The aim of this study is to unveil the role of integrin in influencing the differentiation of PC12 cell grown on PLLA aligned nanofibers by integrative study of cDNA microarray,proteomics technology and microRNA sequencing technology.First,PLLA-aligned nanofibers were prepared by electrospinning,and the chemical composition and surface morphology of the nanofibers were examined.Then,morphology and neurite length of PC12 cells and GRGDS-treated PC12 cells on PLLA-aligned nanofibers(AF and GA group)were measured by high-content analysis system.Subsequently,protein expression profile was analyzed by iTRAQ proteomics technology and 250 differentially expressed proteins were screened in the GA group.Integrative analysis of mRNAmicroRNA-protein data showed that‘MAPK signaling pathway’was the key affected pathway after the function of integrin were interfered by GRGDS.Other seven pathways were also influenced,and the differentiation of PC12 cell grown on PLLA aligned nanofibers was finally inhibited.
The aim of this article is to reveal the influence of aligned/random poly(L-lactic acid)(PLLA)nanofibers on PC12 cell differentiation from the perspective of metabolic level.First,three materials-PLLA aligned nanofibers(PLLA AF),PLLA random nanofibers(PLLA RF)and PLLA films(control)-were prepared by electrospinning and spin coating.Their surface morphologies were characterized.Subsequently,the cell viability,cell morphology and neurite length of PC12 cells on the surface of the three materials were evaluated,indicating more neurites in the PLLA RF groups but the longer average neurite length in the PLLA AF groups.Next,the metabolite profiles of PC12 cells cultured on the surface of the three nanofibers after 12 h,24 h and 36 h showed that,compared with the control,51,48 and 31 types of differential metabolites were detected at the three time points among the AF groups,respectively;and 56,45 and 41 types among the RF groups,respectively.Furthermore,the bioinformatics analysis of differential metabolites identified two pathways and three metabolites critical to PC12 cell differentiation influenced by the nanofibers.In addition,the verification experiment on critical metabolites and metabolic pathways were performed.The integrative analysis combining cytology,metabolomics and bioinformatics approaches revealed that though both PLLA AF and RF were capable of stimulating the synthesis of neurotransmitters,the PLLA AF were more beneficial for PC12 cell differentiation,whereas the PLLA RF were less effective.
Xiaoman SuYan HuangRong ChenYiwen ZhangMeichen HeXiaoying Lu
