Preparation method of an injectable porous microchip and its multi-fraction time-delivery carrier
A microchip and microchip technology, applied in prostheses, drug delivery, pharmaceutical formulations, etc., can solve the problems of low cell migration rate, low yield, waste of scaffold materials, etc., and achieve high practical value and small geometric size. , good uniformity
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Embodiment 1
[0045] Embodiment 1 Teflon plate (Teflon plate) prepares alginate-based porous microchip (MC)
[0046] The prepared 1.5% (w / v) sodium alginate (Sigma-Aldrich, USA) solution was sucked into a 1mL syringe (Hongda Medical Instrument Co., Ltd., Jiangxi), and then figure 1 As shown in A, the composite solution was extruded through a 32G stainless steel needle to form micro-droplets, and quickly placed on a pre-cooled Teflon plate (for the purpose of pre-cooling, to slow down the evaporation rate of micro-droplets).
[0047] After the micro-droplets are dripped, put the Teflon plate (Kexin Plastic Products Factory, Guangdong) in a refrigerator at 4°C for 1 hour, and then put it in a refrigerator at -20°C for 12 hours; 50 ℃, vacuum degree 2 ~ 10Pa, until completely freeze-dried (about 12h). Add an appropriate amount of 2% calcium chloride (w / v; Sigma-Aldrich) cross-linking agent dropwise to the microchips on the Teflon plate after lyophilization, completely soak the microchips, fu...
Embodiment 2
[0052] The MC double-layer scaffold system (G / MC) of embodiment 2 hydrogel encapsulation is constructed
[0053] Utilize residual Ca on the surface of MC 2+ Self-cross-linked with sodium alginate solution to form a hydrogel, and coated on the outside of MC. The specific steps are as follows: Wet MC with PBS and fill it into a 2mL syringe, the volume of which is about half of the volume of the syringe, and then carry out conventional freeze-drying. .
[0054] Then add PBS to the lyophilized syringe to rewet the MC; then use another syringe filled with an appropriate amount of 2% (w / v) sodium alginate aqueous solution (volume about half the volume of the MC) to pass through the Luer Pipe joints (female-female; Zhongchi Plastic Parts Co., Ltd., Shaanxi) connected the two syringes, pushed the piston to fully mix MC and sodium alginate aqueous solution, and obtained the hydrogel-wrapped MC double-layer scaffold system (G / MC), the preparation process can be found in figure 2...
Embodiment 3
[0059] Example 3 Material characterization of MC and G / MC and exploration of injectability of G / MC
[0060] The morphology, geometric size, porous structure and pore size of MC were characterized by SEM.
[0061] The sample preparation method is as follows: directly stick the completely freeze-dried MC to the conductive adhesive, and spray gold for 20 seconds before shooting. Note that the MC must be completely dry.
[0062] To evaluate the degradation rate of the G / MC bilayer structure, we labeled the outer gel (G layer) and the inner MC (MC layer) with rhodamine and FITC fluorophores, respectively, and monitored the two layers separately by confocal laser microscopy the law of degradation.
[0063] The specific method is: use fluorescently labeled sodium alginate to prepare G layer and MC layer respectively, and the preparation method is the same as that described in "Example 2 Construction of Hydrogel-wrapped MC Bilayer Scaffold System (G / MC)". Then the fluorescent G / ...
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