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. Recently, a novel method for codepositing this smart polymer with a sol-gel, tetraethyl orthosilicate (TEOS), was developed. In this work, we adapt this technique for applications in mammalian cell attachment/detachment. In addition, we compare the effects of the pNIPAM/TEOS ratio to functionality using surface analysis techniques (XPS and contact angles). We found the optimal ratio to be 0.35 wt % pNIPAM/TEOS. Cell detachment from these substrates indicate that they would be ideal for applications that do not require intact cell sheets, such as biofouling prevention and protein separation, as this technique is a simple and affordable technique for pNIPAM deposition.

Summary Poly(N-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. The films generated are characterized using X-ray photoelectron spectroscopy (XPS), contact angles, cell culture, and interferometry. We find that a plasma with a decreasing series of power settings (i.e., from 100W to 1W) at a pressure of 140 millitorr yields the most favorable results.

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. However, detachment of confluent cell sheets from these pNIPAM films can take well over an hour to achieve potentially impacting cellular behavior. In this work, pNIPAM mats were prepared via electrospinning (i.e., espNIPAM) by a previously described technique that the authors optimized for cell attachment and rapid cell detachment. Several electrospinning parameters were varied (needle gauge, collection time, and molecular weight of the polymer) to determine the optimum parameters. The espNIPAM mats were then characterized using Fourier-transform infrared, x-ray photoelectron spectroscopy, and scanning electron microscopy. The espNIPAM mats showing the most promise were seeded with mammalian cells from standard cell lines (MC3T3-E1) as well as cancerous tumor (EMT6) cells. Once confluent, the temperature of the cells and mats was changed to ∼25 °C, resulting in the extremely rapid swelling of the mats. The authors find that espNIPAM mats fabricated using small, dense fibers made of high molecular weight pNIPAM are extremely well-suited as a rapid release method for cell sheet harvesting.

When poly(N-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. Previously, we compared two methods of deposition (plasma polymerization, and co-deposition with a sol-gel). We proved that although both were technically appropriate for obtaining thermoresponsive pNIPAM films, the surfaces that were co-deposited with a sol-gel caused some disruption in cell activity. The variation of cell behavior could be due to the delamination of pNIPAM films leaching toxic chemicals into solution. In this work, we assessed the stability of these pNIPAM films by manipulating the storage conditions and analyzing the surface chemistry using X-ray photoelectron spectroscopy (XPS) and contact angle measurements over the amount of time required to obtain confluent cell sheets. From XPS, we demonstrated that ppNIPAM (plasma polymerized NIPAM) films remains stable across all storage conditions while sol-gel deposition show large deviations after 48 h of storage. Cell response of the deposited films was assessed by investigating the cytotoxicity and biocompatibility. The 37°C and high humidity storage affects sol-gel deposited films, inhibiting normal cell growth and proper thermoresponse of the film. Surface chemistry, thermoresponse and cell growth remained similar for all ppNIPAM surfaces, indicating these substrates are more appropriate for mammalian cell culture applications.

Poly(N-isopropyl acrylamide) or pNIPAM is a thermoresponsive polymer that is widely studied for use in bioengineering applications. The interest in this polymer lies in the polymer’s unique capability to undergo a sharp property change near physiological temperatures, which aids in the spontaneous release of biological cells from substrates. Currently, there are many methods to deposit pNIPAM onto substrates, including atom transfer radical polymerization (ATRP) and electron beam ionization. Each method yields pNIPAM coated substrates with different surface characteristics which can influence cell behavior. In this work, we compare two methods of pNIPAM deposition: plasma deposition and co-deposition with a sol gel. The resulting pNIPAM films were analyzed for use as substrates for mammalian cell culture based on surface characterization (XPS, ToF-SIMS, AFM, contact angles), cell attachment/detachment studies, and an analysis of exocytosis function using carbon-fiber microelectrode amperometry (CFMA). We find that, although both methods are useful for the deposition of functional pNIPAM films, plasma deposition is much preferred for cell-sheet engineering applications due to the films’ thermoresponse, minimal change in cell density, and maintenance of supported cell exocytosis function.