Pressure Combined with Ischemia/Reperfusion Injury Induces Deep Tissue Injury via Endoplasmic Reticulum Stress in a Rat Pressure Ulcer Model

Pressure ulcer is a complex and significant health problem in long-term bedridden patients, and there is currently no effective treatment or efficient prevention method. Furthermore, the molecular mechanisms and pathogenesis contributing to the deep injury of pressure ulcers are unclear. The aim of the study was to explore the role of endoplasmic reticulum (ER) stress and Akt/GSK3β signaling in pressure ulcers. A model of pressure-induced deep tissue injury in adult Sprague-Dawley rats was established. Rats were treated with 2-h compression and subsequent 0.5-h release for various cycles. After recovery, the tissue in the compressed regions was collected for further analysis. The compressed muscle tissues showed clear cellular degenerative features. First, the expression levels of ER stress proteins GRP78, CHOP, and caspase-12 were generally increased compared to those in the control. Phosphorylated Akt and phosphorylated GSK3β were upregulated in the beginning of muscle compression, and immediately significantly decreased at the initiation of ischemia-reperfusion injury in compressed muscles tissue. These data show that ER stress may be involved in the underlying mechanisms of cell degeneration after pressure ulcers and that the Akt/GSK3β signal pathway may play an important role in deep tissue injury induced by pressure and ischemia/reperfusion.     

Identification of potential biophysical and molecular signalling mechanisms underlying hyaluronic acid enhancement of cartilage formation

This study determined the effects of exogenous hyaluronic acid (HA) on the biomechanical and biochemical properties of self-assembled bovine chondrocytes, and investigated biophysical and genetic mechanisms underlying these effects. The effects of HA commencement time, concentration, application duration and molecular weight were examined using histology, biomechanics and biochemistry. Additionally, the effects of HA application on sulphated glycosaminoglycan (GAG) retention were assessed. To investigate the influence of HA on gene expression, microarray analysis was conducted. HA treatment of developing neocartilage increased compressive stiffness onefold and increased sulphated GAG content by 35 per cent. These effects were dependent on HA molecular weight, concentration and application commencement time. Additionally, applying HA increased sulphated GAG retention within self-assembled neotissue. HA administration also upregulated 503 genes, including multiple genes associated with TGF-β1 signalling. Increased sulphated GAG retention indicated that HA could enhance compressive stiffness by increasing the osmotic pressure that negatively charged GAGs create. The gene expression data demonstrate that HA treatment differentially regulates genes related to TGF-β1 signalling, revealing a potential mechanism for altering matrix composition. These results illustrate the potential use of HA to improve cartilage regeneration efforts and better understand cartilage development.     

The relationship between numbers of bacteria on surfaces and in deep tissues of mechanically tenderized beef

The objective of the study was to identify factors affecting the fractions of the bacteria naturally present on surfaces of beef cuts that are carried into deep tissues when the meat is mechanical tenderized by piercing with banks of thin blades. The surfaces and ten strips of meat from the deep tissues of beef primal cuts tenderized first and last on each of five days at a retail store meat fabrication facility were sampled for enumeration of total aerobic counts. Each strip was excised from the whole thickness of a cut after surfaces were sterilized. The mean log numbers of total aerobes recovered from the surfaces of cuts tenderized first or last each day were 2.18 and 1.57 log cfu cm⁻², respectively. The mean log numbers recovered per strip from individual cuts tenderized first or last each day ranged from 0.30 to 1.45 and from 0.03 to 1.04 log cfu, respectively. These findings indicate that bacteria from the tenderizing equipment augmented the numbers of aerobes on the surfaces of cuts tenderized first each day, with some of the additional aerobes being carried into deep tissues. Subsequently, pieces of cuts stored in air at 2 °C were tenderized at a laboratory using commercial equipment. Each cut was divided into three pieces with one piece being not treated, one being sprayed with water and one being sprayed with 5% lactic acid. The mean log numbers of total aerobes recovered from the surfaces of not treated pieces of cut stored for 0, 2, 4, 6, 10 or 14 days were 0.6, 0.8, 2.6, 4.2, 8.5 and 8.9 log cfu cm⁻², respectively. No aerobes were recovered from the deep tissues of any of the pieces of cuts tenderized on day 0. Mean log numbers recovered from the deep tissues of not treated tenderized pieces of cuts stored for 2, 4, 6, 10 or 14 days were 0.3, 0.3, 2.2, 7.8 and 8.1 log cfu per strip, respectively. Spraying with 5% lactic acid reduced the mean log numbers of aerobes on pieces of cuts stored for 2, 4, or 6 days by 1, 2 or 2 log units, respectively, but did not reduce the numbers on pieces of cuts stored for 10 or 14 days. Mean log numbers recovered from the deep tissues of tenderized pieces of cuts sprayed with 5% lactic acid were not significantly different (P > 0.05) from the mean log numbers recovered from the corresponding, tenderized not treated pieces of cuts. These findings showed that the fraction of the total aerobes on cut surfaces that are carried into deep tissues during mechanical tenderizing can vary with the stage of growth of the spoilage flora; and that reduction of numbers of aerobes on the surface by treatment with lactic acid before tenderizing does not necessarily reduce the numbers carried into deep tissues during tenderizing.     

