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[1]于淑娴,刘颖,叶静,等.α-淀粉酶联合D-精氨酸对牙周炎相关致病菌混合生物膜的分散和抑制作用[J].天津医科大学学报,2025,31(06):527-535.[doi:10.20135/j.issn.1006-8147.2025.06.0527]
 YU Shuxian,LIU Ying,YE Jing,et al.Dispersion and inhibition effect of α-amylase combined with D-arginine against multispecies biofilms of periodontitis-associated pathogenic bacteria[J].Journal of Tianjin Medical University,2025,31(06):527-535.[doi:10.20135/j.issn.1006-8147.2025.06.0527]
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α-淀粉酶联合D-精氨酸对牙周炎相关致病菌混合生物膜的分散和抑制作用(PDF)

《天津医科大学学报》[ISSN:1006-8147/CN:12-1259/R]

卷:
31卷
期数:
2025年06期
页码:
527-535
栏目:
基础医学
出版日期:
2025-11-20

文章信息/Info

Title:
Dispersion and inhibition effect of α-amylase combined with D-arginine against multispecies biofilms of periodontitis-associated pathogenic bacteria
文章编号:
1006-8147(2025)06-0527-09
作者:
于淑娴1刘颖1叶静2温特2马贲2
(1.天津医科大学口腔医院牙体牙髓科,天津 300070;2.天津市天津医院口腔科,天津 300211)
Author(s):
YU Shuxian1 LIU Ying1 YE Jing2 WEN Te2 MA Ben2
(1.Department of Endodontics, Hospital of Stomatology, Tianjin Medical University,Tianjin 300070, China; 2.Department of Stomatology, Tianjin Hospital, Tianjin 300211, China)
关键词:
α-淀粉酶D-精氨酸牙周炎致病菌生物膜
Keywords:
α-amylase D-arginine periodontitis pathogenic bacteria biofilm
分类号:
R781.4
DOI:
10.20135/j.issn.1006-8147.2025.06.0527
文献标志码:
A
摘要:
目的:探究α-淀粉酶和D-精氨酸联合应用对牙周致病菌及混合生物膜的分散和抑制作用。方法:通过结晶紫染色分别测定α-淀粉酶和D-精氨酸对牙周主要致病菌(戈登链球菌、具核梭杆菌和牙龈卟啉单胞菌)的最小生物膜清除浓度(MBEC)和最小生物膜抑制浓度(MBIC)。通过微量棋盘稀释法确定与0.2%氯己定作用等效的联合用药浓度,并结合激光共聚焦显微镜(CLSM)评估其对多菌种生物膜的分散与抑制效果。结果:α-淀粉酶对单菌种和多菌种生物膜的MBEC、MBIC均为10 g/L。D-精氨酸对单菌种MBEC和MBIC均为128 mmol/L,对多菌种生物膜MBEC为128 mmol/L,MBIC为256 mmol/L。联合应用1.0 g/L α-淀粉酶和32 mmol/L D-精氨酸的分散作用以及0.5 g/L α-淀粉酶和32 mmol/L D-精氨酸的抑制作用,均与0.2% 氯己定效果相当。在CLSM检测的分散和抑制研究中,与单独用药相比,联合应用α-淀粉酶和D-精氨酸处理后的样本中活菌量最少(F=781.3、362.5,P<0.001),且与0.2% 氯己定组相比无统计学差异(P>0.05)。结论: α-淀粉酶联合D-精氨酸可显著提高对牙周致病菌混合生物膜的分散和抑制能力。
Abstract:
Objective: To evaluate the dispersion and inhibition effect of α-amylase in combination with D-arginine on periodontal pathogens and multispecies biofilms. Methods: Minimum biofilm clearance concentration (MBEC) and minimum biofilm inhibitory concentration (MBIC) of α-amylase and D-arginine against the major periodontal pathogens (Streptococcus gordonii, Fusobacteriumnucleatum, and Porphyromonas gingivalis) were determined by crystalline violet staining. The micro checkerboard dilution method was applied to determine the concentration of drug combination that was equivalent to the effect of 0.2% chlorhexidine (CHX). Combined with confocal laser scanning microscopelaser (CLSM), its dispersion and inhibition effect on multispecies biofilms was assessed. Results: The MBEC and MBIC of α-amylase were 10 g/L for both single-species biofilm and multispecies biofilm. D-Arginine had MBEC and MBIC values of 128 mmol/L for single-species biofilm. D-Arginine had MBEC values of 128 mmol/L, and MBIC values of 256 mmol/L for multispecies biofilm. The dispersion effect of the combined application of 1.0 g/L α-amylase with 32 mmol/L D-arginine and the inhibition effect of 0.5 g/L α-amylase with 32 mmol/L D-arginine were comparable to the effect of 0.2% CHX. In the dispersion and inhibition study of CLSM assay, the combined application of α-amylase and D-arginine resulted in the lowest viable bacterial count compared to either agent alone (F=781.3, 362.5, P<0.001), with no statistically significant difference from the 0.2% CHX group (P>0.05). Conclusion: The combination of α-amylase and D-arginine significantly enhances the dispersion and inhibition efficacy against multispecies biofilms of periodontal pathogens.

