Concentrated growth factor gel dressing based on broad-spectrum reactive oxygen species scavenging nanoparticles and methods of making and using the same

Abstract

The present application relates to the technical field of gel dressing, in particular to a concentrated growth factor gel dressing based on broad-spectrum active oxygen cluster scavenging nanoparticles and a preparation method and application thereof. The present application uses 4-hydroxy-2,2,6,6-tetramethylpiperidine nitroxide and 4-hydroxymethylphenylboronic acid pinacol ester to modify and combine a skeleton molecule such as glucose to construct degradable nanoparticles, so that the nanoparticles can be combined with concentrated growth factors derived from platelets to form a gel, effectively scavenge different types of active oxygen such as free radicals, superoxide anions, hydrogen peroxide and hypochlorite generated in a refractory wound environment, and promote wound healing in combination with the repair-promoting effect of CGF cytokines. The present technical solution can solve the technical problem of the lack of platelet glue containing broad-spectrum active oxygen cluster scavenging materials with good compatibility and therapeutic effect in the prior art, and can be used to prepare active oxygen scavenging drugs, anti-inflammatory drugs and drugs for promoting wound healing, and has a good application prospect.

Description

Technical Field

[0001] This invention relates to the field of gel dressing technology, specifically to concentrated growth factor gel dressings based on broad-spectrum reactive oxygen species scavenging nanoparticles, their preparation methods, and applications. Background Technology

[0002] Concentrated growth factor (CGF) is a new generation of platelet concentrate, obtained by centrifuging whole blood. After activation, the α-granules of the concentrated platelets release various bioactive factors, such as platelet-derived growth factor (PDGF), transforming growth factor-β (TGF-β), insulin-like growth factor (IGF), and epidermal growth factor (EGF). The gel-like substance formed after activation is called platelet gel (PG), whose bioactive molecules stimulate the proliferation of fibroblasts, smooth muscle cells, and osteoblasts, attracting various cellular components to participate in tissue repair. It plays an important role in anti-infection, hemostasis, coagulation, wound healing, and bone growth. The preparation of antioxidant gel dressings based on platelet-concentrated growth factor has significant practical value. Difficult-to-heal wounds are often complicated by infection, especially when reactive oxygen species are released, causing the wound to be in a state of excessive inflammation, preventing the formation of epidermis and granulation tissue for a long time. Effectively removing reactive oxygen species from the wound surface is one of the key issues in the repair of refractory wounds.

[0003] The applicant previously developed a broad-spectrum reactive oxygen species (ROS) scavenging material (CN108794656A). This material synthesizes a novel biodegradable material (TPCD) with broad-spectrum ROS scavenging properties by covalently bonding 4-hydroxy-2,2,6,6-tetramethylpiperidine nitride (TEMPOL, Tpl) and 4-hydroxymethylphenylboronic acid pinacol ester (PBAP) to the cyclic polysaccharide molecule β-cyclodextrin (β-CD). In vitro experiments have confirmed that the nanomaterial has low cytotoxicity. Furthermore, the applicant has combined the aforementioned broad-spectrum ROS scavenging material with a thermosensitive gel material to form a multifunctional supramolecular hydrogel (CN111632203A) for preventing postoperative tissue adhesions. This gel material can be applied topically during laminectomy to prevent epidural fibrosis adhesions and other postoperative tissue adhesions. Currently, there are no gel dressings that combine platelet-concentrated growth factor with antioxidants, making it impossible to effectively eliminate reactive oxygen species and suppress inflammatory responses during wound healing. There is an urgent need to develop a broad-spectrum reactive oxygen species scavenging material with good compatibility with platelet-concentrated growth factor to improve the repair of refractory wounds. Summary of the Invention

[0004] The present invention aims to provide a concentrated growth factor gel dressing based on broad-spectrum reactive oxygen species scavenging nanoparticles, in order to solve the technical problem of the lack of platelet gel containing broad-spectrum reactive oxygen species scavenging materials with ideal compatibility and therapeutic effect in the prior art.

