Hemoglobin Protects Enamel against Intrinsic Enamel Erosive Demineralization.

IF 2.9 2区医学 Q1 DENTISTRY, ORAL SURGERY & MEDICINE Caries Research Pub Date : 2024-01-01 Epub Date: 2024-01-10 DOI:10.1159/000536200

Thamyris Souza Carvalho, Tamara Teodoro Araújo, Talita Mendes Oliveira Ventura, Aline Dionizio, João Victor Frazão Câmara, Samanta Mascarenhas Moraes, Júlia Chaparro Leme, Larissa Tercilia Grizzo, Edson Crusca, Priscila Yumi Tanaka Shibao, Reinaldo Marchetto, Flavio Henrique-Silva, Juliano Pelim Pessan, Marília Afonso Rabelo Buzalaf

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Abstract

Introduction: This study investigated the changes in the acquired enamel pellicle (AEP) proteome when this integument is formed in vivo after treatment with sugarcane-derived cystatin (CaneCPI-5), hemoglobin (HB), and a statherin-derived peptide (StN15), or their combination and then exposed to an intrinsic acid challenge. The effectiveness of these treatments in preventing intrinsic erosion was also evaluated.

Methods: Ten volunteers, after prophylaxis, in 5 crossover phases, rinsed with the following solutions (10 mL, 1 min): control (deionized water-H2O) - group 1, 0.1 mg/mL CaneCPI-5 - group 2, 1.0 mg/mL HB - group 3, 1.88 × 10-5M StN15 - group 4, or a blend of these - group 5. Following this, AEP formation occurred (2 h) and an enamel biopsy (10 µL, 0.01 m HCl, pH 2.0, 10 s) was conducted on one incisor. The biopsy acid was then analyzed for calcium (Arsenazo method). The vestibular surfaces of the other teeth were treated with the same acid. Acid-resistant proteins in the residual AEP were then collected and analyzed quantitatively via proteomics.

Results: Compared to control, treatment with the proteins/peptide, mixed or isolated, markedly enhanced acid-resistant proteins in the AEP. Notable increases occurred in pyruvate kinase PKM (11-fold, CaneCPI-5), immunoglobulins and submaxillary gland androgen-regulated protein 3B (4-fold, StN15), Hb, and lysozyme C (2-fold, StN15). Additionally, a range of proteins not commonly identified in the AEP but known to bind calcium or other proteins were identified in groups treated with the tested proteins/peptide either in isolation or as a mixture. The mean (SD, mM) calcium concentrations released from enamel were 3.67 ± 1.48a, 3.11 ± 0.72a, 1.94 ± 0.57b, 2.37 ± 0.90a, and 2.38 ± 0.45a for groups 1-5, respectively (RM-ANOVA/Tukey, p < 0.05).

Conclusions: Our findings demonstrate that all treatments, whether using a combination of proteins/peptides or in isolation, enhanced acid-resistant proteins in the AEP. However, only HB showed effectiveness in protecting against intrinsic erosive demineralization. These results pave the way for innovative preventive methods against intrinsic erosion, using "acquired pellicle engineering" techniques.

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血红蛋白能保护珐琅质免受内在珐琅质侵蚀性脱矿的影响。

简介本研究调查了获得性釉质小柱（AEP）蛋白质组在体内形成时的变化，这种小柱是用甘蔗衍生的胱抑素（CaneCPI-5）、血红蛋白（HB）和石炭酸衍生的多肽（StN15）或它们的组合处理后形成的，然后暴露于内在酸的挑战下。同时还评估了这些治疗方法在预防内在酸蚀方面的效果：10 名志愿者在预防性治疗后，分 5 个交叉阶段用以下溶液（10 毫升，1 分钟）冲洗：对照组（去离子水-H2O）- 第 1 组，0.1 毫克/毫升 CaneCPI-5- 第 2 组，1.0 毫克/毫升 HB- 第 3 组，1.之后，AEP 形成（2 小时），并在一颗门牙上进行釉质活检（10µL，0.01M HCl，pH 2.0，10 秒）。然后对活检酸液进行钙分析（砒霜法）。其他牙齿的前庭表面也用同样的酸进行处理。然后收集残留 AEP 中的抗酸蛋白质，并通过蛋白质组学进行定量分析：结果：与对照组相比，用混合或分离的蛋白质/肽处理后，AEP 中的抗酸蛋白明显增加。丙酮酸激酶 PKM（11 倍，CaneCPI-5）、免疫球蛋白和颌下腺雄激素调节蛋白 3B（4 倍，StN15）、血红蛋白和溶菌酶-C（2 倍，StN15）的含量显著增加。此外，在用受测蛋白质/肽单独或混合处理的组中，还发现了一系列在 AEP 中不常见但已知能结合钙或其他蛋白质的蛋白质。1-5组从釉质中释放的钙浓度平均值（标度，毫摩尔）分别为3.67±1.48a、3.11±0.72a、1.94±0.57b、2.37±0.90a和2.38±0.45a（RM-ANOVA/Tukey，p结论）：我们的研究结果表明，所有治疗方法，无论是使用蛋白质/肽的组合还是单独使用，都能增强 AEP 中的抗酸蛋白质。然而，只有 HB 能有效防止内在侵蚀性脱矿化。这些结果为利用 "获得性胶粒工程 "技术，采用创新性预防方法防止内在侵蚀铺平了道路。

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来源期刊

Caries Research 医学-牙科与口腔外科

CiteScore

6.30

自引率

7.10%

发文量

审稿时长

6-12 weeks

期刊介绍： ''Caries Research'' publishes epidemiological, clinical and laboratory studies in dental caries, erosion and related dental diseases. Some studies build on the considerable advances already made in caries prevention, e.g. through fluoride application. Some aim to improve understanding of the increasingly important problem of dental erosion and the associated tooth wear process. Others monitor the changing pattern of caries in different populations, explore improved methods of diagnosis or evaluate methods of prevention or treatment. The broad coverage of current research has given the journal an international reputation as an indispensable source for both basic scientists and clinicians engaged in understanding, investigating and preventing dental disease.