氨基酸/羟基磷灰石复合材料应用于酸蚀牛牙釉质体外再矿化

doi:10.15541/jim20190027

摘要/Abstract

摘要：

早期牙釉质龋病的再矿化在龋齿的预防和修复中起着至关重要的作用。基于牙釉质基质中所含的主要氨基酸, 在10 mmol/L甘氨酸、10 mmol/L L-丝氨酸和5 mmol/L L-天冬氨酸的存在下制备了氨基酸/羟基磷灰石(AA/HAP)复合材料, 表征了其物理、化学和生物学性质, 并评估了其对酸蚀牛牙釉质的再矿化作用。与不含氨基酸的羟基磷灰石(HAP)相比, 在氨基酸的抑制作用下, AA/HAP复合材料具有更低的结晶度和更高的生物相容性。在人工唾液中用AA/HAP对酸蚀牛牙釉质进行体外再矿化。再矿化后, 分别表征了牛牙釉质样品的表面和横截面形态、成分和力学性能。结果表明, AA/HAP可以诱导表面和深层牙釉质病变的修复。复合材料中释放的氨基酸可以吸附在有机基质残基上并诱导平行排列的HAP晶体的形成, 从而使牙釉质表面显微硬度(SMH)得到显著恢复。最后, 讨论了AA/HAP复合材料对酸蚀牛牙釉质的再矿化机制。

关键词: 再矿化, 牙釉质, 羟基磷灰石, 氨基酸

Abstract:

Remineralization of early enamel caries lesions plays a crucial role in prevention and restoration of dental caries. Based on the crucial amino acids contained in enamel matrix, amino acids /hydroxyapatite (AA/HAP) composite was synthesized in the presence of 10 mmol/L glycine, 10 mmol/L L-serine and 5 mmol/L L-aspartic acid. The physical, chemical and biological properties of the composite were characterized. And its remineralization effect on acid-etched bovine enamel was evaluated. Under the inhibition properties of amino acids, the composite has lower crystallinity and higher biocompatibility compared to the hydroxyapatite (HAP) without amino acids. In vitro remineralization of acid-etched bovine enamel using AA/HAP was conducted in artificial saliva. After remineralization, the surface and cross-section morphology, the components, and the mechanical property of bovine enamel samples were characterized, respectively. The results showed that the AA/HAP could induce the restoration of both surface and subsurface enamel lesions. The amino acids released from the composite could adsorb on the organic matrix residues and induce the formation of parallel arranged HAP crystals, eventually formed significant surface microhardness (SMH) recovering.

Key words: remineralization, enamel, hydroxyapatite, amino acid

中图分类号:

R318

刘继涛, 钏定泽, 杨泽斌, 陈希亮, 颜廷亭, 陈庆华. 氨基酸/羟基磷灰石复合材料应用于酸蚀牛牙釉质体外再矿化[J]. 无机材料学报, 2019, 34(11): 1222-1230.

LIU Ji-Tao, CHUAN Ding-Ze, YANG Ze-Bin, CHEN Xi-Liang, YAN Ting-Ting, CHEN Qing-Hua. In Vitro Remineralization of Acid-etched Bovine Enamel with Amino Acids/Hydroxyapatite Composite[J]. Journal of Inorganic Materials, 2019, 34(11): 1222-1230.

图/表 10

Fig. 1 FT-IR spectra of the HAP and AA/HAP composite

Fig. 2 XRD patterns of the HAP and AA/HAP composite

Fig. 3 TEM images of the HAP (a1) and AA/HAP (b1); HRTEM of the HAP (a2) and AA/HAP (b2) with inserts showing their corresponding FTT images

Fig. 4 CCK-8 assay of the HAP and AA/HAP at different concentrations

Fig. 5 Ca2+ concentration in the supernatant of remineralization solution

Fig. 6 Surface and cross-section FESEM images of the enamel: sound enamel (a1) and (a2), acid-etched enamel (b1) and (b2) with insert in each image showing corresponding enlarged view

