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Volume 39 Issue 2
Feb.  2023
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Article Navigation >  CHINESE JOURNAL OF BURNS AND WOUNDS  > 2023  >  39(2): 165-174
Jin RH,Zhang ZZ,Xu PQ,et al.Effects of three-dimensional bioprinting antibacterial hydrogel on full-thickness skin defect wounds in rats[J].Chin J Burns Wounds,2023,39(2):165-174.DOI: 10.3760/cma.j.cn501120-20210809-00274.
Citation: Jin RH,Zhang ZZ,Xu PQ,et al.Effects of three-dimensional bioprinting antibacterial hydrogel on full-thickness skin defect wounds in rats[J].Chin J Burns Wounds,2023,39(2):165-174.DOI: 10.3760/cma.j.cn501120-20210809-00274.
shu

Effects of three-dimensional bioprinting antibacterial hydrogel on full-thickness skin defect wounds in rats

doi: 10.3760/cma.j.cn501120-20210809-00274
  • 1.

    Department of Burns and Wound Repair, the Second Affiliated Hospital of Zhejiang University School of Medicine, Zhejiang Key Laboratory for Diagnosis and Treatment of Severe Trauma and Burn, Hangzhou 310009, China

  • 2.

    Department of Burns, First People's Hospital of Hangzhou Xiaoshan District, Xiaoshan 311201, China

Funds:

National Key Research and Development Program of China 2016YFC1100800

Zhejiang Basic Public Welfare Research Program of China LGF19H150008

More Information
  • Corresponding author: Han Chunmao, Email: zrssk@zju.edu.cn
  • Received Date: 2021-08-09
    Abstract
  •   Objective  To explore the effects of three-dimensional (3D) bioprinting gelatin methacrylamide (GelMA) hydrogel loaded with nano silver on full-thickness skin defect wounds in rats.  Methods  The experimental research method was adopted. The morphology, particle diameter, and distribution of silver nanoparticles in nano silver solution with different mass concentrations and the pore structure of silver-containing GelMA hydrogel with different final mass fractions of GelMA were observed by scanning electron microscope and the pore size was calculated. On treatment day 1, 3, 7, and 14, the concentration of nano silver released from the hydrogel containing GelMA with final mass fraction of 15% and nano silver with final mass concentration of 10 mg/L was detected by mass spectrometer. At 24 h of culture, the diameters of inhibition zone of GelMA hydrogel containing final mass concentration of 0 (no nano silver), 25, 50, and 100 mg/L nano silver against Staphylococcus aureus and Escherichia coli were detected. Fibroblasts (Fbs) and adipose stem cells (ASCs) were isolated respectively by enzymatic digestion using the discarded prepuce after circumcision from a 5-year-old healthy boy who was treated in the Department of Urology of the Second Affiliated Hospital of Zhejiang University School of Medicine in July 2020, and the discarded fat tissue after liposuction from a 23-year-old healthy woman who was treated in the Department of Plastic Surgery of the Hospital in July 2020. The Fbs were divided into blank control group (culture medium only), 2 mg/L nano sliver group, 5 mg/L nano sliver group, 10 mg/L nano sliver group, 25 mg/L nano sliver group, and 50 mg/L nano sliver group, which were added with the corresponding final mass concentrations of nano sliver solution, respectively. At 48 h of culture, the Fb proliferation viability was detected by cell counting kit 8 method. The Fbs were divided into 0 mg/L silver-containing GelMA hydrogel group, 10 mg/L silver-containing GelMA hydrogel group, 50 mg/L silver-containing GelMA hydrogel group, and 100 mg/L silver-containing GelMA hydrogel group and then were correspondingly treated. On culture day 1, 3, and 7, the Fb proliferation viability was detected as before. The ASCs were mixed into GelMA hydrogel and divided into 3D bioprinting group and non-printing group. On culture day 1, 3, and 7, the ASC proliferation viability was detected as before and cell growth was observed by live/dead cell fluorescence staining. The sample numbers in the above experiments were all 3. Four full-thickness skin defect wounds were produced on the back of 18 male Sprague-Dawley rats aged 4 to 6 weeks. The wounds were divided into hydrogel alone group, hydrogel/nano sliver group, hydrogel scaffold/nano sliver group, and hydrogel scaffold/nano sliver/ASC group, and transplanted with the corresponding scaffolds, respectively. On post injury day (PID) 4, 7, 14, and 21, the wound healing was observed and the wound healing rate was calculated (n=6). On PID 7 and 14, histopathological changes of wounds were observed by hematoxylin eosin staining (n=6). On PID 21, collagen deposition of wounds was observed by Masson staining (n=3). Data were statistically analyzed with one-way analysis of variance, analysis of variance for repeated measurement, Bonferroni correction, and independent sample t test.  Results  The sliver nano particles in nano silver solution with different mass concentrations were all round, in scattered distribution and uniform in size. The silver-containing GelMA hydrogels with different final mass fractions of GelMA all showed pore structures of different sizes and interconnections. The pore size of silver-containing GelMA hydrogel with 10% final mass fraction was significantly larger than that of silver-containing GelMA hydrogels with 15% and 20% final mass fractions (with P values both below 0.05). On treatment day 1, 3, and 7, the concentration of nano silver released from silver-containing GelMA hydrogel in vitro showed a relatively flat trend. On treatment day 14, the concentration of released nano silver in vitro increased rapidly. At 24 h of culture, the diameters of inhibition zone of GelMA hydrogel containing 0, 25, 50, and 100 mg/L nano silver against Staphylococcus aureus and Escherichia coli were 0, 0, 0.7, and 2.1 mm and 0, 1.4, 3.2, and 3.3 mm, respectively. At 48 h of culture, the proliferation activity of Fbs in 2 mg/L nano silver group and 5 mg/L nano silver group was both significantly higher than that in blank control group (P<0.05), and the proliferation activity of Fbs in 10 mg/L nano silver group, 25 mg/L nano silver group, and 50 mg/L nano silver group was all significantly lower than that in blank control group (P<0.05). Compared with the that of Fbs in 0 mg/L silver-containing GelMA hydrogel group, the proliferation activity of Fbs in 50 mg/L silver-containing GelMA hydrogel group and 100 mg/L silver-containing GelMA hydrogel group was all significantly decreased on culture day 1 (P<0.05); the proliferation activity of Fbs in 50 mg/L silver-containing GelMA hydrogel group was significantly increased (P<0.05), while the proliferation activity of Fbs in 100 mg/L silver-containing GelMA hydrogel group was significantly decreased on culture day 3 (P<0.05); the proliferation activity of Fbs in 100 mg/L silver-containing GelMA hydrogel group was significantly decreased on culture day 7 (P<0.05). The proliferation activity of ASCs in 3D bioprinting group show no statistically significant differences to that in non-printing group on culture day 1 (P>0.05). The proliferation activity of ASCs in 3D bioprinting group was significantly higher than that in non-printing group on culture day 3 and 7 (with t values of 21.50 and 12.95, respectively, P<0.05). On culture day 1, the number of dead ASCs in 3D bioprinting group was slightly more than that in non-printing group. On culture day 3 and 5, the majority of ASCs in 3D bioprinting group and non-printing group were living cells. On PID 4, the wounds of rats in hydrogel alone group and hydrogel/nano sliver group had more exudation, and the wounds of rats in hydrogel scaffold/nano sliver group and hydrogel scaffold/nano sliver/ASC group were dry without obvious signs of infection. On PID 7, there was still a small amount of exudation on the wounds of rats in hydrogel alone group and hydrogel/nano sliver group, while the wounds of rats in hydrogel scaffold/nano sliver group and hydrogel scaffold/nano sliver/ASC group were dry and scabbed. On PID 14, the hydrogels on the wound surface of rats in the four groups all fell off. On PID 21, a small area of wounds remained unhealed in hydrogel alone group. On PID 4 and 7, the wound healing rates of rats in hydrogel scaffold/nano sliver/ASC group were significantly higher than those of the other three groups (P<0.05). On PID 14, the wound healing rate of rats in hydrogel scaffold/nano sliver/ASC group was significantly higher than the wound healing rates in hydrogel alone group and hydrogel/nano sliver group (all P<0.05). On PID 21, the wound healing rate of rats in hydrogel alone group was significantly lower than that in hydrogel scaffold/nano sliver/ASC group (P<0.05). On PID 7, the hydrogels on the wound surface of rats in the four groups remained in place; on PID 14, the hydrogel in hydrogel alone group was separated from the wounds of rats, while some hydrogels still existed in the new tissue of the wounds of rats in the other three groups. On PID 21, the collagen arrangement in the wounds of rats in hydrogel alone group was out of order, while the collagen arrangement in the wounds of rats in hydrogel/nano sliver group, and hydrogel scaffold/nano sliver/ASC group was relatively orderly.  Conclusions  Silver-containing GelMA hydrogel has good biocompatibility and antibacterial properties. Its three-dimensional bioprinted double-layer structure can better integrate with new formed tissue in the full-thickness skin defect wounds in rats and promote wound healing.

