Ex Vivo Toxicity of Commonly Used Topical Antiseptics and Antibiotics on Human Chondrocytes.

Topical povidone-iodine, chlorhexidine, bacitracin, and vancomycin are commonly used antiseptic and antimicrobial agents to reduce risk and treat surgical site infections in numerous orthopedic procedures. Chondrocytes potentially may be exposed to these agents during operative procedures. The impact of these topical agents on chondrocyte viability is unclear.

How to select the appropriate method(s) of cytotoxicity analysis of mammalian cells at biointerfaces: A tutorial.

Many individuals perform cell viability assays as a measure of biocompatibility whether the focus of their research is on novel drug discovery, development of novel biomedical devices, or the study of biointerfacial phenomena. In this tutorial paper, the most commonly used methods available to users to perform biocompatibility testing are discussed. This includes a brief introduction into the benefits and drawbacks of the techniques, including which are best used as screening assays, which are better suited to experienced users, the relative cost of the assays per unit, and what detection techniques are most appropriate for use in conjunction with the assays.

Exploring the anomalous cytotoxicity of commercially-available poly(-isopropyl acrylamide) substrates.

Poly(-isopropyl acrylamide) (pNIPAM) is a stimulus-responsive polymer that has been of great interest to the bioengineering community. When the temperature is lowered below its lower critical solution temperature (∼32 °C), pNIPAM rapidly hydrates, and adherent cells detach as intact cell sheets. This cell-releasing behavior in a physiologically relevant temperature range has led to NIPAM's use for engineered tissues and other devices.

Effect of substrate storage conditions on the stability of "Smart" films used for mammalian cell applications.

When poly(-isopropyl acrylamide) (pNIPAM) is tethered to a surface, it can induce the spontaneous release of a sheet of mammalian cells. The release of cells is a result of the reversible phase transition the polymer undergoes at its lower critical solution temperature (LCST). Many techniques are used for the deposition of pNIPAM onto cell culture substrates.

Optimization of electrospun poly(N-isopropyl acrylamide) mats for the rapid reversible adhesion of mammalian cells.

Poly(N-isopropyl acrylamide) (pNIPAM) is a "smart" polymer that responds to changes in altering temperature near physiologically relevant temperatures, changing its relative hydrophobicity. Mammalian cells attach to pNIPAM at 37 °C and detach spontaneously as a confluent sheet when the temperature is shifted below the lower critical solution temperature (∼32 °C). A variety of methods have been used to create pNIPAM films, including plasma polymerization, self-assembled monolayers, and electron beam ionization.

Skin irritation testing of antimicrobial conjugated electrolytes.

Each year, the United States spends about $20 billion to treat people who have been infected with antibiotic resistant bacteria. Even so, the development of new antibiotics has slowed considerably since the mid-20th century. As a result, researchers are looking into developing synthetic compounds and materials with antimicrobial activities such as those made by the Schanze and Whitten groups [ACS Appl.

Poly(N-isopropyl acrylamide)-coated surfaces: Investigation of the mechanism of cell detachment.

Although there is a great deal of research focused on cell sheet engineering from polymers such as poly(N-isopropyl acrylamide) (pNIPAM), the biocompatibility of pNIPAM surfaces and the nature of cellular detachment from this polymer is still unclear. The most extensive study of the mechanism of detachment proposed a two-step process, with a first (passive) phase involving hydration of pNIPAM chains, and the second (active) phase involving cellular metabolism. However, a number of studies performed successful cell sheet detachment from pNIPAM-grafted surfaces at low temperatures which calls this hypothesis into question.

Synthesis and optimization of fluorescent poly(N-isopropyl acrylamide)-coated surfaces by atom transfer radical polymerization for cell culture and detachment.

Although there are many stimulus-responsive polymers, poly(N-isopropyl acrylamide) (pNIPAM) is of special interest due to the phase change it undergoes in a physiologically relevant temperature range that leads to the release of cells and proteins. The nondestructive release of cells opens up a wide range of applications, including the use of pNIPAM for cell sheet and tissue engineering. In this work, pNIPAM surfaces were generated that can be distinguished from the extracellular matrix.

Assessment of cytotoxicity of (N-isopropyl acrylamide) and poly(N-isopropyl acrylamide)-coated surfaces.

