In vitro growth and differentiated activities of human periodontal ligament fibroblasts cultured on salmon collagen gel.
Marine-derived collagen is expected to be a much safer alternative to calf collagen, which in medical applications carries the risk of bovine spongiform encephalopathy. In this study, acid-soluble collagen was extracted from salmon skin and crosslinked with 1-ethyl-3-(3-dimethylaminopropyl)-carbodiimide during fibril formation to produce a crosslinked salmon collagen (SC) gel. The growth rates and the differentiated functions of human periodontal ligament fibroblasts (HPdLFs) cultured on the SC gel were investigated. Growth was faster on the SC gel than on porcine collagen (PC) gel. In addition, the HPdLFs cultured on the SC gel exhibited higher alkaline phosphatase (ALP) activity than those cultured on the PC gel. Quantitative RT-PCR revealed higher mRNA expression of type I collagen, ALP, and osteocalcin in the HPdLFs cultured on the SC gel. HPdLFs had a flat shape on the SC gel and a spindle shape on the PC gel, as revealed by observation with scanning electron microscopy and immunostaining with cytoskeletal protein and vinculin. The results showed that HPdLFs could grow and show highly differentiated activity on the SC gel as well as on the PC gel.
Nagai
Nobuhiro
N
Creative Research Initiative "Sousei" (CRIS), Hokkaido University, N21-W10, Kita-ku, Sapporo, Hokkaido 001-0021, Japan. nagai@cris.hokudai.ac.jp
Mori
Kazuo
K
Satoh
Yasuharu
Y
Takahashi
Noritaka
N
Yunoki
Shunji
S
Tajima
Kenji
K
Munekata
Masanobu
M
D000469:Alkaline Phosphatase / Q000235:genetics / Q000378:metabolism; D000818:Animals; D001483:Base Sequence; D001672:Biocompatible Materials / Q000302:isolation & purification; D002454:Cell Differentiation; D049109:Cell Proliferation; D048430:Cell Shape; D002478:Cells, Cultured; D003094:Collagen / Q000302:isolation & purification; D024042:Collagen Type I / Q000235:genetics; D017931:DNA Primers / Q000235:genetics; D005347:Fibroblasts / Q000166:cytology / Q000378:metabolism; D005782:Gels; D006801:Humans; D066298:In Vitro Techniques; D008422:Materials Testing; D008855:Microscopy, Electron, Scanning; D015675:Osteocalcin / Q000235:genetics; D010513:Periodontal Ligament / Q000166:cytology* / Q000378:metabolism; D012333:RNA, Messenger / Q000235:genetics / Q000378:metabolism; D012474:Salmon; D013552:Swine
D001672:Biocompatible Materials; D024042:Collagen Type I; D017931:DNA Primers; D005782:Gels; D012333:RNA, Messenger; D015675:Osteocalcin; D003094:Collagen; D000469:Alkaline Phosphatase