Effects of varied ionic calcium and phosphate on the proliferation, osteogenic differentiation and mineralization of human periodontal ligament cells in vitro

Background and Objective: A number of bone filling materials containing calcium (Ca++) and phosphate (P) ions have been used in the repair of periodontal bone defects; however, the effect that local release of Ca++ and P ions have on biological reactions is not fully understood. In this study, we investigated the effects of various levels of Ca++ and P ions on the proliferation, osteogenic differentiation, and mineralization of human periodontal ligament cells (hPDLCs). Materials and Methods: hPDLCs were obtained using an explant culture method. Defined concentrations and ratios of ionic Ca++ to inorganic P were added to standard culture and osteogenic induction media. The ability of hPDLCs to proliferate in these growth media was assayed using the Cell Counting Kit-8 (CCK-8). Cell apoptosis was evaluated by FITC-Annexin V/PI double staining method. Osteogenic differentiation and mineralization were investigated by morphological observations, alkaline phosphatase (ALP) activity, and Alizarin red S/von Kossa staining. The mRNA expression of osteogenic related markers was analyzed using a reverse transcriptase polymerase chain reaction (RT-PCR). Results: Within the ranges of Ca++ and P ions concentrations tested, we observed that increased concentrations of Ca++ and P ions enhanced cell proliferation and formation of mineralized matrix nodules; whereas ALP activity was reduced. The RT-PCR results showed that elevated concentrations of Ca++ and P ions led to a general increase of Runx2 mRNA expression and decreased ALP mRNA expression, but gave no clear trend on OCN mRNA levels. Conclusion: The concentrations and ratios of Ca++ and P ions could significantly influence proliferation, differentiation, and mineralization of hPDLCs. Within the range of concentrations tested, we found that the combination of 9.0 mM Ca++ ions and 4.5 mM P ions were the optimum concentrations for proliferation, differentiation, and mineralization in hPDLCs.

Calcium ions promote osteogenic differentiation and mineralization of human dental pulp cells : implication for pulp capping materials

Calcium (Ca) is the main element of most pulp capping materials and plays an essential role in mineralization. Different pulp capping materials can release various concentrations of Ca ions leading to different clinical outcomes. The purpose of this study was to investigate the effects of various concentrations of Ca ions on the growth and osteogenic differentiation of human dental pulp cells (hDPCs). Different concentrations of Ca ions were added to growth culture medium and osteogenic inductive culture medium. A Cell Counting Kit-8 (CCK-8) was used to determine the proliferation of hDPCs in growth culture medium. Osteogenic differentiation and mineralization were measured by alkaline phosphatase (ALP) assay, Alizarin red S/von kossa staining, calcium content quantitative assay. The selected osteogenic differentiation markers were investigated by quantitative real-time polymerase chain reaction (qRT-PCR). Within the range of 1.8–16.2 mM, increased concentrations of Ca ions had no effect on cell proliferation, but led to changes in osteogenic differentiation. It was noted that enhanced mineralized matrix nodule formation was found in higher Ca ions concentrations; however, ALP activity and gene expression were reduced. qRT-PCR results showed a trend towards down-regulated mRNA expression of type I collagen (COL1A2) and Runx2 at elevated concentrations of Ca ions, whereas osteopontin (OPN) and osteocalcin (OCN) mRNA expression was significantly up-regulated. Ca ions content in the culture media can significantly influence the osteogenic properties of hDPCs, indicating the importance of optimizing Ca ions release from dental pulp capping materials in order to achieve desirable clinical outcomes.

Stability of hydrotalcites formed from Bayer refinery environmental control processes

