Volume 39 Issue 1
Jan.  2023
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Tang XY,Liu CY,Chu GP,et al.Effects of porcine urinary bladder matrix on motility and polarization of bone marrow-derived macrophages in mice[J].Chin J Burns Wounds,2023,39(1):25-34.DOI: 10.3760/cma.j.cn501225-20220516-00187.
Citation: Tang XY,Liu CY,Chu GP,et al.Effects of porcine urinary bladder matrix on motility and polarization of bone marrow-derived macrophages in mice[J].Chin J Burns Wounds,2023,39(1):25-34.DOI: 10.3760/cma.j.cn501225-20220516-00187.

Effects of porcine urinary bladder matrix on motility and polarization of bone marrow-derived macrophages in mice

doi: 10.3760/cma.j.cn501225-20220516-00187
Funds:

Clinical Frontier Technology Program of Social Development of Science and Technology Agency of Jiangsu Province of China BE2018626

Cultivation Plan of "Double Hundred" Medical Youth Tip-Top Professionals Program of Wuxi City HB2020050

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  •   Objective  To explore the effects of porcine urinary bladder matrix (UBM) on the motility and polarization of bone marrow-derived macrophages in mice, so as to provide evidence for the rational selection of stent in clinical wound repair.  Methods  The method of experimental research was used. The microstructure of porcine UBM and absorbable dressing was observed under scanning electron microscope. Polyacrylamide gel electrophoresis was used to observe the protein distribution of the two stent extracts. The primary macrophages were induced from bone marrow-derived cells isolated from six 6-8-week-old male C57BL/6J mice (mouse age, sex, and strain, the same below) and identified. Three batches of macrophages were divided into porcine UBM extract group and absorbable dressing extract group. The cells in each group were cultured with Dulbecco's modified Eagle medium/F12 medium containing the corresponding extracts. The cell migration rate was detected and calculated on 1, 3, and 7 d after scratching by scratch test. The number of migrated cells at 12 and 24 h of culture was detected by Transwell experiment. The percentages of CD206 and CD86 positive cells at 24 h of culture was detected by flow cytometer. The numbers of sample in the above cell experiments were all 3. An incision was prepared on the left and right back of twelve mice, respectively. The left incision of each mouse was included in porcine UBM group and the right incision was included in absorbable dressing group, and the corresponding stents were implanted into the incisions respectively. On post operation day (POD) 7 and 14, the number of inflammatory cells infiltrated in the stent was detected by hematoxylin-eosin staining; the number of F4/80, transforming growth factor-β1 (TGF-β1), vascular endothelial growth factor (VEGF), and matrix metalloprotein-9 (MMP-9) positive cells and type Ⅰ collagen deposition in stents were observed by immunohistochemistry; the percentages of F4/80, CD86, and CD206 positive cells were observed by immunofluorescence staining. The numbers of sample in the above animal experiments were all 6. Data were statistically analyzed with analysis of variance for factorial design, analysis of variance for repeated measurement, and independent sample t test.  Results  Porcine UBM has a dense basement membrane structure on one side and porous propria containing a fibrous structures on the other. Both sides of the absorbable dressing had three-dimensional porous structure. In the molecular weight range of (50-70)×103, multiple non-type Ⅰ collagen bands appeared in the lanes of porcine UBM extract, while no obvious bands appeared in the lanes of absorbable dressing extract. It had been identified that mouse bone marrow-derived cells had been successfully induced into macrophages. The cell migration rates in porcine UBM extract group were significantly higher than those in absorbable dressing extract group on 1, 3, and 7 d after scratching (with t values of 15.31, 19.76, and 20.58, respectively, P<0.05). The numbers of migrated cells in porcine UBM extract group were significantly more than those in absorbable dressing extract group at 12 and 24 h of culture (with t values of 12.20 and 33.26, respectively, P<0.05). At 24 h of culture, the percentage of CD86 positive cells in porcine UBM extract group ((1.27±0.19)%) was significantly lower than (7.34±0.14)% in absorbable dressing extract group (t=17.03, P<0.05);the percentage of CD206 positive cells in porcine UBM extract group was (73.4±0.7)%, significantly higher than (32.2±0.5)% in absorbable dressing extract group (t=119.10, P<0.05). On POD 7 and 14, the numbers of inflammatory cells infiltrated in the stents in porcine UBM group was significantly more than those in absorbable dressing group (with t values of 6.58 and 10.70, respectively, P<0.05). On POD 7 and 14, the numbers of F4/80, TGF-β1, VEGF, and MMP-9 positive cells in the stents in porcine UBM group were significantly more than those in absorbable dressing group (with t values of 46.11, 40.69, 13.90, 14.15, 19.79, 32.93, 12.16, and 13.21, respectively, P<0.05); type Ⅰ collagen deposition in the stents in porcine UBM group was more pronounced than that in absorbable dressing group; the percentages of CD206 positive cells in the stents in porcine UBM group were significantly higher than those in absorbable dressing group (with t values of 5.05 and 4.13, respectively, P<0.05), while the percentages of CD86 positive cells were significantly lower than those in absorbable dressing group (with t values of 20.90 and 19.64, respectively, P<0.05), and more M2-type macrophages were seen in the stents in porcine UBM group and more M1-type macrophages were seen in the stents in absorbable dressing group.  Conclusions  Porcine UBM can enhance macrophage motility, induce M2 polarization and paracrine function, create a microenvironment containing growth factors such as TGF-β1 and MMP-9 tissue remodeling molecules, and promote tissue regeneration and extracellular matrix remodeling in mice.

