Injectable biological active factor composition and preparation method and application thereof
A bioactive factor and composition technology, which is applied to the injectable bioactive factor composition and the fields of its preparation and application, can solve the problems of reducing the strength of hydrogels, and achieves the improvement of gel strength, easy availability of materials, and shortening of gels. The effect of setting time
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Embodiment 1
[0046] Chitosan is dissolved in the acetic acid solution, is mixed with 3% chitosan solution, slowly adds in the oil phase liquid paraffin containing emulsifier (the emulsifier is the mixture of Span-80 and Tween-80, both volume ratios are 2 : 1, the volume ratio of emulsifier and oil phase is 1: 100.), stirring to form a W / O emulsion, adding 20% (w / w) sulfuric acid solution for cross-linking, forming sulfuric acid cross-linked chitosan microspheres, and then Wash with acetone, absolute ethanol and distilled water, respectively. Then sulfuric acid cross-linked chitosan microspheres are dispersed in water, and the sodium hydroxide solution with a concentration of 2% (w / v) is added dropwise under slow stirring until the final pH value of the reaction system is 7.6; After fully washing with ethanol and water, the chitosan microspheres are obtained after freeze-drying. figure 1 It is the infrared spectrogram of chitosan microspheres, which shows that the ion-crosslinked chitosa...
Embodiment 2
[0048] The chitosan was fully dissolved in 0.08 mol / L hydrochloric acid solution and the insoluble matter was filtered off to prepare a 1.8% (w / w) chitosan solution (pH value 5.8). Dissolve 10 mg of recombinant human bone morphogenetic protein-2 in 10 mL of chitosan solution, then slowly drop 0.5 mL of 45% (w / w) sodium glycerophosphate solution at 4°C to form an isotonic solution (300 mOsm); then Slowly add 50mg of chitosan microspheres into the above homogeneous solution to make recombinant human bone morphogenetic protein-2 composition, its pH value is 7.0, it is a flowable liquid at room temperature, increase the temperature to 37°C for 5 minutes form stable semi-solid gels, such as image 3 shown. Using scanning electron microscope to observe the dry gel, the results are as follows: Figure 4 As shown, the pore size of the xerogel is between 100-200 μm.
Embodiment 3
[0050] Firstly, 2-week-old Kunming mice were anesthetized by intraperitoneal injection of pentobarbital sodium at 3% of their body weight, fixed on the operating board in supine position, shaved and disinfected the posterior side of the right thigh, and injected with 1 mL syringe containing 100 μg of recombinant Composition 100 μL of human bone morphoprotein-2. After the operation, put them into the feeding box and feed them with normal clean standard feed. The mice were sacrificed 4 weeks after the operation, and one of the mice was quickly taken out of the implanted material and its surrounding soft tissue, immersed in 10% formalin solution for fixation, sectioned, and hematoxylin-eosin staining to observe the osteogenesis. observation, such as Figure 5 As shown, bone tissue, including trabecular bone and bone marrow, was found in the striated muscle tissue. The other 3 mice dissected their right legs to remove new long bones, as Image 6 As shown, the average fresh weig...
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