Investigation of mouse amniotic fluid for stimulating ability of keratinocyte differentiation depending on the fetal stage

Kuribayashi, Miki; Kawaguchi, Yusuke; Teshima, Hirofumi; Yamaguchi, Hisateru; Tatsukawa, Hideki; Hitomi, Kiyotaka

doi:10.1016/j.abb.2021.109003

Cited by 2 publications

(3 citation statements)

References 27 publications

Supporting

Mentioning

Contrasting

Order By: Relevance

“…During all the intrauterine life, foetal keratinocytes are subjected to peculiar chemical–physical conditions including their continuous exposure to amniotic fluid—a complex and dynamic milieu with emerging roles in foetal skin formation and function ( 60–62 ). Interestingly, the environmental stimuli challenging keratinocytes during foetal development are recognized to play a pathogenetic role in other congenital skin diseases, such as self-healing collodion baby, characterized by a dramatic amelioration of phenotype shortly after birth ( 63–65 ).…”

Section: Discussionmentioning

confidence: 99%

Proteasome-mediated degradation of keratins 7, 8, 17 and 18 by mutant KLHL24 in a foetal keratinocyte model: Novel insight in congenital skin defects and fragility of epidermolysis bullosa simplex with cardiomyopathy

Logli

Marzuolo

D'agostino

et al. 2021

Human Molecular Genetics

View full text Add to dashboard Cite

Epidermolysis bullosa simplex (EBS) with cardiomyopathy (EBS-KLHL24) is an EBS subtype caused by dominantly inherited, gain-of-function mutations in the gene encoding for the ubiquitin-ligase KLHL24, which addresses specific proteins to proteasomal degradation. EBS-KLHL24 patients are born with extensive denuded skin areas and skin fragility. Whilst skin fragility rapidly ameliorates, atrophy and scarring develop over time, accompanied by life-threatening cardiomyopathy. To date, pathogenetic mechanisms underlying such a unique disease phenotype are not fully characterized. The basal keratin 14 (K14) has been indicated as a KLHL24 substrate in keratinocytes. However, EBS-KLHL24 pathobiology cannot be determined by the mutation-enhanced disruption of K14 alone, as K14 is similarly expressed in foetal and postnatal epidermis and its protein levels are preserved both in vivo and in vitro disease models. In this study, we focused on foetal keratins as additional KLHL24 substrates. We showed that K7, K8, K17 and K18 protein levels are markedly reduced via proteasome degradation in normal foetal keratinocytes transduced with the mutant KLHL24 protein (ΔN28-KLHL24) as compared to control cells expressing the wild-type form. In addition, heat stress led to keratin network defects and decreased resilience in ΔN28-KLHL24 cells. The KLHL24-mediated degradation of foetal keratins could contribute to congenital skin defects in EBS-KLHL24. Furthermore, we observed that primary keratinocytes from EBS-KLHL24 patients undergo accelerated clonal conversion with reduced colony forming efficiency (CFE) and early replicative senescence. Finally, our findings pointed out a reduced CFE in ΔN28-KLHL24-transduced foetal keratinocytes as compared to controls, suggesting that mutant KLHL24 contributes to patients’ keratinocyte clonogenicity impairment.

show abstract

Section: Discussionmentioning

confidence: 99%

Proteasome-mediated degradation of keratins 7, 8, 17 and 18 by mutant KLHL24 in a foetal keratinocyte model: Novel insight in congenital skin defects and fragility of epidermolysis bullosa simplex with cardiomyopathy

Logli

Marzuolo

D'agostino

et al. 2021

Human Molecular Genetics

View full text Add to dashboard Cite

show abstract

“…[42][43][44][45] For instance, an innovative evaluation revealed that the amniotic fluid of mice improved the levels of keratogenic differentiation markers. 46 Additionally, possessing several biological properties like anti-microbial, anti-inflammatory, anti-scarring, as well as low immunogenicity, AM can be blended with PCL, and the fabricated structure may be a good scaffolding option for skin repair. 25,47 There exist several methods to produce threedimensional (3D) scaffolds like as electrospinning and decellularization method.…”

