CN112641957A - Method for encapsulating gold nanoparticles by self-assembly of segmented copolymer PCL-b-PEO based on micro-fluidic chip - Google Patents
Method for encapsulating gold nanoparticles by self-assembly of segmented copolymer PCL-b-PEO based on micro-fluidic chip Download PDFInfo
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Abstract
The invention relates to a block copolymer PCL-b-PEO self-assembly encapsulated gold nanoparticles field. The method for encapsulating gold nanoparticles by self-assembly of a segmented copolymer PCL-b-PEO based on a microfluidic chip comprises the following steps of firstly, preparing the microfluidic chip; step two, connecting an air inlet with argon or nitrogen, respectively arranging a first liquid inlet, a second liquid inlet and a third liquid inlet in three liquid inlets of the five-way valve, adding N, N' -methylene bisacrylamide DMF solution of dodecyl mercaptan into the AuNPs colloidal fluid, stirring uniformly, and then adding PCL-bIntroducing a mixed solution formed by uniformly stirring PEO into a first liquid inlet, connecting a second liquid inlet with a DMF solution, and connecting a third liquid inlet with a mixed solution of DMF and water; the flow rates of the three liquid inlets are controlled to be the same, and the flow rate of the gas in the gas inlet is the same as the total flow rate of the three liquid inlets.
Description
Technical Field
The invention relates to a block copolymer PCL-b-PEO self-assembly encapsulated gold nanoparticles field.
Background
Currently, theranostics have been the core of cancer research, and cancer cells prevent their proliferation during the diagnostic process, and the growth of cancer is hindered. X-ray Computed Tomography (CT) is the most widely used diagnostic technique in clinical applications, with iodine as a contrast agent increasing the sensitivity of CT images, however, rapid renal clearance of iodine shortens the imaging time, large volume scatter and small molecule vascular permeability can lead to side effects such as nausea and vomiting, whereas gold is considered to be an excellent CT sensitizer due to its higher atomic number and high X-ray attenuation (Inose t., Oikawa t., Tokunaga m., et al.
Nanoparticles avoid high permeability and retention effects due to long circulation time and accumulation at tumor sites, especially gold nanoparticles (AuNPs) are considered as the most promising sensitizers and drug delivery vehicles for cancer diagnosis due to their non-toxicity, long biological half-life, stability, morphological diversity, good biocompatibility and easy functionalization (Xiong d., Zhang x., Peng s., et al.B,2018,163:29）
The amphiphilic block copolymer self-assembles to form a polymer micelle with a hydrophilic shell and a hydrophobic core. The hydrophobic core may encapsulate gold and hydrophobic anticancer drugs, etc. to achieve diagnosis and treatment. Lin et al (Lin w., Zhang x., Qian l., et al.Biomacromolecules,2017,18: 3869) prepares a pH-sensitive 21-arm star polymer [ beta-cyclodextrin- (polycaprolactone-polyethyl methacrylate-polyethylene glycol methyl ether methacrylate) [ beta-CD- (PCL-PAEMA-PPEGMA)21]The monomolecular micelle encapsulates gold nanoparticles and anticancer drug Doxorubicin (DOX) as a nanocarrier for effective early stage CT imaging and cancer treatment. However, the amphiphilic block copolymer is complex and has low gold coating rate.
Disclosure of Invention
The technical problem to be solved by the invention is as follows: how to increase the encapsulation rate and gold loading rate of the micelle to AuNPs through a microfluidic chip.
The technical scheme adopted by the invention is as follows: the method for encapsulating gold nanoparticles by self-assembly of block copolymer PCL-b-PEO based on the microfluidic chip comprises the following steps of preparing the microfluidic chip according to the following steps, wherein the microfluidic chip comprises a substrate layer and a channel layer, and the substrate layer is prepared by mixing polydimethylsiloxane and a curing agent by the weight ratio of 20: 1, the channel layer is bonded on the substrate layer, and the channel layer is prepared by mixing polydimethylsiloxane and a curing agent in a ratio of 7: 1, a reaction channel is arranged in the channel layer, a liquid inlet of the reaction channel is connected with a five-way joint, the five-way joint is provided with three liquid inlets, one gas inlet and one liquid outlet, the liquid outlet is connected with the liquid inlet of the reaction channel, and a liquid outlet of the reaction channel is used for flowing out the block copolymer PCL-b-PEO self-assembly encapsulated gold nanoparticles;
connecting an air inlet of the five-way valve with argon or nitrogen, respectively serving as a first liquid inlet, a second liquid inlet and a third liquid inlet in the three liquid inlets of the five-way valve, adding N, N' -methylene bisacrylamide DMF solution of dodecyl mercaptan into the AuNPs colloidal fluid, stirring uniformly, and then adding PCL-bIntroducing a mixed solution formed by uniformly stirring PEO into a first liquid inlet, connecting a second liquid inlet with a DMF solution, and connecting a third liquid inlet with a mixed solution of DMF and water;
step three, controlling the flow rates of the three liquid inlets to be the same, controlling the flow rate of the gas at the gas inlet to be the same as the total flow rate of the three liquid inlets, and controlling the content of water in the mixed solution of DMF and water to be PCL-bThe critical water content CWC +2wt% to CWC +10 wt% of the mixed solution of PEO and AuNPs.
