Biomedical subjects

Delphine Dean

Publications and source records attributed to Delphine Dean.

6 recordsLinked to original sources

PubMed2006

Lateral nanomechanics of cartilage aggrecan macromolecules.

To explore the role of the brush-like proteoglycan, aggrecan, in the shear behavior of cartilage tissue, we measured the lateral resistance to deformation of a monolayer of chemically end-attached cartilage aggrecan on a microcontact printed surface in aqueous NaCl solutions via lateral force microscopy. The effects of bath ionic strength (IS, 0.001-1.0 M) and lateral displacement rate (approximately 1-100 microm/s) were studied using probe tips functionalized with neutral hydroxyl-terminated self-assembled alkanethiol monolayers. Probe tips having two different end-radii (R approximately 50 nm and 2.5 microm) enabled access to different length-scales of interactions (nano and micro). The measured lateral force was observed to depend linearly on the applied normal force, and the lateral force to normal force proportionality constant, mu, was calculated. The value mu increased (from 0.03 +/- 0.01 to 0.11 +/- 0.01) with increasing bath IS (0.001-1.0 M) for experiments using the microsized tip due to the larger compressive strain of aggrecan that resulted from increased IS at constant compressive force. With the nanosized tip, mu also increased with IS but by a smaller amount due to the fewer number of aggrecan involved in shear deformation. The variations in lateral force as a function of applied compressive strain epsilon(n) and changes in bath IS suggested that both electrostatic and nonelectrostatic interactions contributed significantly to the shear deformational behavior of the aggrecan layers. While lateral force did not vary with lateral displacement rate at low IS, where elastic-like electrostatic interactions between aggrecan dominated, lateral force increased significantly with displacement rate at physiological and higher IS, suggestive of additional viscoelastic and/or poroelastic interactions within the aggrecan layer. These data provide insights into molecular-level deformation of aggrecan macromolecules that are important to the understanding of cartilage behavior.

Aggrecans↗

PubMed2006

Silicon addition to hydroxyapatite increases nanoscale electrostatic, van der Waals, and adhesive interactions.

The normal intersurface forces between nanosized probe tips functionalized with COO(-)-terminated alkanethiol self-assembling monolayers and dense, polycrystalline silicon-substituted synthetic hydroxyapatite (SiHA) and phase pure hydroxyapatite (HA) were measured via a nanomechanical technique called chemically specific high-resolution force spectroscopy. A significantly larger van der Waals interaction was observed for the SiHA compared to HA; Hamaker constants (A) were found to be A(SiHA) = 35 +/- 27 zJ and A(HA) = 13 +/- 12 zJ. Using the Derjaguin-Landau-Verwey-Overbeek approximation, which assumes linear additivity of the electrostatic double layer and van der Waals components, and the nonlinear Poisson-Boltzmann surface charge model for electrostatic double-layer forces, the surface charge per unit area, sigma (C/m(2)), was calculated as a function of position for specific nanosized areas within individual grains. SiHA was observed to be more negatively charged than HA with sigma(SiHA) = -0.024 +/- 0.013 C/m(2), two times greater than sigma(HA) = -0.011 +/- 0.006 C/m(2). Additionally, SiHA was found to have increased surface adhesion (0.7 +/- 0.3 nN) compared to HA (0.5 +/- 0.3 nN). The characterization of the nanoscale variations in surface forces of SiHA and HA will enable an improved understanding of the initial stages of bone-biomaterial bonding.

Biocompatible Materials↗

PubMed2005

Compressive nanomechanics of opposing aggrecan macromolecules.

In this study, we have measured the nanoscale compressive interactions between opposing aggrecan macromolecules in near-physiological conditions, in order to elucidate the molecular origins of tissue-level cartilage biomechanical behavior. Aggrecan molecules from fetal bovine epiphyseal cartilage were chemically end-grafted to planar substrates, standard nanosized atomic force microscopy (AFM) probe tips (R(tip) approximately 50 nm), and larger colloidal probe tips (R(tip) approximately 2.5 microm). To assess normal nanomechanical interaction forces between opposing aggrecan layers, substrates with microcontact printed aggrecan were imaged using contact mode AFM, and aggrecan layer height (and hence deformation) was measured as a function of solution ionic strength (IS) and applied normal load. Then, using high-resolution force spectroscopy, nanoscale compressive forces between opposing aggrecan on the tip and substrate were measured versus tip-substrate separation distance in 0.001-1M NaCl. Nanosized tips enabled measurement of the molecular stiffness of 2-4 aggrecan while colloidal tips probed the nanomechanical properties of larger assemblies (approximately 10(4) molecules). The compressive stiffness of aggrecan was much higher when using a densely packed colloidal tip than the stiffness measured for using the nanosized tip with a few aggrecan, demonstrating the importance of lateral interactions to the normal nanomechanical properties. The measured stress at 0.1M NaCl (near-physiological ionic strength) increased sharply at aggrecan densities under the tip of approximately 40 mg/ml (physiological densities are approximately 20-80 mg/ml), corresponding to an average inter-GAG spacing of 4-5 Debye lengths (4-5 nm); this characteristic spacing is consistent with the onset of significant electrostatic interactions between GAG chains of opposing aggrecan molecules. Comparison of nanomechanical data to the predictions of Poisson-Boltzmann-based models further elucidated the regimes over which electrostatic and nonelectrostatic interactions affect aggrecan stiffness in compression. The most important aspects of this study include: the incorporation of experiments at two different length scales, the use of microcontact printing to enable quantification of aggrecan deformation and the corresponding nanoscale compressive stress vs. strain curve, the use of tips of differing functionality to provide insights into the molecular mechanisms of deformation, and the comparison of experimental data to the predictions of three increasingly refined Poisson-Boltzmann (P-B)-based theoretical models for the electrostatic double layer component of the interaction.