[0005] To achieve the above objectives, the present invention adopts the following technical solution:

[0006] The concentrated growth factor gel dressing based on broad-spectrum reactive oxygen species scavenging nanoparticles is characterized in that its raw materials include TPG nanoparticles and concentrated platelet growth factor; the TPG nanoparticles are obtained by self-assembly of carrier material TPG, and the chemical formula of carrier material TPG is shown in formula (1).

[0007]

[0008] Wherein, R2 is H, or a group as shown in formula (2), or a group as shown in formula (3).

[0009] This invention also provides a method for preparing a concentrated growth factor gel dressing based on broad-spectrum reactive oxygen species scavenging nanoparticles, characterized by comprising the following steps performed sequentially:

[0010] S1 Preparation of TPG nanoparticles: TPG carrier material was dissolved in methanol to obtain an organic phase; lecithin and PEG-modified distearate phosphatidylethanolamine were dissolved in deionized water to obtain an aqueous phase; under stirring conditions, the organic phase was added dropwise to the aqueous phase, and then washed and redispersed to obtain TPG nanoparticles.

[0011] S2 is used to prepare concentrated platelet growth factor;

[0012] S3 is used to prepare gel excipients.

[0013] This solution also provides an application of a concentrated growth factor gel dressing based on broad-spectrum reactive oxygen species scavenging nanoparticles in the preparation of reactive oxygen species scavenging drugs, anti-inflammatory drugs, or drugs that promote wound healing.

[0014] Furthermore, the carrier material TPG is prepared by the following method:

[0015] 4-Hydroxy-2,2,6,6-Tetramethylpiperidine N-oxide was dissolved in dichloromethane, and N,N'-carbonyldiimidazole was added, followed by reaction to obtain Tpl-CDI; Tpl-CDI, the backbone molecule, and 4-dimethylaminopyridine were dissolved in dimethyl sulfoxide, and followed by reaction to obtain Glu-Tpl; 4-hydroxymethylphenylboronic acid pinacol ester was dissolved in dichloromethane, and N,N'-carbonyldiimidazole was added, followed by reaction to obtain PBAP-CDI; PBAP-CDI, Glu-Tpl, and DMAP were dissolved in dimethyl sulfoxide, and after reaction and purification, the carrier material TPG was obtained; the backbone molecule is a sugar or sugar alcohol containing a polyhydroxy structure.

[0016] Furthermore, the carrier material TPG is formed into TPG nanoparticles by the following method: TPG carrier material is dissolved in methanol to obtain an organic phase; lecithin and PEG-modified distearate phosphatidylethanolamine are dissolved in deionized water to obtain an aqueous phase; under stirring conditions, the organic phase is added dropwise to the aqueous phase, and then washed and redispersed to obtain TPG nanoparticles.

[0017] Furthermore, the concentrated platelet growth factor is prepared by the following method: platelet suspension is obtained by separating it from whole blood; then, the platelet suspension is centrifuged at 277g for 6min at 22℃-24℃, and the upper platelet layer is taken to obtain the concentrated platelet growth factor.

[0018] Furthermore, the concentrated growth factor gel dressing is obtained by mixing and solidifying TPG nanoparticles, concentrated platelet growth factor, and a calcium gluconate solution containing thrombin.

[0019] Further, in S1, the carrier material TPG is prepared by the following method: 4-hydroxy-2,2,6,6-tetramethylpiperidine nitride and 4-hydroxymethylphenylboronic acid pinacol ester are activated with N,N'-carbonyldiimidazole to obtain Tpl-CDI and PBAP-CDI, respectively; Tpl-CDI is reacted with the backbone molecule to obtain Glu-Tpl; Glu-Tpl is reacted with PBAP-CDI to obtain the carrier material TPG; wherein the backbone molecule is a sugar or sugar alcohol containing a polyhydroxy structure.

