scholarly journals Differential regulation of galectin expression/reactivity during wound healing in porcine skin and in cultures of epidermal cells with functional impact on migration

2009 ◽  
pp. 873-884
Author(s):  
J Klíma ◽  
L Lacina ◽  
B Dvořánková ◽  
D Herrmann ◽  
JW Carnwath ◽  
...  
Keyword(s):  
Wound Healing ◽  
Mammalian Cells ◽  
Human Keratinocytes ◽  
Differential Regulation ◽  
Epidermal Cells ◽  
Porcine Skin ◽  
New Information ◽  
Galectin 3 ◽  
Endogenous Lectins

The glycophenotyping of mammalian cells with plant lectins maps aspects of the glycomic profile and disease-associated alterations. A salient step toward delineating their functional dimension is the detection of endogenous lectins. They can translate sugar-encoded changes into cellular responses. Among them, the members of the lectin family of galectins are emerging regulators of cell adhesion, migration and proliferation. Focusing on galectins-1, -3 and -7, we addressed the issue whether their expression is regulated during wound healing in porcine skin as model. A conspicuous upregulation is detected for galectin-1 in the dermis and a neoexpression in the epidermis, where an increased level of galectin-7 was also found. Applying biotinylated tissue lectins as probes, the signal intensities for accessible binding sites decreased, intimating an interaction of the cell lectin with reactive sites. In contrast, galectin-3 parameters remained rather constant. Of note, epidermal cells in culture also showed an increase in expression/presence of galectin-1, measured on the levels of mRNA and protein, in this case by Western blotting and quantitative immunocytochemistry. Used as matrix, galectin-1 conferred resistance to trypsin treatment to attached human keratinocytes and reduced migration into scratch-wound areas in vitro. This report thus presents new information on endogenous lectins in wound healing and differential regulation among the three tested cases.

scholarly journals Achyrocline satureioides (Lam.) DC (Asteraceae) Extract-Loaded Nanoemulsions as a Promising Topical Wound Healing Delivery System: In Vitro Assessments in Human Keratinocytes (HaCaT) and HET-CAM Irritant Potential

Pharmaceutics ◽  
2021 ◽  
Vol 13 (8) ◽  
pp. 1241
Author(s):  
Lucélia Albarello Balestrin ◽  
Tainá Kreutz ◽  
Flávia Nathiely Silveira Fachel ◽  
Juliana Bidone ◽  
Nicolly Espindola Gelsleichter ◽  
...  
Keyword(s):  
Wound Healing ◽  
Hacat Cell ◽  
Chorioallantoic Membrane ◽  
Human Keratinocytes ◽  
Scratch Assay ◽  
Hen’S Egg ◽  
And Migration ◽  
Achyrocline Satureioides ◽  
Proliferation And Migration

Achyrocline satureioides (Lam.) DC Asteraceae extracts (ASEs) have been investigated for the treatment of various skin disorders. This study reports the effects of ASE-loaded nanoemulsions (NEASE) on the cellular viability, death by necrosis, and migration of immortalized human keratinocytes (HaCaT cell line), as well as the irritant potential through the hen’s egg chorioallantoic membrane test (HET-CAM). NEASE exhibited a polydispersity index above 0.12, with a droplet size of 300 nm, ζ-potential of −40 mV, and content of flavonoids close to 1 mg/mL. No cytotoxicity of the ASE was observed on HaCaT by MTT assay (up to 10 µg/mL). A significant increase of HaCaT viability was observed to NEASE (up to 5 μg/mL of flavonoids), compared to treatment with the ASE. The necrosis death evaluation demonstrated that only NEASE did not lead to cell death at all the tested concentrations. The scratch assay demonstrated that NEASE was able to increase the cell migration at low flavonoid concentrations. Finally, the HET-CAM test proved the non-irritative potential of NEASE. Overall, the results indicate the potential of the proposed formulations for topical use in wound healing, in view of their promising effects on proliferation and migration in keratinocytes, combined with an indication of the absence of cytotoxicity and non-irritating potential.

scholarly journals Evolution of Pseudomonas aeruginosa Virulence as a Result of Phage Predation

2013 ◽  
Vol 79 (19) ◽  
pp. 6110-6116 ◽  
Author(s):  
Zeinab Hosseinidoust ◽  
Theo G. M. van de Ven ◽  
Nathalie Tufenkji
Keyword(s):  
Pseudomonas Aeruginosa ◽  
Mammalian Cells ◽  
Bacterial Infections ◽  
Membrane Damage ◽  
Human Keratinocytes ◽  
Host Population ◽  
Resistant Bacteria ◽  
Content Type ◽  
Metabolic Action

