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Volume 38 Issue 7
Jul.  2022
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Article Navigation >  CHINESE JOURNAL OF BURNS AND WOUNDS  > 2022  >  38(7): 640-649
Chen XJ,Wu X,Lin HH,et al.Effects of methacrylic anhydride gelatin hydrogel loaded with silver and recombinant human basic fibroblast growth factor on deep partial-thickness burn wounds in rabbits[J].Chin J Burns Wounds,2022,38(7):640-649.DOI: 10.3760/cma.j.cn501120-20210726-00260.
Citation: Chen XJ,Wu X,Lin HH,et al.Effects of methacrylic anhydride gelatin hydrogel loaded with silver and recombinant human basic fibroblast growth factor on deep partial-thickness burn wounds in rabbits[J].Chin J Burns Wounds,2022,38(7):640-649.DOI: 10.3760/cma.j.cn501120-20210726-00260.
shu

Effects of methacrylic anhydride gelatin hydrogel loaded with silver and recombinant human basic fibroblast growth factor on deep partial-thickness burn wounds in rabbits

doi: 10.3760/cma.j.cn501120-20210726-00260

  • Department of Burn Orthopaedics, the 969th Hospital of PLA Joint Logistic Support Force, Hohhot 010051, China

Funds:

Natural Science Foundation of Inner Mongolia Autonomous Region of China 2018MS08002

