<p>Study on Adenovirus Infection in vitro with Nanoself-Assembling Peptide as Scaffolds for 3D Culture</p>

Abstract

To construct a three-dimensional (3D) culture model of adenovirus in vitro using the nanoself-assembling peptide RADA16-I as a 3D cell culture scaffold combined with virology experimental technology to provide a novel research method for virus isolation and culture, pathogenesis research, antiviral drug screening and vaccine preparation. Methods: The nanoself-assembling peptide RADA16-I was used as a 3D scaffold material for 293T cell culture, and adenovirus was cultured in the cells. The growth, morphological characteristics and pathological effects of 3D-cultured 293T cells after adenovirus infection were observed with an inverted microscope and MTS. The proliferation of adenovirus in 293T cells was observed by TEM and detected by qPCR. The levels of TNF-α and IL-8 secreted by adenovirus-infected 293T cells in the RADA16-I 3D culture system were detected by ELISA. Results: The 293T cells grew well in the RADA16-I 3D culture system for a prolonged period of time. The adenovirus infection persisted for a long time with multiple proliferation peaks, which closely resembled those of in vivo infections. The adenovirus virions amplified in the 3D system remained infectious. There were multiple secretion peaks of TNF-α and IL-8 secretion levels in adenovirus-infected 293T cells cultured in 3D culture systems. Conclusion: The nanoself-assembling peptide RADA16-I can be used as a 3D scaffold for adenovirus isolation, culture and research. The 3D culture system shows more realistic in vivo effects than two-dimensional (2D) culture.

A Novel Airway-Organoid Model Based on a Nano-Self-Assembling Peptide: Construction and Application in Adenovirus Infection Studies

“…Research has shown that adenovirus can continue to proliferate in a 3D culture model. 27 , 28 However, viral infection in vivo is a complex process, and the target organ of infection is often composed of a variety of cells. Respiratory bronchioles originate from the endoderm and are the target organ of adenovirus infection.…”

Recent research progress of biologically active peptides

“…Especially in recent years, the research on bioactive peptides has gradually become active, and the prospects are becoming more and more promising. The current research and development directions of BAPs are roughly as follows: (1) The discovery of new active peptides, structural analysis, and activity research, and so forth 19 ; (2) Development of targeted enzymatic hydrolysis technology for protein preparation of active peptides, including high‐efficiency and specific enzyme‐producing strains breeding, research on the joint action mechanism of complex enzyme systems, improvement of enzymatic hydrolysis technology, and so forth 20 ; (3) Separation of functional peptides and development of analytical techniques, especially high‐sensitivity, simple, and easy target peptide activity analysis and detection systems and analytical techniques 21 ; (4) Research on functional biological evaluation of peptides 22 ; (5) Development of bioactive peptide medicines and functional foods 23 ; (6) Engineering research on low‐cost, high‐purity, and mass production of bioactive peptides 24 ; (7) Application research of bioengineering technology in the preparation of active peptides, such as protein engineering, 25 enzyme engineering, 26 genetic engineering, 27 cell culture technology, 28 and so forth.…”

“…(3) Separation of functional peptides and development of analytical techniques, especially high-sensitivity, simple, and easy target peptide activity analysis and detection systems and analytical techniques 21 ; (4) Research on functional biological evaluation of peptides 22 ; (5) Development of bioactive peptide medicines and functional foods 23 ; (6) Engineering research on low-cost, highpurity, and mass production of bioactive peptides 24 ; (7) Application research of bioengineering technology in the preparation of active peptides, such as protein engineering, 25 enzyme engineering, 26 genetic engineering, 27 cell culture technology, 28 and so forth.…”