参考文献/References:

[1] ZHANG M, LIU Y, AFZALI H, et al. An update on periodontal inflammation and bone loss[J]. Front Immunol, 2024, 15: 1385436.
[2] DI STEFANO M, POLIZZI A, SANTONOCITO S, et al. Impact of oral microbiome in periodontal health and periodontitis: acritical review on prevention and treatment[J]. Int J of Mol Sci, 2022, 23(9): 5142.
[3] WU X, WANG L, LU Y, et al. A microenvironment-responsive graphdiyne-iron nanozyme hydrogel with antibacterial and anti-inflammatory effect for periodontitis treatment[J]. Adv Healthc Mater, 2024, 20: e2403683.
[4] GRAZIANI F, KARAPETSA D, ALONSO B, et al. Nonsurgical and surgical treatment of periodontitis: how many options for one disease?[J]. Periodontol 2000, 2017, 75(1): 152-188.
[5] POPPOLO DEUS F, OUANOUNOU A. Chlorhexidine in dentistry: pharmacology, uses, and adverse effects[J]. Int Dent J, 2022, 72(3): 269-277.
[6] TIAN S, SU L, LIU Y, et al. Self-targeting, zwitterionic micellar dispersants enhance antibiotic killing of infectious biofilms-an intravital imaging study in mice[J]. Sci Adv, 2020, 6(33): eabb1112.
[7] ZANATTA F B, ANTONIAZZI R P, R?魻SING C K. The effect of 0.12% chlorhexidine gluconate rinsing on previously plaque-free and plaque-covered surfaces: a randomized, controlled clinical trial[J]. J Periodontol, 2007, 78(11): 2127-2134.
[8] REDMAN W K, WELCH G S, WILLIAMS A C, et al. Efficacy and safety of biofilm dispersal by glycoside hydrolases in wounds[J]. Biofilm, 2021, 3: 100061.
[9] PRAKASH O, JAISWAL N. Alpha-amylase: an ideal representative of thermostable enzymes[J]. App Biochem Biotechnol, 2010, 160(8): 2401-2414.
[10] FULAZ S, VITALE S, QUINN L, et al. Nanoparticle-biofilm interactions: the role of the EPS matrix[J]. Trends Microbiol, 2019, 27(11): 915-926.
[11] KOLODKIN-GAL I, ROMERO D, CAO S, et al. D-amino acids trigger biofilm disassembly[J]. Science, 2010, 328(5978): 627-629.
[12] LAM H, OH D C, CAVA F, et al. D-amino acids govern stationary phase cell wall remodeling in bacteria[J]. Science, 2009, 325(5947): 1552-1555.
[13] JAKUBOVICS N S, GOODMAN S D, MASHBURN-WARREN L, et al. The dental plaque biofilm matrix[J]. Periodontol,2000, 2021, 86(1): 32-56.
[14] CHOO S W, MOHAMMED W K, MUTHA N V R, et al. Transcriptomic responses to coaggregation between Streptococcus gordonii and Streptococcus oralis[J]. App Environ Microbiol, 2024, 87(22): e0155821.
[15] BRENNAN C A, GARRETT W S. Fusobacterium nucleatum — symbiont, opportunist and oncobacterium[J]. Nat Rev Microbiol, 2019, 17(3): 156-166.
[16] WANG S, YAN T, ZHANG B, et al. Porphyromonas gingivalis vaccine: antigens and mucosal adjuvants[J]. Vaccines, 2024, 12(6): 619.
[17] BREGAINT S, BOYER E, FONG S B, et al. Porphyromonas gingivalis outside the oral cavity[J]. Odontology, 2022, 110(1): 1-19.
[18] ABDULKAREEM A A, AL-TAWEEL F B, AL-SHARQI AJB, et al. Current concepts in the pathogenesis of periodontitis: from symbiosis to dysbiosis[J]. J Oral Microbiol, 2023, 15(1): 2197779.
[19] PINTO R M, SOARES F A, REIS S, et al. Innovative strategies toward the disassembly of the EPS matrix in bacterial biofilms[J]. Front Microbiol, 2020, 11: 952.
[20] FLEMMING H C, WINGENDER J. The biofilm matrix[J]. Nat Rev Microbiol, 2010, 8(9): 623-633.
[21] LI B, CAI Q, WANG Z, et al. D-arginine enhances the effect of alpha-amylase on disassembling actinomyces viscosusbiofilm[J]. Front Bioeng Biotechnol, 2022, 10: 864012.
[22] FLEMING D, CHAHIN L, RUMBAUGH K. Glycoside hydrolases degrade polymicrobial bacterial biofilms in wounds[J]. Antimicrob Agents Chemother, 2017, 61(2): e0199816.
[23] WATTERS C M, BURTON T, KIRUI D K, et al. Enzymatic degradation of in vitro Staphylococcus aureus biofilms supplemented with human plasma[J]. Infect Drug Resist, 2016, 9: 71.
[24] QI H, LI B, WANG H, et al. Effects of d-valine on periodontal or peri-implant pathogens: Porphyromonas gingivalis biofilm[J]. J Periodontol, 2018, 89(3): 303-314.
[25] ZHANG Z, LI B, CAI Q, et al. Synergistic effects of D-arginine, D-methionine and D-histidine against Porphyromonas gingivalis biofilms[J]. Biofouling, 2021, 37(2): 222-234.
[26] LI Y Y, LI B S, LIU W W, et al. Effects of D-arginine on Porphyromonas gingivalis biofilm[J]. J Oral Sci, 2020, 62(1): 57-61.
[27] CAVA F, LAM H, DE PEDRO M A, et al. Emerging knowledge of regulatory roles of D-amino acids in bacteria[J]. Cell Mol Life Sci, 2010, 68(5): 817-831.

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备注/Memo

备注/Memo:
基金项目 天津市教委科研计划项目(自然科学,2021KJ245);天津市天津医院科技基金(TJYY2404)
作者简介 于淑娴(1999-),女,硕士在读,研究方向:牙体牙髓和牙周病的治疗;通信作者:马贲,E-mail:18604520557@163.com。
更新日期/Last Update: 2025-11-20