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Objective @#To investigate the effect of the combination of stem cell leukemia(SCL)recombinant lentivirus and connexin 43 ( Cx43 ) recombinant lentivirus on the function of diabetic cystipathy ( DCP) in guinea pigs.@*Methods @#After 90 healthy guinea pigs were fed normally for 1 week, streptozotocin(STZ)was injected intraperito⁃ neally at 200 mg/kg in a single dose, and random blood glucose was monitored weekly. After 12 weeks of normal feeding, 40 guinea pigs meeting the criteria were screened by urodynamic examination and randomly divided into 4 groups: diabetic group, SCL group, Cx43 group, and SCL + Cx43 group. In the diabetic group, 0. 2 ml of empty lentivirus without gene was instilled into the bladder through the urethra, in the SCL and Cx43 groups, 0. 2 ml of SCL lentivirus and Cx43 lentivirus were instilled using the same method, in the SCL + Cx43 lentivirus group, 0. 2 ml of each SCL and Cx43 lentivirus was instilled through the urethra. After 14 days of transfection, urodynamic examination was performed, and then the guinea pigs were executed after the examination, and the bladders were quickly removed for frozen bladder sections and fluorescent double staining. @*Results @#There were no differences in urodynamic examinations between the SCL and Cx43 groups compared to the diabetic group( P > 0. 05 ) . Urodynamic examination in the SCL + Cx43 group showed improvement in detrusor pressure, abdominal pressure and bladder pressure compared to the diabetic group(P < 0. 05) . In laser confocal experiments, the number of interstitial cells of Cajal (ICC) was reduced in the diabetic group, and the spindle structure and cell protrusions were obviously destroyed and appeared in a state of cell lysis. In the SCL and Cx43 groups, there was an improvement in the spindle structure and cell protrusion of ICC⁃like cells, and there was no significant change in the number of cells. In the SCL + Cx43 group, there was an increase in the number of ICC⁃like cells, a significant improvement in the spindle structure and cell protrusion, and the formation of ICC⁃dimers.@*Conclusion @#Transurethral co⁃infusion of SCL and Cx43 gene recombinant lentivirus can be successfully transfected in guinea pig DCP bladder, which can restore the number and structure of damaged ICC cells and form ICC dimer structure, improve the pressure of diabetic bladder forced urinary muscle, improve the weakness of urination, ventral pressure voiding and other characteristics. It provides a new direction for the treatment of DCP.
RESUMEN
BACKGROUND:Periodontal tissue engineering technology provides new ideas and new ways for periodontitis-induced bone defect repair. OBJECTIVE:To develop a culture model for the periodontal ligament cells of miniature swine, which was constructed with hydroxyapatite, to investigate the biocompatibility with hydroxyapatite. METHODS:Periodontal ligament cells from miniature swine were harvested by using tissue explant method. Immunofluorescence was used to detect the expression of stromal cel antigen 1 in the periodontal ligament cells of miniature swine. The third passage cells were co-cultured with a three-dimension hydroxyapatite scaffold, and the biological characteristics of the cells were observed under a scanning electron microscope at days 1, 3, 7 of co-culture. RESULTS AND CONCLUSION:The pirmary miniature swine periodontal ligament cells grew wel , and they were positive for stromal cel antigen 1. Under the scanning electron microscope, the periodontal ligament cells of miniature swine grew wel on the hydroxyapatite scaffold at days 1, 3, 7 of co-culture. These prove that the miniature swine periodontal ligament cells, which can be separated using tissue explant method and cultured successful y in vitro, can grow wel on the hydroxyapatite scaffold.