ABSTRACTThe rapid increase in the emergence of antibiotic-resistant bacteria has attracted attention to bacteriophages for treating and preventing bacterial infections. Bacteriophages can drive the diversification ofPseudomonas aeruginosa, giving rise to phage-resistant variants with different phenotypes from their ancestral hosts. In this study, we sought to investigate the effect of phage resistance on cytotoxicity of host populations toward cultured mammalian cells. The library of phage-resistantP. aeruginosaPAO1 variants used was developed previously via experimental evolution of an isogenic host population using phages PP7 and E79. Our results presented herein indicate that the phage-resistant variants developed in a heterogeneous phage environment exhibit a greater ability to impede metabolic action of cultured human keratinocytes and have a greater tendency to cause membrane damage even though they cannot invade the cells in large numbers. They also show a heightened resistance to phagocytosis by model murine macrophages. Furthermore, all isolates produced higher levels of at least one of the secreted virulence factors, namely, total proteases, elastase, phospholipase C, and hemolysins. Reverse transcription-quantitative PCR (RT-qPCR) revealed upregulation in the transcription of a number of genes associated with virulence ofP. aeruginosafor the phage-resistant variants. The results of this study indicate a significant change in thein vitrovirulence ofP. aeruginosafollowing phage predation and highlight the need for caution in the selection and design of phages and phage cocktails for therapeutic use.

scholarly journals Human Keratinocyte Growth and Differentiation on Acellular Porcine Dermal Matrix in relation to Wound Healing Potential

2012 ◽  
Vol 2012 ◽  
pp. 1-8 ◽  
Author(s):  
Robert Zajicek ◽  
Vaclav Mandys ◽  
Ondrej Mestak ◽  
Jan Sevcik ◽  
Radana Königova ◽  
...  
Keyword(s):  
Wound Healing ◽  
Human Keratinocytes ◽  
Dermal Matrix ◽  
Keratinocyte Proliferation ◽  
Proliferation And Differentiation ◽  
A Cell ◽  
Cell Culture Assay ◽  
Air Liquid Interface

A number of implantable biomaterials derived from animal tissues are now used in modern surgery. Xe-Derma is a dry, sterile, acellular porcine dermis. It has a remarkable healing effect on burns and other wounds. Our hypothesis was that the natural biological structure of Xe-Derma plays an important role in keratinocyte proliferation and formation of epidermal architecturein vitroas well asin vivo. The bioactivity of Xe-Derma was studied by a cell culture assay. We analyzed growth and differentiation of human keratinocytes culturedin vitroon Xe-Derma, and we compared the results with formation of neoepidermis in the deep dermal wounds treated with Xe-Derma. Keratinocytes cultured on Xe-Derma submerged in the culture medium achieved confluence in 7–10 days. After lifting the cultures to the air-liquid interface, the keratinocytes were stratified and differentiated within one week, forming an epidermis with basal, spinous, granular, and stratum corneum layers. Immunohistochemical detection of high-molecular weight cytokeratins (HMW CKs), CD29, p63, and involucrin confirmed the similarity of organization and differentiation of the cultured epidermal cells to the normal epidermis. The results suggest that the firm natural structure of Xe-Derma stimulates proliferation and differentiation of human primary keratinocytes and by this way improves wound healing.

scholarly journals IN VITRO ASSESSMENT OF THE BIOLOGICAL ACTIVITY OF A NEW REGENERATIVE AGENT PREPARED FROM THE CONCENTRATE OF DEPROTEINIZED DERMAL LAYER OF PORCINE SKIN

2020 ◽  
pp. 12-22
Author(s):  
Natalia Bezdieniezhnykh ◽  
Aleksandra Lykhova ◽  
Hennadii Borschevskyi ◽  
Kateryna Dyakun ◽  
Ievgen Kruglov
Keyword(s):  
Biological Activity ◽  
Cell Line ◽  
Mammalian Cells ◽  
Therapeutic Agent ◽  
Biological Effects ◽  
Mdbk Cell ◽  
Porcine Skin ◽  
Bovine Kidney ◽  
Dermal Layer

