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Volume 38 Issue 10
Oct.  2022
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Article Navigation >  CHINESE JOURNAL OF BURNS AND WOUNDS  > 2022  >  38(10): 914-922
Zhang QR,Chen CY,Xu N,et al.Effect of P311 microspheres-loaded thermosensitive chitosan hydrogel on the wound healing of full-thickness skin defects in rats[J].Chin J Burns Wounds,2022,38(10):914-922.DOI: 10.3760/cma.j.cn501225-20220414-00135.
Citation: Zhang QR,Chen CY,Xu N,et al.Effect of P311 microspheres-loaded thermosensitive chitosan hydrogel on the wound healing of full-thickness skin defects in rats[J].Chin J Burns Wounds,2022,38(10):914-922.DOI: 10.3760/cma.j.cn501225-20220414-00135.
shu

Effect of P311 microspheres-loaded thermosensitive chitosan hydrogel on the wound healing of full-thickness skin defects in rats

doi: 10.3760/cma.j.cn501225-20220414-00135
  • 1.

    Department of Burns and Plastic Surgery, Affiliated Hospital of Nantong University, Nantong 226001, China

  • 2.

    State Key Laboratory of Trauma, Burns and Combined Injury, Institute of Burn Research, the First Affiliated Hospital of Army Medical University (the Third Military Medical University), Chongqing Key Laboratory for Disease Proteomics, Chongqing 400038, China

  • 3.

    Department of Burns and Plastic Surgery, the First Affiliated Hospital of Wannan Medical College, Wuhu 241001, China

  • 4.

    Beijing Jayyalife Biological Technology Company, Beijing 100094, China

Funds:

Funds for International Cooperation and Exchange of the National Natural Science Foundation of China 81920108022

Nantong Science and Technology Project JC2021178, HS2020006

Industry-university-research Innovation Fund for Chinese Universities 2021JH033

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    Abstract
  •   Objective  To explore the effect of P311 microspheres-loaded thermosensitive chitosan hydrogel on the wound healing of full-thickness skin defects in rats.  Methods  The method of experimental study was adopted. The polyvinyl alcohol/sodium alginate microspheres (simple microspheres), P311 microspheres, and bovine serum albumin labeled with fluorescein isothiocyanate (FITC-BSA) microspheres were prepared by water-in-oil emulsification, and then their morphology was observed under a light microscope/inverted fluorescence microscope. Chitosan solution was prepared, chitosan solution and β-glycerol phosphate disodium hydrate were mixed to prepare simple thermosensitive hydrogels, and thermosensitive hydrogels loaded with simple microspheres or P311 microspheres were prepared by adding corresponding substances in simple thermosensitive hydrogels. The morphological changes of the prepared four liquids in the state of tilt was observed at 37 ℃. After being freeze-dried, the micromorphology of the prepared four liquids was observed under a scanning electron microscope. Eighteen 3-4-week-old male Sprague-Dawley rats were divided into normal group without any treatment, dressing group, chitosan group, hydrogel alone group, simple microspheres-loaded hydrogel group, and P311 microspheres-loaded hydrogel group, which were inflicted with one full-thickness skin defect wound on both sides of the back spine and were dealt correspondingly, with 3 rats in each group. Rats with full-thickness skin defects in the five groups were collected, the wound healing was observed on post injury day (PID) 0 (immediately), 5, 10, and 15, and the wound healing rates on PID 5, 10, and 15 were calculated. The wound and wound margin tissue of rats with full-thickness skin defects in the five groups on PID 15 and normal skin tissue in the same site of rats in normal group were collected, hematoxylin and eosin staining was conducted to observe the histological changes, immunohistochemical staining was performed to observe the expressions of CD31 and vascular endothelial growth factor (VEGF), and Western blotting was conducted to detect the protein expressions of CD31 and VEGF. The number of samples was all three. Data were statistically analyzed with one-way analysis of variance, analysis of variance for repeated measurement, and Bonferroni correction.  Results  Simple microspheres were spherical, with loose and porous surface. The surfaces of P311 microspheres and FITC-BSA microspheres were smooth without pores, and the FITC-BSA microspheres emitted uniform green fluorescence. The diameters of the three microspheres were basically consistent, being 33.1 to 37.7 μm. Compared with chitosan solution and simple thermosensitive hydrogel, the structures of the two microspheres-loaded hydrogels were more stable in the state of tilt at 37 ℃. The two microspheres-loaded hydrogels had denser network structures than those of chitosan solution and simple thermosensitive hydrogel, and in the cross section of which microspheres with a diameter of about 30 μm could be seen. Within PID 15, the wounds of rats in the five groups were healed to different degrees, and the wound healing of rats in P311 microspheres-loaded hydrogel group was the best. On PID 5, 10, and 15, the wound healing rates of rats in dressing group and chitosan group were (26.6±2.4)%, (38.5±3.1)%, (50.9±1.5)%, (47.6±2.0)%, (58.5±3.6)%, and (66.7±4.1)%, respectively, which were significantly lower than (59.3±4.8)%, (87.6±3.2)%, (97.2±1.0)% in P311 microspheres-loaded hydrogel group (P<0.05 or P<0.01). The wound healing rates of rats in hydrogel alone group on PID 10 and 15, and in simple microspheres-loaded hydrogel group on PID 15 were (76.0±3.3)%, (84.5±3.6)%, and (88.0±2.6)%, respectively, which were significantly lower than those in P311 microspheres-loaded hydrogel group (P<0.05). The epidermis, hair follicles, and sebaceous glands could be seen in the normal skin of rats in normal group, without positive expressions of CD31 or VEGF. The wounds of rats in P311 microspheres-loaded hydrogel group on PID 15 were almost completely epithelialized, with more blood vessels, hair follicles, sebaceous glands, and positive expressions of CD31 and VEGF in the wounds than those of rats with full-thickness skin defects in the other four groups, and more protein expressions of CD31 and VEGF than those of rats in the other five groups.  Conclusions  The P311 microspheres-loaded thermosensitive chitosan hydrogel can release the encapsulated drug slowly, prolong the drug action time, and promote wound healing in rats with full-thickness skin defects by promoting wound angiogenesis and re-epithelialization.

