Metal polyphenol nano material based on chlorogenic acid as well as preparation method and application of metal polyphenol nano material

Abstract

The invention discloses a metal polyphenol nano material based on chlorogenic acid and a preparation method and application thereof.The preparation method comprises the steps that chlorogenic acid and distearoyl phosphatidyl ethanolamine-polyethylene glycol 2000 are mixed and dissolved in an anhydrous tetrahydrofuran solvent, and a first mixed solution is prepared; adding a FeCl3 solution into the first mixed solution until a clear and transparent system is formed, so as to obtain a second mixed solution; and removing the anhydrous tetrahydrofuran solvent of the second mixed solution to obtain the metal polyphenol nano material (CA-Fe NPs) based on chlorogenic acid. The metal polyphenol nano material has excellent photothermal conversion performance, and can be applied to in-vitro antibiosis, antioxidation, anti-inflammation and wound healing promotion.

Description

Technical Field

[0001] This invention belongs to the field of nanomaterial preparation technology, specifically relating to a metal polyphenol nanomaterial based on chlorogenic acid, its preparation method, and its application. Background Technology

[0002] The global spread of bacterial resistance (especially methicillin-resistant Staphylococcus aureus, MRSA) poses a serious challenge to traditional antibiotic therapy, urgently requiring the development of novel antibacterial strategies. Photothermal therapy, which uses near-infrared light to excite nanomaterials to generate heat and kill pathogens, has attracted much attention due to its non-invasive nature and low risk of inducing resistance. However, existing photothermal agents suffer from low photothermal conversion efficiency, poor photostability, and lack of biodegradability, and single antibacterial mechanisms are insufficient to cope with complex biofilm environments. Furthermore, oxidative stress and inflammatory cascades in the infection microenvironment can exacerbate tissue damage and delay wound healing, but research on nanomaterials with both antibacterial and microenvironment regulation functions is still in the exploratory stage.

[0003] Chlorogenic acid (CA), as a natural polyphenol compound, is considered an ideal functional molecule due to its broad-spectrum antibacterial, antioxidant, and anti-inflammatory activities. However, the direct application of CA is limited by its poor water solubility, rapid in vivo metabolism, and narrow concentration window. Metal-phenolic networks (MPNs), nanostructures formed by the self-assembly of metal ions and polyphenols, can both protect active molecules and enhance synergistic functional effects. Iron ions (Fe...) 3+ Due to its biocompatibility and photothermal response characteristics, it is often used to construct MPNs, but its coordination stability with polyphenols and the long-term cycling performance of composite materials still need to be optimized. Summary of the Invention

[0004] The purpose of this invention is to solve the above-mentioned technical problems and provide a chlorogenic acid-based metal polyphenol nanomaterial, its preparation method, and its application. This invention combines chlorogenic acid (CA) and Fe... 3+ Chlorogenic acid-based metal polyphenol nanomaterials (CA-FeNPs) were constructed using a self-assembly technique mediated by the amphiphilic polymer disteazylophosphatidylethanolamine-polyethylene glycol (DSPE-PEG).

[0005] To achieve the above objectives, the present invention provides a method for preparing chlorogenic acid-based metal polyphenol nanomaterials, comprising the following steps:

[0006] Step 1: Chlorogenic acid and distearate phosphatidylethanolamine-polyethylene glycol are mixed and added to anhydrous tetrahydrofuran and ultrasonically dispersed to obtain the first mixture;

[0007] Step 2: Add FeCl3 solution to the first mixture and ultrasonically disperse until a clear and transparent system is formed to obtain the second mixture;

[0008] Step 3: Remove anhydrous tetrahydrofuran from the second mixture to obtain a metal polyphenol nanomaterial based on chlorogenic acid.

[0009] Preferably, the mass ratio of chlorogenic acid (CA) to distearylphosphatidylethanolamine-polyethylene glycol (DSPE-PEG) is 1:1 to 3.

[0010] Preferably, the concentration of the FeCl3 solution is 0.022-0.027 mol / L.

[0011] This invention also provides a chlorogenic acid-based metal polyphenol nanomaterial, which is prepared by the above method and has photothermal conversion properties. Specifically, it can absorb light energy of a specific wavelength and convert it into heat energy, which is a core indicator of photothermal therapy.

