TABLE 3.
Biomarkers and assays for nanotoxicity evaluation in 3D organoid models.
| Organoid | Markers | Indication | Analysis |
| Liver | Cell viability, morphology, Albumin, Bile, Cytochrome 450, ALT, GGT and AST, Catalase and carbamoyl phosphate synthetase-1 isoforms, and proinflammatory cytokines | Cytotoxicity, liver secretion, xenobiotic clearance, and oxidative stress | MTT assay, electron microscopy, biochemical, and gene expression study |
| Kidney | Apoptosis, morphology, Blood urea nitrogen (BUN), EPO, γ-glutamyltransferase production, SIX2, NPHS1, SAL1, WT1, PAX2, Bmp4, CITED1, KIM-1, trefoil factor 3 (TFF-3), beta-2 microglobulin, cystatin C, albumin, total protein, clusterin, neutrophil gelatinase-associated lipocalin, IL-18 and osteopontin, proinflammatory cytokines, and ROS generation | Cell viability, Kidney function, secretion, development and regeneration test, and oxidative stress | MTT assay, electron microscopy, biochemical, ELISA, and gene expression study |
| Heart | Alpha-actin, CD45, CD31, LDH, troponin, and heart beat assay | Cardiomyositis, contractile and ion channel function | Biochemical, ELISA, gene expression study, and ECG |
| Brain | Proinflammatory cytokines, ROS generation, CORIN, OTX2, MAP2, and TBR | Oxidative stress, Brain developmental analysis | Gene expression study, immunoassays |
| GIT | CyP3A4,CYP450, Cyp2b10, Cyp3a11, ABCB1/MDR1, and XPGs including alcohol dehydrogenase Adh1, aldehyde dehydrogenase Aldh1a1, and Aldh1a7, secrete gastric intrinsic factor, pepsinogen C, and mucin | GIT function test for detoxification, xenobiotic clearance and enzyme secretion | Gene expression study and ELISA or biochemical assay |
| Lung | KL6, MMP1&7, PFT ABG, and DLCO | Lung function and structural integrity analysis | Biochemical, ELISA, and gene expression study |
| Pancreas | Insulin, amylase, lipase, LDH, PDX1, NKX6.1, Synaptophysin, and islet amyloid polypeptide | Pancreas endocrine function, enzyme assay, and developmental assessment | Biochemical, ELISA and gene expression study |
| Tumor | M2-PK, Wnt Signaling pathways molecules, Her2, cytology, copy number alterations (CNAs), hematoxylin and eosin (H&E) staining, live dead cell, ATP measurement, and Annexin V-FITC/PI staining | Tumor-specific test | Gene expression study, Western blotting, immunohistochemistry, and microscopy |
| Testis | Testosterone and inhibin B | Function test | ELISA, Western blotting, and immunohistochemistry |
| Dental epithelial organoids | α-Amylase activity | Function test | ELISA, Western blotting, and immunohistochemistry |