The extracellular matrix of eggshell displays anti-inflammatory activities through NF-κB in LPS-triggered human immune cells

Abstract

Avian eggshell membrane (ESM) is a natural biomaterial that has been used as an alternative natural bandage on burned and cut skin injuries for >400 years in Asian countries, and is available in large quantities from egg industries. Our aim was to characterize ESM that was separated and processed from egg waste, and to study whether this material possesses anti-inflammatory properties, making it suitable as an ingredient in industrial production of low cost wound healing products. Our results show that the processed ESM particles retain a fibrous structure similar to that observed for the native membrane, and contain collagen, and carbohydrate components such as hyaluronic acid and sulfated glycosaminoglycans, as well as N-glycans, mostly with uncharged structures. Furthermore, both processed ESM powder and the ESM-derived carbohydrate fraction had immunomodulation properties in monocytes and macrophage-like cells. Under inflammatory conditions induced by lipopolysaccharide, the ESM powder and the isolated carbohydrate fraction reduced the activity of the transcription factor nuclear factor-κB. The expression of the immune regulating receptors toll-like receptor 4 and ICAM-1, as well as the cell surface glycoprotein CD44, all important during inflammation response, were down-regulated by these fractions. Interestingly, our experiments show that the two fractions regulated cytokine secretion differently: ESM depressed inflammation by increased secretion of the anti-inflammatory cytokine IL-10 while the carbohydrate fraction reduced secretions of the pro inflammatory cytokines IL-1β and IL-6. Also, the phosphorylation of p65 and p50 subunits of nuclear factor-κB, as well as nuclear localization, differed between processed ESM powder and carbohydrate fraction, suggesting different down-stream regulation during inflammation. In conclusion, processed ESM powder and its soluble carbohydrate components possess anti-inflammatory properties, demonstrating the potential of ESM as a novel biological wound dressing for treatment of chronic inflammatory wounds.

Processed eggshell membrane powder regulates cellular functions and increase MMP-activity important in early wound healing processes

“…Processed eggshell membrane powder (PEP) used in this study was prepared as previously described [ 16 ]. PEP was sterilized by gamma irradiation at 25 kGy before in vitro (cell culture) and in vivo (murine wound model) studies.…”

“…The eggshell membranes are a functional equivalent of ECM, which surrounds the egg white and serves both protective and supportive roles during eggshell mineralization [ 12 ]. Its composition and structural features have been highly characterized [ 13 ]; the meshwork of fibers is approximately 90% proteinaceous [ 14 ], and also contains complex carbohydrates such as glycosaminoglycans (GAGs) [ 15 ] and N-glycans [ 16 ]. Structural proteins such as cysteine-rich eggshell membrane proteins (CREMPS) and collagens (particularly type X) are abundant constituents, together with glycoproteins, Ca-regulatory proteins and enzymes with a disintegrin-like and metalloproteinase domain with thrombospondin type 1 motif (ADAMs) [ 13 , 17 – 20 ].…”

“…Eggshell membrane extracts have been demonstrated to affect MMP mRNA expression, and to be a positive supplier for the extracellular matrix environment of fibroblasts in vitro [ 21 ]. In this study, we have studied processed ESM powder (PEP), a material that mimics the structural properties of intact ECM demonstrated in our previous work [ 16 ]. Our focus has been on the impact of PEP on fibroblasts and MMP-activity in an in vitro cell culture system and an in vivo murine wound healing model.…”

Biomass as a sustainable resource for value‐added modern materials: a review

“…Some studies have reported that scaffolds can be used to release proper doses of drugs that depend on cytotoxicity of silver nanoparticles (AgNPs) 240. Vuong et al 241. reported that processed ESM powder and its soluble carbohydrate components exhibit anti‐inflammatory and immune modulator properties in monocytes and macrophages.…”

Reproductive management of poultry

“…27 Egg shell membranes or egg shell membrane powder have been used as natural bandage for burns and injuries in traditional Asian medicine. Egg shell membrane powder and its carbohydrate constituents have been recently been demonstrated to exert anti-inflammatory effects 119,120 supporting their effectiveness.…”