[0012] Preferably, the chlorogenic acid-based metal polyphenol nanomaterial has a ZETA potential of 18.23 mV ± 0.46.

[0013] The present invention also provides a chlorogenic acid-based metal polyphenol nanomaterial for in vitro antioxidant applications.

[0014] The present invention also provides a chlorogenic acid-based metal polyphenol nanomaterial for in vitro anti-inflammatory applications.

[0015] The present invention also provides a chlorogenic acid-based metal polyphenol nanomaterial for in vitro photothermal sterilization.

[0016] This invention also provides a method for using chlorogenic acid-based metal polyphenol nanomaterials in conjunction with near-infrared light to promote wound healing.

[0017] Compared with the prior art, the beneficial effects of the present invention are as follows:

[0018] This invention involves reacting plant polyphenol chlorogenic acid with metal ions Fe 3+ Metal polyphenol nanomaterials with spherical nanostructures were prepared by ultrasonic mixing and self-assembly. Based on the broad-spectrum antibacterial, antioxidant, and anti-inflammatory activities of chlorogenic acid, these nanomaterials can be directly applied to in vitro antioxidant and anti-inflammatory applications. These nanomaterials also have efficient and stable photothermal conversion properties, absorbing light energy of specific wavelengths and converting it into heat energy. Based on this principle, through the synergistic effect of these nanomaterials and near-infrared lasers, they can be applied to in vitro antibacterial applications and to promote wound healing. Attached Figure Description

[0019] Figure 1This is a schematic diagram of the preparation process of CA-Fe NPs nanomaterials in Example 1;

[0020] Figure 2 This is a transmission electron microscope (TEM) image of the CA-Fe NPs nanomaterial in Example 1;

[0021] Figure 3 This is a surface potential diagram of chlorogenic acid (CA)FeCl3 and CA-Fe NPs nanomaterials from Example 1;

[0022] Figure 4 This is the elemental analysis diagram of Fe, C, O and CA-Fe NPs in Example 1;

[0023] Figure 5 These are infrared thermal images of CA-Fe NPs aqueous solutions of different concentrations in Example 4 under irradiation with an 808nm NIR laser (1.5W / cm2);

[0024] Figure 6 This is a temperature change curve of CA-Fe NPs aqueous solutions of different concentrations under irradiation with an 808nm NIR laser (1.5W / cm2) in Example 4;

[0025] Figure 7 This is the temperature change curve of the 400 μg / ml CA-Fe NPs aqueous solution under NIR laser irradiation of different powers in Example 4;

[0026] Figure 8 This is a test diagram of the photothermal conversion stability of CA-Fe NPs in Example 4;

[0027] Figure 9 These are the maximum temperature change and heat transfer time constant diagrams for the CA-Fe NPs nanomaterials in Example 4; among them, Figure 9 a is the maximum temperature variation graph for CA-Fe NPs. Figure 9 b is the heat transfer time constant of CA-Fe NPs;

[0028] Figure 10 These are infrared thermal images of Staphylococcus aureus, Escherichia coli, and Candida albicans in each group of Example 5;

[0029] Figure 11 This is a bacterial colony distribution diagram of Staphylococcus aureus, Escherichia coli, and Candida albicans in each group of Example 5;

[0030] Figure 12 This is a CFU bacterial count diagram of Staphylococcus aureus, Escherichia coli, and Candida albicans in each group of Example 5;

[0031] Figure 13These are fluorescence micrographs of Staphylococcus aureus, Escherichia coli, and Candida albicans in each group of Example 5, showing the bacterial viability and mortality status of bacteria detected by a bacterial viability kit.

[0032] Figure 14 This is a statistical chart of the cell viability of HK2 cells in each group of experimental group 1 in Example 6;

[0033] Figure 15 This is a statistical chart showing the reactive oxygen species (ROS) status of HK2 cells in each group of experimental group 2 in Example 6, detected by flow cytometry (FCM); among them, Figure 15 a is a graph showing the analysis of reactive oxygen species levels in HK2 cells. Figure 15 b is a statistical result of reactive oxygen species-positive cells in the cells after HK2 cell treatment;

[0034] Figure 16 These are reactive oxygen species fluorescence staining images of HK2 cells in each group of experimental group 2 in Example 6;

[0035] Figure 17 These are flow cytometry (FCM) images of apoptosis in HK2 cells from each group in experimental group 3 of Example 6;

