Injectable nano-network gels for diabetes treatment
A therapeutic agent, nanoparticle technology, applied in the fields of nanotechnology, nanotechnology, nanomedicine, etc., can solve problems such as limiting implantable applications
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example 1
[0082] Example 1 had 3.5 mg total enzymes (GOx and Cat) and 240 mg modified dextran. Those skilled in the art will recognize that the total amount of responsive signaling means will depend on the amount of responsive polymeric matrix and the level of activity of the signaling means. In some embodiments, the responsive polymeric matrix contains an acid-degradable polymer and the GOx / CAT signaling tool described above, wherein preferably the ratio (w / w) of total enzyme to polymer is 1:1000 to 1 :1, preferably 1:500 to 1:2, more preferably 1:100 to 1:15. In principle, the glucose oxidase may be any biocompatible enzyme exhibiting a glucose oxidase activity capable of oxidizing glucose to produce gluconic acid, preferably also peroxide when used in combination with a catalytic enzyme.
[0083] In some embodiments the weight ratio of GOx to CAT is optimized to enhance the ability of the enzyme mixture to respond to changing glucose levels. The weight ratio of GOx to CAT may be 1:...
example
[0169] All chemicals were purchased from Sigma-Aldrich and used as received unless otherwise specified. Human recombinant insulin (Zn salt, 27.5 IU / mg) was purchased from Invitrogen. Deionized water was prepared by a Millipore NanoPure purification system (resistivity higher than 18.2 MΩ·cm-1).
[0170] Example 1. Preparation of m-glucan
[0171] Briefly, 1.0 g of dextran (Mn about 9-11 kDa) was added to a flame-dried round bottom flask and purged with nitrogen. Add 10 mL of anhydrous dimethyl sulfoxide to the flask and stir until the dextran is completely dissolved. Pyridine p-toluenesulfonate (PPTS, 15.6 mg, 0.062 mmol) was added to the solution followed by 2-ethoxypropene (4.16 mL, 37 mmol). The reaction mixture was briefly purged with nitrogen and then sealed with parafilm to prevent evaporation of the reactants. The reaction was stirred at room temperature for 30 minutes to yield m-glucan. At that time, the reaction was quenched by adding 1 mL of triethylamine. The ...
example 2
[0172] Example 2. Preparation of glucose-responsive nanoparticles and nanonetwork gels
[0173] Materials and methods
[0174] Dextran nanoparticles were prepared by a modified double emulsion (water-in-oil-in-water) solvent evaporation / extraction method. Briefly, 0.5 mL containing only 35 mg human recombinant insulin (Invitrogen) or together with 3.5 mg enzyme (glucose oxidase and catalase) by sonication for 45 cycles (1 sec each, with a duty cycle of 60%) The weight ratio: 4:1) of the aqueous phase was emulsified with 5.8 mL of the organic phase (dichloromethane (DCM)) containing 240 mg m-dextran. Immediately thereafter, the primary emulsion was poured into 25 mL of chitosan or alginate aqueous solution (1%) and sonicated for 45 cycles. The double emulsion was then transferred to 150 mL of chitosan (Mn: 612 kDa; degree of deacetylation: 96.1%) or alginate (Mw: 1.6×10 5 ) in aqueous solution (0.2%). The mixed suspension was stirred at room temperature to eliminate DCM b...
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