Dong Ho Lee

Characterization of proteolytically digested Zebrafish chorion as extracellular matrix

Extracellular matrix (ECM) plays pivotal roles in developments, wound healing, migration, angiogenesis, and other tissue developments. Developing new ECM may give us an innovative approach of controlling the cell and tissue behavior, regenerating damaged tissues, and other biotechnology usages. Here we report new ECM like biomaterials, pFZC, obtained from the fertilized Zebrafish chorion (FZC). pFZC was prepared by digesting the fertilized Zebrafish chorion with pronase. Differentiation of P19 embryonic carcinoma cells (P19 EC) on pFZC demonstrated that the level of the differentiation of P19 EC was comparable to cells on gelatin or fibronectin coated cover slide. Analyses of glycosylations of Zebrafish chorions using various Lectins revealed that during the fertilization, structurally organized glycosylations of Zebrafish chorion were increased and FZC was glycosylated with mannose, fucose, n-acetylglucosamine, and sialic acid. In addition, after FZC proteolytic digestion with pronase, the digested FZC proteins, pFZC, displayed similar glycosylation manner. SDS gel electrophoresis and mass spectroscopy analysis of pFZC revealed that the major component of pFZC is ZP2 protein. Newly identified ECM like biomaterials, pFZC, will provide more broad ways of controlling the cell and tissue environments.Extracellular matrix (ECM) plays pivotal roles in developments, wound healing, migration, angiogenesis, and other tissue developments. Developing new ECM may give us an innovative approach of controlling the cell and tissue behavior, regenerating damaged tissues, and other biotechnology usages. Here we report new ECM like biomaterials, pFZC, obtained from the fertilized Zebrafish chorion (FZC). pFZC was prepared by digesting the fertilized Zebrafish chorion with pronase. Differentiation of P19 embryonic carcinoma cells (P19 EC) on pFZC demonstrated that the level of the differentiation of P19 EC was comparable to cells on gelatin or fibronectin coated cover slide. Analyses of glycosylations of Zebrafish chorions using various Lectins revealed that during the fertilization, structurally organized glycosylations of Zebrafish chorion were increased and FZC was glycosylated with mannose, fucose, n-acetylglucosamine, and sialic acid. In addition, after FZC proteolytic digestion with pronase, the digested FZC proteins, pFZC, displayed similar glycosylation manner. SDS gel electrophoresis and mass spectroscopy analysis of pFZC revealed that the major component of pFZC is ZP2 protein. Newly identified ECM like biomaterials, pFZC, will provide more broad ways of controlling the cell and tissue environments.

Blueberry protects LPS-stimulated BV-2 microglia through inhibiting activities of p38 MAPK and ERK1/2

The effects of blueberry extract (BE) were investigated on activities of p38 mitogen-activated protein kinase (MAPK) and extracellular extracellular signal regulated kinase 1/2 (ERK1/2) in mouse BV-2 microglia stimulated lipopolysaccharide (LPS). BV-2 microglia were cultured for 24 h in the presence of 1 μg/mL LPS either with or without BE preincubation for 0, 1, and 12 h. BE relieved repression of cell proliferation, and reduced cell death. These alterations caused by BE addition accompanied reduction of radical oxygen species (ROS). There were also significant decreases in the levels of both transcripts, and proteins of inducible nitric oxide synthase (iNOS) and peroxiredoxin 1 (Prx1) genes in the BE-treated cells, suggesting that BE inhibit the ROS-induced gene expression. The antioxidant effects of BE appeared to involve p38 MAPK and ERK1/2 signaling pathways since BE reduced both kinase activities by inhibiting phosphorylation. Taken these results together, BE protects LPS-stimulated BV-2 microglia by reducing cell death, ROS accumulation, and activities of p38 MAPK and ERK1/2.