Volume 39 Issue 1
Jan.  2023
Turn off MathJax
Article Contents
Ou ZL,Wang J,Shi R,et al.Influence of reactive oxygen species responsive self-assembled nanomicelle loaded with pyroptosis inhibitor on full-thickness skin defects in diabetic rats[J].Chin J Burns Wounds,2023,39(1):35-44.DOI: 10.3760/cma.j.cn501225-20221109-00483.
Citation: Ou ZL,Wang J,Shi R,et al.Influence of reactive oxygen species responsive self-assembled nanomicelle loaded with pyroptosis inhibitor on full-thickness skin defects in diabetic rats[J].Chin J Burns Wounds,2023,39(1):35-44.DOI: 10.3760/cma.j.cn501225-20221109-00483.

Influence of reactive oxygen species responsive self-assembled nanomicelle loaded with pyroptosis inhibitor on full-thickness skin defects in diabetic rats

doi: 10.3760/cma.j.cn501225-20221109-00483
Funds:

International (Regional) Cooperation and Exchange Program of National Natural Science Foundation of China 81920108022

More Information
  • Corresponding author: Luo Gaoxing, Email: logxw@hotmail.com
  • Received Date: 2022-11-09
  •   Objective  To investigate the influence of reactive oxygen species (ROS) responsive self-assembled nanomicelle loaded with pyroptosis inhibitor on full-thickness skin defects in diabetic rats.  Methods  Experimental research methods were employed. A nucleotide-binding oligomerization domain (NOD) 1/2 inhibitor (NOD-IN-1) was encapsulated with nanomicelle polyethylene glycol-block-polypropylene sulfide (PEG-b-PPS), and the resulting product was called PEPS@NOD-IN-1. The morphology and hydration particle size of PEG-b-PPS and PEPS@NOD-IN-1 were observed by transmission electron microscope and particle size analyzer, respectively, and the encapsulation rate and drug loading rate of PEPS@NOD-IN-1 to NOD-IN-1 and the cumulative release rate of NOD-IN-1 by PEPS@NOD-IN-1 in phosphate buffer solution (PBS) alone and hydrogen peroxide-containing PBS within 40 h were measured and calculated by microplate reader, and the sample number was 3. Twenty-four male Sprague-Dawley rats aged 6-7 weeks were injected with streptozotocin to induce type 1 diabetes mellitus. Six full-thickness skin defect wounds were made on the back of each rat. The injured rats were divided into PBS group, NOD-IN-1 group, PEG-b-PPS group, and PEPS@NOD-IN-1 group with corresponding treatment according to the random number table, with 6 rats in each group. The wound healing was observed on post injury day (PID) 3, 7, and 12, and the wound healing rate was calculated. The ROS levels in wound tissue were detected by immunofluorescence method on PID 3. On PID 7, the granulation tissue thickness in wound was assessed by hematoxylin-eosin staining, the mRNA expressions of NOD1 and NOD2 were detected by real-time fluorescence quantitative reverse transcription polymerase chain reaction, and the protein expressions of NOD1, NOD2, and GSDMD-N terminals were detected by Western blotting. Six wounds from different rats in each group were taken for detection of the above indicators. Wound tissue (3 samples per group) was taken from rats in PBS group and PEPS@NOD-IN-1 group on PID 7, and transcriptome sequencing was performed using high-throughput sequencing technology