The curing agent is Dow Corning 184.
The total flow rate of the three liquid inlets is 50-400 mu L/min.
The concentration of N, N' -methylene bisacrylamide DMF solution of dodecyl mercaptan is 3 vol%, the volume is 10-30 mu L, and the concentration of AuNPs colloidal fluid is 3.6 multiplied by 10-8mol/L, volume of 1mL, PCL-bPEO concentration is 1.0 wt%, volume is 45-55 μ L.
The reaction channel is a curved line or a straight line.
The cross sections of the channels of the three liquid inlets and the gas inlet are all 0.015 mu m2The reaction channel is connected with the liquid outlet through a mixing channel, and the cross section of the mixing channel is 0.015 mu m2The cross section of the reaction channel of the curve is 0.03 μm2The cross section of the linear reaction channel was 0.012. mu.m2。
The inventionHas the advantages that: the amphiphilic block copolymer PCL-containing material is prepared by adopting the micro-fluidic chip for the first timebPEO encapsulates gold nanoparticles, and the amphiphilic block copolymer PCL-bThe encapsulation rate and gold coating rate of PEO encapsulated gold nanoparticles provide a new method for mass production of block copolymer gold-coated nanoparticles.
Drawings
FIG. 1 is a schematic view of a curvilinear microfluidic chip;
FIG. 2 is a schematic view of a linear microfluidic chip (a straight line except for a turn);
FIG. 3 is a transmission electron microscope image of a segmented copolymer PCL-b-PEO self-assembly encapsulated gold nanoparticles prepared by a curvilinear microfluidic chip at a flow rate of 400 μ L/min.
Detailed Description
Example 1
Operation of segmented copolymer PCL (polycaprolactone) by adopting microfluidic chipbPEO self-assembly encapsulates gold nanoparticles, argon is injected into the gas inlet, 10. mu.L (3 vol%) of dodecanethiol DMF solution is added into 1mL of 3.6X 10-8Stirring in AuNPs colloidal solution for 1.5 hr, adding 45 μ L PCL105-b-PEO144(1.0 wt%) was stirred for 1 hour and then mixed; the second liquid inlet is filled with a pure DMF solution; the third liquid inlet is filled with a mixed solution of DMF and water, and the content of the water is PCL-b-critical water content of PEO and AuNPs mixed solution 3.8 wt% +2 wt%; the flow rates of the three liquid channels are the same. The flow rate of the gas is the same as the total liquid flow rate, and each channel is 1/3 of the liquid flow rate. When 50 mu L/min of PCL-containing material flows through the linear micro-fluidic chipbThe encapsulation rate of the PEO @ Au nanoparticles to the gold nanoparticles is 13.3 percent, and the gold loading rate is 29.1 percent; when 50 mu L/min of PCL flowing through the curve type micro-fluidic chip is adoptedbThe encapsulation rate of the-PEO @ Au nanoparticles to the gold nanoparticles is 30.4%, and the gold loading rate is 42.5%.
The microfluidic chip consists of a substrate layer and a channel layer, wherein the substrate layer is prepared by mixing polydimethylsiloxane and a curing agent in a ratio of 20: 1, the channel layer is formed by coating a glass plate with a negative photoresist SU-8100 to construct a micro-fluidic chip master mask on a silicon wafer, and the micro-fluidic chip master mask is prepared by mixing polydimethylsiloxane and a curing agent in a ratio of 7: 1, is prepared.
All channels (inlet channels or outlet channels) adopt rectangular channels, the depth of each rectangular channel is 150 micrometers, the width of each of three liquid inlets, one gas inlet and one liquid outlet is 100 micrometers, the width of each mixing channel is 100 micrometers, the width of each curve type micro-fluidic chip is 200 micrometers, and the width of each linear type micro-fluidic chip is 80 micrometers.