[0036] Figure 18 This is a graph showing the statistical results of the recruitment ratio of macrophages in each group of experimental group 1 in Example 7;

[0037] Figure 19 These are flow cytometry (FCM) images of Raw264.7 cells from each group in Experimental Group 2 of Example 7;

[0038] Figure 20 This is a statistical chart showing the proportions of M1 and M2 macrophages in each group of experimental group 2 in Example 7; among them, Figure 20 a is a statistical chart of the proportion of M1 macrophages; Figure 20 b is a statistical chart of the M1 / M2 macrophage ratio;

[0039] Figure 21 This is a schematic diagram of the process for establishing a mouse wound model infected with methicillin-resistant Staphylococcus aureus (MRSA) in Example 8.

[0040] Figure 22 These are infrared thermographic images of mouse wounds in the NIR control group and the CA-Fe NPs+NIR treatment group in Example 8;

[0041] Figure 23 This is a graph showing the ELISA results of serum inflammatory factors (TNF-α, IL-1β, IL-6) in each group of mice in Example 8;

[0042] Figure 24These are hematoxylin-eosin (H&E) staining images and Masson staining images of the mouse wound tissues from each group in Example 8; among them, Figure 24 a is a hematoxylin-eosin (H&E) staining image of mouse wound tissue; Figure 24 b is a Masson staining image of mouse wound tissue;

[0043] Figure 25 These are images showing the healing status of the wounds in each group of mice on days 0, 1, 3, 7, and 11 in Example 8. Detailed Implementation

[0044] The technical solutions in the embodiments of the present invention will be described in detail below with reference to the embodiments and accompanying drawings. The embodiments given are only for illustrating the present invention and are not intended to limit the scope of protection of the present invention.

[0045] This application provides a method for preparing chlorogenic acid-based metal polyphenol nanomaterials, comprising the following steps:

[0046] Step 1: Chlorogenic acid and distearylphosphatidylethanolamine-polyethylene glycol are mixed and added to anhydrous tetrahydrofuran and ultrasonically dispersed to obtain the first mixture; wherein the mass ratio of chlorogenic acid and distearylphosphatidylethanolamine-polyethylene glycol 2000 is 1:1 to 3.

[0047] Step 2: Add FeCl3 solution to the first mixture and disperse it ultrasonically until a clear and transparent system is formed, obtaining the second mixture; wherein the concentration of FeCl3 solution is 0.022-0.027 mol / L. This step ends with the observation of a clear and transparent system. The specific addition process is dropwise, with ultrasonic dispersion required after each drop to ensure sufficient reaction between the FeCl3 solution and the first mixture before adding the next drop.

[0048] Step 3: Remove anhydrous tetrahydrofuran from the second mixture to obtain a metal polyphenol nanomaterial based on chlorogenic acid.

[0049] The metal polyphenol nanomaterials prepared by the above method provided in this application have a near-spherical structure and a Zeta potential of 18.23 mV ± 0.46. Based on the broad-spectrum antibacterial, antioxidant, and anti-inflammatory activities of chlorogenic acid, the metal polyphenol nanomaterials can be directly applied for in vitro antioxidant and anti-inflammatory purposes. The metal polyphenol nanomaterials also possess efficient and stable photothermal conversion properties, absorbing light energy of specific wavelengths and converting it into heat energy. Based on this principle, through the synergistic effect of this nanomaterial and near-infrared laser, it can be applied for in vitro antibacterial and wound healing promotion.

[0050] Example 1

[0051] A method for preparing chlorogenic acid-based metal polyphenol nanomaterials, such as... Figure 1 As shown, it specifically includes:

[0052] Step 1: Accurately weigh 5 mg of chlorogenic acid (CA) and 10 mg of distearate phosphatidylethanolamine-polyethylene glycol 2000 (DSPE-PEG2000), mix them and add them to 2 ml of anhydrous tetrahydrofuran solvent, and sonicate for 5 min until completely dissolved to obtain the first mixture.

[0053] Step 2: Under ultrasonic conditions, add 0.025 mol / L FeCl3 solution dropwise to the first mixture until a clear and transparent system is formed, thus obtaining the second mixture;

[0054] Step 3: Remove anhydrous tetrahydrofuran from the second mixture using a rotary evaporator to prepare a metal polyphenol nanomaterial CA-Fe NPs based on chlorogenic acid.

