The improvement of fibroblast growth on hydrophobic biopolyesters by coating with polyhydroxyalkanoate granule binding protein PhaP fused with cell adhesion motif RGD.
Polyhydroxyalkanoates (PHA), a family of biopolyesters, have been studied as tissue engineering biomaterials due to their adjustable mechanical properties, biodegradability and tissue compatibility. Amphiphilic PHA granule binding protein PhaP has been shown to be able to bind to hydrophobic surfaces of polymers, especially PHA, via strong hydrophobic interaction. Genes of PhaP and RGD peptides, which are a cell adhesion motif recognized by many cell surface receptors, were successfully expressed and obtained as a pure fusion protein PhaP-RGD in Escherichia coli DH5α. When films of poly(3-hydroxybutyrate-co-3-hydroxy- hexanoate) (PHBHHx), poly(3-hydroxybutyrate-co-3-hydroxyvalerate) (PHBV) and polylactic acid (PLA) were coated with PhaP-RGD, their surface hydrophilicities were all increased compared with their corresponding naked (non-coated) films, respectively. Among the three biopolyesters, PHBHHx demonstrated the strongest affinity to PhaP. In vitro study showed that mouse fibroblasts L929 and mouse embryonic fibroblasts NIH/3T3 attached better and grew faster on all three PhaP-RGD coated films compared with their related behaviors on PhaP coated and non-coated films, respectively. Both fibroblasts attached and grew very well on PhaP-RGD coated PHBHHx, PHBV and PLA, even in their serum-free medium, while the non-coated and PhaP coated biopolyesters poorly supported the cell growth if the two fibroblasts were incubated in their serum free medium. These results indicated that PhaP-RGD could be used as a coating material to improve cell growth on hydrophobic biopolyesters for implant tissue engineering purposes.
Dong
Ying
Y
Multidisciplinary Research Center, Shantou University, Shantou 515063, Guangdong, China.
Li
Ping
P
Chen
Chong-bo
CB
Wang
Zhi-hui
ZH
Ma
Ping
P
Chen
Guo-Qiang
GQ
D000818:Animals; D001426:Bacterial Proteins / Q000378:metabolism*; D001681:Biological Assay; D002448:Cell Adhesion / Q000187:drug effects; D049109:Cell Proliferation / Q000187:drug effects; D020099:Coated Materials, Biocompatible / Q000494:pharmacology*; D004268:DNA-Binding Proteins / Q000378:metabolism*; D005347:Fibroblasts / Q000166:cytology* / Q000187:drug effects / Q000378:metabolism / Q000648:ultrastructure; D049452:Green Fluorescent Proteins / Q000378:metabolism; D057927:Hydrophobic and Hydrophilic Interactions / Q000187:drug effects*; D008422:Materials Testing; D051379:Mice; D018625:Microscopy, Atomic Force; D008856:Microscopy, Fluorescence; D009842:Oligopeptides / Q000378:metabolism*; D011091:Polyesters / Q000494:pharmacology*; D054813:Polyhydroxyalkanoates / Q000494:pharmacology*; D011993:Recombinant Fusion Proteins / Q000096:biosynthesis / Q000302:isolation & purification; D012844:Sincalide / Q000494:pharmacology; D017155:Wettability / Q000187:drug effects
D001426:Bacterial Proteins; D020099:Coated Materials, Biocompatible; D004268:DNA-Binding Proteins; D009842:Oligopeptides; C118828:PHAP protein, Bacteria; D011091:Polyesters; D054813:Polyhydroxyalkanoates; D011993:Recombinant Fusion Proteins; C485184:enhanced green fluorescent protein; D049452:Green Fluorescent Proteins; C047981:arginyl-glycyl-aspartic acid; D012844:Sincalide