北京大学学报(医学版) ›› 2022, Vol. 54 ›› Issue (1): 83-88. doi: 10.19723/j.issn.1671-167X.2022.01.013

• 论著 • 上一篇    下一篇

新型大气压冷等离子体射流处理对牙本质胶原纤维交联化的影响

马欣蓉1,朱晓鸣2,李静3,李德利2,李和平3,(),谭建国1,()   

  1. 1.北京大学口腔医学院·口腔医院修复科,国家口腔医学中心,国家口腔疾病临床医学研究中心,口腔数字化医疗技术和材料国家工程实验室,北京 100081
    2.北京大学口腔医学院·口腔医院第二门诊部,北京 100101
    3.清华大学工程物理系,北京 100084
  • 收稿日期:2021-10-11 出版日期:2022-02-18 发布日期:2022-02-21
  • 通讯作者: 李和平,谭建国 E-mail:liheping@tsinghua.edu.cn;kqtanjg@bjmu.edu.cn

Effect of a novel radio-frequency atmospheric-pressure glow discharge plasma jet treatment on crosslinking of dentin collagen

MA Xin-rong1,ZHU Xiao-ming2,LI Jing3,LI De-li2,LI He-ping3,(),TAN Jian-guo1,()   

  1. 1. Department of Prothodontics, Peking University School and Hospital of Stomatology & National Center of Stomatology & National Clinical Research Center for Oral Diseases & National Engineering Laboratory for Digital and Material Technology of Stomatology, Beijing 100081, China
    2. Second Clinical Division, Peking University School and Hospital of Stomatology, Beijing 100101, China
    3. Department of Engineering Physics, Tsinghua University, Beijing 100084, China
  • Received:2021-10-11 Online:2022-02-18 Published:2022-02-21
  • Contact: He-ping LI,Jian-guo TAN E-mail:liheping@tsinghua.edu.cn;kqtanjg@bjmu.edu.cn

摘要:

目的: 研究新型大气压射频辉光放电(radio-frequency atmospheric-pressure glow discharge,RF-APGD)等离子体处理对牙本质胶原纤维交联化的影响。方法: (1)收集20颗新鲜拔除的、完整的第三磨牙,采用低速水冷精密切割机制备平行于牙合面的中层牙本质片,厚度为(1.5±0.1) mm,浸泡于10%(质量分数)H3PO4溶液中16 h获得全脱矿牙本质胶原纤维。将20个牙本质胶原纤维片随机分为5组,对照组无处理,4个实验组采用气体温度为4 ℃的等离子体处理不同时间(20 s、30 s、40 s、50 s)。采用衰减全反射傅里叶变换红外光谱仪测定牙本质胶原纤维结构及交联度,采用扫描电镜观察牙本质胶原纤维表面形貌,采用透射电镜观察牙本质胶原纤维微观结构。(2)收集完整第三磨牙40颗,制备精细牙本质粉5 g,10%H3PO4溶液中完全脱矿,将牙本质胶原纤维粉平均分为5组。对照组无处理,实验组分别采用等离子体处理20 s、30 s、40 s、50 s,采用茚三酮比色法测定各组交联度。(3)收集40颗完整第三磨牙,制备中层牙本质条。将牙本质胶原纤维条随机均分为5组,对照组无处理,实验组采用等离子体处理牙本质胶原纤维条各个轴面20 s、30 s、40 s、50 s,采用万能力学机测定牙本质极限拉伸强度。结果: (1)扫描电镜观察脱矿牙本质表面形貌显示,等离子体处理20 s、 30 s及40 s,脱矿牙本质表面胶原纤维网状结构均能维持蓬松;等离子体处理50 s会出现部分微结构的破坏;透射电镜结果显示等离子体处理20 s、30 s及40 s后,纤维结构蓬松,可见天然Ⅰ型胶原纤维典型的周期性横纹;红外光谱结果显示,等离子体处理后,胶原纤维的二级构象与对照组一致,且处理30 s及40 s后酰胺带强度明显增加。(2)茚三酮交联度测试结果显示等离子体处理30 s组及40 s组交联度最高,差异有统计学意义(P<0.05)。(3)牙本质极限拉伸强度结果显示,对照组为(1.67±0.24) MPa,等离子体处理20 s、30 s、40 s、50 s组分别为(4.21±0.15) MPa、(7.06±0.30) MPa、(7.32±0.27) MPa、(6.87±0.17) MPa,与对照组差异有统计学意义(P<0.05)。结论: 新型RF-APGD等离子体处理可以促进脱矿牙本质胶原纤维交联,同时明显增强了牙本质胶原纤维的机械性能。

