Abstract
The normal function of the airway epithelium is vital for the host’s well-being. Conditions that might compromise the structure and functionality of the airway epithelium include congenital tracheal anomalies, infection, trauma and post-intubation injuries. Recently, the onset of COVID-19 and its complications in managing respiratory failure further intensified the need for tracheal tissue replacement. Thus far, plenty of naturally derived, synthetic or allogeneic materials have been studied for their applicability in tracheal tissue replacement. However, a reliable tracheal replacement material is missing. Therefore, this study used a tissue engineering approach for constructing tracheal tissue. Human respiratory epithelial cells (RECs) were isolated from nasal turbinate, and the cells were incorporated into a calcium chloride-polymerized human blood plasma to form a human tissue respiratory epithelial construct (HTREC). The quality of HTREC in vitro, focusing on the cellular proliferation, differentiation and distribution of the RECs, was examined using histological, gene expression and immunocytochemical analysis. Histological analysis showed a homogenous distribution of RECs within the HTREC, with increased proliferation of the residing RECs within 4 days of investigation. Gene expression analysis revealed a significant increase (p < 0.05) in gene expression level of proliferative and respiratory epithelial-specific markers Ki67 and MUC5B, respectively, within 4 days of investigation. Immunohistochemical analysis also confirmed the expression of Ki67 and MUC5AC markers in residing RECs within the HTREC. The findings show that calcium chloride-polymerized human blood plasma is a suitable material, which supports viability, proliferation and mucin secreting phenotype of RECs, and this suggests that HTREC can be a potential candidate for respiratory epithelial tissue reconstruction.
Highlights
1 from morphology, which sample is known as typical morphology of respiratory epithelial cellsmor[17,33,34]
3A)revealed revealedthat thatthe theexpression expression of of the Ki67 gene had been significantly (p < 0.05) upregulated at day 4 as compared to the Ki67 gene had been significantly (p < 0.05) upregulated at day 4 as compared to day 0 of day 0 of incorporating the respiratory epithelial cells (RECs) into the CaCl2 -polymerised human plasma. This shows a incorporating the RECs into the CaCl2-polymerised human plasma. This shows a significant significant increase in proliferation of residing RECs within the human tissue respiratory epithelial construct (HTREC) and this finding increase in proliferation of residing RECs within the HTREC and this finding is consistent is consistent with the hematoxylin and eosin (H&E) results
In our previous study [32], we proved that RECs from nasal turbinate could be used as a replacement to RECs isolated from the trachea
Summary
Treatments utilizing foreign materials such as titanium mesh [6,7], naturally derived materials such as cellulose [8], nonviable allograft [9], tissue engineering [10] and various types of tracheal transplantation have been tried. Application of such grafts often results in unfavorable outcomes such as chronic inflammation [7], immunogenicity [11]
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