Example 2
Operation of segmented copolymer PCL (polycaprolactone) by adopting microfluidic chipbPEO self-assembly encapsulates gold nanoparticles, argon is injected into the gas inlet, 20. mu.L (3 vol%) of dodecanethiol DMF solution is added into 1mL of 3.6X 10-8Stirring in mol/L AuNPs colloidal solution for 1.5 hours, adding 50 μ L of PCL-bPEO (1.0 wt%) was stirred for 1 hour and mixed; the second liquid inlet is filled with a pure DMF solution; the third liquid inlet is filled with a mixed solution of DMF and water, and the content of the water is PCL-b-critical water content of PEO and AuNPs mixed solution 3.8 wt% +5 wt%; the flow rates of the three liquid channels are the same. The flow rate of the gas is the same as the total liquid flow rate, and each channel is 1/3 of the liquid flow rate. When the material flows through the linear micro-fluidic chip at 100 mu L/min, PCL-bThe encapsulation rate of the PEO @ Au nanoparticles to the gold nanoparticles is 22.1 percent, and the gold loading rate is 35.7 percent; when the liquid flows through the curve type micro-fluidic chip at 100 mu L/min, PCL-bThe encapsulation rate of the PEO @ Au nanoparticles to the gold nanoparticles is 41.3%, and the gold loading rate is 56.5%.
Example 3
Operation of segmented copolymer PCL (polycaprolactone) by adopting microfluidic chipbPEO self-assembly encapsulates gold nanoparticles, argon is injected into the gas inlet, 20. mu.L (3 vol%) of dodecanethiol DMF solution is added into 1mL of 3.6X 10-8Stirring in mol/L AuNPs colloidal solution for 1.5 hours, adding 50 μ L of PCL-bPEO (1.0 wt%) was stirred for 1 hour and mixed; the second liquid inlet is filled with a pure DMF solution; the third liquid inlet is filled with a mixed solution of DMF and water, and the content of the water is PCL-b-critical water content of PEO and AuNPs mixed solution 3.8 wt% +5 wt%; the flow rates of the three liquid channels are the same. The flow rate of the gas is the same as the total liquid flow rate, and each channel is 1/3 of the liquid flow rate. MiningWhen the liquid flows through the linear micro-fluidic chip at 400 mu L/min, PCL-bThe encapsulation rate of the PEO @ Au nanoparticles to the gold nanoparticles is 53.8 percent, and the gold loading rate is 78.2 percent; when the liquid flows through the curve type micro-fluidic chip at 100 mu L/min, PCL-bThe encapsulation rate of the PEO @ Au nanoparticles to the gold nanoparticles is 76.3%, and the gold loading rate is 110.8%.
Example 4
Operation of segmented copolymer PCL (polycaprolactone) by adopting microfluidic chipbPEO self-assembly encapsulates gold nanoparticles, nitrogen is injected into the gas inlet, 20 μ L (3 vol%) of dodecanethiol DMF solution is added into 1mL of 3.6X 10-8Stirring in mol/L AuNPs colloidal solution for 2 hours, and then adding 50 μ L of PCL-bPEO (1.0 wt%) was stirred for 1 hour and mixed; the second liquid inlet is filled with a pure DMF solution; the third liquid inlet is filled with a mixed solution of DMF and water, and the content of the water is PCL-b-critical water content of PEO and AuNPs mixed solution 3.8 wt% +10 wt%; the flow rates of the three liquid channels were the same and the gas was argon. The flow rate of the gas is the same as the total liquid flow rate, and each channel is 1/3 of the liquid flow rate. When 200 mu L/min of PCL-containing material flows through the linear micro-fluidic chipbThe encapsulation rate of the PEO @ Au nanoparticles to the gold nanoparticles is 33.7 percent, and the gold loading rate is 47.9 percent; when the liquid flows through the curve type micro-fluidic chip at the speed of 200 mu L/min, PCL-bThe encapsulation rate of the-PEO @ Au nanoparticles to the gold nanoparticles is 56.4%, and the gold loading rate is 81.8%.