关键词: 等离子体, 牙本质, 胶原, 拉伸强度

Abstract:

Objective: To investigate the effect of a noval radio-frequency atmospheric-pressure glow discharge (RF-APGD) plasma jet on crosslinking of dentin collagen. Methods: (1) Twenty intact third molars were collected. The middle dentin discs were prepared for each tooth by low-speed water-cooled Isomet saw, and then immersed in 10% (mass fraction) H3PO4 solution for 16 h to obtain fully demine-ralized dentin collagen. The twenty dentin discs were then randomly divided into five groups. The control group was untreated while the four experimental groups were treated by plasma jet with gas temperature of 4 ℃ for different times (20 s, 30 s, 40 s, and 50 s). The structure and crosslinking degree of dentin collagen were characterized by attenuated total reflection-Fourier transform infrared spectroscopy. The surface morphology of demineralized dentin was observed by scanning electron microscope, and the microstructure was observed by transmission electron microscope. (2) Fourty non-caries third molars were collected to prepare 5 g fine dentin powder, then completely demineralized with 10% H3PO4 solution. The control group was untreated, while the four experimental groups were treated by plasma jet for 20 s, 30 s, 40 s and 50 s. The crosslinking degree of each group was determined by ninhydrin colorimetric method. (3) Forty intact third molars were collected to obtain dentin strips. Only two central symmetrical dentin strips (nasty 80) were taken from each tooth and immersed in 10% H3PO4 solution for 16 h to obtain fully demineralized dentin collagen. Eighty dentine collagen fiber strips were randomly divided into five groups. The control group was untreated and the axial surfaces of dentin collagen fiber strips in the expe-rimental groups were treated with the plasma jet for 20 s, 30 s, 40 s and 50 s. The ultimate tensile strength of dentin was measured by universal mechanical machine. Results: (1) The surface morphology of demineralized dentin observed by scanning electron microscope showed that the network structure of collagen fibers on the surface of demineralized dentin treated with the plasma jet for 20 s, 30 s and 40 s could maintain the uncollapsed three-dimensional structure, and part of the microstructure was destroyed after plasma jet treated for 50 s. After being treated by plasma jet for 20 s, 30 s and 40 s, the three-dimensional structure was uncollapsed, and the typical periodic transverse pattern of natural type Ⅰ collagen fiber could be seen. The results of infrared spectra showed that the secondary conformation of dentin collagen fibers was the same as that of the control group after being treated with the plasma jet, and the intensity of amide band was significantly increased after being treated for 30 s and 40 s. (2) The results of ninhydrin crosslinking test showed that the crosslinking ratio of dentin collagen treated by plasma jet for 30 s and 40 s was the highest, and the difference was statistically significant (P<0.05). (3) The results of dentin ultimate tensile strength showed that the control group was (1.67±0.24) MPa, and the plasma jet treated 20 s, 30 s, 40 s and 50 s groups were (4.21±0.15) MPa, (7.06±0.30) MPa, (7.32±0.27) MPa, and (6.87±0.17) MPa, which were significantly different from that of the control group (P<0.05). Conclusion: The novel RF-APGD plasma jet treatment can promote the crosslinking degree of demineralized dentin collagen and improve their ultimate tensile strength.

