Stem cell regulatory gene expression in human adult dental pulp and periodontal ligament cells undergoing odontogenic/osteogenic differentiation

Introduction During development and regeneration, odontogenesis and osteogenesis are initiated by a cascade of signals driven by several master regulatory genes. Methods In this study, we investigated the differential expression of 84 stem cell–related genes in dental pulp cells (DPCs) and periodontal ligament cells (PDLCs) undergoing odontogenic/osteogenic differentiation. Results Our results showed that, although there was considerable overlap, certain genes had more differential expression in PDLCs than in DPCs. CCND2, DLL1, and MME were the major upregulated genes in both PDLCs and DPCs, whereas KRT15 was the only gene significantly downregulated in PDLCs and DPCs in both odontogenic and osteogenic differentiation. Interestingly, a large number of regulatory genes in odontogenic and osteogenic differentiation interact or crosstalk via Notch, Wnt, transforming growth factor β (TGF-β)/bone morphogenic protein (BMP), and cadherin signaling pathways, such as the regulation of APC, DLL1, CCND2, BMP2, and CDH1. Using a rat dental pulp and periodontal defect model, the expression and distribution of both BMP2 and CDH1 have been verified for their spatial localization in dental pulp and periodontal tissue regeneration. Conclusions This study has generated an overview of stem cell–related gene expression in DPCs and PDLCs during odontogenic/osteogenic differentiation and revealed that these genes may interact through the Notch, Wnt, TGF-β/BMP, and cadherin signalling pathways to play a crucial role in determining the fate of dental derived cell and dental tissue regeneration. These findings provided a new insight into the molecular mechanisms of the dental tissue mineralization and regeneration

Enhanced human bone marrow stromal cell affinity for modified poly(L-lactide) surfaces by the upregulation of adhesion molecular genes

To enhance and regulate cell affinity for poly (l-lactic acid) (PLLA) based materials, two hydrophilic ligands, poly (ethylene glycol) (PEG) and poly (l-lysine) (PLL), were used to develop triblock copolymers: methoxy-terminated poly (ethylene glycol)-block-poly (l-lactide)-block-poly (l-lysine) (MPEG-b-PLLA-b-PLL) in order to regulate protein absorption and cell adhesion. Bone marrow stromal cells (BMSCs) were cultured on different composition of MPEG-b-PLLA-b-PLL copolymer films to determine the effect of modified polymer surfaces on BMSC attachment. To understand the molecular mechanism governing the initial cell adhesion on difference polymer surfaces, the mRNA expression of 84 human extracellular matrix (ECM) and adhesion molecules was analysed using quantitative reverse transcriptase polymerase chain reaction (qRT-PCR). It was found that down regulation of adhesion molecules was responsible for the impaired BMSC attachment on PLLA surface. MPEG-b-PLLA-b-PLL copolymer films improved significantly the cell adhesion and cytoskeleton expression by upregulation of relevant molecule genes significantly. Six adhesion genes (CDH1, ITGL, NCAM1, SGCE, COL16A1, and LAMA3) were most significantly influenced by the modified PLLA surfaces. In summary, polymer surfaces altered adhesion molecule gene expression of BMSCs, which consequently regulated cell initial attachment on modified PLLA surfaces.

Integrated Diagnosis and Prognosis Model for High Pressure LNG Pump

In condition-based maintenance (CBM), effective diagnostics and prognostics are essential tools for maintenance engineers to identify imminent fault and to predict the remaining useful life before the components finally fail. This enables remedial actions to be taken in advance and reschedules production if necessary. This paper presents a technique for accurate assessment of the remnant life of machines based on historical failure knowledge embedded in the closed loop diagnostic and prognostic system. The technique uses the Support Vector Machine (SVM) classifier for both fault diagnosis and evaluation of health stages of machine degradation. To validate the feasibility of the proposed model, the five different level data of typical four faults from High Pressure Liquefied Natural Gas (HP-LNG) pumps were used for multi-class fault diagnosis. In addition, two sets of impeller-rub data were analysed and employed to predict the remnant life of pump based on estimation of health state. The results obtained were very encouraging and showed that the proposed prognosis system has the potential to be used as an estimation tool for machine remnant life prediction in real life industrial applications.