[0055] In this embodiment, the mass ratio of chlorogenic acid (CA) to distearylphosphatidylethanolamine-polyethylene glycol 2000 (DSPE-PEG2000) is 1:2.

[0056] Figure 2 This is a transmission electron microscope (TEM) image of CA-Fe NPs nanomaterials. The image shows that CA-Fe NPs nanomaterials have a near-spherical structure and the nanoparticles are evenly distributed.

[0057] Figure 3 The diagram shows the surface potential of chlorogenic acid (CA), FeCl3, and CA-Fe NPs nanomaterials. The surface zeta potential of CA-Fe NPs nanomaterials is 18.23 mV ± 0.46, which is between that of CA and FeCl3.

[0058] Figure 4 This is an elemental analysis diagram of Fe, C, O, and CA-Fe NPs, from Figure 4 It can be seen that C, O, and Fe are uniformly distributed in the CA-Fe NPs nanomaterial, proving that the metal polyphenol nanomaterial CA-Fe NPs was successfully synthesized in this embodiment.

[0059] Example 2

[0060] A method for preparing metal polyphenol nanomaterials based on chlorogenic acid:

[0061] Step 1: Accurately weigh 5 mg of chlorogenic acid (CA) and 5 mg of distearate phosphatidylethanolamine-polyethylene glycol 2000 (DSPE-PEG2000), mix them and add them to 2 ml of anhydrous tetrahydrofuran solvent, and sonicate for 5 min until completely dissolved to obtain the first mixture.

[0062] Step 2: Under ultrasonic conditions, add 0.025 mol / L FeCl3 solution dropwise to the first mixture until a clear and transparent system is formed, thus obtaining the second mixture;

[0063] Step 3: Remove anhydrous tetrahydrofuran from the second mixture using a rotary evaporator to prepare a metal polyphenol nanomaterial CA-Fe NPs based on chlorogenic acid.

[0064] In this embodiment, the mass ratio of chlorogenic acid (CA) to distearylphosphatidylethanolamine-polyethylene glycol 2000 (DSPE-PEG2000) is 1:1.

[0065] Example 3

[0066] A method for preparing metal polyphenol nanomaterials based on chlorogenic acid:

[0067] Step 1: Accurately weigh 5 mg of chlorogenic acid (CA) and 15 mg of distearate phosphatidylethanolamine-polyethylene glycol 2000 (DSPE-PEG2000), mix them and add them to 2 ml of anhydrous tetrahydrofuran solvent, and sonicate for 5 min until completely dissolved to obtain the first mixture.

[0068] Step 2: Under ultrasonic conditions, add 0.025 mol / L mM FeCl3 solution dropwise to the first mixture until a clear and transparent system is formed, thus obtaining the second mixture;

[0069] Step 3: Remove anhydrous tetrahydrofuran from the second mixture using a rotary evaporator to prepare a metal polyphenol nanomaterial CA-Fe NPs based on chlorogenic acid.

[0070] In this embodiment, the mass ratio of chlorogenic acid (CA) to distearylphosphatidylethanolamine-polyethylene glycol (DSPE-PEG2000)2000 is 1:3.

[0071] Example 4

[0072] Photothermal conversion performance testing of chlorogenic acid-based metal polyphenol nanomaterials:

[0073] The CA-Fe NPs obtained in Example 1 were used to prepare CA-Fe NPs aqueous solutions with concentrations of 100 μg / ml, 200 μg / ml, and 400 μg / ml, respectively.

[0074] Test 1: A blank control group (0 μg / ml), a 100 μg / ml CA-Fe NPs aqueous solution group, a 200 μg / ml CA-Fe NPs aqueous solution group, and a 400 μg / ml CA-Fe NPs aqueous solution group were set up. An 808 nm near-infrared laser (NIR, 1.5 W / cm²) was used. 2Irradiate for 5 minutes and record infrared thermal images of each group at different times. Figure 5 and Figure 6 Different concentrations of CA-Fe NPs aqueous solutions were subjected to an 808 nm NIR laser (1.5 W / cm²). 2 Infrared thermal images and temperature change curves under illumination, from Figure 5 and Figure 6 It can be seen that the higher the concentration of the CA-Fe NPs aqueous solution, the higher the heating efficiency.

