Chitosan-hyaluronan: promotion of mucociliary differentiation of respiratory epithelial cells and development of olfactory receptor neurons

Abstract

Developing a biomaterial that promotes regeneration of both respiratory epithelium (RE) and olfactory neuroepithelium (ON) improves the surgical outcome of endoscopic sinus surgery. Although chitosan (CS) inhibits mucociliary differentiation of RE, it has been reported to regenerate ON. In addition, hyaluronic acid (HA) has been demonstrated to promote regeneration of RE. Whether the composite CS þ HA would simultaneously benefit RE and ON remains unexplored. Human nasal respiratory epithelial cells (RECs) and olfactory neuroepithelial cells (ONCs) are respectively obtained from the RE and the ON. They are cultured in vitro and divided into groups undergoing four treatments, control, CS, HA, and CS þ HA and assessed by scanning electron microscope, immunocytochemistry, and Western blots following indicated growth conditions. RECs keep polygonal morphology with mucociliary differentiation in the CS þ HA group. The levels of E-cadherin, zonula occludens-1, mucin 5AC, and forkhead box protein J1 are significantly higher in the CS þ HA group than in the CS alone group. In addition, ONCs express lower cytokeratin 18 (CK18) and higher olfactory marker protein (OMP) in the CS þ HA group than in HA alone group. ONCs express more signal transduction apparatuses, adenylate cyclase 3, in CS and CS þ HA groups than in HA and controls. Chitosan-hyaluronan plays a part in promoting differentiation of ORNs and facilitating mucociliary differentiation of RECs. This composite is a promising biomaterial for the sinonasal application.

Evaluating Feasibility of Human Tissue Engineered Respiratory Epithelium Construct as a Potential Model for Tracheal Mucosal Reconstruction

“…The primary culture of human RECs was successfully established at passage 1 from a nasal turbinate sample (Figure 1). The RECs maintained well-defined polygonal morphology, which is known as typical morphology of respiratory epithelial cells [17,33,34]. Furthermore, the cells were found to be actively proliferating, evidenced by their shining borders, which could be more clearly seen under higher magnification.…”

New Opportunity to Formulate Intranasal Vaccines and Drug Delivery Systems Based on Chitosan

“…Initially, the polymer interacts with the mucosa, and is then deposited on the membrane [ 35 ]. Chitosan inhibits mucociliary development and differentiation by increasing factor TGF-b1 and interferes with the formation of tight junctions in nasal respiratory epithelial cells [ 55 , 56 ].…”

“…This is a physiological defense mechanism of the respiratory system which eliminates external substances by covering them with a layer of mucus, reducing the residence time of drugs administered intranasally [ 62 ]. However, the mucoadhesive property of chitosan is also responsible for the decrease of mucociliary clearance, because it adheres to the nasal mucosa and inhibits mucociliary development and differentiation by increasing the factor TGF-b1 [ 55 , 56 ]. The degree of this decrease is also influenced by the viscosity of the formulations [ 59 ].…”

“…An in vitro study on nasal respiratory epithelial cells showed that cell differentiation took place on the control sample and the cells had a polygonal morphology, whereas the morphology of the cells in the chitosan sample was irregular, without cilia formation. The results also showed that chitosan decreases mucus secretion, which blocks the access of pathogens through epithelial tissue by inhibiting mucus-secreting cells and ciliated cells [ 55 ].…”

“…Initially, the polymer interacts with the mucosa, and is then deposited on the membrane [35]. Chitosan inhibits mucociliary development and differentiation by increasing factor TGF-b1 and interferes with the formation of tight junctions in nasal respiratory epithelial cells [55,56]. The mucoadhesive property of the polymer is directly proportional to the size of the chain and its flexibility; the more flexible the chain is, the easier it is to form a network with the mucus, increasing penetration [35].…”