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CN-122005912-A - Medical composite material for synergistically regulating cholesterol and oxidative stress

CN122005912ACN 122005912 ACN122005912 ACN 122005912ACN-122005912-A

Abstract

The invention discloses a medical composite material for synergistically regulating cholesterol and oxidative stress, which comprises an Fe 2 -N 6 /NC carrier, cholesterol oxidase loaded on the carrier and a hyaluronic acid targeting layer modified on the surface, wherein the carrier is a nitrogen-doped carbon material loaded with iron diatomic active sites, the distance between adjacent iron atoms is 0.311+/-0.009 nm, the iron content is 0.393wt%, the average particle size of the composite material is 242.5nm, the zeta potential is-23.71 mV, the preparation method comprises the steps of mechanically-chemically assisted in-situ MOF encapsulation to synthesize Fe 2 -N 6 /NC, ultrasonic loading cholesterol oxidase and hyaluronic acid modification, and the material has cholesterol degradation activity (degradation rate of 33.94%) and active oxygen removal activity (clearance rate of 78.8%), and can be used for preparing implant materials for joint repair or medical materials for joint cavity injection, so as to realize the synergistic regulation of cholesterol metabolic disturbance and oxidative stress of osteoarthritis.

Inventors

  • LI GUODONG
  • ZHANG YIWEI
  • LU HENGLI
  • XU TIANYANG
  • YANG DONG
  • DUAN ZHENGWEI
  • WANG SEN
  • GUAN YONGHAO

Assignees

  • 上海市第十人民医院

Dates

Publication Date
20260512
Application Date
20260415

Claims (10)

  1. 1. A medical composite material for synergistically regulating cholesterol and oxidative stress is characterized by comprising the following components: 1) Fe 2 -N 6 /NC carrier, wherein the carrier is nitrogen-doped carbon material loaded with iron diatomic active sites, iron is dispersed in a diatomic form, and the distance between adjacent iron atoms is 0.311+/-0.009 nm; 2) Cholesterol oxidase, supported on the Fe 2 -N 6 /NC carrier; 3) And hyaluronic acid is modified on the surface of the composite material to form a targeting layer.
  2. 2. The medical composite material for synergistically controlling cholesterol and oxidative stress according to claim 1, wherein the content of iron element in the Fe 2 -N 6 /NC carrier is 0.3-0.5%.
  3. 3. The medical composite material for synergistically regulating cholesterol and oxidative stress according to claim 1, wherein the loading amount of cholesterol oxidase is 10-14wt%.
  4. 4. The medical composite material for synergistically regulating cholesterol and oxidative stress according to claim 1, wherein the molecular weight of hyaluronic acid is 1.0-2.0kDa and the modified density is 10-15 μg/mg.
  5. 5. The medical composite material for synergistically regulating cholesterol and oxidative stress according to claim 1, wherein the average particle size of the composite material is 242.5nm, and the zeta potential is-23.71 mV.
  6. 6. The method for preparing the medical composite material for synergistically regulating and controlling cholesterol and oxidative stress according to any one of claims 1 to 5, comprising the following steps: 1) Synthesis of Fe 2 -N 6 /NC Carrier Mixing 0.4875g of zinc source, 0.0096g of iron diatomic precursor, 1.95g of organic ligand, 15mg of mechanochemical reaction regulating agent and 450 mu L of N, N-dimethylformamide, ball milling, centrifugal washing, drying and grinding, and pyrolyzing under inert atmosphere to obtain Fe 2 -N 6 /NC; 2) Cholesterol oxidase-loaded Dispersing the Fe 2 -N 6 /NC obtained in the step 1) in deionized water, carrying out ultrasonic treatment, adding a Cholesterol Oxidase (COD) solution, carrying out ultrasonic treatment, stirring, centrifuging and collecting to obtain C@Fe 2 -N 6 /NC; 3) Modified hyaluronic acid Dissolving hyaluronic acid in water, then dripping the solution into C@Fe 2 -N 6 /NC obtained in the step 2), and centrifugally collecting H/C@Fe 2 -N 6 /NC by stirring.
  7. 7. The method for preparing the medical composite material for synergistically controlling cholesterol and oxidative stress according to claim 6, wherein in the step 1), the iron diatomic precursor is cyclopentadienyl dicarbonyl iron dimer, the zinc source is zinc oxide, the organic ligand is 2-methylimidazole 2-MI, the mechanochemical reaction controlling agent is ammonium nitrate, the ball milling speed is 400-600rpm, the time is 1-3h, the pyrolysis temperature is 800-1000 ℃ and the time is 2-4h, the centrifugation is 2-3 times (5000 rmp,15 min), and the drying is carried out at 70 ℃.
  8. 8. The preparation method of the medical composite material for cooperatively regulating and controlling cholesterol and oxidative stress according to claim 6, wherein in the step 2), the mass-volume ratio of Fe 2 -N 6 /NC to deionized water is 0.005:5 (g/mL), the 1 st ultrasonic time is 180-300min, the concentration of the cholesterol oxidase COD solution is 0.005mg/mL, the mass ratio of mixed Fe 2 -N 6 /NC to COD is 1:1, the 2 nd ultrasonic time is 20-40min, the stirring time is 22-26h, the centrifugal rotating speed is 10000-14000rpm, and the time is 10-20min.
  9. 9. The method for preparing the medical composite material for cooperatively regulating and controlling cholesterol and oxidative stress according to claim 6, wherein the concentration of the hyaluronic acid solution in the step 3) is 4-6mg/mL, the stirring time is 10-14h, the centrifugal speed is 10000-14000rpm, and the time is 10-20min.
  10. 10. The use of the medical composite material according to any one of claims 1 to 5 for preparing an implant material for joint repair or a medical material for joint cavity injection, wherein the material has cholesterol degradation activity of 33.94% and has an iron diatomic nano-enzyme carrier with a clearance of 78.8%.