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  • Corresponding author: Chen Xiangjun, Email: 1301579255@qq.com
  • Received Date: 2021-07-26
    Abstract
  •   Objective  To investigate the effects of methacrylic anhydride gelatin (GelMA) hydrogel loaded with silver and recombinant human basic fibroblast growth factor (rh-bFGF) on deep partial-thickness burn wounds in rabbits.  Methods  The experimental research method was adopted. Low-concentration GelMA materials, medium-concentration GelMA materials and high-concentration GelMA materials containing different concentrations of methacrylic anhydride (MA) were prepared, after adding photoinitiator, low-concentration GelMA hydrogels, medium-concentration GelMA hydrogels, and high-concentration GelMA hydrogels were obtained, respectively. The nuclear magnetic resonance spectroscopy was performed to detect the hydrogen nuclear magnetic resonance spectra of the above-mentioned three concentrations of GelMA materials, and to calculate the degree of substitution according to the spectrum diagram. The three-dimensional microstructure and pore size of 3 types of above-mentioned GelMA hydrogels were detected by field emission scanning electron microscopy (FESEM), with 9 samples measured. According to the selected concentration of MA, ten kinds of solutions of GelMA with different concentration of silver (silver-containing GelMA) were synthesized, and the silver-containing GelMA solution of each concentration was divided into three parts, and then exposed to ultraviolet light lasting for 20, 25, and 35 s, respectively. After adding photoinitiator,the corresponding silver-containing GelMA hydrogels were obtained. The residual degradation rate of silver-containing GelMA hydrogel with different photocrosslinking times was detected by collagenase degradation method at degradation of 12, 24, 36, and 48 h; and the time required for complete degradation was detected, and the sample number was 5. The inhibition zone diameter of GelMA hydrogel under above screened photocrosslinking times containing 10 concentrations of silver against Staphylococcus aureus was measured to reflect its antibacterial ability, and the sample numbers were all 5. The silver-containing GelMA hydrogel with statistical significance compared with the antibacterial circle diameter of the silver-containing GelMA hydrogel containing the lowest concentration (no silver) was considered as having antibacterial activity. The three-dimensional microstructure and pore size of the silver-containing GelMA hydrogels with antibacterial activity and the lowest drug concentration selected were detected by FESEM, and the sample numbers were all 9. The freeze-dried alone GelMA hydrogel and the freeze-dried silver-containing GelMA hydrogel were soaked in phosphate buffer solution for 24 h, respectively, then the swelling rate of the two GelMA hydrogel were calculated and compared by weighing method, and the sample number was 5. GelMA hydrogel containing silver and rh-bFGF, namely compound hydrogel for short, was prepared according to the preliminary experiment and the above experimental results. The appearance of the composite hydrogel was observed in general, and its three-dimensional microstructure and pore size were detected by FESEM. The deep partial-thickness burn wound was made on the back of 30 rabbits (aged 4-6 months, female half and half). Meanwhile, with the rabbit head as the benchmark, the wounds on the left side of the spine were treated as composite hydrogel treatment group, and the wounds on the right side were treated as gauze control group, and which were treated accordingly. On post injury day (PID) 3, 7, 14, 21, and 28, the healing of wounds in the two groups was observed. On PID 7, 14, 21, and 28, the wound healing area was recorded and the healing rate was calculated, with a sample number of 30. Data were statistically analyzed with analysis of variance for repeated measurement, one-way analysis of variance, and independent sample t test.  Results  The substitution degree among low-concentration GelMA materials, medium-concentration GelMA materials, and high-concentration GelMA materials was significantly different (F=1 628.00, P<0.01). The low-concentration GelMA hydrogel had a loose and irregular three-dimensional spatial network structure with a pore size of (60±17) μm; the medium-concentration GelMA hydrogel had a relatively uniform three-dimensional spatial network and pore size with a pore size of (45±13) μm; the high-concentration GelMA hydrogel had the dense and disordered three-dimensional spatial network with a pore size of (25±15) μm, the pore sizes of 3 types of GelMA hydrogels were significantly differences (F=12.20, P<0.01), and medium concentration of MA was selected for the concentration of subsequent materials. The degradability of silver-containing GelMA hydrogels with different concentrations of the same photocrosslinking time was basically same. The degradation residual rates of silver-containing GelMA hydrogels with 20, 25, and 35 s crosslinking time at 12 h were (74.2±1.7)%, (85.3±0.9)%, and (93.2±1.2)%, respectively; the residual rates of degradation at 24 h were (58.3±2.1)%, (65.2±1.8)%, and (81.4±2.6)%, respectively; the residual rates of degradation at 36 h were (22.4±1.9)%, (45.2±1.7)%, and (68.1±1.4)%, respectively; the residual rates of degradation at 48 h were (8.2±1.7)%, (32.4±1.3)%, and (54.3±2.2)%, respectively, and 20, 25, and 30 s photocrosslinking time required for complete degradation of silver-containing GelMA hydrogels were (50.2±2.4), (62.4±1.4), and (72.2±3.2) h, and the difference was statistically significant (F=182.40, P<0.01), 25 s were selected as the subsequent photocrosslinking time. The antibacterial diameters of 10 types of silver-containing GelMA hydrogels against Staphylococcus aureus from low to high concentrations were (2.6±0.4), (2.5±0.4), (3.2±0.4), (12.1±0.7), (14.8±0.7), (15.1±0.5), (16.2±0.6), (16.7±0.5), (16.7±0.4), and (16.7±0.6) mm, respectively, and which basically showed a concentration-dependent increasing trend, and the overall difference was statistically significant (F=428.70, P<0.01). Compared with the silver-containing GelMA hydrogel with the lowest concentration, the antibacterial circle diameters of other silver-containing GelMA hydrogels with antibacterial ability from low to high concentration were significantly increased (with t values of 26.35, 33.84, 43.65, 42.17, 49.24, 55.74, and 43.72, respectively, P<0.01). The silver-containing GelMA hydrogel with the antibacterial diameter of (12.1±0.7) mm had the lowest antibacterial activity against Staphylococcus aureus and the lowest drug loading concentration, and the concentration of silver was selected for the concentration of subsequent materials. The microscopic morphology of the silver-containing GelMA hydrogel containing silver element with a pore size of (45±13) μm had a regular and linear strip-like structure. After soaking for 24 h, the swelling ratio of silver-containing GelMA hydrogel was similar to that of alone GelMA hydrogel. The composite hydrogel was colorless, clear and transparent, and its three-dimensional microstructure was a regular and uniform grid, with a filament network structure inside, and the pore size of (40±21) μm. On PID 3, a large amount of necrotic tissue and exudate of rabbit wound in composite hydrogel group were observed, and scattered scabs, a small amount of necrotic tissue and exudate of rabbit wound in gauze control group were observed. On PID 7, the area of rabbit wound in composite hydrogel group was significantly reduced, and adhesion of rabbit wound and gauze in gauze control group was observed. On PID 14, In composite hydrogel group, the rabbit wound surface was ruddy, and the growth of granulation tissue was observed, and in gauze control group, the rabbit wound base was pale, and the blood supply was poor. On PID 21, the rabbit wounds in composite hydrogel group healed completely, and rabbit wound in gauze control group had healing trend. On PID 28, new hair could be seen on rabbit wound surface in composite hydrogel group; oval wound of rabbit in gauze control group still remained. On PID 7, 14, 21, and 28, the wound healing areas of rabbit in composite hydrogel group were significantly larger than those in gauze control group (with t values of 2.24, 4.43, 7.67, and 7.69, respectively, P<0.05 or P<0.01).  Conclusions  The medium-concentration GelMA hydrogel has good physical and chemical properties in terms of swelling and degradability. The screened silver-containing GelMA hydrogels had the lowest antibacterial activity and the lowest drug loading concentration. Composite hydrogel can significantly shorten the healing time of deep partial-thickness burn wounds in rabbits.

