleviation of radiation-induced bone injury. Interfering with the METTL3/Clip3 axis can antagonize the effect of CDs, indicating that CDs mediate radiation-induced bone injury via the METTL3/Clip3 axis. Taken together, CDs from L. barbarum alleviate radiation-induced bone injury by inhibiting senescence via regulation of m6A modification of Clip3. The present study paves a new pathway for the management of radiation-induced bone injury.