| Budget Amount *help |
¥1,900,000 (Direct Cost: ¥1,900,000)
Fiscal Year 1995: ¥500,000 (Direct Cost: ¥500,000)
Fiscal Year 1994: ¥600,000 (Direct Cost: ¥600,000)
Fiscal Year 1993: ¥800,000 (Direct Cost: ¥800,000)
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| Research Abstract |
We treated pancreatic stones just after removal each from pancreatic lithiasis patients to clarify the components of the organic matrix by immunohistochemistry. Stones were fixed in 4% buffered formaldehyde for 24 hours, then decalcified stationary in 0.5N HCI with 0.1M EDTA at room temperature for up to 60 minutes. All stones were decalcified with bubble and firm sponge-like matrices were obtained. Standard staining included H-E,PAS,and alcian blue was made. Immunohistochemical staining with specific fibronectin, laminin, collagen IV,lactoferrin (DAKO), and PSP-reg antibodies were carried out by avidin-biotin complex methods. Control staining with non-immune serum was negative. Standard staining revealed that organic component formed matrix throughout the stone and matrix was heterogeneous in texture such as reticular, lamellar, and hyalin-like including altered cellular constituent. In all seven matrices, there were wide areas intensely stained for fibronectin and laminin. All specimens had positive areas for collagen IV,lactoferrin and PSP-reg, but the degree of intensity and the pattern of distribution were different in each matrix. Process of decalcification in this study is suitable for immunohistochemistry. Fragmentation of matrix occured when decalcification was carried out in EDTA with constant stirring. Organic matrix was found throughout the pancreatic stone. Lactoferrin and PSP-reg were positive in the matrix as were expected. Fibronectin, laminin and collagen IV were positive in the matrix, so extracellular matrix and basement membrane components play an important role in the pathogenesis of the pancreatic stones.
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