title: Multi-biofunction of antimicrobial peptide-immobilized silk fibroin nanofiber membrane: Implications for wound healing.
abstract: An antimicrobial peptide motif (Cys-KR12) originating from human cathelicidin peptide (LL37) was immobilized onto electrospun SF nanofiber membranes using EDC/NHS and thiol-maleimide click chemistry to confer the various bioactivities of LL37 onto the membrane for wound care purposes. Surface characterizations revealed that the immobilization density of Cys-KR12 on SF nanofibers could be precisely controlled with a high reaction yield. The Cys-KR12-immobilized SF nanofiber membrane exhibited antimicrobial activity against four pathogenic bacterial strains (Staphylococcus aureus, Staphylococcus epidermidis, Escherichia coli, and Pseudomonas aeruginosa) without biofilm formation on the membrane surface. It also facilitated the proliferation of keratinocytes and fibroblasts and promoted the differentiation of keratinocytes with enhanced cell-cell attachment. In addition, immobilized Cys-KR12 significantly suppressed the LPS-induced TNF-α expression of monocytes (Raw264.7) cultured on the membrane. These results suggest that a Cys-KR12-immobilized SF nanofiber membrane, which has multiple biological activities, would be a promising candidate as a wound dressing material.
  
  
  
  This research article reports various bioactivities of an antimicrobial peptide on electrospun silk fibroin nanofiber membrane. Recently, human cathelicidin peptide LL37 has been extensively explored as an alternative antibiotic material. It has not only a great antimicrobial activity but also a wide variety of bioactivities which can facilitate wound healing process. Especially, many studies on immobilization of LL37 or its analogues have shown the immobilization technique could improve performance of wound dressing materials or tissue culture matrices. Nevertheless, so far studies have only focused on the bactericidal effect of immobilized peptide on material surface. On the other hand, we tried to evaluate multi-biofunction of immobilized antimicrobial peptide Cys-KR12, which is the shortest peptide motif as an analogue of LL37. We fabricated silk fibroin nanofiber membrane as a model wound dressing by electrospinning and immobilized the antimicrobial peptide. As a result, we confirmed that the immobilized peptide can play multi-role in wound healing process, such as antimicrobial activity, facilitation of cell proliferation and keratinocyte differentiation, and inhibition of inflammatory cytokine expression. These findings have not been reported and can give an inspiration in wound-care application.
authors.affiliation: Department of Biosystems and Biomaterials Science and Engineering, Seoul National University, Seoul 08826, Republic of Korea.
authors.forename: Dae Woong
authors.initials: DW
authors.lastname: Song
authors.affiliation: Product Tech Transfer Team, Ajinomoto Genexine Corporation, Incheon 21991, Republic of Korea.
authors.forename: Shin Hwan
authors.initials: SH
authors.lastname: Kim
authors.affiliation: Department of Biosystems and Biomaterials Science and Engineering, Seoul National University, Seoul 08826, Republic of Korea.
authors.forename: Hyung Hwan
authors.initials: HH
authors.lastname: Kim
authors.affiliation: Department of Biosystems and Biomaterials Science and Engineering, Seoul National University, Seoul 08826, Republic of Korea.
authors.forename: Ki Hoon
authors.initials: KH
authors.lastname: Lee
authors.affiliation: Department of Biosystems and Biomaterials Science and Engineering, Seoul National University, Seoul 08826, Republic of Korea. Electronic address: ki.cs@snu.ac.kr.
authors.forename: Chang Seok
authors.initials: CS
authors.lastname: Ki
authors.affiliation: Department of Biosystems and Biomaterials Science and Engineering, Seoul National University, Seoul 08826, Republic of Korea; Center for Food and Bioconvergence, Seoul National University, Seoul 08826, Republic of Korea; Research Institute of Agriculture and Life Sciences, Seoul National University, Seoul 08826, Republic of Korea. Electronic address: nfchempf@snu.ac.kr.
authors.forename: Young Hwan
authors.initials: YH
authors.lastname: Park
mesh_terms: D000818:Animals; D023181:Antimicrobial Cationic Peptides / Q000737:chemistry*; D001419:Bacteria / Q000254:growth & development*; D012831:Bombyx; D005349:Fibroins / Q000737:chemistry*; D008070:Lipopolysaccharides / Q000633:toxicity; D008264:Macrophages / Q000378:metabolism; D008567:Membranes, Artificial; D051379:Mice; D057139:Nanofibers / Q000737:chemistry*; D000067996:RAW 264.7 Cells; D014409:Tumor Necrosis Factor-alpha / Q000096:biosynthesis
chemical_list: D023181:Antimicrobial Cationic Peptides; D008070:Lipopolysaccharides; D008567:Membranes, Artificial; D014409:Tumor Necrosis Factor-alpha; D005349:Fibroins
keywords: Antimicrobial peptide; Immobilization; Nanofiber; Silk fibroin; Wound dressing