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The sorptions of three basic dyes, Astrazon((R)) Blue FGRL (AB), Astrazon((R)) Red GTLN (AR), and methylene blue (MB) onto green macroalga Caulerpa lentillifera were investigated. The results were compared to the sorption performance of a commercial activated carbon (CARBON). The results revealed that the alga exhibited greater sorption capacities than activated carbon for the three basic dyes investigated in this work. The sorption process for all mixture systems (ALGA/AB, ALGA/AR, ALGA/MB, CARBON/AB, CARBON/AR, and CARBON/MB) obeyed the pseudo-second order kinetic model. C. lentillifera could more rapidly sequester AR when compared with activated carbon, but was more slowly in the sorption of AB. For the sorption of MB, both ALGA and CARBON seemed to have the same sorption rate. The sorption processes were initially controlled by both film and pore-diffusion, and only were limited by pore diffusion in the later stage. The isotherms followed Langmuir model which suggested that the sorption was monolayer coverage.

The in vitro responses of Schwann cells (RT4-D6P2T, a schwannoma cell line derived from a chemically induced rat peripheral neurotumor) on various types of electrospun fibrous scaffolds of some commercially available biocompatible and biodegradable polymers, i.e., poly(3-hydroxybutyrate) (PHB), poly(3-hydroxybutyrate-co-3-hydroxyvalerate) (PHBV), polycaprolactone (PCL), poly(l-lactic acid) (PLLA), and chitosan (CS), were reported in comparison with those of the cells on corresponding solution-cast film scaffolds as well as on a tissue-culture polystyrene plate (TCPS), used as the positive control. At 24 h after cell seeding, the viability of the attached cells on the various substrates could be ranked as follows: PCL film > TCPS > PCL fibrous > PLLA fibrous > PHBV film > CS fibrous approximately CS film approximately PLLA film > PHB film > PHBV fibrous > PHB fibrous. At day 3 of cell culture, the viability of the proliferated cells on the various substrates could be ranked as follows: TCPS > PHBV film > PLLA film > PCL film > PLLA fibrous > PHB film approximately PCL fibrous > CS fibrous > CS film > PHB fibrous > PHBV fibrous. At approximately 8 h after cell seeding, the cells on the flat surfaces of all of the film scaffolds and that of the PCL nanofibrous scaffold appeared in their characteristic spindle shape, while those on the surfaces of the PHB, PHBV, and PLLA macrofibrous scaffolds also appeared in their characteristic spindle shape, but with the cells being able to penetrate to the inner side of the scaffolds.

In the present contribution, the potential for use of the ultrafine electrospun fiber mats of poly(3-hydroxybutyrate) (PHB) and poly(3-hydroxybutyrate-co-3-hydroxyvalerate) (PHBV) as scaffolding materials for skin and nerve regeneration was evaluated in vitro using mouse fibroblasts (L929) and Schwann cells (RT4-D6P2T) as reference cell lines. Comparison was made with PHB and PHBV films that were prepared by solution-casting technique. Indirect cytotoxicity assessment of the as-spun PHB and PHBV fiber mats with mouse fibroblasts (L929) and Schwann cells (RT4-D6P2T) indicated that the materials were acceptable to both types of cells. The attachment of L929 on all of the fibrous scaffolds was significantly better than that on both the film scaffolds and tissue-culture polystyrene plate (TCPS), while RT4-D6P2T appeared to attach on the flat surfaces of TCPS and the film scaffolds much better than on the rough surfaces of the fibrous scaffolds. For L929, all of the fibrous scaffolds were superior in supporting the cell proliferation to the film counterparts, but inferior to TCPS at days 3 and 5, while, for RT4-D6P2T, the rough surfaces of the fibrous scaffolds appeared to be very poor in supporting the cell proliferation when comparing with the smooth surfaces of TCPS and the film scaffolds. Scanning electron microscopy was also used to observe the behavior of both types of cells that were cultured on both the fibrous and the film scaffolds and glass substrate for 24 h.