Bauxite refinery residues (red mud) are derived from the Bayer process by the digestion of crushed bauxite in concentrated sodium hydroxide at elevated temperatures and pressures. This slurry residue, if untreated, is unsuitable for discharge directly into the environment and is usually stored in tailing dams. The liquid portion has the potential for discharge, but requires pre-treatment before this can occur. The seawater neutralisation treatment facilitates a significant reduction in pH and dissolved metal concentrations, through the precipitation of hydrotalcite-like compounds and some other Mg, Ca, and Al hydroxide and carbonate minerals. The hydrotalcite-like compounds, precipitated during seawater neutralisation, also remove a range of transition metals, oxy-anions and other anionic species through a combination of intercalation and adsorption reactions: smaller anions are intercalated into the hydrotalcite matrix, while larger molecules are adsorbed on the particle surfaces. A phenomenon known as ‘reversion’ can occur if the seawater neutralisation process is not properly controlled. Reversion causes an increase in the pH and dissolved impurity levels of the neutralised effluent, rendering it unsuitable for discharge. It is believed that slow dissolution of components of the red mud residue and compounds formed during the neutralisation process are responsible for reversion. This investigation looked at characterising natural hydrotalcite (Mg6Al2(OH)16(CO3)∙4H2O) and ‘Bayer’ hydrotalcite (synthesised using the seawater neutralisation process) using a variety of techniques including X-ray diffraction, infrared and Raman spectroscopy, and thermogravimetric analysis. This investigation showed that Bayer hydrotalcite is comprised of a mixture of 3:1 and 4:1 hydrotalcite structures and exhibited similar chemical characteristic to the 4:1 synthetic hydrotalcite. Hydrotalcite formed from the seawater neutralisation of Bauxite refinery residues has been found not to cause reversion. Other components in red mud were investigated to determine the cause of reversion and this investigation found three components that contributed to reversion: 1) tricalcium aluminate, 2) hydrocalumite and 3) calcium hydroxide. Increasing the amount of magnesium in the neutralisation process has been found to be successful in reducing reversion.

Active regulation of the epidermal calcium profile

A distinct calcium profile is strongly implicated in regulating the multi-layered structure of the epidermis. However, the mechanisms that govern the regulation of this calcium profile are currently unclear. It clearly depends on the relatively impermeable barrier of the stratum corneum (passive regulation) but may also depend on calcium exchanges between keratinocytes and extracellular fluid (active regulation). Using a mathematical model that treats the viable sublayers of unwounded human and murine epidermis as porous media and assumes that their calcium profiles are passively regulated, we demonstrate that these profiles are also actively regulated. To obtain this result, we found that diffusion governs extracellular calcium motion in the viable epidermis and hence intracellular calcium is the main source of the epidermal calcium profile. Then, by comparison with experimental calcium profiles and combination with a hypothesised cell velocity distribution in the viable epidermis, we found that the net influx of calcium ions into keratinocytes from extracellular fluid may be constant and positive throughout the stratum basale and stratum spinosum, and that there is a net outflux of these ions in the stratum granulosum. Hence the calcium exchange between keratinocytes and extracellular fluid differs distinctly between the stratum granulosum and the underlying sublayers, and these differences actively regulate the epidermal calcium profile. Our results also indicate that plasma membrane dysfunction may be an early event during keratinocyte disintegration in the stratum granulosum.

Improved sugar cane juice clarification by understanding calcium oxide-phosphate-sucrose systems

It is accepted that the efficiency of sugar cane clarification is closely linked with sugar juice composition (including suspended or insoluble impurities), the inorganic phosphate content, the liming condition and type, and the interactions between the juice components. These interactions are not well understood, particularly those between calcium, phosphate, and sucrose in sugar cane juice. Studies have been conducted on calcium oxide (CaO)/phosphate/sucrose systems in both synthetic and factory juices to provide further information on the defecation process (i.e., simple liming to effect impurity removal) and to identify an effective clarification process that would result in reduced scaling of sugar factory evaporators, pans, and centrifugals. Results have shown that a two-stage process involving the addition of lime saccharate to a set juice pH followed by the addition of sodium hydroxide to a final juice pH or a similar two-stage process where the order of addition of the alkalis is reversed prior to clarification reduces the impurity loading of the clarified juice compared to that of the clarified juice obtained by the conventional defecation process. The treatment process showed reductions in CaO (27% to 50%) and MgO (up to 20%) in clarified juices with no apparent loss in juice clarity or increase in residence time of the mud particles compared to those in the conventional process. There was also a reduction in the SiO2 content. However, the disadvantage of this process is the significant increase in the Na2O content.