Key words: Plasma, Dentin, Collagen, Tensile strength

中图分类号: 

  • R783.1

图1

对照组及新型RF-APGD等离子体处理组牙本质胶原纤维扫描电镜结果"

图2

对照组及新型RF-APGD等离子体处理组牙本质胶原纤维透射电镜结果"

图3

对照组及新型RF-APGD等离子体处理组牙本质胶原纤维红外光谱结果"

图4

新型RF-APGD等离子体处理组牙本质胶原纤维交联度结果"

表1

对照组及新型RF-APGD等离子体处理组牙本质胶原纤维极限拉伸结果"

Group Ultimate tensile strength/MPa
Control 1.67±0.24d
RF-APGD 20 s 4.21±0.15c
RF-APGD 30 s 7.06±0.30b
RF-APGD 40 s 7.32±0.27a
RF-APGD 50 s 6.87±0.17b
[1] 刘明月, 谭建国. 一步一步做好牙体缺损修复体类型的选择[J]. 中华口腔医学杂志, 2021, 56(7):720-725.
[2] De MJ, Van LK, Peumans M, et al. A critical review of the durability of adhesion to tooth tissue: Methods and results[J]. J Dent Res, 2005, 84(2):118-132.
doi: 10.1177/154405910508400204
[3] Nakabayashi N, Kojima K, Masuhara E. The promotion of adhesion by the infiltration of monomers into tooth substrates[J]. J Biomed Mater Res, 1982, 16(3):265-273.
doi: 10.1002/(ISSN)1097-4636
[4] Yao S, Xu Y, Shao C, et al. Collagen: Methods and protocols[M]. New York: Springer New York, 2019: 39-54.
[5] Ma YX, Hoff SE, Huang XQ, et al. Involvement of prenucleation clusters in calcium phosphate mineralization of collagen[J]. Acta Biomater, 2021, 120(7):213-223.
doi: 10.1016/j.actbio.2020.07.038
[6] Wang Y, Spencer P. Hybridization efficiency of the adhesive/dentin interface with wet bonding[J]. J Dent Res, 2003, 82(2):141-145.
pmid: 12562889
[7] Tjäderhane L, Nascimento FD, Breschi L, et al. Optimizing dentin bond durability: Control of collagen degradation by matrix metalloproteinases and cysteine cathepsins[J]. Dent Mater, 2013, 29(1):116-135.
doi: 10.1016/j.dental.2012.08.004 pmid: 22901826
[8] Stammers M, Niewczas IS, Segonds-Pichon A, et al. Mechanical stretching changes crosslinking and glycation levels in the collagen of mouse tail tendon[J]. J Biol Chem, 2020, 295(31):10572-10580.
doi: 10.1074/jbc.RA119.012067 pmid: 32546479
[9] Zhou J, Chiba A, Scheffel DL, et al. Cross-linked dry bonding: A new etch-and-rinse technique[J]. Dent Mater, 2016, 32(9):1124-1132.
doi: 10.1016/j.dental.2016.06.014
[10] Shinkai K, Takada M, Kawashima S, et al. Effects of the percen-tage of air/water in spray on the efficiency of tooth ablation with erbium, chromium: yttrium-scandium-gallium-garnet (Er,Cr:YSGG) laser irradiation[J]. Lasers Med Sci, 2019, 34(1):99-105.
doi: 10.1007/s10103-018-2607-5
[11] Zhu XM, Guo H, Zhou JF, et al. Influences of the cold atmospheric plasma jet treatment on the properties of the demineralized dentin surfaces[J]. Plasma Sci and Tech, 2018, 20(4):044010.
doi: 10.1088/2058-6272/aaa6be
[12] Zhu XM, Zhou JF, Guo H, et al. Effects of a modified cold atmospheric plasma jet treatment on resin-dentin bonding[J]. Dent Mater J, 2018, 37(5):798-804.
doi: 10.4012/dmj.2017-314
[13] Li HP, Wang ZB, Ge N, et al. Studies on the physical characte-ristics of the radio-frequency atmospheric-pressure glow discharge plasmas for the genome mutation of methylosinus trichosporium[J]. IEEE Trans Plasma Sci IEEE Nucl Plasma Sci Soc, 2012, 40(11):2853-2860.
[14] Nanci A. Ten Cate’s oral histology[M]. 9th ed. St. Louis: Mosby, 2016: 157-192.