Under great pressure, a diamond is being formed: The SBS over time

This article observes a paradox in the recent history of the Special Broadcasting Service. It is argued that, in contrast to the Australian Broadcasting Corporation, the role and general direction of SBS were not extensively debated as part of the ‘culture wars’ that occurred during the years of the Howard government. While that made SBS a less fraught space during that period, it may now be a factor in the comparative lack of support being given by the Rudd Labor government to SBS in comparison with the ABC, as some of the ‘special’ status of SBS has been blunted by its drift towards more mainstream programming and a mixed economy of commercial advertising, as well as government funding.

Initiation of embryogenic cell suspensions of taro (Colocasia esculenta var. esculenta) and plant regeneration

Embryogenic callus was initiated by culturing in vitro taro corm slices on agar-solidified half-strength MS medium containing 2.0 mg/L 2,4-dichlorophenoxyacetic acid (2,4-D) for 20 days followed by transfer to 1.0 mg/L thidiazuron (TDZ). Callus was subsequently proliferated on solid medium containing 1.0 mg/L TDZ, 0.5 mg/L 2,4- D and 800 mg/L glutamine before transfer to liquid medium containing the same components but with reduced glutamine (100 mg/L). After 3 months in liquid culture on an orbital shaker, cytoplasmically dense cell aggregates began to form. Somatic embryogenesis was induced by plating suspension cells onto solid media containing reduced levels of hormones (0.1 mg/L TDZ, 0.05 mg/L 2,4-D), high concentrations of sucrose (40–50 g/L) and biotin (1.0 mg/L). Embryo maturation and germination was then induced on media containing 0.05 mg/L benzyladenine (BA) and 0.1 mg/L indole-3-acetic acid (IAA). Histological studies of the developing embryos revealed the presence of typical shoot and root poles suggesting that these structures were true somatic embryos. The rate of somatic embryos formation was 500–3,000 per mL settledcell volume while approximately 60% of the embryos regenerated into plants.

Malnutrition and pressure ulcer risk in adults in Australian health care facilities

Aims To determine the effect of nutritional status on the presence and severity of pressure ulcers in statewide? public healthcare facilities, in Queensland, Australia. Research Methods A multicentre, cross sectional audit of nutritional status of a convenience sample of subjects was carried out as part of a large audit of pressure ulcers in a sample of state based public healthcare facilities in 2002 and 2003. Dietitians in 20 hospitals and six residential aged care facilities conducted single day nutritional status audits of 2208 acute and 839 aged care subjects using the Subjective Global Assessment. The effect of nutritional status on the presence, highest stage and number of pressure ulcers was determined by logistic regression in a model controlling for age, gender, medical specialty and facility location. The potential clustering effect of facility was accounted for in the model using an analysis of correlated data approach. Results Subjects with malnutrition had an adjusted odds risk of 2.6 (95% CI 1.8-3.5, p<0.001) of having a pressure ulcer in acute facilities and 2.0 (95% CI 1.5-2.7, p<0.001) for residential aged care facilities. There was also increased odds risk of having a pressure ulcer, having a higher stage pressure ulcer and a higher number of pressure ulcers with increased severity of malnutrition. Conclusion Malnutrition was associated with at least twice the odds risk of having a pressure ulcer of in public healthcare facilities in Queensland. Action must be taken to identify, prevent and treat malnutrition, especially in patients at risk of pressure ulcer.

Discovery and characterization of IGFBP-mediated endocytosis in the human retinal pigment epithelial cell line ARPE-19