     

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    • References(30)
  • [1]
    陈跃华,徐俊,徐兰举,等.水凝胶敷料对糖尿病足创面的促愈合作用研究进展[J].中华烧伤与创面修复杂志,2022,38(1):95-98.DOI: 10.3760/cma.j.cn501120-20200827-00393.
    [2]
    董云青, 李琳琳, 朱宣儒, 等. 含银黏性水凝胶的制备及其在小鼠细菌定植全层皮肤缺损创面愈合中的作用[J]. 中华烧伤杂志, 2021,37(11):1036-1047. DOI: 10.3760/cma.j.cn501120-20210906-00304.
    [3]
    KumarSSD, RajendranNK, HoureldNN,et al.Recent advances on silver nanoparticle and biopolymer-based biomaterials for wound healing applications[J].Int J Biol Macromol,2018,115:165-175.DOI: 10.1016/j.ijbiomac.2018.04.003.
    [4]
    ChenHL, LanGQ, RanLX, et al. A novel wound dressing based on a Konjac glucomannan/silver nanoparticle composite sponge effectively kills bacteria and accelerates wound healing[J]. Carbohydr Polym, 2018,183:70-80.DOI: 10.1016/j.carbpol.2017.11.029.
    [5]
    有传刚,韩春茂,王新刚,等.纳米银的抗菌和毒性机制及临床研究进展[J].中华烧伤杂志,2011,27(3):243-246.DOI: 10.3760/cma.j.issn.1009-2587.2011.03.031.
    [6]
    YouCG,LiQ,WangXG,et al.Silver nanoparticle loaded collagen/chitosan scaffolds promote wound healing via regulating fibroblast migration and macrophage activation[J].Sci Rep,2017,7(1):10489.DOI: 10.1038/s41598-017-10481-0.
    [7]
    MurphySV,AtalaA.3D bioprinting of tissues and organs[J].Nat Biotechnol,2014,32(8):773-785.DOI: 10.1038/nbt.2958.
    [8]
    NgWL,WangS,YeongWY,et al.Skin bioprinting: impending reality or fantasy?[J].Trends Biotechnol,2016,34(9):689-699.DOI: 10.1016/j.tibtech.2016.04.006.
    [9]
    陈浩娇, 吴攀, 王新刚, 等. 三维生物打印在皮肤组织工程中的应用研究进展[J]. 中华烧伤杂志, 2018,34(6):422-426.DOI: 10.3760/cma.j.issn.1009-2587.2018.06.026.
    [10]
    Gungor-OzkerimPS,InciI,ZhangYS,et al.Bioinks for 3D bioprinting: an overview[J].Biomater Sci,2018,6(5):915-946.DOI: 10.1039/c7bm00765e.
    [11]
    GopinathanJ,NohI.Recent trends in bioinks for 3D printing[J].Biomater Res,2018,22:11.DOI: 10.1186/s40824-018-0122-1.
    [12]
    ZhaoX,LangQ,YildirimerL,et al.Photocrosslinkable gelatin hydrogel for epidermal tissue engineering[J].Adv Healthc Mater,2016,5(1):108-118.DOI: 10.1002/adhm.201500005.
    [13]
    KozickiM,KołodziejczykM,SzynkowskaM,et al.Hydrogels made from chitosan and silver nitrate[J].Carbohydr Polym,2016,140:74-87.DOI: 10.1016/j.carbpol.2015.12.017.
    [14]
    LiuY,SunY,XuYX,et al.Preparation and evaluation of lysozyme-loaded nanoparticles coated with poly-γ-glutamic acid and chitosan[J].Int J Biol Macromol,2013,59:201-207.DOI: 10.1016/j.ijbiomac.2013.04.065.
    [15]
    MirzajaniF,GhassempourA,AliahmadiA,et al.Antibacterial effect of silver nanoparticles on Staphylococcus aureus[J].Res Microbiol,2011,162(5):542-549.DOI: 10.1016/j.resmic.2011.04.009.
    [16]
    YuanYC,DingLP ,ChenY,et al.Nano-silver functionalized polysaccharides as a platform for wound dressings: a review[J].Int J Biol Macromol,2022,194:644-653.DOI: 10.1016/j.ijbiomac.2021.11.108.
    [17]
    ZhangJ, WangF, YalamartySSK,et al.Nano silver-induced toxicity and associated mechanisms[J].Int J Nanomedicine,2022,17:1851-1864.DOI: 10.2147/IJN.S355131.