Description

Medical composite material for synergistically regulating cholesterol and oxidative stress Technical Field The invention belongs to the field of material synthesis and application, and particularly relates to a medical composite material for synergistically regulating cholesterol and oxidative stress. Background Osteoarthritis (Osteoarthritis, OA) is a chronic degenerative disease characterized mainly by degeneration of articular cartilage, synovial inflammation and joint dysfunction, and its pathogenesis is complex involving interactions of multiple factors such as biomechanics, metabolism, inflammation, etc. In recent years, research shows that cholesterol metabolic disorder and oxidative stress enhancement are two major key pathological factors driving OA progression, namely abnormal accumulation of cholesterol in joint cavities can induce cartilage cell lipid toxicity injury, and excessive Reactive Oxygen Species (ROS) can cause mitochondrial dysfunction, extracellular matrix degradation and inflammatory cascade amplification. The two are interwoven to form vicious circle, which promotes the disease course development of OA together. Therefore, the development of an intervention strategy capable of simultaneously regulating cholesterol metabolism and oxidative stress has important significance for realizing source treatment of OA. The nano-enzyme is used as a novel biological material with the physicochemical properties of the nano-material and the catalytic activity of the enzyme, and has wide application prospect in the aspects of scavenging ROS and relieving oxidative stress. In the prior art, there have been attempts by researchers to apply nanoenzymes to OA treatment. For example, chinese patent No. CN118403068a discloses a preparation and application of a Fe monoatomic nano-enzyme for targeted delivery of small interfering RNA, where the nano-enzyme uses a two-dimensional nitrogen doped carbon nano-sheet as a carrier to load Fe monoatomic, and implements targeted delivery of chondrocytes and down-regulation of inflammatory factors by modifying WYRGRL targeted peptide and loading siMMP13, but the core mechanism is still focused on scavenging ROS and gene silencing, and cholesterol metabolic regulation is not involved. In addition, the Chinese patent application with publication number of CN120459055A discloses a preparation method and application of FeMn diatomic nano-enzyme wrapped by M2 type macrophage membrane, which utilizes the synergistic catalytic effect of FeMn diatomic sites to enhance the activities of SOD and CAT enzyme and realizes the inflammation targeting through the biomimetic coating of the macrophage membrane, but the treatment strategy is also limited to scavenging ROS and can not interfere with the abnormal cholesterol metabolism. In the catalytic mechanism research of diatomic nanoenzymes, the literature (Hao Zhang et al, ANGEWANDTE CHEMIE International Edition, 2023) has reported that adjacent iron diatomic sites (Fe 2 -SAzyme) have significantly enhanced Catalase (CAT) like activity and higher CAT/Peroxidase (POD) selectivity compared to Shan Yuanzi iron sites (Fe 1 -SAzyme), and are able to efficiently decompose H 2O2 into O 2 instead of generating more toxic hydroxyl radicals (·oh). The research reveals the key influence of atomic distance regulation on catalytic selectivity, but the application of the method is still limited to basic catalytic research, does not belong to the biomedical field, and does not propose a technical scheme for combining with functional enzymes (such as cholesterol oxidase). Despite the advances made in nanoenzyme catalytic activity optimization or targeted delivery strategies in the above prior art, the following deficiencies still remain: The treatment dimension is single, the prior art focuses on scavenging ROS to relieve oxidative stress, and intervention cannot be carried out on cholesterol metabolic disorder, which is another core pathological factor of OA. A single antioxidant strategy is difficult to block the vicious circle between cholesterol accumulation and oxidative stress, and has limited therapeutic effects. The catalytic selectivity is to be improved, and although the monoatomic iron nano enzyme has enzyme-like activity, the inherent POD-like activity is easy to catalyze and generate OH, and secondary damage can be caused to chondrocytes. Although diatomic iron nanoenzymes have theoretical advantages in catalytic selectivity, they have not been reported for use in combination with functional enzymes and for use in OA therapy. The lack of multifunctional collaborative design is that the existing targeting strategy (such as polypeptide modification or cell membrane bionic) can realize enrichment of inflammatory sites, but the modification layer has no therapeutic function. Hyaluronic acid is used as a main component of joint synovial fluid, has not only CD44 targeting capability, but also joint lubricating and anti-inflammatory activ