Standardization of research methods employed in assessing the interaction metallic-based nanoparticles and the blood-brain barrier: present and future perspectives

peer-reviewedThe full text of this article will not be available until the embargo expires on the 18/01/2020.Treating diseases of the central nervous system (CNS) is complicated by the presence of the blood-brain barrier (BBB), a semipermeable boundary layer protecting the CNS from toxins and homeostatic disruptions. However, this layer also excludes almost 100% of therapeutics, impeding the treatment of CNS diseases. The advent of nanoparticles, in particular metallic-based nanoparticles, presents the potential to overcome this barrier and transport drugs into the CNS. Recent interest in metallic-based nanoparticles has generated an immense array of information pertaining to nanoparticles of different materials, sizes, morphologies, and surface properties. Nanoparticles with different physico-chemical properties lead to distinct nanoparticle-host interactions; yet, comprehensive characterization is often not completed. Similarly, in vivo testing has involved a mixed evaluation of parameters, including: BBB permeability, integrity, biodistribution, and toxicity. The methods applied to assess these parameters are inconsistent; this complicates the comparison of different nanoparticle-host system responses. A systematic review was conducted to investigate the methods by which metallic-based nanoparticles are characterized and assessed in vivo. The introduction of a standardized approach to nanoparticle characterization and in vivo testing is crucial if research is to transition to a clinical setting. The approach suggested, herein, is based on equipment and techniques that are accessible and informative to facilitate the routine incorporation of this standardized, informative approach into different research settings. Thorough characterization could lead to improved interpretation of in vivo responses, which could clarify nanoparticle properties that result in favorable in vivo outcomes whilst exposing nanoparticle-specific weaknesses. Only then will researchers successfully identify nanoparticles capable of delivering life-saving therapeutics across the blood-brain barrier

Regional mechanical and biochemical properties of the porcine cortical meninges

peer-reviewedThe meninges are pivotal in protecting the brain against traumatic brain injury (TBI), an ongoing issue in most mainstream sports. Improved understanding of TBI biomechanics and pathophysiology is desirable to improve preventative measures, such as protective helmets, and advance our TBI diagnostic/prognostic capabilities. This study mechanically characterised the porcine meninges by performing uniaxial tensile testing on the dura mater (DM) tissue adjacent to the frontal, parietal, temporal, and occipital lobes of the cerebellum and superior sagittal sinus region of the DM. Mechanical characterisation revealed a significantly higher elastic modulus for the superior sagittal sinus region when compared to other regions in the DM. The superior sagittal sinus and parietal regions of the DM also displayed local mechanical anisotropy. Further, fatigue was noted in the DM following ten preconditioning cycles, which could have important implications in the context of repetitive TBI. To further understand differences in regional mechanical properties, regional variations in protein content (collagen I, collagen III, fibronectin and elastin) were examined by immunoblot analysis. The superior sagittal sinus was found to have significantly higher collagen I, elastin, and fibronectin content. The frontal region was also identified to have significantly higher collagen I and fibronectin content while the temporal region had increased elastin and fibronectin content. Regional differences in the mechanical and biochemical properties along with regional tissue thickness differences within the DM reveal that the tissue is a non-homogeneous structure. In particular, the potentially influential role of the superior sagittal sinus in TBI biomechanics warrants further investigation

On the mechanical behaviour of carotid artery plaques:the influence of curve-fitting experimental data on numerical model results

Computational models of diseased arteries are advancing rapidly, and a need exists to develop more accurate material models of human atherosclerotic plaques. However, intact samples for in vitro mechanical testing are not readily available. Most plaque samples are harvested from carotid endarterectomies where the geometries are not suitable for the boundary parameters necessary for classical uniaxial tensile testing. Experimental studies of biological tissue, particularly human plaque tissue, have not specified the minimum width-to-length (WL) ratio necessary for appropriate tensile testing. This study proposes either tensile or planar shear testing on whole specimen samples depending on the WL ratio. However, a “grey-area” of WL ratios exists which are unsuitable for either test, between 0.5:1 and 4:1 WL ratio. Eighteen plaque samples are investigated in this study, and according to classical approaches, two of the plaque samples have WL ratios suitable for tensile testing and four are suitable for planar shear testing. The remaining twelve samples fall in the grey-area of WL ratio. The study analyses which test method is suitable for the samples in this grey-area and what effect using the incorrect test method has on results from a computational model. The study highlights that tissues above a WL ratio of 2:1 are suitable for planar shear testing, and samples below 1:1 are more suited for tensile testing. Therefore, the “grey-area” can be reduced with certain limitations applied by the minor strain assumption which need to be taken into account during experimental testing. This study also demonstrates the influence of curve-fitting experimental results using tensile- and planar shear–based boundary parameters from eighteen plaque samples