Example 5
Operation of segmented copolymer PCL (polycaprolactone) by adopting microfluidic chipbPEO self-assembly encapsulates gold nanoparticles, argon is injected into the gas inlet, 20. mu.L (3 vol%) of dodecanethiol DMF solution is added into 1mL of 3.6X 10-8Stirring in mol/L AuNPs colloidal solution for 1.5 hours, and then adding 45 μ L of PCL-bPEO (1.0 wt%) mixed liquid after stirring for 1 hour; the second liquid inlet is filled with a pure DMF solution; the third liquid inlet is filled with a mixed solution of DMF and water, and the content of the water is PCL-b-critical water content of PEO and AuNPs mixed solution 4.5 wt% +8 wt%; the flow rates of the three liquid channels are the same. The flow rate of the gas is the same as the total liquid flow rate, and each channel is 1/3 of the liquid flow rate. When 200 mu L/min of the liquid flows through the linear micro-fluidic chip，PCL-bThe encapsulation rate of the PEO @ Au nanoparticles to the gold nanoparticles is 30.9 percent, and the gold loading rate is 42.3 percent; when the liquid flows through the curve type micro-fluidic chip at the speed of 200 mu L/min, PCL-bThe encapsulation rate of the PEO @ Au nanoparticles to the gold nanoparticles is 50.8%, and the gold loading rate is 72.8%.
Claims (6)
1. Segmented copolymer PCL (polycaprolactone) based on microfluidic chipb-a method for self-assembly encapsulation of gold nanoparticles by PEO, characterized in that: the method comprises the following steps
Step one, preparing a microfluidic chip, wherein the microfluidic chip is composed of a substrate layer and a channel layer, and the substrate layer is prepared by mixing polydimethylsiloxane and a curing agent in a ratio of 20: 1, the channel layer is bonded on the substrate layer, and the channel layer is prepared by mixing polydimethylsiloxane and a curing agent in a ratio of 7: 1, a reaction channel is arranged in the channel layer, a liquid inlet of the reaction channel is connected with a five-way joint, the five-way joint is provided with three liquid inlets, one gas inlet and one liquid outlet, the liquid outlet is connected with the liquid inlet of the reaction channel, and a liquid outlet of the reaction channel is used for flowing out the block copolymer PCL-b-PEO self-assembly encapsulated gold nanoparticles;
connecting an air inlet of the five-way valve with argon or nitrogen, respectively serving as a first liquid inlet, a second liquid inlet and a third liquid inlet in the three liquid inlets of the five-way valve, adding N, N' -methylene bisacrylamide DMF solution of dodecyl mercaptan into the AuNPs colloidal fluid, stirring uniformly, and then adding PCL-bIntroducing a mixed solution formed by uniformly stirring PEO into a first liquid inlet, connecting a second liquid inlet with a DMF solution, and connecting a third liquid inlet with a mixed solution of DMF and water;
step three, controlling the flow rates of the three liquid inlets to be the same, controlling the flow rate of the gas at the gas inlet to be the same as the total flow rate of the three liquid inlets, and controlling the content of water in the mixed solution of DMF and water to be PCL-bThe critical water content CWC +2wt% to CWC +10 wt% of the mixed solution of PEO and AuNPs.
2. The microfluidic chip-based method for self-assembling encapsulation of gold nanoparticles by using segmented copolymer PCL-b-PEO according to claim 1, wherein the method comprises the following steps: the curing agent is Dow Corning 184.
3. The microfluidic chip-based method for self-assembling encapsulation of gold nanoparticles by using segmented copolymer PCL-b-PEO according to claim 1, wherein the method comprises the following steps: the total flow rate of the three liquid inlets is 50-400 mu L/min.
4. The microfluidic chip-based method for self-assembling encapsulation of gold nanoparticles by using segmented copolymer PCL-b-PEO according to claim 1, wherein the method comprises the following steps: the concentration of N, N' -methylene bisacrylamide DMF solution of dodecyl mercaptan is 3 vol%, the volume is 10-30 mu L, and the concentration of AuNPs colloidal fluid is 3.6 multiplied by 10-8mol/L, volume of 1mL, PCL-bPEO concentration is 1.0 wt%, volume is 45-55 μ L.
5. The microfluidic chip-based method for self-assembling encapsulation of gold nanoparticles by using segmented copolymer PCL-b-PEO according to claim 1, wherein the method comprises the following steps: the reaction channel is a curved line or a straight line.
6. The microfluidic chip-based method for self-assembling encapsulation of gold nanoparticles by using segmented copolymer PCL-b-PEO according to claim 5, wherein the method comprises the following steps: the cross sections of the channels of the three liquid inlets and the gas inlet are all 0.015 mu m2The reaction channel is connected with the liquid outlet through a mixing channel, and the cross section of the mixing channel is 0.015 mu m2The cross section of the reaction channel of the curve is 0.03 μm2The cross section of the linear reaction channel was 0.012. mu.m2。
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