     

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  • [1]
    TangRF, ZhouXZ, NiuL, et al. Type Ⅰ collagen scaffold with WNT5A plasmid for in situ cartilage tissue engineering[J]. Biomed Mater Eng, 2022, 33(1):65-76. DOI: 10.3233/BME-211277.
    [2]
    GunatillakePA, AdhikariR. Biodegradable synthetic polymers for tissue engineering[J]. Eur Cell Mater, 2003, 5:1-16; discussion 16. DOI: 10.22203/ecm.v005a01.
    [3]
    SadtlerK, SommerfeldSD, WolfMT, et al. Proteomic composi-tion and immunomodulatory properties of urinary bladder matrix scaffolds in homeostasis and injury[J]. Semin Immunol, 2017, 29:14-23. DOI: 10.1016/j.smim.2017.05.002.
    [4]
    PokrywczynskaM, GubanskaI, DrewaG, et al. Application of bladder acellular matrix in urinary bladder regeneration: the state of the art and future directions[J]. Biomed Res Int, 2015, 2015:613439. DOI: 10.1155/2015/613439.
    [5]
    AndersonSR, GuptaN, JohnsonRM. Nonreplantable total scalp avulsion reconstruction with acellular biologic matrix and long-term outcome[J]. J Craniofac Surg, 2022, 33(4):e445-e446. DOI: 10.1097/SCS.0000000000008556.
    [6]
    HsuKF, ChiuYL, ChiaoHY, et al. Negative-pressure wound therapy combined with artificial dermis (Terudermis) followed by split-thickness skin graft might be an effective treatment option for wounds exposing tendon and bone: a retrospective observation study[J]. Medicine (Baltimore), 2021, 100(14):e25395. DOI: 10.1097/MD.0000000000025395.
    [7]
    王士源,袁光海,鲁尧,等.负荷缓释VEGF的人工真皮对大鼠深Ⅱ度烧伤创面修复及细胞凋亡的影响[J]. 临床和实验医学杂志, 2021,20(21):2259-2262. DOI: 10.3969/j.issn.1671-4695.2021.21.006.
    [8]
    MahonOR, BroweDC, Gonzalez-FernandezT, et al. Nano-particle mediated M2 macrophage polarization enhances bone formation and MSC osteogenesis in an IL-10 dependent manner[J]. Biomaterials,2020,239:119833. DOI: 10.1016/j.biomaterials.2020.119833.
    [9]
    VarelaP, SartoriS, ViebahnR, et al. Macrophage immunomodulation: an indispensable tool to evaluate the performance of wound dressing biomaterials[J]. J Appl Biomater Funct Mater, 2019, 17(1):2280800019830355. DOI: 10.1177/2280800019830355.
    [10]
    TangXY, YangFB, ChuGP, et al. Characterizing the inherent activity of urinary bladder matrix for adhesion, migration, and activation of fibroblasts as compared with collagen-based synthetic scaffold[J/OL]. J Biomater Appl, 2022:8853282221130883[2022-12-28]. https://pubmed.ncbi.nlm.nih.gov/36177498/. DOI: 10.1177/08853282221130883.[published online ahead of print].
    [11]
    VaquetteC, Cooper-WhiteJJ. Increasing electrospun scaffold pore size with tailored collectors for improved cell penetration[J]. Acta Biomater, 2011, 7(6):2544-2557. DOI: 10.1016/j.actbio.2011.02.036.
    [12]
    JiangJ, LiZR, WangHJ, et al. Expanded 3D nanofiber scaffolds: cell penetration, neovascularization, and host response[J]. Adv Healthc Mater, 2016, 5(23):2993-3003. DOI: 10.1002/adhm.201600808.
    [13]
    AhmedT, MarçalH, JohnsonS, et al. Coalescence of extracellular matrix (ECM) from porcine urinary bladder (UBM) with a laser-activated chitosan-based surgical adhesive[J]. J Biomater Sci Polym Ed, 2012, 23(12):1521-1538. DOI: 10.1163/092050611X585431.
    [14]
    JiangD, HuangJW, ShaoHL, et al. Characterization of bladder acellular matrix hydrogel with inherent bioactive factors[J]. Mater Sci Eng C Mater Biol Appl, 2017, 77:184-189. DOI: 10.1016/j.msec.2017.03.222.
    [15]
    MinuttiCM, KnipperJA, AllenJE, et al. Tissue-specific contribution of macrophages to wound healing[J]. Semin Cell Dev Biol, 2017, 61:3-11. DOI: 10.1016/j.semcdb.2016.08.006.
    [16]