The effect of serum starvation on tight junctional proteins and barrier formation in Caco-2 cells

Assessing the ability of pharmaceutics to cross biological barriers and reach the site-of-action requires faithful representation of these barriers in vitro. Difficulties have arisen in replicating in vivo resistance in vitro. This paper investigated serum starvation as a method to increase Caco-2 barrier stability and resistance. The effect of serum starvation on tight junction production was examined using transwell models; specifically, transendothelial electrical resistance (TEER), and the expression and localization of tight junction proteins, occludin and zonula occludens-1 (ZO-1), were studied using western blotting and immunofluorescence. Changing cells to serum-free media 2 days post-seeding resulted in TEER readings of nearly 5000 Ω cm2 but the TEER rapidly declined subsequently. Meanwhile, exchanging cells to serum-free media 4–6 days post-seeding produced barriers with resistance readings between 3000 and 4000 Ω cm2, which could be maintained for 18 days. This corresponded to an increase in occludin levels. Serum starvation as a means of barrier formation is simple, reproducible, and cost-effective. It could feasibly be implemented in a variety of pre-clinical pharmaceutical assessments of drug permeability across various biological barriers with the view to improving the clinical translation of novel therapeutic

Drug delivery across the blood-brain barrier: recent advances in the use of nanocarriers

The blood-brain barrier (BBB) has a significant contribution to homeostasis and protection of the CNS. However, it also limits the crossing of therapeutics and thereby complicates the treatment of CNS disorders. To overcome this limitation, the use of nanocarriers for drug delivery across the BBB has recently been exploited. Nanocarriers can utilize different physiological mechanisms for drug delivery across the BBB and can be modified to achieve the desired kinetics and efficacy. Consequentially, several nanocarriers have been reported to act as functional nanomedicines in preclinical studies using animal models for human diseases. Given the rapid development of novel nanocarriers, this review provides a comprehensive insight into the most recent advancements made in nanocarrier-based drug delivery to the CNS, such as the development of multifunctional nanomedicines and theranostics

Urinary bladder vs gastrointestinal tissue: a comparative study of their biomechanical properties for urinary tract reconstruction

OBJECTIVE To evaluate the mechanical properties of gastrointestinal (GI) tissue segments and to compare them with the urinary bladder for urinary tract reconstruction. METHODS Urinary bladders and GI tissue segments were sourced from porcine models (n = 6, 7 months old [5 male; 1 female]). Uniaxial planar tension tests were performed on bladder tissue, and Cauchy stress-stretch ratio responses were compared with stomach, jejunum, ileum, and colonic GI tissue. RESULTS The biomechanical properties of the bladder differed significantly from jejunum, ileum, and colonic GI tissue. Young modulus (kPa—measure of stiffness) of the GI tissue segments was on average 3.07-fold (±0.21 standard error) higher than bladder tissue (P < .01), and the strain at Cauchy stress of 50 kPa for bladder tissues was on average 2.27-fold (±0.20) higher than GI tissues. There were no significant differences between the averaged stretch ratio and Young modulus of the horizontal and vertical directions of bladder tissue (315.05 ± 49.64 kPa and 283.62 ± 57.04, respectively, P = .42). However, stomach tissues were 1.09- (±0.17) and 0.85- (±0.03) fold greater than bladder tissues for Young modulus and strain at 50 kPa, respectively. CONCLUSION An ideal urinary bladder replacement biomaterial should demonstrate mechanical equivalence to native tissue. Our findings demonstrate that GI tissue does not meet these mechanical requirements. Knowledge on the biomechanical properties of bladder and GI tissue may improve development opportunities for more suitable urologic reconstructive biomaterials