This work aimed to investigate the effects of the bioreactor configurations and their design variables on the cultivation of vegetative cells Haematococcus pluvialis to achieve sustainable high cell density. The addition of vitamin B to F1 growth medium could appreciably enhance the final cell density. Employing this medium, the cultivation in the airlift bioreactor was demonstrated to outperform the bubble column at the same operating conditions. Aeration was crucial for a proper growth of the alga in the airlift bioreactor, but it must be maintained at low level to minimize shear stress. The most appropriate aeration velocity (superficial velocity) was at the lower limit of the pump, i.e. 0.4 cm s(-1) and a smaller riser was shown to have positive influence on the cell growth. A 1% CO(2) supplement to the air supply considerably enhanced the growth rate of H. pluvialis and the most suitable light intensity for the growth was at 20 micromol photon m(-2) s(-1). The semi-continuous culture was successfully implemented with the optimal airlift bioreactor design and under optimal conditions the harvest could be performed every four days with the specific growth rate of 0.31 d(-1).

The sorption of Cu2+, Cd2+, Pb2+, and Zn2+ by a dried green macroalga Caulerpa lentillifera was investigated. The removal efficiency increased with pH. The analysis with FT-IR indicated that possible functional groups involved in metal sorption by this alga were O-H bending, N-H bending, N-H stretching, C-N stretching, C-O, SO stretching, and S-O stretching. The sorption of all metal ions rapidly reached equilibrium within 20min. The sorption kinetics of these metals were governed by external mass transfer and intraparticle diffusion processes. The sorption isotherm followed the Langmuir isotherm where the maximum sorption capacities was Pb2+>Cu2+>Cd2+>Zn2+.

This study investigated the use of ultrasound-assisted extraction (UAE) to improve the extraction efficiency of the classical solvent extraction techniques such as maceration and soxhlet extraction to extract anti-oxidant activity compounds, anthraquinones, from the root of Morinda citrifolia. The effects of different extraction conditions were determined, i.e., temperature of (25, 45, 60 degrees C), ultrasonic power, solvent types, and compositions of ethanol in ethanol-water mixtures. The results show that the yield increases with increasing extraction times and extraction temperatures. The percent recovery of anthraquinones using ultrasound was found to be highly dependent on the type of solvents (acetone > acetonitrile > methanol > ethanol). Furthermore, the use of ethanol-water solution as extraction solvent increased the yield of anthraquinones due to the relative polarity, the swelling effect of plant tissue matrix by water, and increased sound absorption. To achieve the same recovery as that achieved by UAE, soxhlet extraction and maceration required much longer time.

Raw water from the Bangkok (Thailand) main municipal water supply canal was examined for its natural organic composition by fractionation with adsorption resins. DAX-8 resin was the first resin employed to fractionate the hydrophobic fractions. Fractionation at neutral pH resulted in the separation of the hydrophobic neutral components; at a high pH level (approx. 10) separation of the hydrophobic base components occurred; and at a low pH level (approx. 2) the hydrophobic acid components were separated. AG-MP-50 cationic resin was then used to separate the hydrophilic base components, and WA-10, a weak anionic resin, was applied finally to fractionate the hydrophilic acid and neutral components. Subsequently, each fraction was tested for its chlorine disinfection by-product (DBP) formation potential. The HAA formation tests demonstrated that the various organic fractions had different reactivity levels for the formation of haloacetic acids (HAAs). For this source water, the hydrophilic neutral fraction dominated over the other five fractions in being the main organic component and the most significant precursor of HAAs formation. On the other hand, in terms of specific HAA formation potential (FP), the hydrophobic and hydrophilic base fractions were the most reactive precursors to the formation of HAAs. In all cases, the quantity of HAAs formed depended linearly upon the amount of organic constituents in the water sample.

