Immobilized RGD concentration and proteolytic degradation synergistically enhance vascular sprouting within hydrogel scaffolds of varying modulus.

Insufficient vascularization limits the volume and complexity of engineered tissue. The formation of new blood vessels (neovascularization) is regulated by a complex interplay of cellular interactions with biochemical and biophysical signals provided by the extracellular matrix (ECM) necessitating the development of biomaterial approaches that enable systematic modulation in matrix properties. To address this need poly(ethylene) glycol-based hydrogel scaffolds were engineered with a range of decoupled and combined variations in integrin-binding peptide (RGD) ligand concentration, elastic modulus and proteolytic degradation rate using free-radical polymerization chemistry. The modularity of this system enabled a full factorial experimental design to simultaneously investigate the individual and interaction effects of these matrix cues on vascular sprout formation in 3 D culture. Enhancements in scaffold proteolytic degradation rate promoted significant increases in vascular sprout length and junction number while increases in modulus significantly and negatively impacted vascular sprouting. We also observed that individual variations in immobilized RGD concentration did not significantly impact 3 D vascular sprouting. Our findings revealed a previously unidentified and optimized combination whereby increases in both immobilized RGD concentration and proteolytic degradation rate resulted in significant and synergistic enhancements in 3 D vascular spouting. The above-mentioned findings would have been challenging to uncover using one-factor-at-time experimental analyses.

Can we assess the age of bruises? An attempt to develop an objective technique.

AIMS: The aims of this study were (A) to investigate whether the number of years of forensic experience affected the accuracy with which forensic experts (FEs) were able to age bruises and (B) to identify the properties and colours of a bruise that were utilized by FEs in their assessment of bruise age. The study then investigated the possibility of using a more objective technique. It was decided to use readily available digital photography and software to objectively assess changes in bruise colouration and to investigate if this can be used to age bruises. METHODS: Twenty-three FEs were shown 25 photographs of bruises of varying but known ages and asked to estimate the ages. In part two of the study, bruises were inflicted on volunteers using a vacuum pump and photographs taken of the bruise daily from infliction to resolution. The images were analysed using Adobe Photoshop. Red, green and blue (RGB) values were recorded for each bruise and analyses carried out comparing the values over time between subjects and within subjects. RESULTS: This study both enhanced and supported a previous conclusion that visual assessment of photographs is an unreliable method for ageing bruises. Additionally, it found that the degree of forensic experience had no effect on accuracy. It also identified that colour (particularly yellow, red and purple) and intensity of colour were the most commonly used properties of a bruise in the assessment of its age. The RGB method proved to be a reliable technique with which to measure bruise colour, but its validity in the assessment of bruise age was poor. CONCLUSIONS: Visual assessment of bruises is unreliable and the accuracy of ageing was not improved by the degree of forensic experience. The RGB method gave highly reproducible results, but did not accurately assess bruise age. However, results within subjects suggested that there may be individual variation in haemoglobin metabolism.