Poly(N-isopropyl acrylamide) (pNIPAM) is one of the most popular stimulus-responsive polymers for research. It is especially of great interest in the field of tissue engineering. While it is known that the NIPAM monomer is toxic, there is little conclusive research on the cytotoxicity of the polymer.

In vitro cytotoxicity of antimicrobial conjugated electrolytes: interactions with mammalian cells.

An estimated 19 000 deaths and $3-4 billion in health care costs per year in the United States are attributed to methicillin-resistant Staphlococcus aureus (MRSA) infections. Certain conjugated phenylene ethynylene (CPE)-based polymers (PPE) and oligomers (OPE) have been demonstrated to exhibit dark and light-activated antimicrobial activity. Until recently, the relative cytotoxicity of these PPEs and OPEs toward mammalian cells haas been unknown, limiting the applications for which they may be used (e.

ARGET-ATRP synthesis and characterization of PNIPAAm brushes for quantitative cell detachment studies.

Stimuli responsive (or "smart") polymer brushes represent a non-toxic approach for achieving release of biofouling layers. Thermo-responsive poly(N-isopropylacrylamide) (PNIPAAm) polymer brushes have been shown to modulate bacterial adhesion and release through transition between temperatures above and below the lower critical solution temperature (LCST ~32 °C) of PNIPAAm in water. In this article, we describe a convenient method to synthesize grafted PNIPAAm brushes over large areas for biological studies using a relatively simple and rapid method which allows atom transfer radical polymerization (ATRP) in presence of air using the activator regenerated electron transfer (ARGET) mechanism.

Fabrication and Characterization of Thermoresponsive Films Deposited by an RF Plasma Reactor.

Poly(-isopropyl acrylamide) (pNIPAM) undergoes a sharp property change in response to a moderate thermal stimulus at physiological temperatures. In this work, we constructed a radio frequency (RF) plasma reactor for the plasma polymerization of pNIPAM. RF deposition is a method that coats surfaces of any geometry producing surfaces that are sterile and uniform, making this technique useful for forming biocompatible films.

Biological cell detachment from poly(N-isopropyl acrylamide) and its applications.

Over the past two decades, poly(N-isopropyl acrylamide) (pNIPAM) has become widely used for bioengineering applications. In particular, pNIPAM substrates have been used for the nondestructive release of biological cells and proteins. In this feature article, we review the applications for which pNIPAM substrates have been used to release biological cells, including for the study of the extracellular matrix (ECM), for cell sheet engineering and tissue transplantation, the formation of tumorlike spheroids, the study of bioadhesion and bioadsorption, and the manipulation or deformation of individual cells.

A low-cost, rapid deposition method for "smart" films: applications in mammalian cell release.

The "smart" polymer poly (N-isopropyl acrylamide), or pNIPAM, has been studied for bioengineering applications. The polymer's abrupt change in hydrophobicity near physiologic temperatures makes it ideal for use as a substrate in many applications, including protein separation and prevention of biofouling. To tether pNIPAM, many techniques such as plasma deposition, have been utilized, but most are expensive and require long equipment calibration or fabrication periods.

Altered calcium dynamics in cardiac cells grown on silane-modified surfaces.

Chemically defined surfaces were created using self-assembled monolayers (SAMs) of hydrophobic and hydrophilic silanes as models for implant coatings, and the morphology and physiology of cardiac myocytes plated on these surfaces were studied in vitro. We focused on changes in intracellular Ca(2+) because of its essential role in regulating heart cell function. The SAM-modified coverslips were analyzed using X-ray Photoelectron Spectroscopy to verify composition.

Understanding the force-vs-distance profiles of terminally attached poly(N-isopropyl acrylamide) thin films.

In this work, we examine the interaction between thin films composed of terminally anchored poly(N-isopropyl acrylamide) (PNIPAAm) immersed in water and test surfaces. Understanding this force of interaction can be important when using PNIPAAm surfaces in biotechnological applications such as biological cell cultures. The two novel contributions that are presented here are (1) the use of a recently developed self-consistent field (SCF) theory to predict the force-vs-distance profiles, and (2) the use of a modified polymer scaling theory to estimate the wet film thickness from experimental force-vs-distance profiles.