Peptide‐functionalized reduced graphene oxide as a bioactive mechanically robust tissue regeneration scaffold

Bioactive, synthetic materials represent next‐generation composites for tissue regeneration. Design of contemporary materials attempts to recapitulate the complexities of native tissue; however, few successfully mimic the order in nature. Recently, graphene oxide (GO ) has emerged as a scaffold due to its potential for bioactive functionalization and long‐range order instilled by the self‐assembly of graphene sheets. Chemical reduction of GO results in a more compatible material with enhanced properties but compromises the ability to functionalize the graphenic backbone. However, using Johnson–Claisen rearrangement chemistry, functionalization is achieved that is not liable to reduction. From reduced Claisen graphene, we polymerized short homopeptides from α ‐amino acid N ‐carboxyanhydride monomers of glutamate and lysine to result in functionalized graphenes (pGlu‐rCG and pLys‐rCG ) that are cytocompatible, degradable, and bioactive. Exposure to NIH‐3T3 fibroblasts and RAW 264.7 macrophages revealed that the materials are cytocompatible and do not alter important sub‐cellular compartments. Powders were hot pressed to form mechanically stiff (E ′: 41 and 49 MPa ), strong (UCS : 480 and 140 MPa ), and tough (U _T: 2898 and 584 J m^?3 × 10⁴) three‐dimensional constructs (pGlu‐rCG and pLys‐rCG, respectively). Overall, we report a robust chemistry and processing strategy for facile bioactive functionalization of compatible, reduced Claisen graphene for three‐dimensional biomedical applications. © 2017 Society of Chemical Industry     

In vitro evaluation of tissue adhesives composed of hydrophobically modified gelatins and disuccinimidyl tartrate

Abstract

The effect of the hydrophobic group content in gelatin on the bonding strength of novel tissue–penetrating tissue adhesives was evaluated. The hydrophobic groups introduced into gelatin were the saturated hexanoyl, palmitoyl, and stearoyl groups, and the unsaturated oleoyl group. A collagen casing was employed as an adherend to model soft tissue for the in vitro determination of bonding strength of tissue adhesives composed of various hydrophobically modified gelatins and disuccinimidyl tartrate. The adhesive composed of stearoyl-modified gelatin (7.4% stearoyl; 10Ste) and disuccinimidyl tartrate showed the highest bonding strength. The bonding strength of the adhesives decreased as the degree of substitution of the hydrophobic groups increased. Cell culture experiments demonstrated that fluorescein isothiocyanate-labeled 10Ste was integrated onto the surface of smooth muscle cells and showed no cytotoxicity. These results suggest that 10Ste interacted with the hydrophobic domains of collagen casings, such as hydrophobic amino acid residues and cell membranes. Therefore, 10Ste–disuccinimidyl tartrate is a promising adhesive for use in aortic dissection.     

