Leão Juliane Caroline, Mazzu Thiago, Leão Vitor, Souza Paola Gomes, Moura Nathalya Maria Vilela, Carrilho Emanuel, Taba Mario
Department of Oral and Maxillofacial Surgery and Traumatology and Periodontology, School of Dentistry of Ribeirao Preto, University of Sao Paulo, Ribeirao Preto 14040-904, Sao Paulo, Brazil.
Bioanalytics, Microfabrication and Separations Group, Chemistry Institute of Sao Carlos, University of Sao Paulo, Sao Carlos 13566-590, Sao Paulo, Brazil.
Dent J (Basel). 2025 Jul 15;13(7):321. doi: 10.3390/dj13070321.
To develop a microfluidic paper-based analytical device (μPAD) that identifies myeloperoxidase (MPO) levels in the saliva of healthy patients and those with periodontal disease. A platform similar to a 96-well plate was printed on Watman chromatography paper to run the experimental analysis with unstimulated saliva samples were collected from two groups of patients: those with periodontal health (H, n = 15) and established periodontitis (PD, n = 15). Then, three types of chromophore substrates were pipetted into the wells of the prototype: (1) Guaiacol; (2) Guaiacol, 4,4 '-diaminodifenilsulfon (DAB) and hydrogen peroxide in Tris-HCl buffer; and (3) 3,3',5,5'-Tetramethylbenzidine (TMB), followed by saliva samples. The reaction images were analyzed by numbering according to the intensity scale. The comparative results of the reactions using μPAD demonstrated that both the H and PD groups were compatible with each other without differences among the chromophore substrates ( > 0.05). However, the protocol with TMB showed a faster reaction and better color difference when comparing 15.62 ng/mL and 7.81 ng/mL of MPO in the plate embedded with Guaiacol; 1000 ng/mL and 62.5 ng/mL on the Guaiacol and DAB plate; and 62.5 ng/mL of TMB. The average detectable concentrations of MPO in saliva using TMB were H = 21.2 ± 10.4 ng/mL and PD = 28.9 ± 12.8 ng/mL ( = 0.08). The developed microfluidic paper-based analytical device has been tested for identifying the myeloperoxidase saliva levels of healthy patients and those with periodontal disease. This rapid test demonstrated its possible applicability mainly when associated with the TMB chromophore, but further studies are required with different biomarkers to explore this promising diagnostic platform.
开发一种基于微流控纸的分析装置（μPAD），用于识别健康患者和牙周病患者唾液中的髓过氧化物酶（MPO）水平。在沃特曼层析纸上打印一个类似于96孔板的平台，以便对从未受刺激的唾液样本进行实验分析，这些样本来自两组患者：牙周健康组（H，n = 15）和确诊的牙周炎组（PD，n = 15）。然后，将三种类型的发色底物移液到原型的孔中：（1）愈创木酚；（2）愈创木酚、4,4'-二氨基二苯砜（DAB）和Tris-HCl缓冲液中的过氧化氢；（3）3,3',5,5'-四甲基联苯胺（TMB），随后加入唾液样本。根据强度标度对反应图像进行编号分析。使用μPAD进行反应的比较结果表明，H组和PD组相互兼容，发色底物之间无差异（>0.05）。然而，与愈创木酚包埋板中15.62 ng/mL和7.81 ng/mL的MPO相比，TMB方案显示出更快的反应和更好的色差；在愈创木酚和DAB板上为1000 ng/mL和62.5 ng/mL；在TMB板上为62.5 ng/mL。使用TMB检测唾液中MPO的平均可检测浓度为H = 21.2±10.4 ng/mL，PD = 28.9±12.8 ng/mL（P = 0.08）。所开发的基于微流控纸的分析装置已用于识别健康患者和牙周病患者唾液中的髓过氧化物酶水平。这种快速检测方法主要在与TMB发色团相关时显示出其可能的适用性，但需要对不同的生物标志物进行进一步研究，以探索这个有前景的诊断平台。
