Salivary diagnostics on paper microfluidic devices and their use as wearable sensors for glucose monitoring

Microfluidic paper-based devices (μPADs) and wearable devices have been highly studied to be used as diagnostic tools due to their advantages such as simplicity and ability to provide instrument-free fast results. Diseases such as periodontitis and diabetes mellitus can potentially be detected through these devices by the detection of important biomarkers. This study describes the development of μPADs through craft cutter printing for glucose and nitrite salivary diagnostics. In addition, the use of μPADs integrated into a mouthguard as a wearable sensor for glucose monitoring is also presented. μPADs were designed to contain two detection zones for glucose and nitrite assays and a sampling zone interconnected by microfluidic channels. Initially, the analytical performance of the proposed μPADs was investigated and it provided linear behavior (r2 ≥ 0.994) in the concentration ranges between 0 to 2.0 mmol L−1 and 0 to 400 μmol L−1 for glucose and nitrite, respectively. Under the optimized conditions, the limits of detection achieved for glucose and nitrite were 27 μmol L−1 and 7 μmol L−1, respectively. Human saliva samples were collected from healthy individuals and patients previously diagnosed with periodontitis or diabetes and then analyzed on the proposed μPADs. The results found using μPADs revealed higher glucose concentration values in saliva collected from patients diagnosed with diabetes mellitus and greater nitrite concentrations in saliva collected from patients diagnosed with periodontitis, as expected. The results obtained on μPADs did not differ statistically from those measured by spectrophotometry. With the aim of developing paper-based wearable sensors, μPADs were integrated, for the first time, into a silicone mouthguard using a 3D-printed holder. The proof of concept was successfully demonstrated through the monitoring of the glucose concentration in saliva after the ingestion of chocolate. According to the results reported herein, paper-based microfluidic devices offer great potential for salivary diagnostics, making their integration into a silicone mouthguard possible, generating simple, low-cost, instrument-free, and powerful wearable sensors.

[1]  Yixiang Duan,et al.  Saliva: a potential media for disease diagnostics and monitoring. , 2012, Oral oncology.

[2]  Wendell K T Coltro,et al.  Highly sensitive colorimetric detection of glucose and uric acid in biological fluids using chitosan-modified paper microfluidic devices. , 2016, The Analyst.

[3]  P. Clarys,et al.  Evaluation and review of body fluids saliva, sweat and tear compared to biochemical hydration assessment markers within blood and urine , 2017, European Journal of Clinical Nutrition.

[4]  C. Henry,et al.  Uncovering the Formation of Color Gradients for Glucose Colorimetric Assays on Microfluidic Paper-Based Analytical Devices by Mass Spectrometry Imaging. , 2018, Analytical chemistry.

[5]  Gokhan Demirel,et al.  Vapor-phase deposition of polymers as a simple and versatile technique to generate paper-based microfluidic platforms for bioassay applications. , 2014, The Analyst.

[6]  T. Kavin,et al.  Diabetes and periodontal disease , 2012, Journal of pharmacy & bioallied sciences.

[7]  Bowei Li,et al.  Portable paper‐based device for quantitative colorimetric assays relying on light reflectance principle , 2014, Electrophoresis.

[8]  Prashant Kumar,et al.  Saliva: A tool in assessing glucose levels in Diabetes Mellitus. , 2014, Journal of international oral health : JIOH.

[9]  S. Shevkoplyas,et al.  Integrated separation of blood plasma from whole blood for microfluidic paper-based analytical devices. , 2012, Lab on a chip.

[10]  Xu Li,et al.  A perspective on paper-based microfluidics: Current status and future trends. , 2012, Biomicrofluidics.

[11]  Bingcheng Lin,et al.  A rapid, straightforward, and print house compatible mass fabrication method for integrating 3D paper‐based microfluidics , 2013, Electrophoresis.

[12]  Wenyue Li,et al.  Smartphone quantifies Salmonella from paper microfluidics. , 2013, Lab on a chip.

[13]  L. Fu,et al.  Microfluidic paper-based platform for whole blood creatinine detection , 2018, Chemical Engineering Journal.

[14]  C. Henry,et al.  Development of Paper-Based Analytical Devices for Minimizing the Viscosity Effect in Human Saliva , 2018, Theranostics.

[15]  C. Culbertson,et al.  Paper-based microfluidic devices for analysis of clinically relevant analytes present in urine and saliva , 2010, Analytical and bioanalytical chemistry.

[16]  Shruti Gupta,et al.  Comparison of Salivary and Serum Glucose Levels in Diabetic Patients , 2014, Journal of Diabetes Science and Technology.

[17]  Ali Bijani,et al.  Diagnostic Role of Salivary and GCF Nitrite, Nitrate and Nitric Oxide to Distinguish Healthy Periodontium from Gingivitis and Periodontitis , 2014 .

[18]  Emanuel Carrilho,et al.  Determination of nitrite in saliva using microfluidic paper-based analytical devices. , 2014, Analytica chimica acta.

