Assessing the responses of cellular proteins induced by hyaluronic acid-modified surfaces utilizing a mass spectrometry-based profiling system: over-expression of CD36, CD44, CDK9, and PP2A.
The cell responses to biopolymer surface at the early adhesion stages can be critical for cell survival. The purpose of this research was to assess formation of hyaluronic acid (HA) biopolymer surface, the fibroblasts were used as an experimental model to evaluate the responses of cellular proteins induced by biopolymer materials using a mass spectrometry-based profiling system. Surfaces were covered by multi-walled carbon nanotubes (CNT), chitosan (CS), and HA to increase the surface area, enhance the adhesion of biopolymer and promote the rate of cell proliferation. The amount of adhered fibroblasts on CNT/CS/HA electrodes of quartz crystal microbalance (QCM) were greatly exceeded those on other surfaces that were consistent with cell-count technique. Moreover, analyzing differential protein expressions of adhered fibroblasts on those biopolymer surfaces by proteomic approaches identified CD36, CD44, PP2A, and CDK9 as key proteins. To validate the influences of those four proteins on adhesions of fibroblasts on biopolymers, the cells were blocked by antibodies of the proteins and the adhesions of cells on the tested biopolymer surfaces were examined using a QCM technique, flow cytometric analysis and morphological observations. The results of significantly decreasing the weights and densities of the blocked fibroblasts adhering to CNT/CS/HA surfaces were obtained, and validate those proteins found by proteomic approaches. Utilizing mass spectrometry-based proteomics to evaluate cell adhesions on biopolymers is proposed.
Department of Chemical and Materials Engineering, National Yunlin University of Science and Technology, 123 University Road, Section 3, Douliou, Yunlin, 64002 Taiwan, ROC.
Ming-Hui
MH
Yang
Shiang-Bin
SB
Jong
Chi-Yu
CY
Lu
Yu-Fen
YF
Lin
Pei-Wen
PW
Chiang
Yu-Chang
YC
Tyan
Tze-Wen
TW
Chung
D000595:Amino Acid Sequence; D001973:Bromodeoxyuridine / Q000378:metabolism; D018955:CD36 Antigens / Q000737:chemistry / Q000378:metabolism; D002460:Cell Line; D049109:Cell Proliferation / Q000187:drug effects; D048271:Chitosan / Q000737:chemistry; D042863:Cyclin-Dependent Kinase 9 / Q000737:chemistry / Q000378:metabolism; D004566:Electrodes; D005347:Fibroblasts / Q000166:cytology / Q000187:drug effects / Q000378:metabolism; D005786:Gene Expression Regulation / Q000187:drug effects*; D006801:Humans; D018960:Hyaluronan Receptors / Q000737:chemistry / Q000378:metabolism; D006820:Hyaluronic Acid / Q000737:chemistry / Q000494:pharmacology*; D013058:Mass Spectrometry; D008969:Molecular Sequence Data; D037742:Nanotubes, Carbon / Q000737:chemistry; D054648:Protein Phosphatase 2 / Q000737:chemistry / Q000378:metabolism; D040901:Proteomics / Q000379:methods*; D013499:Surface Properties
D018955:CD36 Antigens; D018960:Hyaluronan Receptors; D037742:Nanotubes, Carbon; D006820:Hyaluronic Acid; D048271:Chitosan; D042863:Cyclin-Dependent Kinase 9; D054648:Protein Phosphatase 2; D001973:Bromodeoxyuridine