Background. Presently, a prospective direction for the development of regenerative medicine in the world is the search for regulatory molecules and the identification of molecular targets to stimulate the body's endogenous regenerative potential. The concentrate of the deproteinized dermal layer of porcine skin (СDDLPS) is a new therapeutic agent with restorative properties, the action of which is directed on the induction of the self resources of cells. Aim. The assessment of the effect of СDDLPS on the proliferative activity of mammalian cells of different histogenesis in vitro. Materials and Methods. To determine the amino acid composition of the СDDLPS liquid chromatography and biochemical methods were used. The biological effects and mechanisms of action of the drug were investigated by cell culture and molecular biological methods. The research was carried out using stable cell lines: human keratinocytes (HaCaT cell line), porcine endothelial cells (PAE cell line), bovine kidney cells (MDBK cell line) and mouse fibroblasts (3T3A31 cell line). Results. The cells of the bovine kidney MDBK cell line were the most sensitive to the effect of the CDDLPS. Also, the obtained results suggest that, depending on the concentration, the drug not only stimulates cell proliferation by 10–30 %, but also significantly enhances biosynthetic processes in cells, in particular, protein synthesis by 20–40 %. Conclusions. CDDLPS is an effective and affordable therapeutic agent with restorative properties, the biological activity of which manifests itself in the activation of cell biosynthetic and proliferative potentials and is comparable to effects of some growth factors, in particular epidermal growth factor

scholarly journals Antibacterial Multi-Layered Nanocellulose-Based Patches Loaded with Dexpanthenol for Wound Healing Applications

Nanomaterials ◽  
2020 ◽  
Vol 10 (12) ◽  
pp. 2469
Author(s):  
Daniela F. S. Fonseca ◽  
João P. F. Carvalho ◽  
Verónica Bastos ◽  
Helena Oliveira ◽  
Catarina Moreirinha ◽  
...  
Keyword(s):  
Wound Healing ◽  
Mechanical Performance ◽  
In Vitro Release ◽  
Human Keratinocytes ◽  
Controlled Drug Release ◽  
Release Mechanism ◽  
Layer By Layer ◽  
Number Of Layers ◽  
Migration Capacity

Antibacterial multi-layered patches composed of an oxidized bacterial cellulose (OBC) membrane loaded with dexpanthenol (DEX) and coated with several chitosan (CH) and alginate (ALG) layers were fabricated by spin-assisted layer-by-layer (LbL) assembly. Four patches with a distinct number of layers (5, 11, 17, and 21) were prepared. These nanostructured multi-layered patches reveal a thermal stability up to 200 °C, high mechanical performance (Young’s modulus ≥ 4 GPa), and good moisture-uptake capacity (240–250%). Moreover, they inhibited the growth of the skin pathogen Staphylococcus aureus (3.2–log CFU mL−1 reduction) and were non-cytotoxic to human keratinocytes (HaCaT cells). The in vitro release profile of DEX was prolonged with the increasing number of layers, and the time-dependent data imply a diffusion/swelling-controlled drug release mechanism. In addition, the in vitro wound healing assay demonstrated a good cell migration capacity, headed to a complete gap closure after 24 h. These results certify the potential of these multi-layered polysaccharides-based patches toward their application in wound healing.

Prostaglandin I1 analogues, SM-10902 and SM-10906, affect human keratinocytes and fibroblasts in vitro in a manner similar to PGE1: therapeutic potential for wound healing

1995 ◽  
Vol 287 (6) ◽  
pp. 539-545 ◽  
Author(s):  
F. Kaneko ◽  
J. -Z. Zhang ◽  
K. Maruyama ◽  
Y. Nihei ◽  
I. Ono ◽  
...  
Keyword(s):  
Wound Healing ◽  
Therapeutic Potential ◽  
Human Keratinocytes

scholarly journals A Novel Three-Polysaccharide Blend In Situ Gelling Powder for Wound Healing Applications

Pharmaceutics ◽  
2021 ◽  
Vol 13 (10) ◽  
pp. 1680
Author(s):  
Chiara Amante ◽  
Tiziana Esposito ◽  
Pasquale Del Del Gaudio ◽  
Veronica Di Di Sarno ◽  
Amalia Porta ◽  
...  
Keyword(s):  
Wound Healing ◽  
Wound Repair ◽  
Healing Process ◽  
Antimicrobial Properties ◽  
Human Keratinocytes ◽  
Wound Healing Process ◽  
Host Matrix ◽  
In Situ Gelling

In this paper, alginate/pectin and alginate/pectin/chitosan blend particles, in the form of an in situ forming hydrogel, intended for wound repair applications, have been successfully developed. Particles have been used to encapsulate doxycycline in order to control the delivery of the drug, enhance its antimicrobial properties, and the ability to inhibit host matrix metalloproteinases. The presence of chitosan in the particles strongly influenced their size, morphology, and fluid uptake properties, as well as drug encapsulation efficiency and release, due to both chemical interactions between the polymers in the blend and interactions with the drug demonstrated by FTIR studies. In vitro antimicrobial studies highlighted an increase in antibacterial activity related to the chitosan amount in the powders. Moreover, in situ gelling powders are able to induce a higher release of IL-8 from the human keratinocytes that could stimulate the wound healing process in difficult-healing. Interestingly, doxycycline-loaded particles are able to increase drug activity against MMPs, with good activity against MMP-9 even at 0.5 μg/mL over 72 h. Such results suggest that such powders rich in chitosan could be a promising dressing for exudating wounds.