    ChenYN, HuMR, LeiW, et al. Macrophage M1/M2 polarization[J]. Eur J Pharmacol, 2020,877:173090. DOI: 10.1016/j.ejphar.2020.173090.
    [17]
    DasA, DattaS, RocheE, et al. Novel mechanisms of collagenase santyl ointment (CSO) in wound macrophage polarization and resolution of wound inflammation[J]. Sci Rep, 2018, 8(1):1696. DOI: 10.1038/s41598-018-19879-w.
    [18]
    XinLB, LinXN, ZhouF, et al. A scaffold laden with mesenchymal stem cell-derived exosomes for promoting endometrium regeneration and fertility restoration through macrophage immunomodulation[J]. Acta Biomater, 2020, 113:252-266. DOI: 10.1016/j.actbio.2020.06.029.
    [19]
    KazimierczakP, KoziolM, PrzekoraA. The chitosan/agarose/nanoHA bone scaffold-induced M2 macrophage polarization and its effect on osteogenic differentiation in vitro[J]. Int J Mol Sci, 2021, 22(3):1109. DOI: 10.3390/ijms22031109.
    [20]
    ZhukauskasR, FischerDN, DeisterC, et al. A comparative study of porcine small intestine submucosa and cross-linked bovine type Ⅰ collagen as a nerve conduit[J]. J Hand Surg Glob Online, 2021, 3(5):282-288. DOI: 10.1016/j.jhsg.2021.06.006.
    [21]
    YangB, ZhangYF, ZhouLH, et al. Development of a porcine bladder acellular matrix with well-preserved extracellular bioactive factors for tissue engineering[J]. Tissue Eng Part C Methods, 2010, 16(5):1201-1211. DOI: 10.1089/ten.TEC.2009.0311.
    [22]
    FengC, ShanMJ, XiaYJ, et al. Single-cell RNA sequencing reveals distinct immunology profiles in human keloid[J].Front Immunol, 2022, 13: 940645. DOI: 10.3389/fimmu.2022.940645.
    [23]
    YinJL, WuY, YuanZW, et al. Advances in scarless foetal wound healing and prospects for scar reduction in adults[J]. Cell Prolif, 2020, 53(11):e12916. DOI: 10.1111/cpr.12916.
    [24]
    XinLB,LinXN,PanYB, et al. A collagen scaffold loaded with human umbilical cord-derived mesenchymal stem cells facilitates endometrial regeneration and restores fertility[J].Acta Biomater, 2019, 92: 160-171. DOI: 10.1016/j.actbio.2019.05.012.
    [25]
    BrowneS, JhaAK, AmeriK, et al. TGF-β1/CD105 signaling controls vascular network formation within growth factor sequestering hyaluronic acid hydrogels[J]. PLoS One, 2018, 13(3):e0194679. DOI: 10.1371/journal.pone.0194679.
    [26]
    CastroAB, CortelliniS, TemmermanA, et al. Characterization of the leukocyte- and platelet-rich fibrin block: release of growth factors, cellular content, and structure[J]. Int J Oral Maxillofac Implants, 2019, 34(4):855-864. DOI: 10.11607/jomi.7275.
    [27]
    XiaoB, YangWJ, LeiD, et al.PGS scaffolds promote the in vivo survival and directional differentiation of bone marrow mesenchymal stem cells restoring the morphology and function of wounded rat uterus[J]. Adv Healthc Mater, 2019, 8(5):e1801455. DOI: 10.1002/adhm.201801455.
    [28]
    XuL, DingLJ, WangL, et al.Umbilical cord-derived mesenchymal stem cells on scaffolds facilitate collagen degradation via upregulation of MMP-9 in rat uterine scars[J]. Stem Cell Res Ther, 2017, 8(1):84. DOI: 10.1186/s13287-017-0535-0.
    [29]
    StacyMR, NaitoY, MaxfieldMW, et al. Targeted imaging of matrix metalloproteinase activity in the evaluation of remodeling tissue-engineered vascular grafts implanted in a growing lamb model[J]. J Thorac Cardiovasc Surg, 2014, 148(5):2227-2233. DOI: 10.1016/j.jtcvs.2014.05.037.
    [30]
    TianWM, KyriakidesTR. Matrix metalloproteinase-9 deficiency leads to prolonged foreign body response in the brain associated with increased IL-1β levels and leakage of the blood-brain barrier[J]. Matrix Biol, 2009, 28(3):148-159. DOI: 10.1016/j.matbio.2009.02.002.
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