[0020] Further, in S2, platelet suspension is separated from whole blood; then, the platelet suspension is centrifuged at 277g for 6 minutes at 22℃-24℃, and the upper platelet layer is taken to obtain concentrated platelet growth factor.

[0021] Furthermore, in S3, TPG nanoparticles, concentrated platelet growth factor, and calcium gluconate solution containing thrombin are mixed and solidified to obtain a concentrated growth factor gel dressing; the solute concentration of the calcium gluconate solution is 10%; and the enzyme activity of thrombin is 500U.

[0022] The principle and beneficial effects of this technical solution are as follows:

[0023] Previously, the inventors synthesized a novel biodegradable material (TPCD) with broad-spectrum reactive oxygen species (ROS) scavenging properties by covalently bonding 4-hydroxy-2,2,6,6-tetramethylpiperidine nitride (TEMPOL, Tpl) and 4-hydroxymethylphenylboronic acid pinacol ester (PBAP) to the cyclic polysaccharide molecule β-cyclodextrin (β-CD). In vitro experiments confirmed that the nanomaterial had low cytotoxicity. However, the compatibility of this material with platelet-concentrated growth factor (PCGF) was limited, making it difficult to achieve ideal wound repair effects. In this technical solution, the inventors replaced β-cyclodextrin with sugars or sugar alcohols containing multiple hydroxyl groups, such as glucose, and covalently modified the above-mentioned backbone molecule with Tpl and PBAP to construct biodegradable nanoparticles (TPG). The TPG of this solution retains the ROS scavenging and anti-inflammatory effects of the original TPCD while improving the compatibility between the molecule and platelets. Furthermore, molecules such as glucose are themselves in vitro platelet protectants, which can enhance the stability and efficacy of PCGF. TPG can bind with platelet-derived CGF to form a gel, effectively scavenging various types of reactive oxygen species (ROS) generated in refractory wound environments, such as free radicals, superoxide anions, hydrogen peroxide (H2O2), and hypochlorite. Combined with the repair-promoting effects of CGF cytokines, it promotes wound healing. This concentrated growth factor gel dressing based on broad-spectrum ROS-scavenging nanoparticles can be applied in multiple medical practices, such as the preparation of ROS-scavenging drugs, anti-inflammatory drugs, and drugs that promote wound healing.

[0024] This solution utilizes broad-spectrum reactive oxygen species (ROS) scavenging nanoparticles that effectively scavenge a wide range of ROS, exhibiting good biocompatibility in vivo, biodegradability, and non-toxic degradation products. The Tpl and PBAP used in this solution are widely used nitric oxide radicals in biological experiments and demonstrate excellent ROS scavenging effects. Animal experiments have shown that nitric oxide can effectively alleviate acute and chronic inflammatory diseases, such as acute lung injury, atherosclerosis, pancreatitis, and colitis. The formation of nanoparticles and gels addresses the issues of non-specific distribution, short half-life, and low bioavailability of small-molecule ROS scavenging drugs in vivo, expanding their clinical applications. Furthermore, the preparation of functionalized small-molecule antioxidants overcomes the poor water solubility of small-molecule compounds, enhancing the efficacy of small-molecule anti-inflammatory and ROS scavenging agents.

[0025] Since the biological effects of CGF depend on the levels of various growth factors released by platelets, the bioactivity of platelets determines the biological effects of CGF. Because centrifugation is essential in the preparation of CGF, the levels of various growth factors in platelet gels can be affected by many factors, including differences in the type of centrifuge used, the concentration employed, and the preparation method. This method uses a modified platelet enrichment technique to obtain sufficient CGF. In vitro studies have confirmed that it is rich in growth factors, with a proportion close to normal physiological ratios. It also contains fibrin, possessing unique properties that improve and enhance tissue regeneration, thus effectively leveraging the synergistic effects among growth factors. The CGF prepared using this method has high internal tensile strength, strong stretching and ductility, and short hemostasis time. Furthermore, CGF is non-toxic, non-immunogenic, eliminates the possibility of immune reactions, and is easy to prepare, with short operation time, minimal trauma, and low cost. Attached Figure Description

[0026] Figure 1 This is a schematic diagram of the synthesis of TPG, the carrier material in Example 1.

