ABSTRACT
Self-assembled monolayers (SAMs) on gold containing a thermo-responsive poly( N-isopropylacrylamide)-poly(ethylene glycol)-thiol copolymer were formed. These layers show considerable potential for inducing enzyme-free and gentle detachment of cultivated cells. In an effort to optimize detachment of cells, including strongly adhering ones, two approaches are presented. First, two thermo-responsive copolymers with different poly(ethylene glycol) (PEG) contents of 15 wt % ("P15") and 19 wt % ("P19") were grafted to Au surfaces. Second, mixed monolayers were formed containing P19 and various concentrations of thiol bearing PEG. X-ray photoelectron spectroscopy (XPS) on pure and mixed P19 containing layers confirmed the expected layer compositions. Contact angle measurements showed good functionality of all surfaces prepared. Upon a temperature decrease below the lower critical solution temperature (LCST), the duration until cultivated fibroblasts detached from pure P19 surfaces was half of the one determined on P15. Strongly adherent human osteosarcoma cells could not be detached from pure P19 layers. Through co-adsorption of P19 and thiol-bearing PEG of a molar composition of 1:6, layers were formed that allowed good spreading of osteosarcoma cells above LCST and their efficient detachment below LCST.
Subject(s)
Gold/chemistry , Osteosarcoma/metabolism , Polymers/chemistry , Adhesiveness , Animals , Cell Adhesion , Cell Line, Tumor , Humans , Mice , Microscopy, Phase-Contrast , Polyethylene Glycols/chemistry , Spectrometry, X-Ray Emission/methods , Sulfhydryl Compounds , Surface Properties , TemperatureABSTRACT
The control of cell adhesion is crucial in many procedures in cellular biotechnology. A thermo-responsive poly(N-isopropylacrylamide)-poly(ethylene glycol)-thiol (PNIPAAm-PEG-thiol) copolymer was synthesized for the formation of self-assembled monolayers (SAM) that allow the control of adhesion of cells on gold substrates. The contact angle of water on these layers varies between 65 degrees at a temperature of 45 degrees C and 54 degrees at 25 degrees C. This behaviour is consistent with a transition of the polymer chains from an extended and highly hydrated to a collapsed coil-like state. At 37 degrees C, cultivated fibroblasts adhere and spread normally on this surface and detach by reducing the temperature below the lower critical solution temperature (LCST). Layers can repeatedly be used without loss of their functionality. In order to quantify the capability of the copolymer layer to induce cell detachment, defined shear forces are applied to the cells. For this purpose, the laminar flow in a microfluidic device is used. Our approach provides a strategy for the optimization of layer properties that is based on establishing a correlation between a functional parameter and molecular details of the layers.
Subject(s)
Acrylic Resins/chemistry , Cell Adhesion/physiology , Cell Culture Techniques/instrumentation , Fibroblasts/physiology , Gold/chemistry , Microfluidic Analytical Techniques/instrumentation , Micromanipulation/instrumentation , Animals , Cell Culture Techniques/methods , Cell Line , Coated Materials, Biocompatible/chemistry , Equipment Design , Equipment Failure Analysis , Fibroblasts/cytology , Mice , Microfluidic Analytical Techniques/methods , Micromanipulation/methods , TemperatureABSTRACT
The surface characteristics of intravenously administered particulate drug carriers decisively influence the protein adsorption that is regarded as a key factor for the in vivo fate of the carriers. We labeled surface-modified polymer particles with the gamma-emitting radioisotope 99mTc in order to test their properties in blood and follow their in vivo fate. The biodistribution was different in various types of polymer particles. As expected, labeled particles were found in the mononuclear phagocyte system in a large scale but markedly different biodistribution for some particles were also shown.
Subject(s)
Drug Carriers/chemistry , Microspheres , Technetium/chemistry , Technetium/pharmacokinetics , Animals , Drug Carriers/administration & dosage , Drug Stability , Female , Injections, Intravenous , Isotope Labeling/methods , Metabolic Clearance Rate , Organ Specificity , Radiopharmaceuticals/administration & dosage , Radiopharmaceuticals/chemistry , Radiopharmaceuticals/pharmacokinetics , Rats , Rats, Wistar , Surface Properties , Technetium/administration & dosage , Tissue DistributionABSTRACT
The surface characteristics of intravenously administered particulate drug carriers decisively influence the protein adsorption that is regarded as a key factor for the in vivo fate of the carriers. Latex nanoparticles were synthesized to study the influence of different basic and acidic functional groups on particulate surfaces on the protein adsorption from human serum. The protein mass adsorbed to the particles was assessed by BCA protein assay, the protein adsorption patterns were analyzed by two-dimensional electrophoresis. Considerable differences in the protein adsorption with regard to preferential adsorbed proteins were detectable for the different functional groups. Possible correlations between the surface characteristics and the protein adsorption are shown and discussed. The knowledge concerning the interactions of proteins and nanoparticles can be used for a rational development of particulate drug carriers and can also be useful for an optimized design of medical devices, e.g., hemodialysis membranes or implants.
Subject(s)
Polystyrenes/chemistry , Proteins/chemistry , Adsorption , Electrophoresis, Gel, Two-Dimensional , Emulsions , Gels , Glycoproteins/chemistry , Hemoglobins/chemistry , Humans , Immunoglobulin Light Chains/chemistry , Latex/chemistry , Microspheres , Models, Biological , Surface Properties , beta 2-Glycoprotein IABSTRACT
Plasma protein adsorption is regarded as a key factor for the in vivo organ distribution of intravenously administered colloidal drug carriers, and strongly depends on their surface characteristics, e.g. surface hydrophobicity or charge. A range of polymeric nanoparticles with a steep variation of the surface charge density was synthesized as model drug carriers. Physicochemical parameters, i.e. particle size, surface charge density, hydrophobicity and surface topography were determined. Two-dimensional electrophoresis (2-DE) was employed for determination of particle interactions with human plasma proteins. Increasing surface charge density showed an increase in plasma protein adsorption, but did not show differences in the detected protein species. For the first time it was possible to show plasma protein adsorption patterns on a range of nanoparticles with variation of only one parameter, i.e. the charge, while size and surface hydrophobicity remain practically unchanged. The knowledge about the interactions of proteins with particulate surfaces can be exploited for the future controlled design of colloidal drug carriers and possibly in the controlled creation of biocompatible surfaces of other devices that come into contact with proteins (e.g. microparticles and implants).
