Olfactory epithelium, which detects and transmits odor signals, is critical for the function of olfactory system. Olfactory epithelium is able to recover spontaneously after injury under normal circumstances, but this ability is dampened in certain diseases or senility, which causes olfactory dysfunction. The olfactory epithelium consists of basal cells, sustentacular cells and olfactory sensory neurons. In order to develop an olfactory epithelial organoid containing multiple olfactory cell types in vitro, we used three-dimensional culture model and small molecules screening. This organoid system consists of horizontal basal-like cells, globose basal-like cells, sustentacular-like cells and olfactory sensory neurons-like cells. Through statistical analysis of clone diameter, immunofluorescence staining and qPCR detection of the expression level of related marker genes. We identified a series of growth factors and small molecule compounds that affected the proliferation, composition and gene expression of the organoids. CHIR-99021, an activator of Wnt signaling pathway, increased the colony formation and proliferation rate of olfactory epithelial organoids and the expression level of marker genes of olfactory sensory neurons-like cells. In addition, each factor in the culture system increased the proportion of c-Kit-positive globose basal-like cell colonies in organoids. Moreover, EGF and vitamin C were both beneficial to the expression of horizontal basal-like cell marker genes in organoids. The established olfactory epithelial organoid system mimicked the process of olfactory epithelial stem cells differentiating into various olfactory epithelial cell types, thus providing a research model for studying olfactory epithelial tissue regeneration, the pathological mechanism of olfactory dysfunction and drug screening for olfactory dysfunction treatment.
Humans ; Olfactory Mucosa/metabolism* ; Epithelial Cells ; Organoids/metabolism* ; Olfaction Disorders/metabolism*

OBJECTIVE: To study the expression of PCNA and LNGFr in olfactory epithelium of patients suffering from dysosmia caused by chronic sinusitis, and the function of LNGFr. METHOD: Forty-six patients undergoing FESS were chosen. Before operation, their olfactory functions were examined with CCCRC. According to their CCCRC scores, they were divided into three groups. Group A: Patients with chronic sinusitis and dysosmia 25 cases; Group B: Patients with chronic sinusitis and a normal olfactory function 10 cases; Group C: Patients with deviation of nasal septum and a normal olfactory function 11 cases. The expressions of PCNA and LNGFr were measured in olfactory mucosas of the three groups by immunohistochemistry. RESULT: In basal cells, the expression of PCNA and LNGFr in group A was higher than that in group B (P < 0.01). and in group C (P < 0.01). There was negative correlation between positive cells of PCNA and CCCRC score in basal cells of group A (r = -0.7441, P < 0.01); There was negative correlation between integral optical density of LNGFr and CCCRC score in basal cells of group A (r = -0.4407, P < 0.05). There was positive correlation between positive cells of PCNA and integral optical density of LNGFr in basal cells of group A (r = 0.5317, P < 0.01). CONCLUSION Basal cells proliferated dramatically in patients suffering from dysosmia caused by chronic sinusitis. The proliferating capacity of basal cells was related to up-regulation of LNGFr expression.
Chronic Disease ; Humans ; Immunohistochemistry ; Nerve Tissue Proteins ; metabolism ; Olfaction Disorders ; metabolism ; Olfactory Mucosa ; metabolism ; Proliferating Cell Nuclear Antigen ; metabolism ; Receptors, Nerve Growth Factor ; metabolism ; Sinusitis ; complications