A scanning electron microscopy study of projectile entry fractures in cortical bone; genesis and microarchitectural features.

The present paper presents a scanning electron microscope (SEM) analysis of the genesis and microarchitecture of experimentally induced cortical entry fractures in porcine scapulae impacted at velocities ranging from 54 to 897 m/s. SEM observation was conducted on polyurethane replicas cast from negative silicone moulds. Analysis of the sequence of fracture processes operative during projectile impact revealed the presence of ring cracks at the site of impact, confirming that penetration in sandwich bones is achieved by cone crack propagation. Despite impulsive loading, two forms of plastic deformation were identified in the cortical bone surrounding the entry fracture up to a maximum velocity of 871 m/s. Microscopic radial and concentric cracks were associated with projectile impact, and the role of pores and pits as stress concentrators was captured. Possible underlying mechanisms for the observed plastic deformation are described, and the diagnostic utility of SEM analysis is presented.

Crack propagation through sandwich bones due to low-velocity projectile impact.

Projectile impact in sandwich bones typically results in formation of conoidal wounds exhibiting a larger region of damage on the inner cortical plate termed the bevel. To date, a number of hypotheses have been put forward to explain the formation of this wound type. The plug and spall hypothesis suggests that the conoidal morphology is produced by a two-phase mechanism of shear plug formation followed by internal bevel production during projectile exit. In contrast, the cone crack hypothesis suggests that such wounds are produced by cone crack propagation through the three laminae of the sandwich bone, resulting in the formation of bioceramic conoids consisting of all three bone laminae. In order to test these hypotheses, 28 non-human sandwich bones were impacted with 6-mm carbon steel spheres at velocities ranging from 26 to 96 metres per second (m/s). Impacts were filmed utilizing high-speed videography and fracture morphology analysed using micro-computerized tomography (µ-CT). Sequential increase in velocity successfully captured the genesis of conoidal wounds. Low-velocity impact produced circular depressed fractures in the outer cortex exhibiting angulated cortical fracture edges. An increase in velocity resulted in translaminar fracture and production of one intact and three fragmentary bioceramic conoids. At the highest velocities, conoids were fragmented and lost in the ejecta plume, with attached fragments undergoing dynamic movement during and after perforation. Significantly, projectile exit was not required for bevel production. The implications of these findings in wound interpretation are discussed.

A novel hypothesis for the formation of conoidal projectile wounds in sandwich bones.

When perforated by a projectile, sandwich bones typically exhibit wounds with a distinct conoidal morphology that is widely utilised both in wound diagnosis and trajectory determinations. However, the dynamic fracture mechanisms underlying this intriguing wound type have yet to be experimentally verified. The most frequently quoted hypothesis for their formation, plug and spall, is difficult to reconcile with the conoidal morphology exhibited by such wounds. The present article carries out a high-speed videographic and micro-computerised tomographic (µ-CT) analysis of perpendicularly produced projectile wounds induced from 139.15 to 896.84 metres per second (m/s) in pig scapulae. Fundamental data on energy absorption, wound shape and bevel symmetry are presented. Cross-sectional fracture morphology revealed by µ-CT raises the novel hypothesis that tensile stresses induced by the projectile in the outer cortex elicit cone crack formation and that this cone crack then propagates catastrophically through the entire sandwich structure. This process results in the momentary formation of a bioceramic conoid, a conoidal volume of bone consisting of all three sandwich bone layers separated from the parent bone by the internal bevel. Fragmentation of the separated volume leaves the conoidal wound behind as its counterpart. The significance of this hypothesis in terms of differential diagnosis and interpretation of bevel shape is discussed.

Mammalian complex III heme dynamics studied with pump-probe spectroscopy and red light illuminations.

The electronic or molecular mechanisms that initiate photobiomodulation (PBM) in cells are not yet fully understood. The porcine complex III (C-III) of the electron transport chain was characterized with transient absorption spectroscopy (TAS). We then applied our recently developed continuous wave laser coupled TAS procedure (CW-TAS) to investigate the effect of red light irradiances on the heme dynamics of C-III in its c1 reduced state. The time constants were found to be 3.3 ± 0.3 ps for vibrational cooling of the oxidized state and 4.9 ± 0.4 ps for rebinding of the photodissociated axial ligand of the c1 reduced state. The analysis of the CW-TAS procedure yielded no significant changes in the C-III heme dynamics. We rule out the possibility of 635 nm CW light at 4.7 mW/cm2 inducing a PBM effect on the heme dynamic of C-III, specifically with the photodissociation of its axial ligand.

Transient absorption spectroscopy to explore cellular pathways to photobiomodulation.

Photobiomodulation (PBM) describes the use of low irradiance light in the red to near-infrared wavelength range to stimulate biological effects in tissue, and many biological and spectroscopic techniques are used to study PBM. However, these techniques focus on the products or downstream effects rather than the electronic transitions that initiate the PBM processes. This study presents a novel approach to studying low irradiance light exposures on individual proteins and/or protein complexes by combining a continuous wave (CW) laser diode with femtosecond transient absorption spectroscopy (TAS), coined here as CW-TAS, and tests the system on reduced cytochrome c (Cyt c) for proof of principle. TAS was conducted using a 532-nm excitation pump beam and a 350-600 nm supercontinuum probe. CW laser diodes with wavelengths of 450 nm, 635 nm, and 808 nm were interchangeably fiber coupled into the HELIOS Fire. Samples of Cyt c were tested by TAS using a pump power of 15 µW, both with and without CW exposure. CW exposures were carried out with irradiances of 1.60 and 3.20 mW/cm2, except for 808 nm, which was only tested at 1.60 mW/cm2. Both kinetic and global analyses were performed on the TAS data and the time constants for sets with and without CW exposures were compared. The TAS data for Cyt c with the full dosage of CW exposures did not alter the TAS data distinguishably from the control data. No new electronic transient signals were observed beyond the background when testing Cyt c with the CW exposures. Kinetic analysis confirmed that existing transients did not deviate beyond uncertainty. Global time constants for Cyt c were calculated to be 0.25 ± 0.03 ps and 5.1 ± 0.3 ps for the control study, and the time constants for the CW exposed Cyt c were not significantly different. This study concludes that CW irradiation, at doses delivered, does not alter the transient absorption data of Cyt c. The CW-TAS method provides a new tool for studying PBM effects in other proteins and protein complexes that might respond to the CW wavelengths, such as Complex IV, in future studies.

Scanning electron microscope analysis of gunshot defects to bone: an underutilized source of information on ballistic trauma.

Recent years have seen increasing involvement by forensic anthropologists in the interpretation of skeletal trauma. With regard to ballistic injuries, there is now a large literature detailing gross features of such trauma; however, less attention has been given to microscopic characteristics. This article presents analysis of experimentally induced gunshot trauma in animal bone (Bos taurus scapulae) using full metal jacket (FMJ), soft point (SP), and captive bolt projectiles. The results were examined using scanning electron microscopy (SEM). Additional analysis was conducted on a purported parietal gunshot lesion in a human cranial specimen. A range of features was observed in these samples suggesting that fibrolamellar bone response to projectile impact is analogous to that observed in synthetic composite laminates. The results indicate that direction of bullet travel can be discerned microscopically even when it is ambiguous on gross examination. It was also possible to distinguish SP from FMJ lesions. SEM analysis is therefore recommended as a previously underexploited tool in the analysis of ballistic trauma.