| Objective:To extract the total flavonoids from Drynaria Rhizoma, prepare the microcapsules by Drynariae total flavonoids/Poly lactic-co-glycolic Acid(DR/PLGA) evaluate the influence of formulation parameters, and to prepare the microcapsules with the concentration and volume of the drug, amount of the polymer, stirring rate of the primary emulsions. To observe the activity of MC3T3-E1by CCK-8after co-cultured with DR/PLGA microcapsules.Methods:1.Referring to the content of total flavonoids, the Drynaria was selected and placed in the reflux extraction device to extract Drynaria total flavonoids.2.The DR/PLGA microcapsules was prepared by emulsion solvent evaporation technique under certain conditions. Optimized the preparation technology with different single factor conditions.the microspheres characteristics, such as the particle size,entrapment efficiency were taken as indexes for evaluating the microspheres. The general morphology of the microcapsules was observed by microscope, the surface morphology of the microcapsules was observed by scanning electron microscope(SEM) and transmission electron microscope(TEM), the distribution width of the microcapsules was observed by the particle size analyzer, encapsulation efficiency of the total flavonoids in microcapsules was measured by UV spectrophotometer. Try to find a DR/PLGA microcapsules with smaller particle size, uniform dispersion and high encapsulation efficiency.3. The cell proliferation by CCK-8was observed after co-cultured the DR/PLGA microcapsules with MC3T3-E1under sterile conditions.Results:1.Detected by the UV spectrophotometer, the content of total flavonoid was9.95%in Drynaria.2. The preparation technology was optimized with different single factor conditions, such as the concentration of PLGA, the stirring speed in colostrum, the emulsification time in colostrum,the water-oil ratio. Observed the microspheres characteristics such as the particle size, entrapment efficiency were taken as indexes for evaluating the microspheres.①Under the observed of gross morphology and size distribution with microscope, We found that the microcapsule dispersion was good, the particle size was dispersion uniformly and distribution narrowly when the PLGA concentration was less than140mg/ml, the stirring speed between2000rpm-4000rpm, the colostrum emulsification time between4minutes to6minutes, the colostrum water oil ratio was greater than1:10.②The microcapsules quality was measured by the total flavonoids encapsulation efficiency of Drynaria witch was measured by the the UV spectrophotometer. The best processing conditions were as follows:PLGA concentration:140mg/ml; stirring speed:2000rpm; Colostrum emulsification time:6minutes; water-oil ratio:1:15.③The microcapsules prepared in the optimization process display a uniform distribution and the particle diameter distribution is narrow under the microscope. The microcapsules performance edge regular round under the scanning electron microscope(SEM) and the transmission electron microscope(TEM). The particle size analyzer shows that the average particle diameter of the microcapsule was789.8±712.3nm, the distribution is relatively narrow, substantially less than5μm. The average encapsulation efficiency was47.72%.3. The DR/PLGA microcapsules and MC3T3-E1were co-cultured under sterile conditions. The cell viability had not increased significantly when the concentration of the microcapsules at O.lmg/ml and0.5mg/mL. When the microcapsule concentration greater than1mg/ml, the absorbance value A450also increases. the data had statistically significant.Conclusion:This experimental extracted the active ingredients of total flavonoids in Drynaria, prepared the DR/PLGA microcapsules, optimized the preparation conditions, prepared the DR/PLGA microcapsulesa with small particle size, uniform disperse, narrow distribution particle size and high encapsulation efficiency... |
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