Aggrecans↗

PubMed2005

Nanoscale variation in surface charge of synthetic hydroxyapatite detected by chemically and spatially specific high-resolution force spectroscopy.

The normal intersurface forces between nanosized probe tips functionalized with COO-- and NH3+-terminated alkanethiol self-assembling monolayers and dense polycrystalline phase pure synthetic hydroxyapatite (HA) were measured via a powerful nanomechanical technique called chemically specific high-resolution force spectroscopy. The data taken on approach of the probe tip to the HA surface was compared to the nonlinear Poisson-Boltzmann-based electrostatic double layer theory to predict the surface charge per unit area of the HA, sigmaHA (C/m2), as a function of ionic strength, position within a variety of grains, and across grain boundaries. The average sigmaHA was found to be approximately -0.02 C/m2 and to vary from -0.0037 to -0.072 C/m2 with nanoscale position in relation to grain boundaries and crystal planes up to -0.19 C/m2/microm. Positional measurement of nanoscale surface properties holds great promise in elucidating the molecular origins of physicochemical processes occurring at the biomaterial interface.

Biocompatible Materials↗

PubMed2005

Nanomechanics of opposing glycosaminoglycan macromolecules.

In this study, the net intermolecular interaction force between a chondroitin sulfate glycosaminoglycan (GAG)-functionalized probe tip and an opposing GAG-functionalized planar substrate was measured as a function of probe tip-substrate separation distance in aqueous electrolyte solutions using the technique of high resolution force spectroscopy. A range of GAG grafting densities as near as possible to native cartilage was used. A long-range repulsive force between GAGs on the probe tip and substrate was observed, which increased nonlinearly with decreasing separation distance between probe tip and substrate. Data obtained in 0.1 M NaCl was well predicted by a recently developed Poisson-Boltzmann-based theoretical model that describes normal electrostatic double layer interaction forces between two opposing surfaces of end-grafted, cylindrical rods of constant volume charge density and finite length, which interdigitate upon compression. Based on these results, the nanomechanical data and interdigitated rod model were used together to estimate the electrostatic component of the equilibrium modulus of cartilage tissue, which was then compared to that of normal adult human ankle cartilage measured in uniaxial confined compression.

Biomechanical Phenomena↗

PubMed2004

Mechanical compression of cartilage explants induces multiple time-dependent gene expression patterns and involves intracellular calcium and cyclic AMP.

Chondrocytes are influenced by mechanical forces to remodel cartilage extracellular matrix. Previous studies have demonstrated the effects of mechanical forces on changes in biosynthesis and mRNA levels of particular extracellular matrix molecules, and have identified certain signaling pathways that may be involved. However, the broad extent and kinetics of mechano-regulation of gene transcription has not been studied in depth. We applied static compressive strains to bovine cartilage explants for periods between 1 and 24 h and measured the response of 28 genes using real time PCR. Compression time courses were also performed in the presence of an intracellular calcium chelator or an inhibitor of cyclic AMP-activated protein kinase A. Cluster analysis of the data revealed four main expression patterns: two groups containing either transiently up-regulated or duration-enhanced expression profiles could each be subdivided into genes that did or did not require intracellular calcium release and cyclic AMP-activated protein kinase A for their mechano-regulation. Transcription levels for aggrecan, type II collagen, and link protein were up-regulated approximately 2-3-fold during the first 8 h of 50% compression and subsequently down-regulated to levels below that of free-swelling controls by 24 h. Transcription levels of matrix metalloproteinases-3, -9, and -13, aggrecanase-1, and the matrix protease regulator cyclooxygenase-2 increased with the duration of 50% compression 2-16-fold by 24 h. Thus, transcription of proteins involved in matrix remodeling and catabolism dominated over anabolic matrix proteins as the duration of static compression increased. Immediate early genes c-fos and c-jun were dramatically up-regulated 6-30-fold, respectively, during the first 8 h of 50% compression and remained up-regulated after 24 h.

ADAM Proteins↗