Abstract
Increased collagen-derived advanced glycation end-products (AGEs) are consistently related to painful diseases, including osteoarthritis, diabetic neuropathy, and neurodegenerative disorders. We have recently developed a model combining a two-dimensional glycated extracellular matrix (ECM-GC) and primary dorsal root ganglion (DRG) that mimicked a pro-nociceptive microenvironment. However, culturing primary cells is still a challenge for large-scale screening studies. Here, we characterized a new model using ECM-GC as a stimulus for human sensory-like neurons differentiated from SH-SY5Y cell lines to screen for analgesic compounds. First, we confirmed that the differentiation process induces the expression of neuron markers (MAP2, RBFOX3 (NeuN), and TUBB3 (β-III tubulin), as well as sensory neuron markers critical for pain sensation (TRPV1, SCN9A (Nav1.7), SCN10A (Nav1.8), and SCN11A (Nav1.9). Next, we showed that ECM-GC increased c-Fos expression in human sensory-like neurons, which is suggestive of neuronal activation. In addition, ECM-GC upregulated the expression of critical genes involved in pain, including SCN9A and TACR1. Of interest, ECM-GC induced substance P release, a neuropeptide widely involved in neuroinflammation and pain. Finally, morphine, the prototype opiate, decreased ECM-GC-induced substance P release. Together, our results suggest that we established a functional model that can be useful as a platform for screening candidates for the management of painful conditions.
Highlights
Collagen glycation is frequently observed in neuroinflammatory and neurodegenerative disorders, such as Parkinson’s and Alzheimer’s diseases, osteoarthritis, diabetic neuropathy, amyloid polyneuropathy, and the aging process [1,2]
Collagen incubation with 200 mM D-ribose, 160 mM D-glucose and 200 mM D-threose for seven days resulted in a significant increase in fluorescence intensities of both total advanced glycation end products (AGEs) and pentosidine compared to normal, non-glycated collagen (Figure 2a,b), confirming that the glycation protocol produced highly modified collagen
Collagen-derived advanced glycation endproducts acting in human sensory neuronlike cells mimic a painful microenvironment that positively responds to analgesics
Summary
Collagen glycation is frequently observed in neuroinflammatory and neurodegenerative disorders, such as Parkinson’s and Alzheimer’s diseases, osteoarthritis, diabetic neuropathy, amyloid polyneuropathy, and the aging process [1,2]. Glycation is an irreversible and spontaneous process in which proteins, such as collagen, bind to sugar molecules, forming advanced glycation end products (AGEs) [3]. AGEs can activate specific cell surface receptors that transduce signals through downstream proteins, leading to the inflammatory process. Other AGE receptors, including AGE-R1/OST-48/p60, AGER2/80K-H/p90, and AGE/Galectin-3, are found in neuronal cells [4] and participate in AGE signaling. Collagen glycation modifies the extracellular matrix (ECM) structure and properties, resulting in matrix stiffening, which is frequently observed in osteoarthritic knee joints, for example. This process may affect the optimal biochemical functions and cell–matrix component interactions, potentially disrupting cellular homeostasis [6]. ECMGC-induced tissue damage and inflammation can result in excitation, neuropeptide release, and peripheral sensitization that contribute to the nociceptive process [7,8]
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