Insulin-like growth factor binding proteins (IGFBPs) are prime regulators of IGF-action in numerous cell types including the retinal pigment epithelium (RPE). The RPE performs several functions essential for vision, including growth factor secretion and waste removal via a phagocytic process mediated in part by vitronectin (Vn). In the course of studying the effects of IGFBPs on IGF-mediated VEGF secretion and Vn-mediated phagocytosis in the RPE cell line ARPE-19, we have discovered that these cells avidly ingest synthetic microspheres (2.0 μm diameter) coated with IGFBPs. Given the novelty of this finding and the established role for endocytosis in mediating IGFBP actions in other cell types, we have explored the potential role of candidate cell surface receptors. Moreover, we have examined the role of key IGFBP structural motifs, by comparing responses to three members of the IGFBP family (IGFBP-3, IGFBP-4 and IGFBP-5) which display overlapping variations in primary structure and glycosylation status. Coating of microspheres (FluoSpheres®, sulfate modified polystyrene filled with a fluorophore) was conducted at 37 °C for 1 h using 20 μg/mL of test protein, followed by extensive washing. Binding of proteins was confirmed using a microBCA assay. The negative control consisted of microspheres treated with 0.1% bovine serum albumin (BSA), and all test samples were post-treated with BSA in an effort to coat any remaining free protein binding sites, which might otherwise encourage non-specific interactions with the cell surface. Serum-starved cultures of ARPE-19 cells were incubated with microspheres for 24 h, using a ratio of approximately 100 microspheres per cell. Uptake of microspheres was quantified using a fluorometer and was confirmed visually by confocal fluorescence microscopy. The ARPE-19 cells displayed little affinity for BSA-treated microspheres, but avidly ingested large quantities of those pre-treated with Vn (ANOVA; p < 0.001). Strong responses were also observed towards recombinant formulations of non-glycosylated IGFBP-3, glycosylated IGFBP-3 and glycosylated IGFBP-5 (all p < 0.001), while glycosylated IGFBP-4 induced a relatively minor response (p < 0.05). The response to IGFBP-3 was unaffected in the presence of excess soluble IGFBP-3, IGF-I or Vn. Likewise, soluble IGFBP-3 did not induce uptake of BSA-treated microspheres. Antibodies to either the transferrin receptor or type 1 IGF-receptor displayed slight inhibitory effects on responses to IGFBPs and Vn. Heparin abolished responses to Vn, IGFBP-5 and non-glycosylated IGFBP-3, but only partially inhibited the response to glycosylated IGFBP-3. Our results demonstrate for the first time IGFBP-mediated endocytosis in ARPE-19 cells and suggest roles for the IGFBP-heparin-binding domain and glycosylation status. These findings have important implications for understanding the mechanisms of IGFBP actions on the RPE, and in particular suggest a role for IGFBP-endocytosis.

The costs arising from pressure ulcers attributable to malnutrition

Aim To estimate the economic consequences of pressure ulcers attributable to malnutrition. Method Statistical models were developed to predict the number of cases of pressure ulcer, associated bed days lost and the dollar value of these losses in public hospitals in 2002/2003 in Queensland, Australia. The following input parameters were specified and appropriate probability distributions fitted • Number of at risk discharges per annum • Incidence rate for pressure ulcer • Attributable fraction of malnutrition in the development of pressure ulcer • Independent effect of pressure ulcer on length of hospital stay • Opportunity cost of hospital bed day One thousand random re-samples were made and the results expressed as (output) probabilistic distributions. Results The model predicts a mean 16060 (SD 5 671) bed days lost and corresponding mean economic cost of AU$12 968 668 (SD AU$4 924 148) (EUROS 6 925 268 SD 2 629 495; US$ 7 288 391 SD 2 767 371) of pressure ulcer attributable to malnutrition in 2002/2003 in public hospitals in Queensland, Australia. Conclusion The cost of pressure ulcer attributable to malnutrition in bed days and dollar terms are substantial. The model only considers costs of increased length of stay associated with pressure ulcer and not other factors associated with care.

Intraocular pressure in keratoconus

Aims: To investigate IOP measurements with the dynamic contour tonometer (DCT) and non contact tonometer (NCT) in subjects with keratoconus. Methods: Twenty keratoconic subjects and 20 age-matched control subjects had IOP measurements taken using DCT and NCT instruments. Central and offcentre measures were taken with the DCT in order to highlight any systematic errors associated with corneal biomechanical factors. Measures of anterior and posterior corneal topography and thickness were also taken for each subject. Results: No significant difference was found between the central and off-centre DCT IOP readings for the keratoconics and age-matched controls (p>0.05). The average DCT IOP for the keratoconics was 14.2 ± 1.4 mmHg and for the agematched controls was 14.2 ± 1.6 mmHg. However, the average NCT readings differed significantly (p<0.001) between the keratoconics (9.2 ± 1.5 mmHg) and age-matched controls (12.9 ± 2.4 mmHg). DCT IOP showed no significant (p>0.05) correlation with the severity of keratoconus, as determined through measures of corneal topography and thickness. NCT IOP was correlated significantly with certain measures of corneal curvature and thickness in the keratoconic population. The difference between DCT and NCT IOP was strongly correlated with measures of corneal topography and thickness, with differences increasing for more advanced keratoconus. Conclusions: The measurements from the DCT do not appear to be dependent upon corneal factors, unlike the NCT. The presence or severity of keratoconus was not correlated with DCT IOP values.