[0075] Test 2: A 400 μg / ml CA-Fe NPs aqueous solution group was set up, using 0.8 W / cm². 2 1.0W / cm 2 1.5W / cm 2 The NIR laser was used to irradiate the sample, and the temperature was recorded at different times. Figure 7 This is the temperature change curve of a 400 μg / ml CA-Fe NPs aqueous solution under NIR laser irradiation of different powers. Figure 7 It can be seen that the higher the NIR laser power of CA-Fe NPs aqueous solution, the higher the heating efficiency.

[0076] Test 3: A 400 μg / ml aqueous solution group was set up, using an 808 nm near-infrared laser (NIR, 1.5 W / cm²). 2 Irradiation was performed, followed by 6 heating-cooling cycles. Figure 8 This is a test chart of the photothermal conversion stability of CA-Fe NPs, from... Figure 8 It can be seen that after 6 heating-cooling cycles within 1 hour, the temperature change curve did not change significantly, indicating that the photothermal conversion performance of CA-Fe NPs is stable. Figure 9 a is the maximum temperature variation graph for CA-Fe NPs. Figure 9 b is the heat transfer time constant of CA-Fe NPs. The photothermal conversion efficiency is calculated based on the maximum heating temperature and the heat transfer time constant. The photothermal conversion efficiency of CA-Fe NPs is as high as 81.6%.

[0077] Example 5

[0078] A metal polyphenol nanomaterial based on chlorogenic acid is used for in vitro photothermal sterilization:

[0079] The CA-Fe NPs prepared in Example 1 were used to set up a blank control group, a NIR control group, a CA-Fe NPs treatment group, and a CA-Fe NPs+NIR treatment group. The bacteria used in the experiment included Staphylococcus aureus, Escherichia coli, and Candida albicans.

[0080] Blank control group: No treatment was given to Staphylococcus aureus, Escherichia coli, or Candida albicans;

[0081] NIR control group: Staphylococcus aureus, Escherichia coli, and Candida albicans were treated with 1.5 W / cm². 2 Near-infrared laser (NIR) irradiation;

[0082] CA-Fe NPs treatment group: CA-Fe NPs aqueous solution was added to Staphylococcus aureus, Escherichia coli and Candida albicans;

[0083] CA-Fe NPs+NIR treatment group: Staphylococcus aureus, Escherichia coli, and Candida albicans were treated with CA-Fe NPs aqueous solution at 1.5 W / cm². 2 Near-infrared laser (NIR) irradiation.

[0084] Figure 10 These are infrared thermal images of Staphylococcus aureus, Escherichia coli, and Candida albicans in each group, from... Figure 10 It can be seen that the temperature of the NIR control group did not change significantly, while the temperature of the CA-Fe NPs+NIR treatment group rose rapidly within 10 minutes and approached 50℃.

[0085] Figure 11 and Figure 12 These are bacterial colony distribution diagrams and CFU bacterial count diagrams for Staphylococcus aureus, Escherichia coli, and Candida albicans in each group. Figure 11 and Figure 12 It can be seen that the bacterial colonies in the CA-Fe NPs treatment group and the CA-Fe NPs+NIR treatment group were significantly reduced, and the CA-Fe NPs+NIR treatment group had the fewest bacterial colonies, indicating that CA-Fe NPs and CA-Fe NPs+NIR have antibacterial effects, and the antibacterial effect is stronger under the synergistic effect of CA-Fe NPs+NIR.

[0086] Figure 13 These are fluorescence micrographs of Staphylococcus aureus, Escherichia coli, and Candida albicans in each group, showing the viability of bacteria as determined by a bacterial viability kit. Green fluorescence indicates viable bacteria, while yellow / red fluorescence indicates dead bacteria. Figure 13 It can be seen that the CA-Fe NPs+NIR treatment group had the highest bacterial death / live ratio.

[0087] Example 6

[0088] A metal polyphenol nanomaterial based on chlorogenic acid is used for in vitro antioxidant applications:

[0089] First group of experiments:

[0090] The CA-Fe NPs prepared in Example 1 were used to set up a blank control group, an H2O2 control group, and a CA-Fe NPs aqueous solution treatment group (5 μg / ml, 10 μg / ml, 20 μg / ml). The cells used in the experiment were human renal tubular epithelial cells (HK2).

