Effect Of Ascorbic Acid On The Mitochondria In Bone Marrow Stromal Cells In Vitro

Subject: The defects of oral and maxillofacial bone tissue have always been one of the common clinical diseases.During the series of studies on the promotion of osteogenesis by nanomaterials,it has been demonstrated that ascorbic acid carbon dots(CDs)promotes osteogenesis.However,it is unclear whether this new type of nanomaterial has toxic effects on cells as other nanomaterials.In this study,from the perspective of subcellular analysis,the effect of ascorbic acid CDs on mitochondria in bone marrow stromal cells(BMSCs)was explored,thus providing basis for the research and application of ascorbic acid CDs in alveolar bone.Methods: Bone marrow stromal cells were extracted from the femurs of Wistar rat and cultured in vitro.The 3rd to 5th generation cells were co-cultured with ascorbic acid CDs for 0 h,24 h,48 h,and 72 h.1,select the best experimental concentration: MTT detection of different concentrations of ascorbic acid CDs on the proliferation of BMSCs,and select the best experimental concentration;2,detection of intracellular ROS content: CM-H2 DCFDA staining for detection of intracellular ROS;3.Detection of mitochondrial morphology: observation of mitochondrial morphology by transmission electron microscopy(TEM);4.Detection of the effect on mitochondrial structure: After staining with JC-1,the mitochondrial structure was observed under a microscope;5.Detection of effects on mitochondrial function: Western Blot was used to detect the expression of AMPK?,and ATP test kit was used to detect ATP production.Result:1.With the increase of ascorbic acid CDs concentration,the proliferation ability of BMSCs decreased;and the optimal experimental concentration was 40ug/ml;2.With the prolongation of co-culture time,intracellular ROS content increased;3.With the prolongation of co-culture time,the morphology of mitochondria gradually changed into fission,pyknosis and vacuole-like changes.4.With the prolongation of co-culture time,the mitochondrial membrane potential decreased;5.With the prolongation of co-culture time,AMPK? expression increased and ATP production decreased.Conclusions: Low concentration(concentration <40 ?g/m L)of ascorbic acid CDs has little effect on BMSCs;high concentrations(>40 ?g/m L)of ascorbic acid CDs can cause increased ROS content,changes of mitochondrial morphology,loss of structure,and loss of function in BMSCs,with increasing concentrations and prolonged co-culture times.