platform. Differentially expressed genes (DEGs) significantly down-regulated in PEPS@NOD-IN-1 group as compared with PBS group were screened, and the enrichment analysis of Kyoto Encyclopedia of Genes and Genomes (KEGG) was performed. The DEG heatmap of the NOD-like receptor pathway, a pyroptosis-related pathway, was made. Protein-protein interaction (PPI) analysis of DEGs in heatmap was performed through the STRING database to screen key genes of PEPS@NOD-IN-1 regulating the NOD-like receptor pathway. Data were statistically analyzed with analysis of variance for repeated measurement, one-way analysis of variance, and Tukey test.  Results  PEG-b-PPS and PEPS@NOD-IN-1 were in spherical structures of uniform size, with hydration particle sizes of (134.2±3.3) and (143.1±2.3) nm, respectively. The encapsulation rate of PEPS@NOD-IN-1 to NOD-IN-1 was (60±5)%, and the drug loading rate was (15±3)%. The release of NOD-IN-1 from PEPS@NOD-IN-1 in PBS alone was slow, and the cumulative release rate at 40 h was only (12.4±2.3)%. The release of NOD-IN-1 from PEPS@NOD-IN-1 in hydrogen peroxide-containing PBS within 10 h was very rapid, and the cumulative release rate at 10 h reached (90.1±3.6)%. On PID 3 and 7, the wounds of rats in the four groups were gradually healed, and the healing in PEPS@NOD-IN-1 group was better than that in the other three groups. On PID 12, the wound scab area in PBS group was large, the wound epithelialization in NOD-IN-1 group and PEG-b-PPS group was obvious, and the wound in PEPS@NOD-IN-1 group was close to complete epithelialization. Compared with those in PBS group, NOD-IN-1 group, and PEG-b-PPS group, the wound healing rates on PID 7 and 12 in PEPS@NOD-IN-1 group were significantly increased (P<0.05), the level of ROS in wound tissue on PID 3 was significantly decreased (P<0.05), the thickness of granulation tissue in wound on PID 7 was significantly thickened (P<0.05), and the mRNA expressions of NOD1 and NOD2 and the protein expressions of NOD1, NOD2, and GSDMD-N terminals in wound tissue on PID 7 were significantly decreased (P<0.05). KEGG pathway analysis showed that DEGs significantly down-regulated in PEPS@NOD-IN-1 group as compared with PBS group were significantly enriched in NOD-like receptors, hypoxia-inducible factors, mitogen-activated protein kinases, and tumor necrosis factor (TNF) pathways. In the DEG heatmap of NOD-like receptor pathway, the genes regulating pyroptosis mainly involved NOD1, NOD2, NOD-like receptor thermoprotein domain-related protein 3, Jun, signal transduction and transcriptional activator 1 (STAT1), TNF-α-induced protein 3. The PPI results showed that NOD1, NOD2, and STAT1 were the key genes of PEPS@NOD-IN-1 regulating the NOD-like receptor pathway.  Conclusions  PEPS@NOD-IN-1 can down-regulate the level of local ROS in wounds and the expression of NOD1, NOD2, and GSDMD-N terminals, the key regulators of pyroptosis, thereby promoting the repair of full-thickness skin defect wounds in diabetic rats. PEPS@NOD-IN-1 can also significantly down-regulate the pyroptosis, inflammation, and hypoxia-related pathways of wounds, and regulate NOD-like receptor pathways by down-regulating key genes NOD1, NOD2, and STAT1.