Regional mechanical and biochemical properties of the porcine cortical meninges

The meninges are pivotal in protecting the brain against traumatic brain injury (TBI), an ongoing issue in most mainstream sports. Improved understanding of TBI biomechanics and pathophysiology is desirable to improve preventative measures, such as protective helmets, and advance our TBI diagnostic/prognostic capabilities. This study mechanically characterised the porcine meninges by performing uniaxial tensile testing on the dura mater (DM) tissue adjacent to the frontal, parietal, temporal, and occipital lobes of the cerebellum and superior sagittal sinus region of the DM. Mechanical characterisation revealed a significantly higher elastic modulus for the superior sagittal sinus region when compared to other regions in the DM. The superior sagittal sinus and parietal regions of the DM also displayed local mechanical anisotropy. Further, fatigue was noted in the DM following ten preconditioning cycles, which could have important implications in the context of repetitive TBI. To further understand differences in regional mechanical properties, regional variations in protein content (collagen I, collagen III, fibronectin and elastin) were examined by immunoblot analysis. The superior sagittal sinus was found to have significantly higher collagen I, elastin, and fibronectin content. The frontal region was also identified to have significantly higher collagen I and fibronectin content while the temporal region had increased elastin and fibronectin content. Regional differences in the mechanical and biochemical properties along with regional tissue thickness differences within the DM reveal that the tissue is a non-homogeneous structure. In particular, the potentially influential role of the superior sagittal sinus in TBI biomechanics warrants further investigation

Mechanical and morphological characterisation of porcine urethras for the assessment of paediatric urinary catheter safety

Paediatric urinary catheters are often necessary in critical care settings or to address congenital anomalies  affecting the urogenital system. Iatrogenic injuries can occur during the placement of such catheters, highlighting  the need for a safety device that can function in paediatric settings. Despite successful efforts to develop devices  that improve the safety of adult urinary catheters, no such devices are available for use with paediatric catheters.  This study investigates the potential for utilising a pressure-controlled safety mechanism to limit the trauma  experienced by paediatric patients during inadvertent inflation of a urinary catheter anchoring balloon in the  urethra. Firstly, we establish a paediatric model of the human urethra using porcine tissue by characterising the  mechanical and morphological properties of porcine tissue at increasing postnatal timepoints (8, 12, 16 and 30  weeks). We identified that porcine urethras harvested from pigs at postnatal week 8 and 12 exhibit morpho?logical properties (diameter and thickness) that are statistically distinct from adult porcine urethras (postnatal  week 30). We therefore utilise urethra tissue from postnatal week 8 and 12 pigs as a model to evaluate a pressure-controlled approach to paediatric urinary catheter balloon inflation intended to limit tissue trauma during  inadvertent inflation in the urethra. Our results show that limiting catheter system pressure to 150 kPa avoided  trauma in all tissue samples. Conversely, all of the tissue samples that underwent traditional uncontrolled urinary  catheter inflation experienced complete rupture. The findings of this study pave the way for the development of a  safety device for use with paediatric catheters, thereby alleviating the burden of catastrophic trauma and life  changing injuries in children due to a preventable iatrogenic urogenital event.  </p

Development of a platform for studying 3D astrocyte mechanobiology: compression of astrocytes in collagen gels

Glaucoma is a common optic neuropathy characterized by retinal ganglion cell death. Elevated intraocular pressure (IOP), a key risk factor for glaucoma, leads to significant biomechanical deformation of optic nerve head (ONH) cells and tissues. ONH astrocytes respond to this deformation by transforming to a reactive, proliferative phenotype, which has been implicated in the progression of glaucomatous vision loss. However, little is known about the mechanisms of this transformation. In this study, we developed a 3D collagen gel culture system to mimic features of ONH deformation due to elevated IOP. Compressive loading of astrocyte-seeded collagen gels led to cell alignment perpendicular to the direction of strain, and increased astrocyte activation, as assayed by GFAP, vimentin, and s100β levels, as well as MMP activity. This proof-of-concept study shows that this system has potential for studying mechanisms of astrocyte mechanobiology as related to the pathogenesis of glaucoma. Further work is needed to establish the possible interplay of mechanical stimulation, matrix properties, and hypoxia on the observed response of astrocytes