[0091] Blank control group: HK2 cells were not treated in any way;

[0092] H2O2 control group: HK2 cells were supplemented with H2O2 reagent;

[0093] CA-Fe NPs aqueous solution treatment group: H2O2 reagent was added to HK2 cells, and CA-Fe NPs aqueous solution was added at concentrations of 5 μg / ml, 10 μg / ml, and 20 μg / ml, respectively.

[0094] Figure 14 This is a statistical chart of cell viability of HK2 cells in each group, by Figure 14 It can be seen that the aqueous solution of CA-Fe NPs can effectively inhibit the cytotoxicity of H2O2 on HK2 cells and improve cell survival rate. The inhibitory effect is concentration-dependent.

[0095] Second group of experiments:

[0096] The CA-Fe NPs prepared in Example 1 were used to set up a blank control group, an H2O2 control group, and a CA-Fe NPs aqueous solution treatment group (15 μg / ml, 30 μg / ml). The cells used in the experiment were human renal tubular epithelial cells (HK2), and the results were detected by flow cytometry (FCM).

[0097] Blank control group: HK2 cells were not treated in any way;

[0098] H2O2 control group: HK2 cells were supplemented with H2O2 reagent;

[0099] CA-Fe NPs aqueous solution treatment group: H2O2 reagent was added to HK2 cells, and CA-Fe NPs aqueous solution of 15 μg / ml and 30 μg / ml was added respectively.

[0100] Figure 15 This is a statistical chart showing the reactive oxygen species (ROS) status of HK2 cells in each group as detected by flow cytometry (FCM). Figure 15 a is a graph analyzing the level of reactive oxygen species in HK2 cells, from... Figure 15 It can be seen that the reactive oxygen species level of HK2 cells treated with CA-Fe NPs aqueous solution was significantly lower than that of the H2O2 control group; Figure 15 b is a statistical result of reactive oxygen species-positive cells after HK2 cell treatment, from... Figure 15b shows that the number of reactive oxygen species-positive cells decreased significantly relative to the H2O2 control group after treatment with CA-Fe NPs aqueous solution, indicating that CA-Fe NPs have a significant inhibitory effect on H2O2-induced cytotoxicity.

[0101] Figure 16 These are fluorescence staining images of reactive oxygen species (ROS) in HK2 cells from each group. The ROS fluorescence signal appears as green. Figure 16 It can be seen that the reactive oxygen species fluorescence intensity of HK2 cells treated with CA-Fe NPs aqueous solution was significantly weaker than that of the H2O2 control group, indicating that CA-Fe NPs can eliminate the excessive reactive oxygen species generated by the addition of H2O2 in HK2 cells.

[0102] Third group of experiments:

[0103] The CA-Fe NPs prepared in Example 1 were used to set up a blank control group, an H2O2 control group, and a CA-Fe NPs aqueous solution treatment group (10 μg / ml, 20 μg / ml). The cells used in the experiment were human renal tubular epithelial cells (HK2). The apoptosis of HK2 cells after treatment was statistically analyzed by flow cytometry (FCM).

[0104] Blank control group: HK2 cells were not treated in any way;

[0105] H2O2 control group: HK2 cells were supplemented with H2O2 reagent;

[0106] CA-Fe NPs aqueous solution treatment group: H2O2 reagent was added to HK2 cells, and 10 μg / ml and 20 μg / ml CA-Fe NPs aqueous solution were added respectively.

[0107] Figure 17 These are FCM images of apoptosis in HK2 cells from each group, by... Figure 17 It can be seen that the apoptosis rate of cells in the CA-Fe NPs aqueous solution treatment group was significantly reduced, and the apoptosis rate of cells in the 20 μg / ml CA-Fe NPs aqueous solution treatment group was reduced to 6.4%.

[0108] Example 7

[0109] A metal polyphenol nanomaterial based on chlorogenic acid is used for in vitro anti-inflammatory effects:

[0110] First group of experiments:

[0111] The CA-Fe NPs prepared in Example 1 were used to set up a blank control group, an LPS-induced control group, and CA-Fe NPs aqueous solution treatment groups (2.5 μg / ml, 5 μg / ml, 10 μg / ml, 20 μg / ml, 30 μg / ml, 40 μg / ml). Macrophages were used as the experimental cells.

