2006 Fiscal Year Final Research Report Summary
Epithelial wound healing activity of lactic acid bacteria
| Project/Area Number |
17580231
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| Research Category |
Grant-in-Aid for Scientific Research (C)
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| Allocation Type | Single-year Grants |
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| Section | 一般 |
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| Research Field |
Zootechnical science/Grassland science
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| Research Institution | Hirosaki University |
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Principal Investigator |
TOBA Takahiro Hirosaki University, Faculty of Agriculture and Life Science, Professor, 農学生命科学部, 教授 (10108483)
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| Project Period (FY) |
2005 – 2006
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| Keywords | epithelial cell culture / adhesion / acceleration of wound healing / lactobacilli |
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| Research Abstract |
In 2006 about 50 lactobacillar and bifidobacterial strains of human or animal origins were tested for adhesion to extracellular matrix and human enterocyte-like cell lines. Adhesive and well-growing strains were selected among them.
In 2007 wound healing activity was evaluated among each one strain of L. acidophilus, L. crispatus and L. gasseri That is, lactobacillar cells were applied onto the postconfluent Caco-2 cell monolayer, a human enterocyte-like cell line, and incubated under 5% CO_2 atmosphere. After removal of lactobacillar cells and washing with PBS, circular wounds were made by aspiration of a part of cell monolayer (wounding). Then after, wound healing process of injured cell monolayer was observed under CO_2 atmosphere using microscope equipped with digital camera. Wound healing was accelerated by the pre-treatment of the cell monolayer in all of three lactobacillar strains. Next wound healing mechanism was studied on a L. acidophilus strain that showed the strongest activity among three strains tested. When the wound healing activity was compared among living-cells, heat-killed cells and supernatant obtained from the cell suspension, heat-killed cells showed the activity comparable to living cells but the supernatant did not. Therefore we postulated cell surface component is responsible for the activity. Then S-layer protein was extracted and subjected to evaluation of wound healing activities as L. acidophilus is known to expressing an S-layer protein. Activity was observed in insoluble faction, but not in soluble fraction. Events occurred in Caco-2 cells after wounding with or without pre-treatment of lactobacillar cells were observed by fluorescent microscopy. Accumulation of actin filaments was observed in Caco-2 cells pretreated with lactobacillar cells. It is concluded that L.acidophilus cells accelerates wound healing of caco-2 cell monolayer by inducing the accumulation of actin filament.
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