Section: Introductionmentioning

confidence: 99%

“…Studies have shown that the EGF and keratinocyte GF contents of the AM can play a key role in the stem cells' keratinogenic differentiation 42–45 . For instance, an innovative evaluation revealed that the amniotic fluid of mice improved the levels of keratogenic differentiation markers 46 . Additionally, possessing several biological properties like anti‐microbial, anti‐inflammatory, anti‐scarring, as well as low immunogenicity, AM can be blended with PCL, and the fabricated structure may be a good scaffolding option for skin repair 25,47 …”

Section: Introductionmentioning

confidence: 99%

Fabrication and multiscale modeling of polycaprolactone/amniotic membrane electrospun nanofiber scaffolds for wound healing

et al. 2023

View full text Add to dashboard Cite

BackgroundEnhancing the efficiency of cell‐based skin tissue engineering (TE) approaches is possible via designing electrospun scaffolds possessing natural materials like amniotic membrane (AM) with wound healing characteristics. Concentrating on this aim, we fabricated innovative polycaprolactone (PCL)/AM scaffolds through the electrospinning process.MethodsThe manufactured structures were characterized by employing scanning electron microscope (SEM), attenuated total reflection‐Fourier transform infrared (ATR‐FTIR) spectroscopy, tensile testing, Bradford protein assay, etc. In addition, the mechanical properties of scaffolds were simulated by the multiscale modeling method.ResultsAs a result of conducting various tests, it was concluded that the uniformity and distribution of fibers decreased with an increase in the amniotic content. Furthermore, PCL‐AM scaffolds contained amniotic and PCL characteristic bands. In the case of protein release, greater content of AM led to the release of higher amounts of collagen. Tensile testing revealed that scaffolds' ultimate strength increased when the AM content augmented. The multiscale modeling demonstrated that the scaffold had elastoplastic behavior. In order to assess cellular attachment, viability, and differentiation, human adipose‐derived stem cells (ASCs) were seeded on the scaffolds. In this regard, SEM and 3‐(4,5‐dimethylthiazol‐2‐yl)‐2,5‐diphenyl‐2H‐tetrazolium bromide (MTT) assays showed significant cellular proliferation and viability on the proposed scaffolds, and these analyses illustrated that higher cell survival and adhesion could be achieved when scaffolds possessed a larger amount of AM. After 21 days of cultivation, particular keratinocyte markers, such as keratin I and involucrin, were identified through utilizing immunofluorescence and real‐time polymerase chain reaction (PCR) tests. The markers' expressions were higher in the PCL‐AM scaffold with a ratio of 90:10 v v−1 compared with the PCL‐epidermal growth factor (EGF) structure. Moreover, the presence of AM in the scaffolds resulted in the keratinogenic differentiation of ASCs even without employing EGF. Consequently, this state‐of‐the‐art experiment suggests that the PCL‐AM scaffold can be a promising candidate in skin bioengineering.ConclusionThis study showed that mixing AM with PCL, a widely used polymer, in different concentrations can overcome PCL disadvantages such as high hydrophobicity and low cellular compatibility.

show abstract

Investigation of mouse amniotic fluid for stimulating ability of keratinocyte differentiation depending on the fetal stage

Cited by 2 publications

References 27 publications

Proteasome-mediated degradation of keratins 7, 8, 17 and 18 by mutant KLHL24 in a foetal keratinocyte model: Novel insight in congenital skin defects and fragility of epidermolysis bullosa simplex with cardiomyopathy

Proteasome-mediated degradation of keratins 7, 8, 17 and 18 by mutant KLHL24 in a foetal keratinocyte model: Novel insight in congenital skin defects and fragility of epidermolysis bullosa simplex with cardiomyopathy

Fabrication and multiscale modeling of polycaprolactone/amniotic membrane electrospun nanofiber scaffolds for wound healing

Contact Info

Product

Resources

About