[0112] Blank control group: Macrophages were not treated in any way;

[0113] LPS-induced control group: Macrophages were induced by LPS to polarize macrophages towards the M1 type (pro-inflammatory type) and enhance the inflammatory response;

[0114] CA-Fe NPs aqueous solution treatment group: Macrophages were induced with LPS and then treated with CA-Fe NPs aqueous solution at concentrations of 2.5 μg / ml, 5 μg / ml, 10 μg / ml, 20 μg / ml, 30 μg / ml, and 40 μg / ml, respectively.

[0115] Figure 18 This is a graph showing the statistical results of macrophage recruitment ratios in each group. Figure 18 It can be seen that CA-Fe NPs inhibit LPS-induced macrophage recruitment in a concentration-dependent manner.

[0116] Second group of experiments:

[0117] The CA-Fe NPs prepared in Example 1 were used to set up a blank control group, an LPS-induced control group, and a CA-Fe NPs aqueous solution treatment group. The cells used in the experiment were mouse mononuclear macrophage leukemia cells (Raw264.7). Flow cytometry was used to analyze the results of CA-Fe NPs inhibiting LPS-induced macrophage M1 polarization.

[0118] Blank control group: Raw264.7 cells were not treated in any way;

[0119] LPS-induced control group: Raw264.7 cells were induced by LPS to promote M1 polarization (pro-inflammatory type);

[0120] CA-Fe NPs treatment group: Raw264.7 cells were induced with LPS and then CA-Fe NPs were added.

[0121] Figure 19 These are flow cytometry (FCM) images of Raw264.7 cells from each group. Figure 19 It can be seen that CA-Fe NPs inhibit macrophage polarization towards M1 type (pro-inflammatory type) and promote macrophage polarization towards M2 type (anti-inflammatory type).

[0122] Figure 20 This is a statistical chart showing the proportions of M1 and M2 macrophages in each group. Figure 20 a is a statistical graph of the proportion of M1 macrophages. After LPS induction, the proportion of M1 macrophages was significantly higher than that of the blank control group. After treatment with CA-Fe NPs, the proportion of M1 macrophages decreased significantly, indicating that it can inhibit the transformation of macrophages induced by LPS to M1 type. Figure 20b is a statistical chart of the proportion of M1 / M2 macrophages. The proportion of M1 / M2 macrophages in the CA-Fe NPs treatment group was significantly lower than that in the LPS-induced group and the blank group, indicating that CA-Fe NPs have the effect of promoting macrophage polarization towards M2 type.

[0123] Example 8

[0124] A metal polyphenol nanomaterial based on chlorogenic acid, in synergistic effect with near-infrared light, can be used to promote wound healing.

[0125] The CA-Fe NPs obtained in Example 1 were used to set up a blank control group, a NIR control group, a CA-Fe NPs treatment group, and a CA-Fe NPs+NIR treatment group, as follows: Figure 21 As shown, a mouse model of MRSA infection was established, and the effects of each group on wound healing in mice were observed.

[0126] Blank control group: The mouse wound model was not treated in any way;

[0127] NIR control group: 808nm near-infrared laser (NIR, 1.5W / cm²) was used. 2 Irradiation was applied to a mouse wound model;

[0128] CA-Fe NPs treatment group: CA-Fe NPs material was applied to the wounds of a mouse wound model;

[0129] CA-Fe NPs+NIR treatment group: CA-Fe NPs material was applied to the wounds of a mouse wound model, and an 808nm near-infrared laser (NIR, 1.5W / cm²) was used. 2 Irradiation was applied to a mouse wound model.

[0130] Temperature changes were measured in the NIR control group and the CA-Fe NPs+NIR treatment group;

[0131] Serum inflammatory factors were measured in mice of each group using an ELISA kit.

[0132] The wound tissues of mice in each group were stained with hematoxylin and eosin (H&E) and observed.

[0133] Masson staining was performed on the wound tissues of mice in each group, and the results were observed.

[0134] The wound healing status of mice in each group was observed.

[0135] Figure 22These are infrared thermographic images of mouse wounds in the NIR control group and the CA-Fe NPs+NIR treatment group. The wound temperature rise in the CA-Fe NPs+NIR treatment group was 15℃, which was significantly higher than the 4.2℃ temperature rise in the NIR control group.

