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Advances in tissue engineering require biofunctional scaffolds that can not only provide cells with structural support, but also interact with cells in a biological manner. To achieve this goal, a frequently used cell adhesion peptide Arg-Gly-Asp (RGD) was covalently incorporated into poly(ethylene glycol) diacrylate (PEODA) hydrogel and its dosage effect (0.025, 1.25 and 2.5 mm) on osteogenesis of marrow stromal cells in a three-dimensional environment was examined. Expression of bone-related markers, osteocalcin (OCN) and Alkaline phosphatase (ALP), increased significantly as the RGD concentration increased. Compared with no RGD, 2.5 mm RGD group showed a 1344% increase in ALP production and a 277% increase in OCN accumulation in the medium. RGD helped MSCs maintain cbfa-1 expression when shifted from a two-dimensional environment to a three-dimensional environment. Soluble RGD was found to completely block the mineralization of marrow stromal cells, as manifested by quantitative calcium assay, phosphorus elemental analysis and Von Kossa staining. In conclusion, we have demonstrated that RGD-conjugated PEODA hydrogel promotes the osteogenesis of MSCs in a dosage-dependent manner, with 2.5 mm being optimal concentration.

The development of biocompatible photopolymerizing polymers for biomedical and tissue engineering applications has the potential to reduce the invasiveness and cost of biomaterial implants designed to repair or augment tissues. However, more information is needed about the cellular toxicity of the compounds and initiators used in these systems. The current study evaluates the cellular toxicity of three ultraviolet sensitive photoinitiators on six different cell populations that are used for engineering numerous tissues. The photoinitiator 2-hydroxy-1-[4-(hydroxyethoxy)phenyl]-2-methyl-1-propanone (Irgacure 2959) caused minimal toxicity (cell death) over a broad range of mammalian cell types and species. It was also demonstrated that different cell types have variable responses to identical concentrations of the same photoinitiator. While inherent differences in the cell lines may contribute to the variable cytotoxicity, a correlation between cellular proliferation rate (population doubling time) and increased cytotoxicity of the photoinitiator was observed. Cell lines that divided more quickly were more sensitive to photoinitiator-induced cell death. In summary, the photoinitiator Irgacure 2959 is well tolerated by many cell types over a range of mammalian species. Cell photoencapsulation strategies may be optimized to improve cell survival by manipulating proliferation rate.

The effect of anodization on passive dissolution of titanium was studied by measuring titanium levels in peritoneal leukocytes and tissues of laboratory animals with titanium plates implanted into the peritoneal cavity. Fifteen Sprague-Dawley rats were assigned randomly to three treatment groups of five animals. One group served as controls, the other two groups had an anodized or an unanodized implant placed in the left paracolic gutter. Peritoneal lavage samples and blood samples, organ tissues and tissue surrounding the implants, were removed for histologic examination and titanium levels. Titanium was not detected in any distant organs or in the peritoneal lavage fluid. The capsular tissues surrounding the implants contained titanium at levels ranging from 2610 to 16786 ng/g for unanodized plates, and 888-5933 ng/g for anodized plates. The titanium levels within the peritoneal leukocytes of animals with unanodized implants were significantly elevated (P = 0.01) over time, as compared with controls. The level of titanium in the peritoneal leukocytes of animals with anodized implants was not significantly elevated when compared with controls. Titanium levels in the trace range, as measured in the capsular tissues, are likely a result of corrosion. Surface treatment of titanium by anodization reduces passive dissolution.

Frostbite is characterized by acute tissue injury induced by freezing and thawing. Initial complete ischemia is followed by reperfusion and later, tissue necrosis. These vascular events support the hypothesis that free radical-mediated reperfusion injury at thawing might contribute to tissue necrosis after frostbite in a manner similar to that seen after normothermic ischemia. To test this hypothesis, rabbit ears were frozen at -21 degrees C for 30, 60, 90, or 120 s and rewarmed at room temperature (22 degrees C). Rabbits were treated "blindly" with saline alone, highly purified, pharmaceutical grade superoxide dismutase (SOD), allopurinol, or deferoxamine. The area of ear necrosis was determined 3 weeks after frostbite by "blinded" morphometry. The administration of SOD at the time of thawing significantly improved viability in ears frozen for 60 and 90 s, but not in those frozen for 30 or 120 s. Deferoxamine also improved viability in ears frozen for 60 s. Allopurinol did not significantly affect ear survival. Electron micrographs showed the appearance of severe endothelial cell injury beginning during freezing and extending through early reperfusion. Later, neutrophil adhesion, erythrocyte aggregation, and microvascular stasis were seen. These findings suggest that free radical-mediated reperfusion injury has a role in frostbite, and quantitate the proportion of the injury that is due to this mechanism.

Post-traumatic orbital osseous and soft tissue surface evaluation was performed using high resolution computerized tomography and three-dimensional surface reconstructions (3DSR) using specially developed computer software. Four patients with facial fractures involving the orbit are presented as examples of the technique. Orbital volume was measured and surface images prepared using the original CT scans as input to CT computer software developed for this purpose. The results simplify interpretation of the nature and extent of traumatic orbital disruption. The interpretation of serial high resolution CT scans of the orbit (now obtained routinely in fractures involving the orbit) has been augmented by employing three-dimensional surface reconstructions (3DSR).