Cascade enzyme-like nano system for promoting healing of diabetic wounds and preparation method and application thereof
A technology for wound healing and diabetes, which is applied in the field of cascade enzyme-like nanosystems for promoting diabetic wound healing and its preparation, can solve the problems of inability to promote diabetic wound healing and unfavorable wound healing, and achieve excellent in vitro antibacterial effect, high efficiency and rapid Treatment of diabetic wounds, easily accessible results
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[0040] In one embodiment of the present disclosure, a preparation method of a cascade enzyme-like nanosystem for promoting diabetic wound healing is disclosed, comprising: mixing metal oxide nanoparticles with PAH solution, so that a PAH coating is formed on the surface of the metal oxide layer; then, glucose oxidase was added, and glucose oxidase was covalently conjugated to the amino group of PAH.
[0041] In some specific embodiments, the process of mixing the metal oxide nanoparticles with the PAH solution includes: configuring a NaCl solution containing PAH, the concentration of the PAH in the NaCl solution is 5-20 mg / mL, preferably, 10mg / mL.
[0042] In some specific embodiments, the metal oxide and the PAH solution are mixed and stirred at room temperature. After the reaction is completed, the mixed solution is centrifuged to obtain metal oxide / PAH nanoparticles; further, the stirring time is 2-5h , preferably, is 3h.
[0043] In some specific embodiments, before mixi...
Embodiment 1
[0053] A cascade enzyme-like nanosystem for promoting diabetic wound healing: Fe 3 o 4 -GOx nano system preparation method is as follows (principle such as figure 1 shown):
[0054] Material: Iron(III) Chloride Hexahydrate (FeCl 3 ·6H 2 O), sodium oleate, oleic acid, 1-octadecene, polyallylamine hydrochloride (PAH; Mw=15000Da), phosphate buffered saline (PBS), tetramethylammonium hydroxide 1-(3- Dimethylaminopropyl)-3-ethylcarbodiimide hydrochloride (EDC), N-hydroxysulfosuccinimide sodium salt (NHS), 3-(4,5-dimethyl-2 -thiazolyl)-2,5-diphenyl-2H-tetrazolium bromide (MTT) and 2′,7′-dichlorodifluorofluorescein diacetate (DCFH-DA; ≥94%) were purchased from Sigma-Aldrich. 3,3',5,5'-tetramethylbenzidine (TMB), 2,2'-azidobis(3-ethylbenzothiazoline-6-sulfonic acid) diammonium salt (ABTS), o-Phenylenediamine (OPD; 98%), methylene blue (MB), terephthalic acid (TA) and hydrogen peroxide (H 2 o 2 ) analysis kit was purchased from Beyotime Biotechnology. Hydrogen peroxide (≥30%)...
Embodiment 2
[0060] A cascade enzyme-like nanosystem for promoting diabetic wound healing: MnO 2 -GOx nano system preparation method is as follows:
[0061] MnO 2 Preparation of nanoparticles: Put potassium permanganate into alkyl alcohol polyoxyethylene ether in batches at a molar ratio of 1:1 to 3, control the reaction temperature below 60°C, then stir for 3 to 4 hours, add anhydrous Ethanol, continue to stir for 30 minutes, filter out the solids, then reflux with absolute ethanol for 2 to 3 hours, and elute; filter again, filter out the solids as crude manganese dioxide, and then dry them at 75 to 85°C , crystallize the dried manganese dioxide at 400°C for 4 hours to obtain black nano manganese dioxide powder.
[0062] MnO 2 - Preparation of GOx nanosystem: Dissolve 0.1 g PAH in 10 mL of 1 mM NaCl solution, and add 1 mL of prepared MnO 2 nanoparticles. The mixture was stirred at room temperature at 900 rpm for 3 h to obtain MnO 2 / PAH solution. After the reaction is completed, th...
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