Enhanced Cellular Uptake and Sustained Transdermal Delivery of Collagen for Skin Regeneration

Abstract

The present study reports a method for transporting high molecular weight collagen for skin regeneration. An independent engineered enzymatic vehicle that has the ability for efficient transdermal delivery of regenerative biomaterial was developed for tissue regeneration. Collagen has been well recognized as a skin regeneration molecule due to its interaction with the extracellular matrix to stimulate skin cell growth, proliferation, and differentiation. However, the transdermal delivery of collagen poses a significant challenge due to its high molecular weight as well as a lack of efficient approaches. Here, to improve the transdermal delivery efficiency, α-1,4-glycosidic hydrolase was engineered with genetically encoded 3,4-dihydroxy-L-phenylalanine, which enhanced its biological activity as revealed by microscale thermophoresis. The remodeled catalytic pocket resulted in enhanced substrate binding activity of the enzyme with a predominant glycosaminoglycan (chondroitin sulfate) present in the extracellular matrix of the skin. The engineered enzyme rapidly opened up the skin extracellular matrix fiber (15 min) to ferry collagen across the wall, without disturbing the cellular bundle architecture. Confocal microscopy indicated that macromolecules had diffused three times deeper into the engineered enzyme-treated skin than the native enzyme-treated skin. Gene expression, histopathology, and hematology analysis also supported the penetration of macromolecules. Cytotoxicity (mammalian cell culture) and in vivo (Caenorhabditis elegans and Rattus noryegicus) studies revealed that the congener enzyme could potentially be used as a penetration enhancer, which is of paramount importance for the multimillion cosmetic industries. Hence, it offers promise as a pharmaceutical enzyme for transdermal delivery bioenhancement and dermatological applications.

Redirecting the JAK‐STAT signal blocks the SARS‐CoV‐2 replication

“…A dose of 150 µg/mL of polyethylene glycol (PEG)conjugated IFN-β, IFN-β, and IFN-β-A was injected via the intraperitoneal route in animals. Blood samples (~300 µL) were drawn via a jugular catheter in the EDTA-coated collection tube at predetermined time points (2,4,6,12,24,48, 72 h) before and after protein administration. Before analysis, the blood sample was centrifuged to separate the serum.…”

“…All protein-receptor binding studies were performed at 40%−60% MST power. 24 The dissociation constant (K d ) was determined from MO. Binding affinity Nano Temper Analysis 1.2.231 (Nano Temper Technologies) using K d binding model equation based on the Langmuir binding isotherm.…”

Recombinant and genetic code expanded collagen-like protein as a tailorable biomaterial

“…The comprehension of the protein translational machinery of ribosomal biosynthesis provoked researchers to manipulate the orthogonal translational machinery and incorporate UAAs in proteins. [122][123][124] Genetic code expansion strategies are categorized into residue-specific and site-specific UAA incorporation (Fig. 6).…”

Protein‐based metal bio‐cleaner for detoxification of wastewater

“…Synthetic NCAAs offer non‐natural potential with new functional chemistry for proteins with enhanced spectral properties. Frequently, two experimental approaches, namely reassignment of sense (residue‐specific incorporation) and nonsense suppression (site‐specific incorporation), are used for in vivo incorporation of NCAAs into recombinant proteins 17‐25 . In the work reported here, we aimed to introduce donor–acceptor non‐natural chemistry in the GFP chromophore, offering significant Stokes shifts to GFP 11,26 .…”

“…Frequently, two experimental approaches, namely reassignment of sense (residue-specific incorporation) and nonsense suppression (site-specific incorporation), are used for in vivo incorporation of NCAAs into recombinant proteins. [17][18][19][20][21][22][23][24][25] In the work reported here, we aimed to introduce donor-acceptor non-natural chemistry in the GFP chromophore, offering significant Stokes shifts to GFP. 11,26 We aimed to incorporate aminotyrosine as an electron-donating group (amine group) into the GFP chromophore for the development of an efficient red-shifted fluorescent protein-based biosensor that can also act as a bio-cleaner for the removal of Cu 2+ ions from environmental samples.…”