[0027] Figure 2 This is a schematic diagram of the preparation process of the TPG CGF gel in Example 2.

[0028] Figure 3 This is a photograph of the TPG CGF gel from Example 2.

[0029] Figure 4 The results of ROS detection in in vitro platelet preservation in Experiment Example 1 are shown.

[0030] Figure 5 These are micrographs of platelet morphology under different conditions in Experiment Example 1.

[0031] Figure 6 The statistical results show the expression of CD62p on the plasma membrane of platelets stored under different storage conditions for 5 days in Example 1.

[0032] Figure 7 The results show the platelet aggregation rate under different conditions in Example 1.

[0033] Figure 8 The image shows the WB images of αⅡbβ3 protein expression in platelets stored under different storage conditions for 5 days, as shown in Example 1.

[0034] Figure 9 This is a photograph of the wound on the model animal before repair in Example 2.

[0035] Figure 10 This is a photograph of the wound after repair in the model animal of Example 2. Detailed Implementation

[0036] The present invention will be further described in detail below with reference to embodiments, but the implementation of the present invention is not limited thereto. Unless otherwise specified, the technical means used in the following embodiments and experimental examples are conventional means well known to those skilled in the art, and the materials and reagents used are all commercially available.

[0037] Example 1: Preparation and characterization of TPG nanoparticles

[0038] (1) Synthesis of carrier material TPG

[0039] Under nitrogen protection, 501 mg of 4-hydroxy-2,2,6,6-tetramethylpiperidine nitride (TEMPOL, Tpl) was dissolved in 10 mL of anhydrous dichloromethane (DCM), followed by the addition of 955 mg of N,N'-carbonyldiimidazole (CDI). The reaction yielded Tpl activated by CDI. The Tpl-CDI obtained in the previous step, 144 mg of glucose (Glu), and 600 mg of 4-dimethylaminopyridine (DMAP) were dissolved in 10 mL of anhydrous DMSO and stirred at room temperature to obtain Glu-Tpl (TG).

[0040] 555 mg of pinacol 4-hydroxymethylphenylboronic acid (PBAP) was dissolved in 10 mL of DCM, and then 765 mg of CDI was added and stirred to obtain CDI-activated PBAP. Finally, the PBAP-CDI obtained in the previous step, the TG obtained in the previous step, and 503 mg of DMAP were dissolved in 10 mL of anhydrous DMSO and reacted overnight at 30 °C. The product was purified according to standard procedures to obtain the broad-spectrum ROS-scavenging nanomaterial TPG (TPG carrier material). The specific synthetic route is as follows... Figure 1 As shown.

[0041] Through nuclear magnetic resonance spectroscopy ( 1 The structure of TPG was determined by 1H NMR, FT-IR, electron spin resonance (EPR) and ultraviolet (UV) spectra, and the molecular weight and distribution of the material were determined by matrix-assisted laser desorption / ionization time-of-flight mass spectrometry (MALDI-TOF MS) combined with gel permeation chromatography (GPC).

[0042] (2) Preparation and characterization of TPG nanoparticles

[0043] Various nanoparticles were prepared using a nanoprecipitation / self-assembly method. First, 15 mg of the carrier material TPG was dissolved in 1.5 mL of methanol to obtain the organic phase. Then, 1.8 mg of lecithin and 2.7 mg of DSPE-PEG were dissolved in 0.3 mL of deionized water to obtain the aqueous phase. The organic phase was slowly added dropwise to the aqueous phase under vigorous stirring. After centrifugation and washing with water to remove the organic solvent, the nanoparticles (TPG NPs) were redispersed in water by ultrasonication to obtain the nanoparticles.

