| The function damage of salivary glands caused by some diseases can result in severe xerotomia syndrome in clinical that makes the patients so suffering , but there is no effective remedy nowadays. The construction of submandibular glands by submandibular tissue engineering following with the development of tissue engineering provides a new approach to treat the xerotomia by the function damage of salivary glands. And in the study of salivary glands tissue engineering , the emphasis is the extracellular matrix scaffold material which can provide nutrition for the seeding cell adhesion, proliferation and differentiation. Scaffold material such as PGA, PLA may act as the place of cellular metabolism, but that may cause the immune exclusion effect and infection because the material are synthetic polymers which are foreign body. So that, it is attached importance to the finding of natural scaffold materials in the field.The purpose of this study was to get natural bioderived material by foreign submandibular glands after acellular process. The hzstological ingredients after acellular process were analyzed by histological and histochemistry study. The submandibular cells with optimization cultivation were mixed - cultured, and the histocompatibility and cytocompatibility were analyzed of scaffold material by in vivo and in vitro experimental studies. So, the suitable scaffold material for submandibular glands tissue engineering may be found.Methods1. Submandibular glands were cell - extracted by chemical detergent and amylase digestion to obtain natural bioderived scaffold material.2. The effects of different concentrations of TritonX - 100 to cell - extract the submandibular glands were compared.3. The histological ingredients of the bioderived scaffold material were analyzed by histology and histochemistry study. The surface structures were observed by scanning electron microscope, and the diameters of holes were measured, the holes proportion was calculated at the same time.4. The extracting liquid of submandibular gland acellular matrix scaffold material was made, and its toxicity effect to the body was analyzed by acute systemic toxicity test.5. The effect to tissues was observed by implanting test in body, and then the histocompatibility was determined.6. The MTT colorimetric method was used to test the OD value at the 490nm wavelength of the mixed - culturing cells at the 7th day, and the survival rate was obtained. The cytocompatibility was judged by relative proliferation per-cents of cells.7. Submandibular glands cells were isolated and purified by amylase digestion. And then the cells were cultured and subcultured. The shapes and structures were observed by phase contrast microscope.8. The second generation of the subculturing cells were mixed with the submandibular gland acellular matrix (SGAM ) scaffold material to be cultured, and then at the 3th,5th,7th day, both the growth condition of the cells on the scaffold material and the surface shapes and structures of the cells were observed by the scanning electron microscope.Results1. Analysis of the histological ingredients of submandibular gland acellular matrix (SGAM) scaffold materials.(1) Histomorphological observation:Observed by microscope: SGAM after cell - extracted by 1 % TritonX -100, the figures existed, but cells arranged out of order, crowding, and the nucleus were irregular that some concentrated, some broke, the shapes of cells hadno obvious variation. However, SGAM after cell - extracted by 3% TritonX -100, the submandibular glands cellular ingredients disappeared, the shapes couldnt be seen, the pink collagen fibers arranged loosely, the splits were big and appeared network structure.( 2 ) Ultrastructural observationObserved by transmission electron microscope: SGAM after cell - extracted by 1 % TritonX - 100, the cell membranes broke, the cell organs destroyed and the chromatin concentrated, that became some scattered fragments, some nucleus concentrated, the semilunar apoptosis b...
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