Cell-specific response of NSIP- and IPF-derived fibroblasts to the modification of the elasticity, biological properties, and 3D architecture of the substrate

The fibrotic fibroblasts derived from idiopathic pulmonary fibrosis (IPF) and nonspecific interstitial pneumonia (NSIP) are surrounded by specific environments, characterized by increased stiffness, aberrant extracellular matrix (ECM) composition, and altered lung architecture. The presented research was aimed at investigating the effect of biological, physical, and topographical modification of the substrate on the properties of IPF- and NSIP-derived fibroblasts, and searching for the parameters enabling their identification. Soft and stiff polydimethylsiloxane (PDMS) was chosen for the basic substrates, the properties of which were subsequently tuned. To obtain the biological modification of the substrates, they were covered with ECM proteins, laminin, fibronectin, and collagen. The substrates that mimicked the 3D structure of the lungs were prepared using two approaches, resulting in porous structures that resemble natural lung architecture and honeycomb patterns, typical of IPF tissue. The growth of cells on soft and stiff PDMS covered with proteins, traced using fluorescence microscopy, confirmed an altered behavior of healthy and IPF- and NSIP-derived fibroblasts in response to the modified substrate properties, enabling their identification. In turn, differences in the mechanical properties of healthy and fibrotic fibroblasts, determined using atomic force microscopy working in force spectroscopy mode, as well as their growth on 3D-patterned substrates were not sufficient to discriminate between cell lines

Fabrication and impact of fouling-reducing temperature-responsive POEGMA coatings with embedded CaCO3 nanoparticles on different cell lines

In the present work, we have successfully prepared and characterized novel nanocomposite material exhibiting temperature-dependent surface wettability changes, based on grafted brush coatings of non-fouling poly(di(ethylene glycol)methyl ether methacrylate) (POEGMA) with the embedded CaCO3 nanoparticles. Grafted polymer brushes attached to the glass surface were prepared in a three-step process using atom transfer radical polymerization (ATRP). Subsequently, uniform CaCO3 nanoparticles (NPs) embedded in POEGMA-grafted brush coatings were synthesized using biomineralized precipitation from solutions of CaCl2 and Na2CO3. An impact of the low concentration of the embedded CaCO3 NPs on cell adhesion and growth depends strongly on the type of studied cell line: keratinocytes (HaCaT), melanoma (WM35) and osteoblastic (MC3T3-e1). Based on the temperature-responsive properties of grafted brush coatings and CaCO3 NPs acting as biologically active substrate, we hope that our research will lead to a new platform for tissue engineering with modified growth of the cells due to the release of biologically active substances from CaCO3 NPs and the ability to detach the cells in a controlled manner using temperature-induced changes of the brush

Effect of substrate stiffness on physicochemical properties of normal and fibrotic lung fibroblasts

The presented research aims to verify whether physicochemical properties of lung fibroblasts, modified by substrate stiffness, can be used to discriminate between normal and fibrotic cells from idiopathic pulmonary fibrosis (IPF). The impact of polydimethylsiloxane (PDMS) substrate stiffness on the physicochemical properties of normal (LL24) and IPF-derived lung fibroblasts (LL97A) was examined in detail. The growth and elasticity of cells were assessed using fluorescence microscopy and atomic force microscopy working in force spectroscopy mode, respectively. The number of fibroblasts, as well as their shape and the arrangement, strongly depends on the mechanical properties of the substrate. Moreover, normal fibroblasts remain more rigid as compared to their fibrotic counterparts, which may indicate the impairments of IPF-derived fibroblasts induced by the fibrosis process. The chemical properties of normal and IPF-derived lung fibroblasts inspected using time-of-flight secondary ion mass spectrometry, and analyzed complexly with principal component analysis (PCA), show a significant difference in the distribution of cholesterol and phospholipids. Based on the observed distinctions between healthy and fibrotic cells, the mechanical properties of cells may serve as prospective diagnostic biomarkers enabling fast and reliable identification of idiopathic pulmonary fibrosis (IPF)

Effect of tuned elasticity and chemical modification of substrate on fibrotic and healthy lung fibroblasts

Response to substrate elasticity, dependent on mechanical properties of cells, differs for lung fibroblast derived from idiopathic pulmonary fibrosis (IPF) and the healthy ones. These altered interactions might potentially act as a ‘biomarker’ for easy and reliable IPF diagnosis. In this work, systematic studies on the effect of polydimethylsiloxane (PDMS) substrate elasticity, tuned stepwise from 600 kPa to 1.5 MPa on the growth of IPF–derived (LL97A) and healthy (LL24) lung fibroblasts were reported. Additionally, impact of substrate chemistry on both cell lines was studied for fibroblasts cultured on glass substrates modified with three organosilanes – 3-aminopropyltriethoxysilane (APTES), 3-mercaptopropyltriethoxysilane (MPTES) and 3-glycidyloxypropyl trimethoxysilane (GOPS), with different end groups. Finally, the effect of the simultaneous modification of mechanical and chemical properties on the cellular behavior was studied for fibroblast cultured on PDMS substrates covered with silanes. The growth of cells was traced using fluorescence microscopy and analyzed quantitatively by nucleus-cytoplasm ratio, indicating strong, cell-dependent impact of substrate elasticity dominating over effect of chemical modification