[0044] The physicochemical properties of TPG NPs were characterized, including observation and research on particle size, surface potential, morphology, nanoparticle stability, and responsive hydrolysis. Details are as follows:

[0045] Particle size and surface potential: The particle size and distribution of nanoparticles were determined by laser particle size analyzer, and the surface zeta potential was determined by potentiometer.

[0046] Morphology observation: Field emission scanning electron microscopy (FE-SEM), transmission electron microscopy (TEM), and atomic force microscopy (AFM) were used to observe the morphology of the nanoparticles.

[0047] Nanoparticle stability study: Nanoparticles were placed in PBS or fetal bovine serum (FBS), and their size and distribution were measured periodically.

[0048] Response hydrolysis study of nanoparticles: Freshly prepared nanoparticles were hydrolyzed by adding hydrogen peroxide solutions of different concentrations. The transmittance of the solution was measured at specific time points, the hydrolysis percentage was calculated, and hydrolysis kinetic curves were plotted.

[0049] (3) Evaluation of the in vitro biological effects of TPG NP

[0050] (3.1) Determination of the broad-spectrum ROS scavenging performance of TPG NP

[0051] Determination of TPG NP free radical scavenging ability: Different concentrations of TPG NP and 100 μg / mL of 1,1-diphenyl-2-trinitrophenylhydrazine (DPPH·) were co-incubated in the dark. The absorbance at 517 nm was measured using UV-Vis spectroscopy at different time points, and the free radical scavenging ability curve of TPG NP was calculated.

[0052] Determination of TPG NP's ability to scavenge superoxide anions: Different concentrations of TPG NP and excess superoxide anions were co-incubated, and the content of residual superoxide anions in the solution was detected using a kit. Curves of superoxide anion scavenging abilities of different TPG NPs were calculated.

[0053] Determination of TPG NP's ability to scavenge hydrogen peroxide: Different concentrations of TPG NP and excess H2O2 (50 mM) were co-incubated overnight. The concentration of residual hydrogen peroxide in the solution was detected using a kit, and the curves of TPG NP's ability to scavenge H2O2 were calculated.

[0054] Determination of TPG NP hypochlorite scavenging ability: Using the method previously established by our research group, NaClO solution and different concentrations of TPG NP were reacted in a 96-well plate for 15 min. The chemiluminescence intensity was measured by a live imaging system, and the curve of TPG NP hypochlorite scavenging ability was calculated.

[0055] (3.2) Investigate the uptake of TPG NP by platelets

[0056] Platelets were co-incubated with Cy5-labeled TPG NPs. The concentration of nanoparticles or the incubation time were varied, and cells were collected. The fluorescence of Cy5 in various cells was detected by flow cytometry and statistically analyzed to investigate the time- and dose-dependent effects of TPG NP uptake by cells.

[0057] (3.3) Evaluation of the protective effect of TPG NP against H2O2-induced platelet apoptosis

[0058] Platelets were co-incubated with different TPG NPs for 2 hours, then incubated overnight with 200 μM H2O2. Cells were collected, resuspended in Annexin V and PI solutions, and the proportion of apoptotic cells was detected by flow cytometry.

[0059] Example 2: Preparation of TPG CGF Gel

[0060] Platelets are separated from whole blood collected at room temperature and transported within 6 hours of collection, or from whole blood collected at 20℃-24℃ and transported within 24 hours of collection, and suspended in a certain amount of plasma. This technical solution does not involve the blood collection process, but only utilizes the collected blood to separate platelets, and blood products such as blood and plasma have already been commercialized.