    [18]
    LiuYN, LiF, GuoZR, et al. Silver nanoparticle-embedded hydrogel as a photothermal platform for combating bacterial infections[J]. Chem Eng J, 2020,382:122990.DOI: 10.1016/j.cej.2019.122990.
    [19]
    JiangYG,HuangJJ,WuXW,et al.Controlled release of silver ions from AgNPs using a hydrogel based on konjac glucomannan and chitosan for infected wounds[J].Int J Biol Macromol,2020,149:148-157.DOI: 10.1016/j.ijbiomac.2020.01.221.
    [20]
    KonopM, DampsT, MisickaA, et al. Certain aspects of silver and silver nanoparticles in wound care: a minireview[J]. J Nanomater, 2016,2016:1-10.DOI: 10.1155/2016/7614753.
    [21]
    ZhouFF, HongY, LiangRJ, et al. Rapid printing of bio-inspired 3D tissue constructs for skin regeneration[J]. Biomaterials, 2020,258:120287.DOI: 10.1016/j.biomaterials.2020.120287.
    [22]
    GriffinDR,WeaverWM,ScumpiaPO,et al.Accelerated wound healing by injectable microporous gel scaffolds assembled from annealed building blocks[J].Nat Mater,2015,14(7):737-744.DOI: 10.1038/nmat4294.
    [23]
    ChenYC,LinRZ,QiH,et al.Functional human vascular network generated in photocrosslinkable gelatin methacrylate hydrogels[J].Adv Funct Mater,2012,22(10):2027-2039.DOI: 10.1002/adfm.201101662.
    [24]
    有传刚, 张莉萍, 王新刚, 等. 含纳米银的胶原蛋白-丝素蛋白支架对大鼠全层皮肤缺损创面真皮再生的影响[J]. 中华烧伤杂志, 2017,33(2):103-110. DOI: 10.3760/cma.j.issn.1009-2587.2017.02.011.
    [25]
    KimBS,KwonYW,KongJS,et al.3D cell printing of in vitro stabilized skin model and in vivo pre-vascularized skin patch using tissue-specific extracellular matrix bioink: a step towards advanced skin tissue engineering[J].Biomaterials,2018,168:38-53.DOI: 10.1016/j.biomaterials.2018.03.040.
    [26]
    CaiY,LiJY,JiaCS,et al.Therapeutic applications of adipose cell-free derivatives: a review[J].Stem Cell Res Ther,2020,11(1):312.DOI: 10.1186/s13287-020-01831-3.
    [27]
    NolanGS,SmithOJ,JellG,et al.Fat grafting and platelet-rich plasma in wound healing: a review of histology from animal studies[J].Adipocyte,2021,10(1):80-90.DOI: 10.1080/21623945.2021.1876374.
    [28]
    XiaSZ,WengTT,JinRH,et al.Curcumin-incorporated 3D bioprinting gelatin methacryloyl hydrogel reduces reactive oxygen species-induced adipose-derived stem cell apoptosis and improves implanting survival in diabetic wounds[J/OL].Burns Trauma,2022,10:tkac001[2022-11-12]. https://pubmed.ncbi.nlm.nih.gov/35291229/.DOI: 10.1093/burnst/tkac001.
    [29]
    JinRH,CuiYC,ChenHJ,et al.Three-dimensional bioprinting of a full-thickness functional skin model using acellular dermal matrix and gelatin methacrylamide bioink[J].Acta Biomater,2021,131:248-261.DOI: 10.1016/j.actbio.2021.07.012.
    [30]
    MasoodN, AhmedR, TariqM, et al. Silver nanoparticle impregnated chitosan-PEG hydrogel enhances wound healing in diabetes induced rabbits[J]. Int J Pharm, 2019,559:23-36.DOI: 10.1016/j.ijpharm.2019.01.019.
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