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Device Technologies and Biomedical Robotics

Device Technologies and Biomedical Robotics - Poster Session B

Poster I14 - Detection of Childhood Obesity using Adiponectin as a biomarker using electrochemical biosensor: a Proof-of Concept study

Thursday, October 24, 2024
2:45 PM - 3:45 PM EST
Location: Exhibit Hall E, F & G

Introduction:  Childhood obesity is considered a complex chronic disease caused by excess adiposity with persistent weight gain into adulthood, which increases the risk of developing systemic diseases such as diabetes, heart disease, or cancer and early mortality [1]. The prevalence of childhood obesity is 8.24% worldwide, creating a global health concern [2]. However, current diagnostic tools for childhood obesity are invasive and may not be accurate [3]. Adiponectin, a critical adipokine secreted by adipose tissue, has significant antidiabetic and antiatherogenic properties [4]. During obesity conditions, this protein decreases in concentration due to a dysfunction in the adipose tissue and serves as a potential biomarker for early detection of childhood obesity [5]. Previous data has correlated adiponectin levels of known concentrations to biosensor data outputs and for liver cells and cardiomyocytes supernatant. By culturing adipocytes and evaluating them through a biosensor, we can more closely simulate human adipose tissue on an in vitro scale. Through this preliminary study, the efficacy of a biosensor using adiponectin secreted by adipocytes can be evaluated for potential use as a biosensor for childhood obesity detection.

Materials and

Methods:  Preadipocytes (Human Preadipocytes HAPd from Sigma) were grown and, once confluent, were treated with adipocyte differentiation media (Human Adipocytes Differentiation Medium-811D-250 from Sigma) every three days until preadipcytes are differentiated to adipocytes. The presence of adipocytes was confirmed using Oil Red O staining and imaged using phase contrast microscopy [6]. The presence of adiponectin protein in the cells and the spent media was visually verified using confocal microscopy by adding a secondary FITC-conjugated antibody to produce green fluorescence (Invitrogen Goat Anti-Mouse IgG Fc Secondary Antibody, FITC). The spent media of the differentiated adipocytes was further used with a gold screen-printed biosensor (DRP-250AT from DropSens), which was fabricated using adiponectin antibodies to test the efficacy of this non-invasive model. The biosensor is connected to Gamry made Potentiostat (Interface 100) and a specific electrochemical sequence was adopted for the sensor testing. The change in impedance (ΔZ), resistance (R), and capacitance (C) obtained from electrochemical impedance spectroscopy (EIS) and cyclic voltommetry (CV) area from CV plots were used to detect adiponectin on the electrochemical biosensor and compared with the standards of measurements of known adiponectin concentrations developed earlier.

Results, Conclusions, and Discussions: 

Results: Preadipocytes (Figure 1.a-i) were differentiated into mature adipocytes (Figure 1.a-ii) and verified by Oil Red O staining, which detected the oil droplets using phase contrast microscopy (Figure 1.a.-iii). The presence of adiponectin in cells and the supernatant was visually verified by confocal microscopy (Figure 1.b.). The site of storage of adiponectin within the cells is still unknown. The adipocyte spent media within the biosensor indicates the release of adiponectin, an essential component of biosensor development. The spent media was tested on the biosensor, which revealed the presence of adiponectin compared with the standardized concentrations of 5 μg/mL, 8.5 μg/mL and 12 μg/mL (Figure 1.c.).

Discussion: The results indicate the correlation between electrochemical responses and increasing adipocyte concentration. More specifically, the impedance change and CV area change could be considered indicative parameters. It is well established that the electrochemical responses will vary based on the chemistry and structure of the proteomic biomarker. The study's outcome exhibits the presence of adiponectin and its concentration, which can be between 5 μg/mL and 12 μg/mL. However, the study has many limitations, so future work will focus on conducting tests to generate more robust data sets and statistically accepted calibrations and standard curves for the better prediction. In addition, a machine-learning model can be developed to classify adiponectin concentrations based on risk levels in serum and saliva.

Conclusions: Differentiated adipocytes release adiponectin in spent media, which is detected by our electrochemical biosensor. Thus, this electrochemical biosensor is a possible avenue for early diagnosis of childhood obesity using adiponectin as a biomarker.

Acknowledgements (Optional):  UIC Rockford Campus and The Blazer Foundation

Reference:
[1] Morales Camacho et al. (2019). Diabetes/Metabolism Research and Reviews. 35. 10.1002/dmrr.3203.
[2] Ash, T et al. (2017). The international journal of behavioral nutrition and physical activity, 14(1), 113.
[3] Gutin I. (2022). Social science & medicine (1982), 294, 114714.
[4] Nguyen T. M. D. et al (2020).International journal of preventive medicine, 11, 136.
[5] Han Y, et al. Journal of Pharmaceutical Analysis. 2024 May;14(5):100913.
[6] You, J. S. et al (2014). Journal of the science of food and agriculture, 94(3), 568–575.