[0061] The specific process for separating and obtaining concentrated platelet-derived growth factor is as follows: Collect 400ml of fresh whole blood using a disposable six-piece collection bag and place it at room temperature (collection time <10min). Shake the whole blood for 20-30min at a shaking speed of 60 times / min. Centrifuge the whole blood for the first time at 1875g (2600 rpm) at 22℃-24℃ for 15min. Gently remove the blood and place it 2cm above the plasma separator. Separate the upper plasma layer into a plasma transfer bag, leaving approximately 50ml of plasma. Transfer 10ml of the remaining plasma into a platelet storage bag. Transfer the remaining plasma, along with the white blood cell layer and the 1mm thick layer of red blood cells below the white blood cell layer, into a white blood cell bag. Mix the plasma from the platelet storage bag into the white blood cell layer and clean the connecting tubing. Seal the white blood cell bag and platelet storage bag together from the mother bag using heat sealing to obtain a platelet suspension. Check the heat seal to ensure there is no leakage. After standing at room temperature for 1 hour, place the suspension in a platelet storage box and shake for storage. Centrifuge the platelet suspension at 277g (1000 rpm) for 6 minutes at 22℃-24℃. After centrifugation, gently transfer the supernatant platelets into a platelet storage bag to obtain concentrated platelet CGF. Store at 22℃-24℃ with shaking. To use, mix 5mg TPG, 1mL CGF, and 100μL of a 10% calcium gluconate solution containing 500U thrombin powder (commercial reagent) evenly. After coagulation, TPG CGF gel is obtained. See the schematic diagram for the TPG CGF gel preparation process. Figure 2 See the physical photo of the obtained TPG CGF gel. Figure 3 .

[0062] Experimental Example 1: Study on the alleviating effect of TPG nanoparticles on platelet storage damage

[0063] (1) Experimental methods

[0064] (1.1) Storage damage detection: Platelets stored in vitro were sampled and tested on days 1, 3, and 5. The specific tests are as follows:

[0065] Reactive oxygen species (ROS) detection: Platelets from each group were collected to detect ROS levels.

[0066] Morphological observation: Platelet smears from each group were taken and their morphology was observed under a transmission electron microscope.

[0067] Aggregation function test: ADP was added to induce platelet aggregation in each group, and the aggregation rate was measured. The specific procedure was as follows: the platelet concentration for aggregation function testing was adjusted to 300 × 10⁻⁶. 9The maximum aggregation rate was determined by zeroing the instrument with PPP (platelet Poor Plasma) and adding PRP (Platelet Rich Plasma) and an inducer (a mixture of ADP, adenosine diphosphate (12.5 μM) and EPN (epinephrine (750 μM)) at a ratio of 9:1.

[0068] (1.2) Flow cytometry platelet detection: Take a certain amount of platelets, centrifuge at 1000g for 5 minutes, collect the platelets, and dilute each tube to 10. 6 / L, perform the following tests respectively:

[0069] PS detection: PS antibody incubation is performed, and the PS content is measured by flow cytometry.

[0070] GPIb-α and GPV expression levels were determined by incubation with the corresponding monoclonal antibodies and observation under flow cytometry.

[0071] GPⅡbⅢa content determination: The GPⅡbⅢa content was determined by flow cytometry using Pac-1 antibody.

[0072] CD62p expression level determination: CD62p levels were measured using flow cytometry with CD62p antibody. The specific procedure was as follows: the platelet count was adjusted to 200 × 10⁻⁶ using platelet-rich plasma from individuals with anemia. 9 / L, blank tube: 5μL diluted platelets + 5μL IgG-PE, control tube: 5μL diluted platelets + 5μL CD61-FITC, test tube: 5μL diluted platelets + 5μL CD62p-PE, each tube was fixed with 400μL PBS solution containing 1% paraformaldehyde (pH 7.38) for testing.