Comparison of native extracellular matrix with adsorbed protein films using secondary ion mass spectrometry.

In the past decade, the temperature-responsive behavior of poly(N-isopropyl acrylamide) (pNIPAM) has come to be recognized as a convenient method for the nondestructive harvest of confluent cell layers. Recently, we have utilized this nondestructive cell harvest method as a means to ascertain the nature of the extracellular matrix (ECM) secreted from cells. In this work, we compare the ECM obtained after cell liftoff to individual ECM proteins adsorbed directly onto RF-plasma-deposited pNIPAM (ppNIPAM).

Temperature dependent activity and structure of adsorbed proteins on plasma polymerized N-isopropyl acrylamide.

Thorough studies of protein interactions with stimulus responsive polymers are necessary to provide a better understanding of their applications in biosensors and biomaterials. In this study, protein behavior on a thermoresponsive polymer surface, plasma polymerized N-isopropyl acrylamide (ppNIPAM), is investigated using multiple characterization techniques above and below its lower critical solution temperature (LCST). Protein adsorption and binding affinity are probed using radiolabeled proteins.

Surface chemical and mechanical properties of plasma-polymerized N-isopropylacrylamide.

Surface-immobilized poly(N-isopropyl acrylamide) (pNIPAM) is currently used for a wide variety of biosensor and biomaterial applications. A thorough characterization of the surface properties of pNIPAM thin films will benefit those applications. In this work, we present analysis of a plasma-polymerized NIPAM (ppNIPAM) coating by multiple surface analytical techniques, including time-of-flight secondary-ion mass spectrometry (ToF-SIMS), contact angle measurement, atomic force microscopy (AFM), and sum frequency generation (SFG) vibrational spectroscopy.

Cell sheet detachment affects the extracellular matrix: a surface science study comparing thermal liftoff, enzymatic, and mechanical methods.

This work compares the removal of bovine aortic endothelial cell (BAEC) monolayers via 1) low-temperature liftoff from a "smart polymer," plasma polymerized poly(N-isopropyl acrylamide) (ppNIPAM), 2) enzymatic digestion, and 3) mechanical dissociation from ppNIPAM surfaces. We examine the surfaces after cell removal by using X-ray photoelectron spectroscopy (XPS), time-of-flight secondary ion mass spectrometry (ToF-SIMS), immunostaining, and cell adhesion assay. Immunoassay results indicate that low-temperature liftoff nondestructively harvests the cell sheet and most of the underlying extracellular matrix (ECM), whereas enzymatic digestion and mechanical dissociation are damaging to both the cells and ECM.

Evidence of impurities in thiolated single-stranded DNA oligomers and their effect on DNA self-assembly on gold.

The diversity of techniques used in the synthesis, treatment, and purification of the single-stranded DNA oligomers containing a thiol anchor group (SH-ssDNA) has led to a significant variation in the purity of commercially available SH-ssDNA. In this work, we use X-ray photoelectron spectroscopy (XPS) and time-of-flight secondary ion mass spectrometry (ToF-SIMS) to study how the impurities present in commercially synthesized SH-ssDNA oligomers affected the structure of the resulting DNA films on Au. XPS results indicate that two of the purchased SH-ssDNA oligomers contain excess carbon and sulfur.

Surface characterization of the extracellular matrix remaining after cell detachment from a thermoresponsive polymer.

The temperature-responsive behavior of poly(N-isopropyl acrylamide) (pNIPAM) directly affects the attachment and detachment of cells cultured on these surfaces. At culture temperatures, cells behave similarly to those on tissue culture polystyrene (TCPS), while at room temperature, cells cultured on pNIPAM spontaneously detach as a confluent sheet. In comparison, cells grown on TCPS remain attached indefinitely after the temperature drop, requiring enzymatic or mechanical removal.

Quantitative X-ray photoelectron spectroscopy and time-of-flight secondary ion mass spectrometry characterization of the components in DNA.

The great diversity of techniques to synthesize and use DNA microarrays has made them extremely flexible for a variety of applications. This flexibility also has made standardization difficult, leading to problems comparing data from these different systems. In this work, we use the surface science techniques of X-ray photoelectron spectroscopy (XPS) and time-of-flight secondary ion mass spectrometry (TOF-SIMS) to analyze the components of DNA.