The Study Of Mast Cells Of Mucosa Scar After Cleft Palate

Objective:Through the rabbit model of cleft palate made,simultaneously cleft palate repaired, two months later, take the scar tissue, and the normal mucosa tissue for comparative study. cleft palate repair postoperatively,to observe the number of mast cells in the scar tissue:the structure, distribution, and its mast cells related active material change.This study included two parts as follows:Part Ⅰ Histochemical stainingMethods:Choose7Zhou Ling, Japan’s big ears white rabbit12for research samples, raising7days, random only6under general anesthesia jaws made crack, simultaneously cleft surgery to repair; Stay jaws mucosal scarring, take intravenous air embolism method to be put to death, take the experimental group and control group in part in jaws mucosal1cm×0.5cm,4%in darfur-marin liquid fixed. After24hours, paraffin. Paraffin wax biopsy4～5micron, conventional lost wax to water. Bismarck brown, with the iodine green, Martin’s yellow dye combined. Under light microscopy and photography.Results:1. Macroscopic observation group jaws mucous membrane of scar tissue around with the normal mucosa tissue boundaries visible, scar area palatal Zhou Bi disappear, the color is more white, about1.0cm long,0.5cm wide; Quality of a material is less fat, the normal mucosa color pink, a soft, palatal Zhou Bi obvious. The mucous membrane for hoar section or shallow pink; The control group mucosal color pink.2. The mast cells large size, shape irregular;3.The layered each cell is not obvious, part of the mast cells are degranulating; Part of the mast cells still remaining in the particles. The control group mast cells is mainly distributed in the lamina propria,often on the connective tissue of the deep, blood vessels, secretion of tube around, quantity is less, clear-cut, did not find degranulation phenomenon, the cytoplasm less particles. Part Ⅱ Immunohistochemical stainingMaterials:The same to experiment oneMethods:Substance P and VIP vascular active peptide immunohistochemical staining, tissue samples from more than stick in the treatment of the slides together lysine, roast piece for the night. Alcohol with no water benzene,95%alcohol for alcohol respectively, PBS buffer for irrigation. Dry the slice, temperature nourishing, and PBS buffer for irrigation. Dry the slice, normal serum working liquid goat closed at room temperature incubation, respectively, adding the rabbit dilution of people substance P antibody and VIP polyclonal antibody,4℃for the night. Adding biotin marks two of working liquid, horseradish enzyme mark LuanBai element working liquid, and temperature. PBS buffer for flushing, DAB show color. Haematoxylin immersion, differentiation liquid differentiation, Neutral gum sealing piece. Under light microscopy and photography.Results:1. Immunohistochemical stains substance P reactant exists in the cytoplasm in MC,agranular, brown, the nucleus for the negative.2. Brown is rare in the control group particle, particle concentration.3. the more granular brown, and grain scattered in not focused.Conclusion:1. The two kinds of dyeing methods all found markers of mast cells in the organization mucosa number which is in the normal mucosa.2. The combination of dyeing method is more obvious mast cells that form, structure, distribution changes. Show the mast cells in the scar tissue in the normal mucosa and the differences between them.3. Immunohistochemical stain reveals in the scar tissue more granular brown; And scar tissue on mucosa particles. And the normal mucosa brown particles in less, and scattered.4. Neuropeptide substance P in scarring plays an important role in the process, but the relationship with the mast cells, the action mechanism does not clear.