[0136] Figure 23 The image shows the ELISA results of serum inflammatory factors (TNF-α, IL-1β, IL-6) in each group of mice. The CA-FeNPs+NIR treatment group had the lowest levels of inflammatory factors (IL-1β, IL-6, TNF-α), which were significantly lower than those in the blank control group. This indicates that the synergistic effect of CA-FeNPs+NIR can reduce the levels of inflammatory factors (TNF-α, IL-1β, IL-6) in the serum of injured mice, thereby promoting wound healing.

[0137] Figure 24 These are hematoxylin-eosin (H&E) staining images and Masson staining images of wound tissue from each group of mice. Figure 24 a is a hematoxylin-eosin (H&E) staining image of mouse wound tissue. The amount of granulation tissue generated in the CA-Fe NPs treatment group and the CA-Fe NPs+NIR treatment group was significantly higher than that in the NIR control group and the blank control group. Figure 24 b is a Masson staining image of mouse wound tissue, showing that collagen deposition levels were significantly increased in the CA-Fe NPs treatment group and the CA-Fe NPs+NIR treatment group.

[0138] Figure 25 The images show the wound healing status of mice in each group on days 0, 1, 3, 7, and 11. The wound healing rate of mice in the CA-Fe NPs+NIR treatment group was significantly higher than that of other groups.

[0139] Unless otherwise specified, the preparation methods, experimental schemes, or detection methods involved in the embodiments of this invention are all conventional schemes in the prior art, and their names and abbreviations are also common terms in the art, and their uses are clear and well known in the art. Furthermore, the various instruments, equipment, raw materials, or reagents used in the embodiments of this application have no special source requirements and are all conventional products that can be purchased through regular commercial channels. It should be noted that those skilled in the art can freely combine the technical features in the embodiments and make various modifications and improvements without departing from the concept of this invention, and these all fall within the protection scope of this invention. Therefore, all equivalent transformations and modifications made within the scope of the claims of this invention should be included in the protection scope of the claims of this invention.

Claims

1. A method for preparing metal polyphenol nanomaterials based on chlorogenic acid, characterized in that, Includes the following steps: Step 1: Chlorogenic acid and distearate phosphatidylethanolamine-polyethylene glycol are mixed and added to anhydrous tetrahydrofuran and ultrasonically dispersed to obtain the first mixture; Step 2: Add FeCl3 solution to the first mixture and ultrasonically disperse until a clear and transparent system is formed to obtain the second mixture; Step 3: Remove anhydrous tetrahydrofuran from the second mixture to obtain a metal polyphenol nanomaterial based on chlorogenic acid.

2. The method for preparing chlorogenic acid-based metal polyphenol nanomaterials according to claim 1, characterized in that, The mass ratio of chlorogenic acid and distearate phosphatidylethanolamine-polyethylene glycol is 1:1 to 3.

3. The method for preparing chlorogenic acid-based metal polyphenol nanomaterials according to claim 1, characterized in that, The concentration of the FeCl3 solution is 0.022-0.027 mol / L.

4. A metal polyphenol nanomaterial based on chlorogenic acid, characterized in that, The metal polyphenol nanomaterials are prepared by the preparation method described in any one of claims 1 to 3.

5. A metal polyphenol nanomaterial based on chlorogenic acid according to claim 4, characterized in that, The metal polyphenol nanomaterial has a spherical structure.

6. The chlorogenic acid-based metal polyphenol nanomaterial according to claim 4, characterized in that, The ZETA potential of the metal polyphenol nanomaterial is 18.23 mV ± 0.

46.

7. An application of a metal polyphenol nanomaterial based on chlorogenic acid, characterized in that, The chlorogenic acid-based metal polyphenol nanomaterial described in claim 4 is used for in vitro antioxidant applications.

8. An application of a metal polyphenol nanomaterial based on chlorogenic acid, characterized in that, The chlorogenic acid-based metal polyphenol nanomaterial described in claim 4 is used for in vitro anti-inflammatory applications.

9. An application of a metal polyphenol nanomaterial based on chlorogenic acid, characterized in that, The chlorogenic acid-based metal polyphenol nanomaterial described in claim 4 is used for in vitro photothermal sterilization.

10. An application of a metal polyphenol nanomaterial based on chlorogenic acid, characterized in that, The chlorogenic acid-based metal polyphenol nanomaterial described in claim 4 is used in conjunction with near-infrared light to promote wound healing.

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