     

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    • References(28)
  • [1]
    YuP,ZhongW.Hemostatic materials in wound care[J/OL].Burns Trauma,2021,9:tkab019[2022-09-23].https://pubmed.ncbi.nlm.nih.gov/34541007/.DOI: 10.1093/burnst/tkab019.
    [2]
    VarkeyM,VisscherDO,van ZuijlenP,et al.Skin bioprinting: the future of burn wound reconstruction?[J/OL].Burns Trauma,2019,7:4[2022-09-23].https://pubmed.ncbi.nlm.nih.gov/30805375/. DOI: 10.1186/s41038-019-0142-7.
    [3]
    WangS,ZhangX,QianW,et al.P311 deficiency leads to attenuated angiogenesis in cutaneous wound healing[J].Front Physiol,2017,8:1004.DOI: 10.3389/fphys.2017.01004.
    [4]
    YaoZ,LiH,HeW,et al.P311 accelerates skin wound reepithelialization by promoting epidermal stem cell migration through rhoa and rac1 activation[J].Stem Cells Dev,2017,26(6):451-460.DOI: 10.1089/scd.2016.0249.
    [5]
    LiH,YaoZ,HeW,et al.P311 induces the transdifferentiation of epidermal stem cells to myofibroblast-like cells by stimulating transforming growth factor β1 expression[J].Stem Cell Res Ther,2016,7(1):175.DOI: 10.1186/s13287-016-0421-1.
    [6]
    TaylorGA,HudsonE,ResauJH,et al.Regulation of P311 expression by Met-hepatocyte growth factor/scatter factor and the ubiquitin/proteasome system[J].J Biol Chem,2000,275(6):4215-4219.DOI: 10.1074/jbc.275.6.4215.
    [7]
    ZhouHY,JiangLJ,CaoPP,et al.Glycerophosphate-based chitosan thermosensitive hydrogels and their biomedical applications[J].Carbohydr Polym,2015,117:524-536.DOI: 10.1016/j.carbpol.2014.09.094.
    [8]
    OldenkampHF,Vela RamirezJE,PeppasNA.Re-evaluating the importance of carbohydrates as regenerative biomaterials[J].Regen Biomater,2019,6(1):1-12.DOI: 10.1093/rb/rby023.
    [9]
    HanF,DongY,SuZ,et al.Preparation, characteristics and assessment of a novel gelatin-chitosan sponge scaffold as skin tissue engineering material[J].Int J Pharm,2014,476(1/2):124-133.DOI: 10.1016/j.ijpharm.2014.09.036.
    [10]
    SuY,ZhangB,SunR,et al.PLGA-based biodegradable microspheres in drug delivery: recent advances in research and application[J].Drug Deliv,2021,28(1):1397-1418.DOI: 10.1080/10717544.2021.1938756.
    [11]
    FrokjaerS,OtzenDE.Protein drug stability: a formulation challenge[J].Nat Rev Drug Discov,2005,4(4):298-306.DOI: 10.1038/nrd1695.
    [12]
    MoellerEH,JorgensenL.Alternative routes of administration for systemic delivery of protein pharmaceuticals[J].Drug Discov Today Technol,2008,5(2/3):e89-94.DOI: 10.1016/j.ddtec.2008.11.005.
    [13]
    AhmedTA,AlharbyYA,El-HelwAR,et al.Depot injectable atorvastatin biodegradable in situ gel: development, optimization, in vitro, and in vivo evaluation[J].Drug Des Devel Ther,2016,10:405-415.DOI: 10.2147/DDDT.S98078.
    [14]
    周紫萱,姜耀男,肖仕初.原位成形可注射水凝胶特性及其促创面愈合作用研究进展[J].中华烧伤杂志,2021,37(1):82-85.DOI: 10.3760/cma.j.cn501120-20200428-00243.
    [15]
    CheniteA,ChaputC,WangD,et al.Novel injectable neutral solutions of chitosan form biodegradable gels in situ[J].Biomaterials,2000,21(21):2155-2161.DOI: 10.1016/s0142-9612(00)00116-2.