[19]  Yang Hao,et al.  Detecting Vital Signs with Wearable Wireless Sensors , 2010, Sensors.

[20]  Gi-Ja Lee,et al.  Paper-based 3D microfluidic device for multiple bioassays , 2015 .

[21]  Ali Khademhosseini,et al.  Wearables in Medicine , 2018, Advanced materials.

[22]  Peter Griess,et al.  Bemerkungen zu der Abhandlung der HH. Weselsky und Benedikt „Ueber einige Azoverbindungen” , 1879 .

[23]  G. Whitesides,et al.  Measuring markers of liver function using a micropatterned paper device designed for blood from a fingerstick. , 2012, Analytical chemistry.

[24]  Alexander Travis Adams,et al.  EmotionCheck: A Wearable Device to Regulate Anxiety through False Heart Rate Feedback , 2017, GETMBL.

[25]  Hyun C. Yoon,et al.  Paper-based glucose biosensing system utilizing a smartphone as a signal reader , 2014, BioChip Journal.

[26]  Kunnyun Kim,et al.  Flexible heartbeat sensor for wearable device. , 2017, Biosensors & bioelectronics.

[27]  G. Whitesides,et al.  Simple telemedicine for developing regions: camera phones and paper-based microfluidic devices for real-time, off-site diagnosis. , 2008, Analytical chemistry.

[28]  E. Carrilho,et al.  Simultaneous determination of renal function biomarkers in urine using a validated paper-based microfluidic analytical device. , 2018, Analytica chimica acta.

[29]  L. Busch,et al.  Total salivary nitrates and nitrites in oral health and periodontal disease. , 2014, Nitric oxide : biology and chemistry.

[30]  Yunqing Du,et al.  Noninvasive glucose monitoring using saliva nano-biosensor , 2015 .

[31]  Emanuel Carrilho,et al.  Technical aspects and challenges of colorimetric detection with microfluidic paper-based analytical devices (μPADs) - A review. , 2017, Analytica chimica acta.

[32]  Ali K. Yetisen,et al.  A smartphone algorithm with inter-phone repeatability for the analysis of colorimetric tests , 2014 .

[33]  Hye Rim Cho,et al.  Wearable/disposable sweat-based glucose monitoring device with multistage transdermal drug delivery module , 2017, Science Advances.

[34]  Wei Wang,et al.  Bienzyme colorimetric detection of glucose with self-calibration based on tree-shaped paper strip , 2014 .

[35]  Subhas Chandra Mukhopadhyay,et al.  Wearable Sensors for Human Activity Monitoring: A Review , 2015, IEEE Sensors Journal.

[36]  David Sinton,et al.  Turning the Page: Advancing Paper-Based Microfluidics for Broad Diagnostic Application. , 2017, Chemical reviews.

[37]  John Herbert,et al.  Data Management within mHealth Environments: Patient Sensors, Mobile Devices, and Databases , 2012, JDIQ.

[38]  Shalini Prasad,et al.  Lancet-free and label-free diagnostics of glucose in sweat using Zinc Oxide based flexible bioelectronics , 2017 .

[39]  G. Whitesides,et al.  Patterned paper as a platform for inexpensive, low-volume, portable bioassays. , 2007, Angewandte Chemie.

[40]  Cormac J. Sreenan,et al.  A context aware wireless body area network (BAN) , 2009, 2009 3rd International Conference on Pervasive Computing Technologies for Healthcare.

[41]  HerbertJohn,et al.  Data Management within mHealth Environments , 2012 .

[42]  Shen Wei,et al.  Microfluidic paper-based analytical device for the determination of nitrite and nitrate. , 2014, Analytical chemistry.

[43]  Emanuel Carrilho,et al.  Recent advances in low‐cost microfluidic platforms for diagnostic applications , 2014, Electrophoresis.

[44]  Ke Yang,et al.  Novel developments in mobile sensing based on the integration of microfluidic devices and smartphones. , 2016, Lab on a chip.

[45]  R. Bhadra,et al.  NIH Public Access , 2014 .

[46]  Seok Hyun Yun,et al.  Contact Lens Sensors in Ocular Diagnostics , 2015, Advanced healthcare materials.

[47]  A. Batista,et al.  Nitric oxide synthesis and severity of human periodontal disease. , 2002, Oral diseases.

[48]  V. Gupta,et al.  Noninvasive Method for Glucose Level Estimation by Saliva , 2013 .

[49]  R. Faria,et al.  A simple method to produce 2D and 3D microfluidic paper-based analytical devices for clinical analysis. , 2017, Analytica chimica acta.

[50]  Fei Li,et al.  Advances in paper-based point-of-care diagnostics. , 2014, Biosensors & bioelectronics.

[51]  L. Capitán-Vallvey,et al.  Smartphone-based simultaneous pH and nitrite colorimetric determination for paper microfluidic devices. , 2014, Analytical chemistry.