Synergistic rate boosting of collagen fibrillogenesis in heterogeneous mixtures of crowding agents.

The competition for access to space that arises between macromolecules is the basis of the macromolecular crowding phenomenon, known to modulate biochemical reactions in subtle ways. Crowding is a highly conserved physiological condition in and around cells in metazoans, and originates from a mixture of heterogeneous biomolecules. Here, using collagen fibrillogenesis as an experimental test platform and ideas from the theory of nonideal solutions, we show that an entropy-based synergy is created by a mixture of two different populations of artificial crowders, providing small crowders with extra volume occupancy when in the vicinity of bigger crowders. We present the physiological mechanism by which synergistic effects maximize volume exclusion with the minimum amount of heterogeneous crowders, demonstrating how the evolutionarily optimized crowded conditions found in vivo can be reproduced effectively in vitro.

Microhardness of bi-antibiotic-eluting bone cement scaffolds.

Bi-antibiotic-impregnated bone cements (BIBCs) are widely used in orthopaedics as a prophylactic agent (depot) to address post-surgical infections. Although hardness is widely considered a viable index to measure the integrity of the cement structure, there are few specific studies involving changes in hardness characteristics of BIBCs post elution of high doses of two widely used antibiotics: tobramycin and gentamicin. Increased doses of antibiotics and increased duration of elution may also decrease the hardness of polymethyl methacrylate (PMMA) bone cement, thus increasing the chances of shattering, scratching, and deformation.In this project, we have investigated the changes in surface hardness of five different antibiotic-loaded specimens: 0.5 g tobramycin and 0.5 g gentamicin together, 1 g tobramycin, 1 g gentamicin, 5 g tobramycin and 5 g gentamicin together, and 10 g tobramycin (each added to 40 g of PMMA), post elution for various time periods (1, 3, and 21 days). The effect of hydration on the hardness of bone cement was studied to replicate in vivo conditions. The micro-indentation tester (Buehler m5103) was utilized to determine if the increased antibiotic loads would compromise the integrity of the bone cement matrix.The results demonstrated that the amount of drug initially incorporated determined the hardness of the cement post elution. As compared to the control (no antibiotic), specimens containing 1 and 10 g of antibiotic exhibited over 50% and 73% decrease in hardness, respectively. The different treatment durations (post 1 day) as well as the hydration conditions had insignificant effect on the hardness of the cement.

Fractionation and characterization of particles simulating wear of total joint replacement (TJR) following ASTM standards.

Reactions of bone cells to orthopedic wear debris produced by the articulating motion of total joint replacements (TJRs) are largely responsible for the long-term failure of such replacements. Metal and polyethylene (PE) wear particles isolated from fluids from total joint simulators, as well as particles that are fabricated by other methods, are widely used to study such in vitro cellular response. Prior investigations have revealed that cellular response to wear debris depends on the size, shape, and dose of the particles. Hence, to have a better understanding of the wear-mediated osteolytic process it is important that these particles are well characterized and clinically relevant, both qualitatively, and quantitatively. In this study we have fractionated both ultra-high molecular weight polyethylene (UHMWPE) and Ti particles, into micron (1.0-10.0 µm), submicron (0.2-1.0 µm), and nanoparticle (0.01-0.2 µm) fractions, and characterized them based on the following size-shape descriptors as put forth in ASTM F1877: i) equivalent circle diameter (ECD), ii) aspect ratio (AR), iii) elongation (E), iv) roundness (R), and v) form factor (FF). The mean (± SD) ECDs (in µm) for micron, submicron, and nanoparticles of UHMWPE were 1.652 ± 0.553, 0.270 ± 0.180, and 0.061 ± 0.035, respectively, and for Ti were 1.894 ± 0.667, 0.278 ± 0.180, and 0.055 ± 0.029, respectively. The values for other descriptors were similar (no statistically significant difference). The nanofraction particles were found to be more sphere-like (higher R and FF values, and lower E and AR values) as compared to larger particles. Future experiments will involve use of these well characterized particles for in vitro studies.

Identification and characterization of polymeric and metallic wear debris from periprosthetic tissues after total hip revision surgery.

In spite of the growing interest in the field of orthopedic wear debris, there is no standardized technique to simultaneously isolate and analyze both ultra-high-molecular-weight polyethylene (UHMWPE) and metallic debris from periprosthetic tissues. Using a modification of the previously employed base-digestion protocol involving solvent and mechanical treatment, we were able to separate the wear particles from the tissue. Subsequently, using environmental scanning electron microscopy (ESEM) and energy-dispersive spectrometry (EDS), we characterized individual particulate species. Metallic debris, particularly Co, Cr, and Mo, appeared as irregular and amorphous-like structures, whereas UHMWPE and Ti appeared as crystalline-like structures, some as small as 15 nm. The investigation revealed that UHMWPE forms the bulk (~82%), followed by Ti (~8%-9%), Co (~5%-6%), and Cr (~3%-4%), along with many other trace elements, such as Mo, Pb, Fe, Pb, Fe, Si (~1%-2%).