Discrimination between NSIP- and IPF-derived fibroblasts based on multi-parameter characterization of their growth, morphology and physic-chemical properties

Background: The aim of the research presented here was to find a set of parameters enabling discrimination between three types of fibroblasts, i.e., healthy ones and those derived from two disorders mimicking each other: idiopathic pulmonary fibrosis (IPF), and nonspecific interstitial pneumonia (NSIP). Methods: The morphology and growth of cells were traced using fluorescence microscopy and analyzed quantitatively using cell proliferation and substrate cytotoxicity indices. The viability of cells was recorded using MTS assays, and their stiffness was examined using atomic force microscopy (AFM) working in force spectroscopy (FS) mode. To enhance any possible difference in the examined parameters, experiments were performed with cells cultured on substrates of different elasticities. Moreover, the chemical composition of cells was determined using time-of-flight secondary ion mass spectrometry (ToF-SIMS), combined with sophisticated analytical tools, i.e., Multivariate Curve Resolution (MCR) and Principal Component Analysis (PCA). Results: The obtained results demonstrate that discrimination between cell lines derived from healthy and diseased patients is possible based on the analysis of the growth of cells, as well as their physical and chemical properties. In turn, the comparative analysis of the cellular response to altered stiffness of the substrates enables the identification of each cell line, including distinguishing between IPF- and NSIP-derived fibroblasts

Pokrzywka przewlekła jako potencjalny wskaźnik choroby ogólnoustrojowej

Wprowadzenie: Pokrzywka przewlekła stanowi złożony pod względemetiologicznym problem kliniczny, wymagający szerokiego paneludiagnostycznego w celu ustalenia ewentualnego związku z innymischorzeniami, zwłaszcza o charakterze zapalnym. Cel pracy: Ocena prawdopodobnych przyczyn pokrzywki przewlekłeju pacjentów hospitalizowanych w Klinice Chorób Skórnych i WenerycznychPomorskiej Akademii Medycznej. Materiał i metodyka: Analizie poddano 104 pacjentów w wieku od15 do 82 lat, w tym 73 kobiety i 31 mężczyzn. U chorych przeprowadzonobadania diagnostyczne obejmujące między innymi ocenę parametrówstanu zapalnego, wskaźników infekcji przewodu pokarmowegooraz chorób tarczycy i tkanki łącznej. Wykonano również próbyfizykalne oraz diagnostykę alergii IgE-zależnej. Uzyskane wyniki poddanoanalizie statystycznej z użyciem programu Statistica 7.0. Wyniki: Średnia wieku chorych wynosiła 42,5 roku, w każdej grupiewiekowej przeważały kobiety, które stanowiły łącznie 70% badanych.W przypadku 39,4% pacjentów nie znaleziono potencjalnego czynnikawywołującego objawy i rozpoznano pokrzywkę idiopatyczną.U 37,6% osób spośród ogółu badanych wskazano jedną, a u pozostałych23% osób – więcej niż jedną prawdopodobną przyczynę występowaniapokrzywki. Wśród potencjalnych przyczyn przeważałyprzewlekłe stany infekcyjne (34%), zwłaszcza w obrębie przewodupokarmowego. Wykazano również związek z chorobami gruczołutarczowego, głównie o etiologii autoimmunologicznej (16,3%). Podejrzenietła autoimmunologicznego pokrzywki potwierdzono próbąz surowicą autologiczną u 24,3% chorych. Pokrzywkę fizykalnąrozpoznano u 17,3%, a współwystępowanie alergii IgE-zależnejstwierdzono u 15,4% pacjentów. Wnioski: Pokrzywka przewlekła występuje przede wszystkimu kobiet w średnim wieku. Spośród współistniejących z nią choróbogólnoustrojowych szczególne miejsce zajmują zakażenia o przewlekłym,często utajonym lub subklinicznym przebiegu. W naszym ma -teriale liczną grupę stanowili pacjenci, u których stwierdzono współwystępowaniekilku potencjalnych przyczyn pokrzywki

Non-cytotoxic, temperature-responsive and antibacterial POEGMA based nanocomposite coatings with silver nanoparticles

Non-cytotoxic, temperature-responsive and antibacterial poly(di(ethylene glycol)methyl ether methacrylate) – POEGMA188 based nanocomposite coatings attached to a glass surface were successfully prepared using ATRP polymerization. The thickness, morphology and wettability of the resulting coatings were analyzed using ellipsometry, AFM and contact angle measurements, respectively. The strong impact of the thicknesses of the POEGMA188 grafted brush coatings and content of AgNPs on the morphology and temperature-induced wettability changes of the nanocomposite was demonstrated. In addition to the strong temperature-dependent antibacterial activity, the proposed nanocomposite coatings have no significant cytotoxic effect towards normal cells. Moreover, the slight anti-cancer effect of AgNPs may be suggested