[15] Ferracane JL. Resin composite: State of the art[J]. Dent Mater, 2011, 27(1):29-38.
doi: 10.1016/j.dental.2010.10.020 pmid: 21093034
[16] Moradian-Oldak J, George A. Biomineralization of enamel and dentin mediated by matrix proteins[J]. J Dent Res, 2021, 100(10):1020-1029.
doi: 10.1177/00220345211018405
[17] Wang Y, Azaïs T, Robin M, et al. The predominant role of collagen in the nucleation, growth, structure and orientation of bone apatite[J]. Nat Mater, 2012, 11(8):724-733.
doi: 10.1038/nmat3362 pmid: 22751179
[18] Hartwig S, Doll C, Voss JO, et al. Treatment of wound healing disorders of radial forearm free flap donor sites using cold atmospheric plasma: A proof of concept[J]. J Oral Maxillofac Surg, 2017, 75(2):429-435.
doi: 10.1016/j.joms.2016.08.011
[19] Liguori A, Bigi A, Colombo V, et al. Atmospheric pressure non-equilibrium plasma as a green tool to crosslink gelatin nanofibers[J]. Sci Rep, 2016, 6:38542.
doi: 10.1038/srep38542 pmid: 27924840
[20] Chiang YS, Chen YL, Chuang SF, et al. Riboflavin-ultraviolet-A-induced collagen cross-linking treatments in improving dentin bonding[J]. Dent Mater, 2013, 29(6):682-692.
doi: 10.1016/j.dental.2013.03.015
[21] Craciun G, Manaila E, Stelescu MD. New elastomeric materials based on natural rubber obtained by electron beam irradiation for food and pharmaceutical use[J]. Materials, 2016, 9(12):999.
doi: 10.3390/ma9120999
[22] He L, Mu C, Shi J, et al. Modification of collagen with a natural cross-linker, procyanidin[J]. Int J Biol Macromol, 2011, 48(2):354-359.
doi: 10.1016/j.ijbiomac.2010.12.012
[23] Wu L, Shao H, Fang Z, et al. Mechanism and effects of polyphenol derivatives for modifying collagen[J]. ACS Biomaterials Science & Engineering, 2019, 5(9):4272-4284.
[1] 骆池怡,彭楚芳,杨媛,秦满,王媛媛. 3种自酸蚀粘接系统和轻度唾液污染对乳牙釉质及牙本质粘接耐久性的影响[J]. 北京大学学报(医学版), 2021, 53(1): 46-53.
[2] 李秋菊,宫玮玉,董艳梅. 生物活性玻璃预处理对牙本质粘接界面耐久性的影响[J]. 北京大学学报(医学版), 2020, 52(5): 931-937.
[3] 王平,宋婧,方翔宇,李鑫,刘栩,贾园,栗占国,胡凡磊. 成红细胞样Ter细胞在胶原诱导性关节炎发病中的作用[J]. 北京大学学报(医学版), 2019, 51(3): 445-450.
[4] 郑苗,詹凌璐,刘志强,李和平,谭建国. 不同等离子体处理氧化锆对人牙龈成纤维细胞黏附能力的影响[J]. 北京大学学报(医学版), 2019, 51(2): 315-320.
[5] 潘怡湘,李秀花,田福聪,王晓燕. 髓腔内压对树脂水门汀与牙本质粘接强度的影响[J]. 北京大学学报(医学版), 2019, 51(2): 321-326.
[6] 张倩莉,袁重阳,刘力,温世鹏,王晓燕. 胶原静电纺纳米纤维膜对人牙髓细胞生物学行为的影响[J]. 北京大学学报(医学版), 2019, 51(1): 28-34.
[7] 朱晓鸣,齐璇,李德利,张玉玮,李和平,谭建国. 不同温度新型大气压冷等离子体处理对牙本质粘接强度的影响[J]. 北京大学学报(医学版), 2019, 51(1): 43-48.
[8] 李芳,刘洋,刘浩辰,冯海兰. 乳光牙本质患者的基因变异分析及患牙的组织学观察[J]. 北京大学学报(医学版), 2018, 50(4): 666-671.
[9] 臧海玲,王月,梁宇红. 有机溶剂对牙本质表面残留根管封闭剂的清除效果[J]. 北京大学学报(医学版), 2018, 50(1): 63-68.
[10] 贾维茜,赵玉鸣,葛立宏. 人重组转化生长因子β1促进牙髓干细胞的增殖和矿化[J]. 北京大学学报(医学版), 2017, 49(4): 680-681.
[11] 李皓,刘玉华,罗志强. 生物活性玻璃用于缓解活髓牙全冠预备后敏感的效果评价[J]. 北京大学学报(医学版), 2017, 49(4): 709-713.
[12] 信义,王赛楠,崔彩云,董艳梅. 生物活性玻璃和牙本质浸提蛋白对人牙髓细胞的作用[J]. 北京大学学报(医学版), 2017, 49(2): 331-336.
[13] 郭惠杰,高承志, 林斐,刘伟,岳林. 唾液污染对复合树脂间粘接强度的影响[J]. 北京大学学报(医学版), 2017, 49(1): 96-100.
[14] 傅昭然,田福聪,张路,韩冰,王晓燕. 通用型粘接剂对双固化树脂水门汀牙本质粘接强度的影响[J]. 北京大学学报(医学版), 2017, 49(1): 101-104.
[15] 王月,梁宇红. 次氯酸钠溶液表面处理对牙本质粘接强度的影响[J]. 北京大学学报(医学版), 2017, 49(1): 105-109.
Viewed
Full text