The Role of Pericytes in Neurovascular Unit Remodeling in Brain Disorders

Neurons are extremely vulnerable cells that tightly rely on the brain’s highly dynamic and complex vascular network that assures an accurate and adequate distribution of nutrients and oxygen. The neurovascular unit (NVU) couples neuronal activity to vascular function, controls brain homeostasis, and maintains an optimal brain microenvironment adequate for neuronal survival by adjusting blood-brain barrier (BBB) parameters based on brain needs. The NVU is a heterogeneous structure constituted by different cell types that includes pericytes. Pericytes are localized at the abluminal side of brain microvessels and contribute to NVU function. Pericytes play essential roles in the development and maturation of the neurovascular system during embryogenesis and stability during adulthood. Initially, pericytes were described as contractile cells involved in controlling neurovascular tone. However, recent reports have shown that pericytes dynamically respond to stress induced by injury upon brain diseases, by chemically and physically communicating with neighboring cells, by their immune properties and by their potential pluripotent nature within the neurovascular niche. As such, in this paper, we would like to review the role of pericytes in NVU remodeling, and their potential as targets for NVU repair strategies and consequently neuroprotection in two pathophysiologically distinct brain disorders: ischemic stroke and Alzheimer’s disease (AD).     

A Novel Protein Is Lower Expressed in Renal Cell Carcinoma

Engrailed-2 (EN2) has been identified as a candidate oncogene in breast cancer and prostate cancer. It is usually recognized as a mainly nuclear staining in the cells. However, recent studies showed a cytoplasmic staining occurred in prostate cancer, bladder cancer and clear cell renal cell carcinoma. The inconsistency makes us confused. To clarify the localization and expression of EN2 in renal cell carcinoma, anti-EN2 antibody (ab28731) and anti-EN2 antibody (MAB2600) were used for immunohistochemistry (IHC) respectively. Interestingly, we found that EN2 detected by ab28731 was mainly presented in cytoplasm while EN2 detected by MAB2600 was mainly presented in nucleus. To further investigate the different patterns observed above, lysates from full-length EN2 over expression in HEK293T cells were used to identify which antibody the EN2 molecule bound by western blot. Results showed ab28731 did not react with the lysates. For this reason, the novel specific protein detected by ab28731 was not the EN2 molecule and was named nonEN2. Then using the renal carcinoma tissue microarray and renal tissues, we found that the protein expression levels of nonEN2 in kidney tumor tissues was significantly lower than that in kidney normal tissues (p < 0.05), so was in renal cell lines. Taken together, nonEN2 is lower expressed and may play an important role in renal cell carcinoma.     

Short communication: Genetic characterization of digital cushion thickness

Dairy cow lameness is a serious animal welfare issue. It is also a significant cause of economic losses, reducing reproductive efficiency and milk production and increasing culling rates. The digital cushion is a complex structure composed mostly of adipose tissue located underneath the distal phalanx and has recently been phenotypically associated with incidence of claw horn disruption lesions (CHDL); namely, sole ulcers and white line disease. The objective of this study was to characterize digital cushion thickness genetically and to investigate its association with body condition score (BCS), locomotion score (LOCO), CHDL, and milk production. Data were collected from 1 large closely monitored commercial dairy farm located in upstate New York; 923 dairy cows were used. Before trimming, the following data were collected by a member of the research team: BCS, cow height measurement, and LOCO. Presence or not of CHDL (sole ulcer or white line disease, or both) was recorded at trimming. Immediately after the cows were hoof trimmed, they underwent digital sonographic B-mode examination for the measurement of digital cushion thickness. Factors such as parity number, stage of lactation, calving date, mature-equivalent 305-d milk yield (ME305MY), and pedigree information were obtained from the farm’s dairy management software (DairyCOMP 305; Valley Agricultural Software, Tulare, CA). Univariate animal models were used to obtain variance component estimations for each studied trait (CHDL, BCS, digital cushion thickness average, LOCO, height, and ME305MY) and a 6-variate analysis was conducted to estimate the genetic, residual, and phenotypic correlations between the studied traits. The heritability estimate of DCTA was 0.33 ± 0.09, whereas a statistically significant genetic correlation was estimated between DCTA and CHDL (−0.60 ± 0.29). Of the other genetic correlations, significant estimates were derived for BCS with LOCO (−0.49 ± 0.19) and ME305MY (−0.48 ± 0.20). Digital cushion thickness is moderately heritable and genetically strongly correlated with CHDL.