Evaluation of the biological effects of amelogenin on human oral keratinocytes

IF 4.6 1区医学 Q1 DENTISTRY, ORAL SURGERY & MEDICINE Dental Materials Pub Date : 2023-08-26 DOI:10.1016/j.dental.2023.08.176

Iris Frasheri , Maria Paschalidou , Thomas Imhof , Thorsten Steinberg , Thomas Spinell , Reinhard Hickel , Matthias Folwaczny

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Abstract

Objectives

Amelogenins are clinically used in periodontal regeneration as main components of root surface modifying agents, even without specifically preventing the premature colonization of the healing tissue defect by means of a physical barrier membrane. The objective of this study was to investigate the effects of human amelogenin on the proliferation, migration, and morphology of Immortalized Human Oral Keratinocytes (iHOKs).

Methods

Immortalized Human Oral Keratinocytes were expanded in Keratinocyte Growth Medium-2 (KGM-2). Full-length recombinant amelogenin protein was diluted in KGM-2 in five concentrations (10 ng/ml, 100 ng/ml, 1.000 ng/ml, 5.000 ng/ml and 10.000 ng/ml). iHOKs were cultured in medium supplemented with the amelogenin dilutions. Samples without amelogenin served as control. Cell metabolism and cell proliferation together with cell migration were evaluated at day 7, 14, 21.

Results

At day 7, iHOKs treated with 10,000 ng/ml showed a significant decrease in keratinocytes´ proliferation. The metabolic activity at this timepoint was significantly lower for concentrations ≥ 1000 ng/ml. At days 14 and 21, both the addition of 5000 ng/ml and even more 10,000 ng/ml amelogenin reduced significantly the proliferation of keratinocytes. The effects on the metabolic activity for these timepoints were visible already with 100 ng/ml. Treatment of iHOKs with amelogenin of ≥ 1000 ng/ml led to inhibitory effects on cell migration already after 24 h.

Conclusions

The full-length recombinant amelogenin has a significant biological impact on iHOKs. The increasing dose dependent inhibitory effects of amelogenin shown on iHOKs might explain the disruption of the apical migration of the junctional epithelium during regenerative healing.

Clinical significance

Amelogenin, presents time- and dose-dependent inhibitory effects on the growth of keratinocytes, which might explain the biological rationale behind its application in periodontal regeneration.

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淀粉原蛋白对人口腔角质形成细胞生物学效应的评价

目的在牙周再生中，即使没有通过物理屏障膜特异性地阻止愈合组织缺损的过早定植，仍可作为根表面修饰剂的主要成分应用于临床。本研究的目的是探讨人淀粉原蛋白对永生化人口腔角质形成细胞(iHOKs)的增殖、迁移和形态的影响。方法用角化细胞生长培养基-2 (KGM-2)扩增人口腔角化细胞。全长重组淀粉原蛋白在KGM-2中稀释成5个浓度(10 ng/ml、100 ng/ml、1.000 ng/ml、5.000 ng/ml和10.000 ng/ml)。iHOKs在添加淀粉原稀释液的培养基中培养。不含淀粉原蛋白的样品作为对照。在第7、14、21天观察细胞代谢、细胞增殖和细胞迁移情况。结果第7天，1万ng/ml iHOKs处理后，角质形成细胞增殖明显降低。浓度≥1000 ng/ml时，该时间点的代谢活性显著降低。在第14天和第21天，添加5000 ng/ml甚至更多10,000 ng/ml的淀粉原蛋白均可显著降低角质形成细胞的增殖。100 ng/ml对这些时间点的代谢活性的影响已经可见。用≥1000 ng/ml的淀粉原蛋白处理iHOKs, 24 h后就能抑制细胞迁移。结论全长重组淀粉原蛋白对iHOKs具有显著的生物学影响。在iHOKs中显示的越来越多的剂量依赖性抑制作用可能解释了再生愈合过程中连接上皮顶端迁移的中断。临床意义淀粉原蛋白对角质形成细胞的生长具有时间和剂量依赖性的抑制作用，这可能解释了其在牙周再生中的应用背后的生物学原理。

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来源期刊

Dental Materials 工程技术-材料科学：生物材料

CiteScore

9.80

自引率

10.00%

发文量

290

审稿时长

67 days

期刊介绍： Dental Materials publishes original research, review articles, and short communications. Academy of Dental Materials members click here to register for free access to Dental Materials online. The principal aim of Dental Materials is to promote rapid communication of scientific information between academia, industry, and the dental practitioner. Original Manuscripts on clinical and laboratory research of basic and applied character which focus on the properties or performance of dental materials or the reaction of host tissues to materials are given priority publication. Other acceptable topics include application technology in clinical dentistry and dental laboratory technology. Comprehensive reviews and editorial commentaries on pertinent subjects will be considered.