     

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    • References(33)
  • [1]
    PereiraRF, BártoloPJ. Traditional therapies for skin wound healing[J]. Adv Wound Care (New Rochelle), 2016,5(5):208-229. DOI: 10.1089/wound.2013.0506.
    [2]
    罗高兴, 刘梦龙. 应用功能材料促进皮肤创面修复[J].中华烧伤杂志,2021,37(11):1005-1010. DOI: 10.3760/cma.j.cn501120-20210930-00340.
    [3]
    Van Den Bulcke AI, BogdanovB, De RoozeN, et al. Structural and rheological properties of methacrylamide modified gelatin hydrogels[J]. Biomacromolecules, 2000,1(1):31-38. DOI: 10.1021/bm990017d.
    [4]
    OhwadaK. Body surface area of the golden Syrian hamster[J]. Jikken Dobutsu, 1992,41(2):221-224. DOI: 10.1538/expanim1978.41.2_221.
    [5]
    胥杰龙改性明胶/聚丙烯酰胺复合水凝胶及其用于软骨缺损修复的研究成都西南交通大学2016

    胥杰龙.改性明胶/聚丙烯酰胺复合水凝胶及其用于软骨缺损修复的研究[D].成都:西南交通大学,2016.
    [6]
    YueK, Trujillo-de SantiagoG, AlvarezMM, et al. Synthesis, properties, and biomedical applications of gelatin methacryloyl (GelMA) hydrogels[J]. Biomaterials, 2015,73:254-271. DOI: 10.1016/j.biomaterials.2015.08.045.
    [7]
    BessaLJ, FaziiP, Di GiulioM, et al. Bacterial isolates from infected wounds and their antibiotic susceptibility pattern: some remarks about wound infection[J]. Int Wound J, 2015,12(1):47-52. DOI: 10.1111/iwj.12049.
    [8]
    LokCN, HoCM, ChenR, et al. Silver nanoparticles: partial oxidation and antibacterial activities[J]. J Biol Inorg Chem, 2007,12(4):527-534. DOI: 10.1007/s00775-007-0208-z.
    [9]
    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.
    [10]
    XuR, LuoG, XiaH, et al. Novel bilayer wound dressing composed of silicone rubber with particular micropores enhanced wound re-epithelialization and contraction[J]. Biomaterials, 2015,40:1-11. DOI: 10.1016/j.biomaterials.2014.10.077.
    [11]
    Xavier MendesA, Moraes SilvaS, O'ConnellCD, et al. Enhanced electroactivity, mechanical properties, and printability through the addition of graphene oxide to photo-cross-linkable gelatin methacryloyl hydrogel[J]. ACS Biomater Sci Eng, 2021,7(6):2279-2295. DOI: 10.1021/acsbiomaterials.0c01734.
    [12]
    ArmatoU,FreddiG.Editorial: biomaterials for skin wound repair: tissue engineering, guided regeneration, and wound scarring prevention[J]. Front Bioeng Biotechnol, 2021, 9:722327. DOI: 10.3389/fbioe.2021.722327.
    [13]
    XiaoY, ReisLA, FericN, et al. Diabetic wound regeneration using peptide-modified hydrogels to target re-epithelialization[J]. Proc Natl Acad Sci U S A, 2016,113(40):E5792-E5801. DOI: 10.1073/pnas.1612277113.
    [14]
    EkeG, MangirN, HasirciN, et al. Development of a UV crosslinked biodegradable hydrogel containing adipose derived stem cells to promote vascularization for skin wounds and tissue engineering[J]. Biomaterials, 2017,129:188-198. DOI: 10.1016/j.biomaterials.2017.03.021.
    [15]
    ZhaoX, SunX, YildirimerL, et al. Cell infiltrative hydrogel fibrous scaffolds for accelerated wound healing[J]. Acta Biomater, 2017,49:66-77. DOI: 10.1016/j.actbio.2016.11.017.
    [16]
    宋知仁, 郑建锋, 成路, 等. 同型新鲜冰冻血浆联合负压封闭引流在压疮修复中的效果[J].中华烧伤杂志,2017,33(3):171-172. DOI: 10.3760/cma.j.issn.1009-2587.2017.03.009.
    [17]
    SamaniMK, SaberiBV, Ali TabatabaeiSM, et al. The clinical evaluation of platelet-rich plasma on free gingival graft's donor site wound healing[J]. Eur J Dent, 2017,11(4):447-454. DOI: 10.4103/ejd.ejd_76_17.
    [18]
    SridharanK, SivaramakrishnanG. Growth factors for diabetic foot ulcers: mixed treatment comparison analysis of randomized clinical trials[J]. Br J Clin Pharmacol, 2018,84(3):434-444. DOI: 10.1111/bcp.13470.
    [19]
    中国老年医学学会烧创伤分会. 胶原类创面材料临床应用全国专家共识(2018版)[J].感染、炎症、修复,2018,19(4):200-203. DOI: 10.3969/j.issn.1672-8521.2018.04.002.
    [20]
    陈秋东中等纯度重组人表皮生长因子(rhEGF)的分离纯化及在美容产品中的应用杭州浙江大学2003