    [16]
    DengA,KangX,ZhangJ,et al.Enhanced gelation of chitosan/β-sodium glycerophosphate thermosensitive hydrogel with sodium bicarbonate and biocompatibility evaluated[J].Mater Sci Eng C Mater Biol Appl,2017,78:1147-1154.DOI: 10.1016/j.msec.2017.04.109.
    [17]
    KaranS,DebnathS,KuotsuK,et al.In-vitro and in-vivo evaluation of polymeric microsphere formulation for colon targeted delivery of 5-fluorouracil using biocompatible natural gum katira[J].Int J Biol Macromol,2020,158:922-936.DOI: 10.1016/j.ijbiomac.2020.04.129.
    [18]
    ZhouX,HouC,ChangTL,et al.Controlled released of drug from doubled-walled PVA hydrogel/PCL microspheres prepared by single needle electrospraying method[J].Colloids Surf B Biointerfaces,2020,187:110645.DOI: 10.1016/j.colsurfb.2019.110645.
    [19]
    RacV,LevićS,BalančB,et al.PVA Cryogel as model hydrogel for iontophoretic transdermal drug delivery investigations. Comparison with PAA/PVA and PAA/PVP interpenetrating networks[J].Colloids Surf B Biointerfaces,2019,180:441-448.DOI: 10.1016/j.colsurfb.2019.05.017.
    [20]
    StrehlC,GaberT,MauriziL,et al.Effects of PVA coated nanoparticles on human immune cells[J].Int J Nanomedicine,2015,10:3429-3445.DOI: 10.2147/IJN.S75936.
    [21]
    阿拉腾珠拉,胡永飞.褐藻寡糖的制备方法及生物活性研究进展[J].生物工程学报,2022,38(1):104-118.DOI: 10.13345/j.cjb.210377.
    [22]
    JadachB,ŚwietlikW,FroelichA.Sodium alginate as a pharmaceutical excipient: novel applications of a well-known polymer[J].J Pharm Sci,2022,111(5):1250-1261.DOI: 10.1016/j.xphs.2021.12.024.
    [23]
    YaacobEN,GoethalsJ,BajekA,et al.Preparation and characterization of alginate microparticles containing a model protein for oral administration in gnotobiotic european sea bass (dicentrarchus labrax) larvae[J].Mar Biotechnol (NY),2017,19(4):391-400.DOI: 10.1007/s10126-017-9758-4.
    [24]
    RameshbabuAP,BankotiK,DattaS,et al.Silk sponges ornamented with a placenta-derived extracellular matrix augment full-thickness cutaneous wound healing by stimulating neovascularization and cellular migration[J].ACS Appl Mater Interfaces,2018,10(20):16977-16991.DOI: 10.1021/acsami.7b19007.
    [25]
    WangX,LinM,KangY.Engineering porous β-tricalcium phosphate (β-TCP) scaffolds with multiple channels to promote cell migration, proliferation, and angiogenesis[J].ACS Appl Mater Interfaces,2019,11(9):9223-9232.DOI: 10.1021/acsami.8b22041.
    [26]
    路青青,吕国忠,吕强.具有促血管化能力的酸化丝蛋白海绵敷料的细胞相容性及该敷料对大鼠全层皮肤缺损创面愈合的影响[J].中华烧伤杂志,2021,37(1):25-33.DOI: 10.3760/cma.j.cn501120-20200925-00423.
    [27]
    BarrientosS,BremH,StojadinovicO,et al.Clinical application of growth factors and cytokines in wound healing[J].Wound Repair Regen,2014,22(5):569-578.DOI: 10.1111/wrr.12205.
    [28]
    ChengT, YueM, AslamMN, et al. Neuronal protein 3.1 deficiency leads to reduced cutaneous scar collagen deposition and tensile strength due to impaired transforming growth factor-β1 to -β3 translation[J]. Am J Pathol, 2017,187(2):292-303. DOI: 10.1016/j.ajpath.2016.10.004.
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