The purpose of this article is to integrate the transcriptomic analysis and the proteomic profiles and to reveal and compare the different molecular mechanisms of PC12 cell growth on the surface of chitosan films and collagen/chitosan films.First,the chitosan films and the collagen/chitosan films were prepared.Subsequently,the cell viability assay was performed;the cell viability of the PC12 cells cultured on the collagen/chitosan films for 24 h was significantly higher than that on the chitosan films.Then,with cDNA microarray,the numbers of differentially expressed genes of PC12 cells on the surface of chitosan and collagen/chitosan films were 13349 and 5165,respectively.Next,the biological pathway analysis indicated that the differentially expressed genes were involved in 40 pathways directly related to cell adhesion and growth.The integrated transcriptomic and our previous proteomic analysis revealed that three biological pathways-extracellular matrix-receptor interaction,focal adhesion and regulation of actin cytoskeleton-were regulated in the processes of protein adsorption,cell adhesion and growth.The adsorbed proteins on the material surfaces further influenced the expression of important downstream genes by regulating the expression of related receptor genes in these three pathways.In comparison,chitosan films had a strong inhibitory effect on PC12 cell adhesion and growth,resulting in the significantly lower cell viability on its surface;on the contrary,collagen/chitosan films were more conducive to promoting PC12 cell adhesion and growth,resulting in higher cell viability.
The purpose of this paper is to utilize the signaling pathway polymerase chain reaction(PCR)arrays to investigate the activation of two important biological signaling pathways in endothelial cell adhesion and growth mediated by adsorbed serum protein on the surface of bare and titanium nitride(TiN)-coated nickel titanium(NiTi)alloys.First,the endothelial cells were cultured on the bare and TiN-coated NiTi alloys and chitosan films as control for 4 h and 24 h,respectively.Then,the total RNA of the cells was collected and the PCR arrays were performed.After that,the differentially expressed genes in the transforming growth factor beta(TGF-b)signaling pathway and the regulation of actin cytoskeleton pathway were screened out;and the further bioinformatics analyses were performed.The results showed that both TGF-b signaling pathway and regulation of actin cytoskeleton pathway were activated in the cells after 4 h and 24 h culturing on the surface of bare and TiN-coated NiTi alloys compared to the chitosan group.The activated TGF-b signaling pathway promoted cell adhesion;the activated regulation of actin cytoskeleton pathway promoted cell adhesion,spreading,growth and motility.In addition,the activation of both pathways was much stronger in the cells cultured for 24 h versus 4 h,which indicated that cell adhesion and growth became more favorable with longer time on the surface of two NiTi alloy materials.
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
The aim of this article is to apply proteomics in the comparison of the molecular mechanisms of PC12 cell adhesion and growth mediated by the adsorbed serum proteins on the surfaces of chitosan and collagen/chitosan films.First,the chitosan and the collagen/chitosan films were prepared by spin coating;and their surface morphologies were characterized by scanning electron microscopy,X-ray energy dispersive spectroscopy,contact angle measurement and Fourier transform infrared spectroscopy.Subsequently,cell proliferation experiments on two materials were performed and the dynamic curves of protein adsorption on their surfaces were measured.Then,proteomics and bioinformatics were used to analyze and compare the adsorbed serum proteins on the surfaces of two biomaterials;and their effects on cell adhesion were discussed.The results showed that the optimum concentration of chitosan film was 2%w/v.When compared with chitosan film,collagen/chitosan film promoted the growth and proliferation of PC12 cells more significantly.Although the dynamic curves showed no significant difference in the total amount of the adsorbed proteins on both surfaces,proteomics and bioinformatics analyses revealed a difference in protein types:the chitosan surface adsorbed more vitronectin whereas collagen/chitosan surface adsorbed more fibronectin 1 and contained more cell surface receptor binding sites and more Leu-Asp-Val sequences in its surface structure;the collagen/chitosan surface were more conducive to promoting cell adhesion and growth.
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.