    陈秋东.中等纯度重组人表皮生长因子(rhEGF)的分离纯化及在美容产品中的应用[D].杭州:浙江大学,2003.
    [21]
    SubravetiSN, RaghavanSR. A simple way to synthesize a protective "skin" around any hydrogel[J]. ACS Appl Mater Interfaces, 2021,13(31):37645-37654. DOI: 10.1021/acsami.1c09460.
    [22]
    KoH, SuthiwanichK, MaryH, et al. A simple layer-stacking technique to generate biomolecular and mechanical gradients in photocrosslinkable hydrogels[J]. Biofabrication, 2019,11(2):025014. DOI: 10.1088/1758-5090/ab08b5.
    [23]
    SunX,LangQ,ZhangH,et al. Cell scaffolds: electrospun photocrosslinkable hydrogel fibrous scaffolds for rapid in vivo vascularized skin flap regeneration[J]. Adv Funct Mater, 2017,27(2).DOI: 10.1002/adfm.201770008
    [24]
    JannaschM, GroeberF, BrattigNW, et al. Development and application of three-dimensional skin equivalents for the investigation of percutaneous worm invasion[J]. Exp Parasitol, 2015,150:22-30. DOI: 10.1016/j.exppara.2015.01.005.
    [25]
    WuJ, ZhuJ, HeC, et al. Comparative study of heparin- poloxamer hydrogel modified bFGF and aFGF for in vivo wound healing efficiency[J]. ACS Appl Mater Interfaces, 2016,8(29):18710-18721. DOI: 10.1021/acsami.6b06047.
    [26]
    ZhaoL, LiX, ZhaoJ, et al. A novel smart injectable hydrogel prepared by microbial transglutaminase and human-like collagen: its characterization and biocompatibility[J]. Mater Sci Eng C Mater Biol Appl, 2016,68:317-326. DOI: 10.1016/j.msec.2016.05.108.
    [27]
    LiuM, LuoG, WangY, et al. Optimization and integration of nanosilver on polycaprolactone nanofibrous mesh for bacterial inhibition and wound healing in vitro and in vivo[J]. Int J Nanomedicine, 2017,12:6827-6840. DOI: 10.2147/IJN.S140648.
    [28]
    LiuY, Chan-ParkMB. A biomimetic hydrogel based on methacrylated dextran-graft-lysine and gelatin for 3D smooth muscle cell culture[J]. Biomaterials, 2010,31(6):1158-1170. DOI: 10.1016/j.biomaterials.2009.10.040.
    [29]
    HuangR, HuJ, QianW,et al. Recent advances in nano- therapeutics for the treatment of burn wounds[J]. Burns Trauma,2021,9:tkab026[2022-04-02]. https://pubmed.ncbi.nlm.nih.gov/34778468/.DOI: 10.1093/burnst/tkab026.
    [30]
    ZhangX, ShuW, YuQ, et al. Functional biomaterials for treatment of chronic wound[J]. Front Bioeng Biotechnol, 2020,8:516. DOI: 10.3389/fbioe.2020.00516.
    [31]
    ChenG, YuY, WuX, et al. Wound healing: bioinspired multifunctional hybrid hydrogel promotes wound healing[J/OL]. Adv Funct Mater,2021,28(33):1870233[2021-09-30]. https://doi. org/10.1002/adfm.202105749. DOI: 10.1002/adfm.201870233.
    [32]
    焦建强, 李烨, 黄喆, 等. 重组人表皮生长因子凝胶联合纳米银敷料对烧伤后瘢痕的影响[J].中国组织工程研究,2015,(25):4007-4011. DOI: 10.3969/j.issn.2095-4344.2015.25.014.
    [33]
    KamalyN, YameenB, WuJ, et al. Degradable controlled- release polymers and polymeric nanoparticles: mechanisms of controlling drug release[J]. Chem Rev, 2016,116(4):2602-2663. DOI: 10.1021/acs.chemrev.5b00346.
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