Abstract

Cited

  Shared   
  Discussed   
[1] 田增民, 陈涛, Nanbert ZHONG, 李志超, 尹丰, 刘爽. 神经干细胞移植治疗遗传性小脑萎缩的临床研究(英文稿)[J]. 北京大学学报(医学版), 2009, 41(4): 456 -458 .
[2] 汪毅, 刘粹, 王玉凤. 社会技能训练对有行为问题儿童影响的随机对照研究[J]. 北京大学学报(医学版), 2007, 39(3): 315 -318 .
[3] . 消息[J]. 北京大学学报(医学版), 2007, 39(3): 12 .
[4] 王海燕. 办好病例分析类栏目 为提高临床诊疗水平提供新平台[J]. 北京大学学报(医学版), 2007, 39(4): 341 .
[5] 田雨, 刘建湘, 王化虹. 经皮肝穿刺活检确诊无症状原发性肝结核一例[J]. 北京大学学报(医学版), 2007, 39(4): 365 -368 .
[6] 朱炳梅, 李婷, 周玉玲, 宋泉声, 王露. 新的潜在的雄激素受体协同抑制因子CMTM1-v17[J]. 北京大学学报(医学版), 2007, 39(4): 388 -393 .
[7] 袁葛, 赵继宗, 王硕, 许俊, 辛宇. 术中血管造影在脑动静脉畸形手术中的应用[J]. 北京大学学报(医学版), 2007, 39(4): 412 -415 .
[8] 王军, 肖水芳, 秦永, 王全桂, 陈丽. 以面神经麻痹为首诊表现的Wegener肉芽肿病一例[J]. 北京大学学报(医学版), 2007, 39(4): 434 -436 .
[9] 汤坚强, 万远廉, 刘玉村, 戎龙, 汪欣, 吴涛, 潘义生, 朱静. 组织因子/活化凝血Ⅶ因子复合物对大肠癌LoVo细胞系表达基质金属蛋白酶-7的调控作用[J]. 北京大学学报(医学版), 2007, 39(5): 453 -457 .
[10] 朱红, 姚婉贞, 沈宁, 张立强, 王筱宏, 梁岩静, 周庆涛. 北京延庆农村支气管哮喘流行病学调查结果[J]. 北京大学学报(医学版), 2007, 39(5): 494 -497 .