Revision total hip replacement using the cement-in-cement technique for the acetabular component : technique and results for 60 hips

The technique of femoral cement-in-cement revision is well established, but there are no previous series reporting its use on the acetabular side at the time of revision total hip arthroplasty. We describe the surgical technique and report the outcome of 60 consecutive cement-in-cement revisions of the acetabular component at a mean follow-up of 8.5 years (range 5-12 years). All had a radiologically and clinically well fixed acetabular cement mantle at the time of revision. 29 patients died. No case was lost to follow-up. The 2 most common indications for acetabular revision were recurrent dislocation (77%) and to compliment a femoral revision (20%). There were 2 cases of aseptic cup loosening (3.3%) requiring re-revision. No other hip was clinically or radiologically loose (96.7%) at latest follow-up. One case was re-revised for infection, 4 for recurrent dislocation and 1 for disarticulation of a constrained component. At 5 years, the Kaplan-Meier survival rate was 100% for aseptic loosening and 92.2% (95% CI; 84.8-99.6%) with revision for all causes as the endpoint. These results support the use of the cement-in-cement revision technique in appropriate cases on the acetabular side. Theoretical advantages include preservation of bone stock, reduced operating time, reduced risk of complications and durable fixation.

Cement-in-cement revision for selected Vancouver Type B1 femoral periprosthetic fractures : A biomechanical analysis

The aim of this study was to perform a biomechanical analysis of the cement-in-cement (c-in-c) technique for fixation of selected Vancouver Type B1 femoral periprosthetic fractures and to assess the degree of cement interposition at the fracture site. Six embalmed cadaveric femora were implanted with a cemented femoral stem. Vancouver Type B1 fractures were created by applying a combined axial and rotational load to failure. The femora were repaired using the c-in-c technique and reloaded to failure. The mean primary fracture torque was 117 Nm (SD 16.6, range 89–133). The mean revision fracture torque was 50 Nm (SD 16.6, range 29–74), which is above the torque previously observed for activities of daily living. Cement interposition at the fracture site was found to be minimal.

Flexural performance of an innovative Hybrid Composite Floor Plate System comprising Glass–fibre Reinforced Cement, Polyurethane and steel laminate

This study explored the flexural performance of an innovative Hybrid Composite Floor Plate System (HCFPS), comprised of Polyurethane (PU) core, outer layers of Glass-fibre Reinforced Cement (GRC) and steel laminates at tensile regions, using experimental testing and Finite Element (FE) modelling. Bending and cyclic loading tests for the HCFPS panels and a comprehensive material testing program for component materials were carried out. HCFPS test panel exhibited ductile behaviour and flexural failure with a deflection ductility index of 4. FE models of HCFPS were developed using the program ABAQUS and validated with experimental results. The governing criteria of stiffness and flexural performance of HCFPS can be improved by enhancing the properties of component materials. HCFPS is 50-70% lighter in weight when compared to conventional floor systems. This study shows that HCFPS can be used for floor structures in commercial and residential buildings as an alternative to conventional steel concrete composite systems.

Preparation of porous nanofibers from electrospun polyacrylonitrile/calcium carbonate composite nanofibers using porogen leaching technique

Production of nanofibrous polyacrylonitrile/calcium carbonate (PAN/CaCO3) nanocomposite web was carried out through solution electrospinning process. Pore generating nanoparticles were leached from the PAN matrices in hydrochloric acid bath with the purpose of producing an ultimate nanoporous structure. The possible interaction between CaCO3 nanoparticles and PAN functional groups was investigated. Atomic absorption method was used to measure the amount of extracted CaCO3 nanoparticles. Morphological observation showed nanofibers of 270–720 nm in diameter containing nanopores of 50–130 nm. Monitoring the governing parameters statistically, it was found that the amount of extraction (ε) of CaCO3was increased when the web surface area (a) was broadened according to a simple scaling law (ε = 3.18 a0.4). The leaching process was maximized in the presence of 5% v/v of acid in the extraction bath and 5 wt % of CaCO3 in the polymer solution. Collateral effects of the extraction time and temperature showed exponential growth within a favorable extremum at 50°C for 72 h. Concentration of dimethylformamide as the solvent had no significant impact on the extraction level.

Aspects of the kinetics and solubility of silica and calcium oxalate composites in sugar solutions

Fouling of industrial surfaces by silica and calcium oxalate can be detrimental to a number of process streams. Solution chemistry plays a large roll in the rate and type of scale formed on industrial surfaces. This study is on the kinetics and thermodynamics of SiO2 and calcium oxalate composite formation in solutions containing Mg2+ ions, trans-aconitic acid and sucrose, to mimic factory sugar cane juices. The induction time (ti) of silicic acid polymerization is found to be dependent on the sucrose concentration and SiO2 supersaturation ratio (SS). Generalized kinetic and solubility models are developed for SiO2 and calcium oxalate in binary systems using response surface methodology. The role of sucrose, Mg, trans-aconitic acid, a mixture of Mg and trans-aconitic acid, SiO2 SS ratio and Ca in the formation of com- posites is explained using the solution properties of these species including their ability to form complexes.