Fig. 7 Surface and cross-section FESEM images of the enamel: blank group (a1, a2), HAP (b1, b2), and AA/HAP (c1, c2) with insert in each image showing corresponding to its enlarged view

Table 1 Ca/P molar ratio of the enamel surface

Sample	Elemental composition/at%			Ca/P ratio
Sample	O	P	Ca	Ca/P ratio
Polished enamel	19.86	17.97	29.04	1.62
Acid-etched enamel	25.95	19.34	28.59	1.48
Blank group	14.07	17.59	27.85	1.58
HAP group	19.63	16.05	26.20	1.63
AA/HAP group	40.61	17.87	29.49	1.65

Fig. 8 XRD patterns of the enamel surface: sound enamel (a); acid-etched enamel (b); blank group (c); HAP group (d) and AA/HAP group (e)

Fig. 9 Comparison of the SMH before demineralization, after demineralization and after remineralization

参考文献 33

[1]	DUVERGER O, BENIASH E, MORASSO M I . Keratins as components of the enamel organic matrix. Matrix Biology, 2016,52/54:260-265.
[2]	CAO Y, MEI L, LI Q L , et al. Methods for biomimetic mineralisation of human enamel: a systematic review. Materials, 2015,8(6):2873-2886.
[3]	YEOM B, SAIN T, LACEVIC N , et al. A biotic tooth enamel. Nature, 2017,543(7643):95-98.
[4]	HE L H, SWAIN M V . Understanding the mechanical behavior of human enamel from its structural and compositional characteristics. Journal of the Mechanical Behavior of Biomedical Materials, 2008,1(1):18-29.
[5]	WEINER S, ADDADI L . Design strategies in mineralized biological materials. Journal of Materials Chemistry, 1997,7(5):689-702.
[6]	ENSANYA A N, ANAS A, ADAM S , et al. Demineralization- remineralization dynamics in teeth and bone. International Journal of Nanomedicine, 2016,11:4743-4763.
[7]	FEATHERSTONE J D B, LUSSI A . Understanding the chemistry of dental erosion. Monographs in Oral Science, 2006,20:66-76.
[8]	LI X, WANG J, JOINER A , et al. The remineralisation of enamel: a review of the literature. Journal of Dentistry, 2014,42(S1):S12-S20.
[9]	CHEN M, YANG J, LI J , et al. Modulated regeneration of acid-etched human tooth enamel by a functionalized dendrimer that is an analog of amelogenin. Acta Biomaterialia, 2014,10(10):4437-4446.
[10]	RUAN Q, ZHANG Y, YANG X , et al. Amelogenin-chitosan matrix promotes assembly of an enamel-like layer with a dense interface. Acta Biomaterialia, 2013,9(7):7289-7297.
[11]	GUNGORMUS M, OREN E, HORST J A , et al. Cementomimetics- constructing a cementum-like biomineralized microlayer via amelogenin- derived peptides. International Journal of Oral Science, 2012,4(2):69-77.
[12]	CHUNG H Y, LI C, HSU C . Characterization of the effects of 3DSS peptide on remineralized enamel in artificial saliva. Journal of the Mechanical Behavior of Biomedical Materials, 2012,6:74-79.
[13]	LOW I M . Depth-profiling of crystal structure, texture, and microhardness in a functionally graded tooth enamel. Journal of the American Ceramic Society, 2004,87(11):2125-2131.
[14]	AL-OBAIDI R, SALEHI H, DESOUTTER A , et al. Chemical & nano-mechanical study of artificial human enamel subsurface lesions. Scientific Reports, 2018,8(1):4047.
[15]	BERGSTRAND F, TWETMAN S . A review on prevention and treatment of post-orthodontic white spot lesions-evidence-based methods and emerging technologies. The Open Dentistry Journal, 2011,5(1):158-162.