     

  • loading
  • [1]
    MatooriS, VevesA, MooneyDJ. Advanced bandages for diabetic wound healing[J]. Sci Transl Med,2021, 13(585):eabe4839.DOI: 10.1126/scitranslmed.abe4839.
    [2]
    DekoninckS, BlanpainC. Stem cell dynamics, migration and plasticity during wound healing[J]. Nat Cell Biol,2019,21(1):18-24.DOI: 10.1038/s41556-018-0237-6.
    [3]
    WillenborgS, EmingSA. Cellular networks in wound healing[J]. Science,2018,362(6417):891-892.DOI: 10.1126/science.aav5542.
    [4]
    SiesH, JonesDP. Reactive oxygen species (ROS) as pleiotropic physiological signalling agents[J]. Nat Rev Mol Cell Biol,2020,21(7):363-383.DOI: 10.1038/s41580-020-0230-3.
    [5]
    MittlerR,ZandalinasSI,FichmanY,et al.Reactive oxygen species signalling in plant stress responses[J].Nat Rev Mol Cell Biol,2022,23(10):663-679.DOI: 10.1038/s41580-022-00499-2.
    [6]
    NathanC,Cunningham-BusselA.Beyond oxidative stress: an immunologist's guide to reactive oxygen species[J].Nat Rev Immunol,2013,13(5):349-361.DOI: 10.1038/nri3423.
    [7]
    FranchinaDG, DostertC, BrennerD. Reactive oxygen species: involvement in T cell signaling and metabolism[J].Trends Immunol,2018,39(6):489-502.DOI: 10.1016/j.it.2018.01.005.
    [8]
    ChaiQ, YuS, ZhongY, et al. A bacterial phospholipid phosphatase inhibits host pyroptosis by hijacking ubiquitin[J].Science,2022,378(6616):eabq0132.DOI: 10.1126/science.abq0132.
    [9]
    BergsbakenT, FinkSL, CooksonBT. Pyroptosis: host cell death and inflammation[J]. Nat Rev Microbiol,2009,7(2):99-109.DOI: 10.1038/nrmicro2070.
    [10]
    RaoZ, ZhuY, YangP, et al. Pyroptosis in inflammatory diseases and cancer[J]. Theranostics,2022,12(9):4310-4329.DOI: 10.7150/thno.71086.
    [11]
    YuP, ZhangX, LiuN, et al. Pyroptosis: mechanisms and diseases[J]. Signal Transduct Target Ther,2021,6(1):128.DOI: 10.1038/s41392-021-00507-5.
    [12]
    HussainZ,ThuHE,Rawas-QalajiM,et al.Recent developments and advanced strategies for promoting burn wound healing[J]. J Drug Deliv Sci Technol,2022,68:103092.DOI: 10.1016/j.jddst.2022.103092.
    [13]
    NiuY,LiQ,DingY,et al.Engineered delivery strategies for enhanced control of growth factor activities in wound healing[J].Adv Drug Deliv Rev,2019,146:190-208.DOI: 10.1016/j.addr.2018.06.002.
    [14]
    MaX,HaoJ,WuJ,et al.Prussian blue nanozyme as a pyroptosis inhibitor alleviates neurodegeneration[J].Adv Mater,2022,34(15):e2106723.DOI: 10.1002/adma.202106723.
    [15]
    XuN, YuanY, DingL, et al. Multifunctional chitosan/gelatin@tannic acid cryogels decorated with in situ reduced silver nanoparticles for wound healing[J/OL]. Burns Trauma,2022,10:tkac019[2022-11-09].https://pubmed.ncbi.nlm.nih.gov/35910193/.DOI: 10.1093/burnst/tkac019.
    [16]
    ShiR,LiH,JinX,et al.Promoting re-epithelialization in an oxidative diabetic wound microenvironment using self-assembly of a ROS-responsive polymer and P311 peptide micelles[J].Acta Biomater,2022,152:425-439.DOI: 10.1016/j.actbio.2022.09.017.
    [17]
    HuangR,HuJ,QianW,et al.Recent advances in nanotherapeutics for the treatment of burn wounds[J/OL].Burns Trauma,2021,9:tkab026[2022-11-09].https://pubmed.ncbi.nlm.nih.gov/34778468/.DOI: 10.1093/burnst/tkab026.
    [18]
    吴近芳,洪旭东,金剑,等.季铵化壳聚糖-重组组织因子途径抑制物复合物对大鼠碾压撕脱皮瓣的影响[J].中华烧伤杂志,2021,37(12):1158-1165.DOI: 10.3760/cma.j.cn501120-20200914-00409.
    [19]
    YanR,LiuX,XiongJ,et al.pH-responsive hyperbranched polypeptides based on Schiff bases as drug carriers for reducing toxicity of chemotherapy[J].RSC Adv,2020,10(23):13889-13899.DOI: 10.1039/d0ra01241f.
    [20]
    DunnillC,PattonT,BrennanJ,et al.Reactive oxygen species (ROS) and wound healing: the functional role of ROS and emerging ROS-modulating technologies for augmentation of the healing process[J].Int Wound J,2017,14(1):89-96.DOI: 10.1111/iwj.12557.
    [21]