Vitronectin modulates human mesenchymal stem cell response to insulin-like growth factor-I and transforming growth factor beta 1 in a serum-free environment

The use of animal sera for the culture of therapeutically important cells impedes the clinical use of the cells. We sought to characterize the functional response of human mesenchymal stem cells (hMSCs) to specific proteins known to exist in bone tissue with a view to eliminating the requirement of animal sera. Insulin-like growth factor-I (IGF-I), via IGF binding protein-3 or -5 (IGFBP-3 or -5) and transforming growth factor-beta 1 (TGF-beta(1)) are known to associate with the extracellular matrix (ECM) protein vitronectin (VN) and elicit functional responses in a range of cell types in vitro. We found that specific combinations of VN, IGFBP-3 or -5, and IGF-I or TGF-beta(1) could stimulate initial functional responses in hMSCs and that IGF-I or TGF-beta(1) induced hMSC aggregation, but VN concentration modulated this effect. We speculated that the aggregation effect may be due to endogenous protease activity, although we found that neither IGF-I nor TGF-beta(1) affected the functional expression of matrix metalloprotease-2 or -9, two common proteases expressed by hMSCs. In summary, combinations of the ECM and growth factors described herein may form the basis of defined cell culture media supplements, although the effect of endogenous protease expression on the function of such proteins requires investigation.

Microassembly fabrication of tissue engineering scaffolds with customized design

Browse > Journals> Automation Science and Enginee ...> Volume: 5 Issue: 3 Microassembly Fabrication of Tissue Engineering Scaffolds With Customized Design 4468741 abstract Han Zhang; Burdet, E.; Poo, A.N.; Hutmacher, D.W.; GE Global Res. Center Ltd., Shanghai This paper appears in: Automation Science and Engineering, IEEE Transactions on Issue Date: July 2008 Volume: 5 Issue:3 On page(s): 446 - 456 ISSN: 1545-5955 Digital Object Identifier: 10.1109/TASE.2008.917011 Date of Current Version: 02 July 2008 Sponsored by: IEEE Robotics and Automation Society Abstract This paper presents a novel technique to fabricate scaffold/cell constructs for tissue engineering by robotic assembly of microscopic building blocks (of volume 0.5$,times,$0.5$,times,$0.2 ${hbox{mm}}^{3}$ and 60 $mu {hbox{m}}$ thickness). In this way, it becomes possible to build scaffolds with freedom in the design of architecture, surface morphology, and chemistry. Biocompatible microparts with complex 3-D shapes were first designed and mass produced using MEMS techniques. Semi-automatic assembly was then realized using a robotic workstation with four degrees of freedom integrating a dedicated microgripper and two optical microscopes. Coarse movement of the gripper is determined by pattern matching in the microscopes images, while the operator controls fine positioning and accurate insertion of the microparts. Successful microassembly was demonstrated using SU-8 and acrylic resin microparts. Taking advantage of parts distortion and adhesion forces, which dominate at micro-level, the parts cleave together after assembly. In contrast to many current scaffold fabrication techniques, no heat, pressure, electrical effect, or toxic chemical reaction is involved, a critical condition for creating scaffolds with biological agents.

Doped poly(vinylidene fluoride) as solid polymer electrolyte in nanocrystalline dye-sensitized solar cell

A new solid composite polymer electrolyte was reported by incorporating Azino-bis-(3-ethyl benzo thiazoline-6-sulphonate) ion [ABTS] as dopant in poly(vinylidene flouride) along with redox couple (1-/13-). Under certain conditions, the electrolyte composition forms brush like nano-rods while it is doped with Azino-bis-(3-ethly) benzo thiazoline-6-sulphonate) ion [ABTS], a pi-electron donor. The polymer electrolyte forms nanoscale interpenetrating network with the crystalline order of the polymer electrolyte that seems to be a desirable architecture for the active layer of the photoelectrochemical cell. With this new polymer electrolyte, dye-sensitized solar cell was fabricated using N3 dye absorbed over Ti02- nonoparticles (photoanode) and conducting carbon cement coated on the conducting press (FTO, photocathode). This polymer composite has been successfully used as a promising candidate as solid polymer electrolyte in nanocrystalline dye-sensitized solar cell.