scholarly journals A frog cathelicidin peptide effectively promotes cutaneous wound healing in mice

2018 ◽  
Vol 475 (17) ◽  
pp. 2785-2799 ◽  
Author(s):  
Jing Wu ◽  
Jun Yang ◽  
Xiaofang Wang ◽  
Lin Wei ◽  
Kai Mi ◽  
...  
Keyword(s):  
Wound Healing ◽  
Growth Factor ◽  
Mammalian Cells ◽  
Transforming Growth Factor ◽  
Transforming Growth Factor Β1 ◽  
Skin Wound ◽  
Promoting Effect ◽  
Monocyte Chemoattractant Protein 1

Although cathelicidins in mammals have been well characterized, little is known about the function of cathelicidin in amphibians. In the present study, a novel 24-residue peptide (cathelicidin-NV, ARGKKECKDDRCRLLMKRGSFSYV) belonging to the cathelicidin family was identified from the skin of the plateau frog Nanorana ventripunctata. Cathelicidin-NV showed strong wound healing-promoting activity in a murine model with a full-thickness dermal wound. It directly enhanced the proliferation of keratinocyte cells, resulting in accelerated re-epithelialization of the wound site. Cathelicidin-NV also promoted the proliferation of fibroblasts, the differentiation of fibroblasts to myofibroblasts and collagen production in fibroblasts, which are implicated in wound contraction and repair processes. Furthermore, cathelicidin-NV promoted the release of monocyte chemoattractant protein-1, tumor necrosis factor-α, vascular endothelial growth factor and transforming growth factor-β1 in vivo and in vitro, which are essential in the wound-healing processes such as migration, proliferation and differentiation. The MAPK (ERK, JNK and p38) signaling pathways were involved in the wound healing-promoting effect. Additionally, unlike other cathelicidins, cathelicidin-NV did not have any direct effect on microbes and showed no cytotoxicity and hemolytic activity toward mammalian cells at concentrations up to 200 µg/ml. This current study may facilitate the understanding of the cellular and molecular events that underlie quick wound healing in N. ventripunctata. In addition, the combination of these properties makes cathelicidin-NV an excellent candidate for skin wound therapeutics.

Intracellular redistribution of Ku immunoreactivity in response to cell-cell contact and growth modulating components in the medium

1996 ◽  
Vol 109 (7) ◽  
pp. 1937-1946 ◽  
Author(s):  
J.W. Fewell ◽  
E.L. Kuff
Keyword(s):  
Mammalian Cells ◽  
Culture Medium ◽  
Primary Cultures ◽  
Human Keratinocytes ◽  
Inhibitory Effect ◽  
Cell Contact ◽  
Nuclear Component ◽  
Confluent Cell ◽  
Cell Cell

Ku is a heterodimeric protein first recognized as a human autoantigen but now known to be widely distributed in mammalian cells. Analysis of repair-deficient mutant cells has shown that Ku is required for DNA repair, and roles in DNA replication and transcription have also been suggested on the basis of in vitro observations. Ku is generally regarded as a nuclear component. However, in the present paper, we show that a quantitatively significant fraction (half or more) of Ku is located in the cytoplasm of cultured primate cells, and that major changes in epitope accessibility of both nuclear and cytoplasmic Ku components are associated with the transition from sparse to confluent cell densities. The same changes in immunoreactivity were seen in HeLa, 293, CV-1 (monkey) and HPV-transformed keratinocyte cell lines, and in primary cultures of human keratinocytes. The immunostaining pattern of sparsely grown cells could be converted to the ‘confluent’ configuration by re-plating them at the same low density on a monolayer of mouse 3T3 cells. The confluent antigen pattern could also be induced in sparse cells within 15–30 minutes by exposure of the cells to serum- or Ca(2+)-free medium or overnight with 2 mM hydroxyurea. Somatostatin at 0.12 mM blocked the effects of serum/Ca2+ deprivation of Ku p70 antigen distribution in sparse CV-1 cells, and in confluent cultures reversed the usual nuclear concentration of p70 immunoreactivity. However, somatostatin did not alter the expected immunostaining patterns of p86. Preliminary studies indicate that sparse CV-1 cells, but not HeLa cells, respond to as little as 1 pM of TGF-beta 1 in the culture medium by the rapid appearance of nuclear immunoreactivity. TGF-alpha had no apparent effect. These findings are consistent with the participation of Ku in a signal transduction system responsive to the inhibitory effect of cell-cell contact on the one hand and to cytokines and growth-supportive components of the culture medium on the other.

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