    Las HerasK, IgartuaM, Santos-VizcainoE, et al. Chronic wounds: current status, available strategies and emerging therapeutic solutions[J]. J Control Release,2020,328:532-550.DOI: 10.1016/j.jconrel.2020.09.039.
    [22]
    VeithAP,HendersonK,SpencerA,et al.Therapeutic strategies for enhancing angiogenesis in wound healing[J].Adv Drug Deliv Rev,2019,146:97-125.DOI: 10.1016/j.addr.2018.09.010.
    [23]
    张清荣,杨晓,李正,等.活性氧响应性抗菌微针对糖尿病小鼠细菌定植全层皮肤缺损创面的影响[J].中华烧伤杂志,2021,37(11):1024-1035.DOI: 10.3760/cma.j.cn501120-20210831-00299.
    [24]
    YaoY, ZhangH, WangZ, et al. Reactive oxygen species (ROS)-responsive biomaterials mediate tissue microenvironments and tissue regeneration[J]. J Mater Chem B,2019,7(33):5019-5037. DOI: 10.1039/c9tb00847k.
    [25]
    XieW,HuW,HuangZ,et al.Betulinic acid accelerates diabetic wound healing by modulating hyperglycemia-induced oxidative stress, inflammation and glucose intolerance[J/OL]. Burns Trauma,2022,10:tkac007[2022-11-09].https://pubmed.ncbi.nlm.nih.gov/35415192/.DOI: 10.1093/burnst/tkac007.
    [26]
    ShenM,LiH,YaoS,et al.Shear stress and ROS-responsive biomimetic micelles for atherosclerosis via ROS consumption[J].Mater Sci Eng C Mater Biol Appl,2021,126:112164.DOI: 10.1016/j.msec.2021.112164.
    [27]
    LiL, WangY, GuoR, et al. Ginsenoside Rg3-loaded, reactive oxygen species-responsive polymeric nanoparticles for alleviating myocardial ischemia-reperfusion injury[J]. J Control Release,2020,317:259-272. DOI: 10.1016/j.jconrel.2019.11.032.
    [28]
    ZhaoZ,HanZ,NaveenaK,et al.ROS-responsive nanoparticle as a berberine carrier for OHC-targeted therapy of noise-induced hearing loss[J].ACS Appl Mater Interfaces,2021,13(6):7102-7114.DOI: 10.1021/acsami.0c21151.
    [29]
    ShiCX,WangY,ChenQ,et al.Extracellular histone H3 induces pyroptosis during sepsis and may act through NOD2 and VSIG4/NLRP3 pathways[J].Front Cell Infect Microbiol,2020,10:196.DOI: 10.3389/fcimb.2020.00196.
    [30]
    MoulinE,NyrkovaIA,GiusepponeN,et al.Homodyne dynamic light scattering in supramolecular polymer solutions: anomalous oscillations in intensity correlation function[J].Soft Matter,2020,16(12):2971-2993.DOI: 10.1039/c9sm02480h.
    [31]
    ChenYQ, CaoJ, ZhuHY, et al. Synthesis and evaluation of methionine and folate co-decorated chitosan self-assembly polymeric micelles as a potential hydrophobic drug-delivery system[J]. Chin Sci Bull, 2013,58: 2379-2386.DOI: 10.1007/s11434-013-5733-2.
    [32]
    ColganSP,CampbellEL,KominskyDJ.Hypoxia and mucosal inflammation[J].Annu Rev Pathol,2016,11:77-100.DOI: 10.1146/annurev-pathol-012615-044231.
    [33]
    PashenkovMV,BalyasovaLS,DagilYA,et al.The role of the p38-MNK-eIF4E signaling axis in TNF production downstream of the NOD1 receptor[J].J Immunol,2017,198(4):1638-1648.DOI: 10.4049/jimmunol.1600467.
    [34]
    SchäfflerH,GeissD,GittelN,et al.Mutations in the NOD2 gene are associated with a specific phenotype and lower anti-tumor necrosis factor trough levels in Crohn's disease[J].J Dig Dis,2018,19(11):678-684.DOI: 10.1111/1751-2980.12677.
    [35]
    Di CarloS,HäckerG,GentleIE.GM-CSF suppresses antioxidant signaling and drives IL-1β secretion through NRF2 downregulation[J].EMBO Rep,2022,23(8):e54226.DOI: 10.15252/embr.202154226.
    [36]
    Abdul-SaterAA,Saïd-SadierN,PadillaEV,et al.Chlamydial infection of monocytes stimulates IL-1β secretion through activation of the NLRP3 inflammasome[J].Microbes Infect,2010,12(8/9):652-661.DOI: 10.1016/j.micinf.2010.04.008.
  • 加载中

Catalog

    通讯作者: 陈斌, bchen63@163.com
    • 1. 

      沈阳化工大学材料科学与工程学院 沈阳 110142

    1. 本站搜索
    2. 百度学术搜索
    3. 万方数据库搜索
    4. CNKI搜索

    Figures(7)  / Tables(2)

    Article Metrics

    Article views (3051) PDF downloads(41) Cited by()
    Proportional views
    Related

    /

    DownLoad:  Full-Size Img  PowerPoint
    Return
    Return