[0073] (2) Experimental groups and experimental results

[0074] Platelets were obtained according to the concentrated platelet preparation method in Example 2, and the same apheresis platelets were divided into four groups (platelet count > 2.5 × 10⁻⁶). 10 Group A: Standard storage (platelets without any added substances, stored at 22℃); Group B: Platelets + superoxide dismutase (SOD) (SOD dosage 10 mg / mL); Group C: Platelets + novel broad-spectrum reactive oxygen species scavenger nanoparticles (prepared according to Example 1, TPG nanoparticle dosage 5 mg / mL). The concentration of reactive oxygen species and related functions in platelets were measured at 1, 3, and 5 days. Results are shown in [Table missing]. Figure 4 And Table 1. In Figure 4In the figures, A represents the ROS concentration of platelets stored under normal conditions for different time periods; B represents the ROS concentration of platelets stored with added SOD for different time periods; and C represents the intracellular ROS concentration of platelets stored with novel broad-spectrum reactive oxygen species scavengers (★ < 0.05) for different time periods. These experimental results demonstrate that TPG nanoparticles possess excellent free radical scavenging capabilities and can reduce platelet storage damage.

[0075] Table 1: Results of reactive oxygen species detection in platelets after 5 days of storage under different conditions

[0076]

[0077] The morphology of platelets under different preservation conditions was also observed. Figure 5 ), serous membrane CD62p expression ( Figure 6 ), clustering rate ( Figure 7 ), and metabolic level ( Figure 8 Under different storage conditions, all of the above indicators changed. Figure 5 In the figures, A represents the platelet morphology after 5 days of conventional storage; B represents the platelet cell morphology after 5 days of storage with added SOD; and C represents the platelet morphology after 5 days of storage with novel nanoparticles containing broad-spectrum reactive oxygen species scavengers. Figure 7 In the figures, A represents platelet aggregation after 5 days of normal storage; B represents platelet aggregation after 5 days of storage with added SOD; and C represents platelet aggregation after 5 days of storage with added novel nanoparticles containing broad-spectrum reactive oxygen species scavengers.

[0078] Experimental Example 2: In vivo experiment

[0079] Full-thickness skin was removed from the back of the mouse, and a puncture device was used to create a wound on the back, approximately 6 mm in diameter (see [link]). Figure 9 According to the method in Example 2 of this protocol, TPG CGF and a 10% calcium gluconate solution containing 500U thrombin powder (commercial reagent) were mixed evenly to form a gel, which was then applied to the wound. 1 mL was applied to each wound. The gel dressing was changed on days 1, 3, 5, 7, and 14. Wound healing was observed after 14 days (see [link to protocol]). Figure 10 After treatment, the experimental animals' wounds healed, indicating that the TPG CGF gel excipient in this protocol has the effect of promoting wound healing.

[0080] The above descriptions are merely embodiments of the present invention, and common knowledge such as specific technical solutions and / or characteristics are not described in detail here. It should be noted that those skilled in the art can make various modifications and improvements without departing from the technical solutions of the present invention, and these should also be considered within the scope of protection of the present invention. These modifications and improvements will not affect the effectiveness of the implementation of the present invention or the practicality of the patent. The scope of protection claimed in this application should be determined by the content of its claims, and the specific embodiments described in the specification can be used to interpret the content of the claims.

Claims

1. A concentrated growth factor gel dressing based on broad-spectrum reactive oxygen species scavenging nanoparticles, characterized in that, Its raw materials include TPG nanoparticles and concentrated platelet growth factor; the TPG nanoparticles are obtained by self-assembly of the carrier material TPG, and the chemical formula of the carrier material TPG is shown in formula (1); The backbone of the carrier material TPG shown in formula (1) is glucose; R2 is H, or a group as shown in formula (2), or a group as shown in formula (3), and formula (1) contains both a group as shown in formula (2) and a group as shown in formula (3); In equations (2) and (3), This represents the chemical bond that connects the carbonyl carbon to the oxygen atom of formula (1).

2. The concentrated growth factor gel dressing based on broad-spectrum reactive oxygen species scavenging nanoparticles according to claim 1, characterized in that, The carrier material TPG is prepared by the following method: 4-Hydroxy-2,2,6,6-Tetramethylpiperidine N-oxide was dissolved in dichloromethane, and N,N'-carbonyldiimidazole was added, followed by reaction to obtain Tpl-CDI; Tpl-CDI, glucose, and 4-dimethylaminopyridine were dissolved in dimethyl sulfoxide, and followed by reaction to obtain Glu-Tpl; 4-hydroxymethylphenylboronic acid pinacol ester was dissolved in dichloromethane, and N,N'-carbonyldiimidazole was added, followed by reaction to obtain PBAP-CDI; PBAP-CDI, Glu-Tpl, and DMAP were dissolved in dimethyl sulfoxide, and after reaction and purification, the carrier material TPG was obtained.

