Multidisciplinary Department of Medical-Surgical and Dental Specialties, University of Campania Luigi Vanvitelli, Via L. De Crecchio, 6, 80138, Naples, Italy.
Department of Precision Medicine, University of Campania Luigi Vanvitelli, Via L. De Crecchio 7, 80138, Naples, Italy.
J Dent. 2023 Jul;134:104553. doi: 10.1016/j.jdent.2023.104553. Epub 2023 May 17.
The evaluation of salivary biomarkers has been proposed as a simple and non-invasive aid to the conventional periodontal diagnosis based on clinical-radiographic parameters. Matrix metalloproteinase-8 (MMP-8), especially in its active form, is considered one of the most reliable biomarkers of periodontitis, and point-of-care tests (POCTs) have been proposed for its clinical monitoring. In this proof-of-concept study, a novel highly sensitive POCT based on a plastic optical fiber (POF) biosensor exploiting surface plasmon resonance (SPR) to detect salivary MMP-8 is described.
A SPR-POF biosensor was functionalized with a specific antibody to develop a surface-assembled monolayer (SAM) for the detection of total MMP-8. A white light source and a spectrometer connected to the biosensor were used to quantify MMP-8 level in both buffer and real matrix (saliva) by analysing the shift of the resonance wavelength determined by the specific antigen-antibody binding upon the SAM.
Dose-response curves by serial dilutions of human recombinant MMP-8 were realized, obtaining a limit of detection (LOD) of 40 pM (1.76 ng/ml) in buffer and 225 pM (9.9 ng/ml) in saliva and high selectivity compared to interferent analytes (MMP-2 and IL-6).
The proposed optical fiber-based POCT was able to detect and measure total MMP-8 with high selectivity and very low LOD in both buffer and saliva.
The SPR-POF technology may be employed to create highly sensitive biosensors to monitor salivary MMP-8 levels. The possibility of specifically detecting its active, rather than total, form need to be further investigated. If confirmed and clinically validated, such a device may represent a promising tool to make an immediate, highly sensitive and reliable diagnosis of periodontitis, and to carry out a timely and targeted therapy, possibly helping to prevent the onset of local and systemic periodontitis-related complications.
基于临床-影像学参数的传统牙周病诊断,人们提出了评估唾液生物标志物的方法,作为一种简单、非侵入性的辅助手段。基质金属蛋白酶-8(MMP-8),尤其是其活性形式,被认为是牙周炎最可靠的生物标志物之一,并且已经提出了用于其临床监测的即时检测(POCT)。在这项概念验证研究中,描述了一种基于塑料光纤(POF)生物传感器的新型高灵敏度 POCT,该传感器利用表面等离子体共振(SPR)来检测唾液 MMP-8。
通过特定抗体功能化 SPR-POF 生物传感器,开发用于检测总 MMP-8 的表面组装单层(SAM)。使用白光光源和连接到生物传感器的分光光度计,通过分析 SAM 上特定抗原-抗体结合引起的共振波长的位移,定量缓冲液和真实基质(唾液)中的 MMP-8 水平。
通过人重组 MMP-8 的连续稀释实现了剂量反应曲线,在缓冲液中获得了 40 pM(1.76 ng/ml)的检测限(LOD),在唾液中获得了 225 pM(9.9 ng/ml)的 LOD,并且与干扰分析物(MMP-2 和 IL-6)相比具有高选择性。
所提出的基于光纤的 POCT 能够在缓冲液和唾液中以高选择性和非常低的 LOD 检测和测量总 MMP-8。
SPR-POF 技术可用于创建高灵敏度生物传感器,以监测唾液 MMP-8 水平。进一步研究需要确定是否可以特异性地检测其活性形式,而不是总形式。如果得到证实并在临床上得到验证,这种设备可能成为一种有前途的工具,可以立即进行高度敏感和可靠的牙周炎诊断,并进行及时和有针对性的治疗,可能有助于预防局部和全身牙周炎相关并发症的发生。