[16]	ROVERI N, BATTISTELLA E, FOLTRAN I , et al. Synthetic biomimetic carbonate-hydroxyapatite nanocrystals for enamel remineralization. Advanced Materials Research, 2008, 47-50:821-824.
[17]	LI L, PAN H, TAO J , et al. Repair of enamel by using hydroxyapatite nanoparticles as the building blocks. Journal of Materials Chemistry, 2008,18:4079-4084.
[18]	ROBINSON C, LOWE N R, WEATHERELL J A . Amino-acid composition, distribution and origin of “tuft” protein in human and bovine dental enamel. Archives of Oral Biology, 1975,20(1):29-42.
[19]	PAN H, TAO J, XU X , et al. Adsorption processes of Gly and Glu amino acids on hydroxyapatite surfaces at the atomic level. Langmuir, 2007,23(17):8972-8981.
[20]	TAVAFOGHI M, CERRUTI M . The role of amino acids in hydroxyapatite mineralization. Journal of the Royal Society Interface, 2016,13(123):1-12.
[21]	FAN Z J, WANG J Q, WANG Z F , et al. One-pot synthesis of graphene/ hydroxyapatite nanorod composite for tissue engineering. Carbon, 2014,66(1):407-416.
[22]	MATSUMOTO T, OKAZAKI M, INOUE M , et al. Crystallinity and solubility characteristics of hydroxyapatite adsorbed amino acid. Biomaterials, 2002,23(10):2241-2247.
[23]	GONZALEZ-MCQUIRE R, CHANE-CHING J Y, VIGNAUD E , et al. Synthesis and characterization of amino acid-functionalized hydroxyapatite nanorods. Journal of Materials Chemistry, 2004,14(14):2277-2281.
[24]	LI L, MAO C, WANG J , et al. Bio-inspired enamel repair via Glu-directed assembly of apatite nanoparticles: an approach to biomaterials with optimal characteristics. Advanced Materials, 2011,23(40):4695-4701.
[25]	BOANINI E, TORRICELLI P, GAZZANO M , et al. Nanocomposites of hydroxyapatite with aspartic acid and glutamic acid and their interaction with osteoblast-like cells. Biomaterials, 2006,27(25):4428-4433.
[26]	MORO D, ULIAN G, VALDRÈ G . Single molecule investigation of glycine-chlorite interaction by cross-correlated scanning probe microscopy and quantum mechanics simulations. Langmuir, 2015,31(15):4453-4463.
[27]	KRUKOWSKI S, LYSENKO N, KOLODZIEJSKI W . Synthesis and characterization of nanocrystalline composites containing calcium hydroxyapatite and glycine. Journal of Solid State Chemistry, 2018,264:59-67.
[28]	LAURANCE-YOUNG P, BOZEC L, GRACIA L , et al. A review of the structure of human and bovine dental hard tissues and their physicochemical behavior in relation to erosive challenge and remineralisation. Journal of Dentistry, 2011,39(4):266-272.
[29]	CAI F, SHEN P, WALLKER G D , et al. Remineralization of enamel subsurface lesions by chewing gum with added calcium. Journal of Dentistry, 2009,37(10):763-768.
[30]	MATSUMOTO T, OKAZAKI M, INOUE M , et al. Role of acidic amino acid for regulating hydroxyapatite crystal growth. Dental Materials Journal, 2006,25(2):360-364.
[31]	FAN Y, SUN Z, MORADIANOLDAK J . Controlled remineralization of enamel in the presence of amelogenin and fluoride. Biomaterials, 2009,30(4):478-483.
[32]	ZHANG X, LI Y, SUN X , et al. Biomimetic remineralization of demineralized enamel with nano-complexes of phosphorylated chitosan and amorphous calcium phosphate. Journal of Materials Science: Materials in Medicine, 2014,25(12):2619-2628.
[33]	KOULOURIDES T, CUETO H, PIGMAN W . Rehardening of softened enamel surfaces of human teeth by solutions of calcium phosphates. Nature, 1961,189(4760):226-227.