3. The concentrated growth factor gel dressing based on broad-spectrum reactive oxygen species scavenging nanoparticles according to claim 2, characterized in that, TPG nanoparticles were formed from the carrier material TPG by the following method: TPG was dissolved in methanol to obtain an organic phase; lecithin and PEG-modified distearate phosphatidylethanolamine were dissolved in deionized water to obtain an aqueous phase; the organic phase was added dropwise to the aqueous phase under stirring conditions, and then washed and redispersed to obtain TPG nanoparticles.

4. The concentrated growth factor gel dressing based on broad-spectrum reactive oxygen species scavenging nanoparticles according to claim 1, characterized in that, The concentrated platelet growth factor was prepared by the following method: a platelet suspension was obtained by separating it from whole blood; then, the platelet suspension was centrifuged at 277g for 6 minutes at 22℃-24℃, and the upper platelet layer was taken to obtain the concentrated platelet growth factor.

5. The concentrated growth factor gel dressing based on broad-spectrum reactive oxygen species scavenging nanoparticles according to claim 4, characterized in that, The concentrated growth factor gel dressing is obtained by mixing and solidifying TPG nanoparticles, concentrated platelet growth factor, and a calcium gluconate solution containing thrombin.

6. The method for preparing a concentrated growth factor gel dressing based on broad-spectrum reactive oxygen species scavenging nanoparticles according to any one of claims 1-5, characterized in that, The following steps are performed sequentially: S1 Preparation of TPG nanoparticles: TPG carrier material was dissolved in methanol to obtain an organic phase; lecithin and PEG-modified distearate phosphatidylethanolamine were dissolved in deionized water to obtain an aqueous phase; under stirring conditions, the organic phase was added dropwise to the aqueous phase, and then washed and redispersed to obtain TPG nanoparticles. S2 is used to prepare concentrated platelet growth factor; S3 is used to prepare gel dressings.

7. The method for preparing a concentrated growth factor gel dressing based on broad-spectrum reactive oxygen species scavenging nanoparticles according to claim 6, characterized in that, In S1, the carrier material TPG is prepared by activating 4-hydroxy-2,2,6,6-tetramethylpiperidine nitride and 4-hydroxymethylphenylboronic acid pinacol ester with N,N'-carbonyldiimidazole to obtain Tpl-CDI and PBAP-CDI, respectively; reacting Tpl-CDI with glucose to obtain Glu-Tpl; and reacting Glu-Tpl with PBAP-CDI to obtain the carrier material TPG.

8. The method for preparing a concentrated growth factor gel dressing based on broad-spectrum reactive oxygen species scavenging nanoparticles according to claim 7, characterized in that, In S2, platelet suspension is separated from whole blood; then, the platelet suspension is centrifuged at 277g for 6 minutes at 22℃-24℃, and the upper platelet layer is collected to obtain concentrated platelet growth factor.

9. The method for preparing a concentrated growth factor gel dressing based on broad-spectrum reactive oxygen species scavenging nanoparticles according to claim 8, characterized in that, In S3, TPG nanoparticles, concentrated platelet growth factor, and calcium gluconate solution containing thrombin are mixed and solidified to obtain a concentrated growth factor gel dressing; the solute concentration of the calcium gluconate solution is 10%; and the enzyme activity of thrombin is 500U.

10. The use of the concentrated growth factor gel dressing based on broad-spectrum reactive oxygen species scavenging nanoparticles according to any one of claims 1-5 in the preparation of a drug that promotes wound healing.

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