Water and chloride ion penetration resistance of highstrength ultra lightweight cement composite

Durability is a significant issue to focus on for newly developed structural lightweight cement composite (ULCC). This paper presents an experimental study to evaluate the resistance of ULCC to water and chloride ion penetration. Chloride penetrability and sorptivity were evaluated for ULCC (unit weight about 1450 kg/m3) and compared with those of a normal weight concrete (NWC), a lightweight aggregate concrete (LWC), and an ultra lightweight composite with proprietary cementitious binder (DB) (unit weight about 1450 kg/m3) at similar compressive strength of about 60 MPa. Rapid chloride penetrability test, rapid migration test, water absorption (sorptivity) test, and water permeability test were conducted on these mixtures. Results indicate that ULCC and DB had comparable performance. Compared with control LWC and NWC at similar strength level, the ULCC and DB mixtures had higher resistance to chloride ion penetration, lower water absorption and virtually impermeable to water penetration.

Creep and shrinkage of ultra lightweight cement composite

Creep and shrinkage behaviour of an ultra lightweight cement composite (ULCC) up to 450 days was evaluated in comparison with those of a normal weight aggregate concrete (NWAC) and a lightweight aggregate concrete (LWAC) with similar 28-day compressive strength. The ULCC is characterized by low density < 1500 kg/m3 and high compressive strength about 60 MPa. Autogenous shrinkage increased rapidly in the ULCC at early-age and almost 95% occurred prior to the start of creep test at 28 days. Hence, majority of shrinkage of the ULCC during creep test was drying shrinkage. Total shrinkage of the ULCC during the 450-day creep test was the lowest compared to the NWAC and LWAC. However, corresponding total creep in the ULCC was the highest with high proportion attributed to basic creep (≥ ~90%) and limited drying creep. The high creep of the ULCC is likely due to its low E-modulus. Specific creep of the ULCC was similar to that of the NWAC, but more than 80% higher than the LWAC. Creep coefficient of the ULCC was about 47% lower than that of the NWAC but about 18% higher than that of the LWAC. Among five creep models evaluated which tend to over-estimate the creep coefficient of the ULCC, EC2 model gives acceptable prediction within +25% deviations.

Flexural and shear bond characteristics of thin layer polymer cement mortared concrete masonry

This paper presents an experimental investigation of the flexural and shear bond characteristics of thin layer polymer cement mortared concrete masonry. It is well known that the bond characteristics of masonry depend upon the mortar type, the techniques of dispersion of mortar and the surface texture of concrete blocks; there exists an abundance of literature on the conventional 10 mm thick cement mortared masonry bond; however, 1-4 mm thick polymer cement mortared masonry bond is not yet well researched. This paper reports a study on the examination of the effect of mortar compositions, dispersion methods and unit surface textures to the flexural and shear bond characteristics of thin layer mortared concrete masonry. A non-contact digital image correlation method was adopted for the measurement of strains at the unit-mortar interface in this research. All mortar joints have been carefully prepared to ensure achievement of the desired thin layer mortar thickness on average. The results exhibit that the bond strength of thin mortar layered concrete masonry with polymer cement mortar is higher than that of the conventional masonry; moreover the unit surface texture and the mortar dispersion methods are found to have significant influence on the flexural and shear bond characteristics. From the experimental results, a correlation between the flexural and the shear bond strengths has been determined and is presented in this paper.

Modeling the co-precipitation of silica and calcium oxalate in sugar solutions

Solution chemistry plays a significant role in the rate and type of foulant formed on heated industrial surfaces. This paper describes the effect of sucrose, silica (SiO2), Ca2+ and Mg2+ ions, and trans-aconitic acid on the kinetics and solubility of SiO2 and calcium oxalate monohydrate (COM) in mixed salt solutions containing sucrose and refines models previously proposed. The developed SiO2 models show that sucrose and SiO2 concentrations are the main parameters that determine apparent order (n) and apparent rate of reaction (k) and SiO2 solubility over a 24 h period. The calcium oxalate solubility model shows that while increasing [Mg2+] increases COM solubility, the reverse is so with increasing sucrose concentrations. The role of solution species on COM crystal habit is discussed and the appearance of the uncommon (001) face is explained.