The purpose of this study was to compare the early response of human osteoblast-like cells (SaOS-2) on commercially pure titanium (cpTi) and titanium-6-aluminium-7-niobium (Ti-6Al-7Nb) using glass slide as a control. In terms of cell attachment, no significance was observed when cells were seeded on the materials. However, morphological analysis by scanning electron microscope revealed that cells on Ti-6Al-7Nb showed better spreading after 4 hrs. After 48 hrs, both Western analysis and reverse transcription polymerase chain reaction analyses showed that cells cultured on Ti-6Al-7Nb synthesized a higher amount of fibronectin and osteopontin as compared to cells seeded on cpTi or on glass slide. These results suggest that Ti-6Al-7Nb possess a good potential to support SaOS-2 cells on spreading and fibronectin and osteopontin synthesis, therefore, this material may be one of a candidate material used in implant dentistry.

The macroalga Caulerpa lentillifera was found to have adsorption capacity for a basic dye, Astrazon Blue FGRL. For the whole range of concentrations employed in this work (20-1280 mgl(-1)), the adsorption reached equilibrium within the first hour. The kinetic data corresponded well with the pseudo second-order kinetic model where the rate constants decreased as initial dye concentrations increased. At low dye concentrations (20-80 mgl(-1)), an increase in the adsorbent dosage resulted in a higher removal percentage of the dye, but a lower amount of dye adsorbed per unit mass (q). The adsorption isotherm followed both the Langmuir and Freundlich models within the temperature range employed in this work (18-70 degrees C). The highest maximum adsorption capacity (q(m)) was obtained at 50 degrees C. The enthalpy of adsorption was estimated at 14.87 kJmol(-1) suggesting a chemical adsorption mechanism.

Novel bone scaffolding materials were successfully fabricated by electrospinning from polycaprolactone (PCL) solutions containing nanoparticles of calcium carbonate (CaCO3) or hydroxyapatite (HA). The potential use of the electrospun fibrous scaffolds for bone regeneration was evaluated in vitro with human osteoblasts (SaOS2) in terms of attachment, proliferation, and alkaline phosphatase (ALP) activity of the cells that were cultured directly on the scaffolds. The results were compared with those on corresponding solution-cast film scaffolds and tissue-culture polystyrene plate (TCPS). It was found that all of the fibrous scaffolds promoted much better adhesion and proliferation of cells than the corresponding film scaffolds and TCPS. Interestingly, the cells that were seeded on all of the fibrous scaffolds appeared to be well-expanded and attach on the fiber surface very well even only about 1 hr in culture, while those seeded on all of the film scaffolds and the glass substrate were still in round shape. Among the various fibrous scaffolds investigated, the one that was filled with 1.0% HA showed the highest ALP activity. Finally, all of the fibrous scaffolds exhibited much greater tensile strength at yield than all of the corresponding film scaffolds.

Novel bone-scaffolding materials were successfully fabricated by electrospinning from polycaprolactone (PCL) solutions containing nanoparticles of calcium carbonate (CaCO(3)) or hydroxyapatite (HA). The diameters of the as-spun fibers were found to increase with the addition and increasing amounts of the nanoparticles. The observed increase in the diameters of the as-spun fibers with the addition and increasing amounts of the nanoparticulate fillers was responsible for the observed increase in the tensile strength of the obtained fiber mats. An increase in the concentration of the base PCL solution caused the average diameter of the as-spun PCL/HA composite fibers to increase. Increasing applied electrical potential also resulted in an increase in the diameters of the obtained PCL/HA composite fibers. Lastly, indirect cytotoxicity evaluation of the electrospun mats of PCL, PCL/CaCO(3), and PCL/HA fibers based on human osteoblasts (SaOS2) and mouse fibroblasts (L929) revealed that these as-spun mats posed no threat to the cells, a result that implied their potential for utilization as bone-scaffolding materials.

The formation of haloacetic acids (HAAs) from the chlorination of individual and binary mixtures of organic fractions obtained from the intake of Bangkhen Water Treatment Plant in Bangkok, Thailand was investigated. Experimental results revealed that, as an individual fraction, hydrophobic base (HPOB) was the most active in forming HAAs (approx. 200 microg/mg) whereas hydrophilic acid (HPIA) was the least (approx. 40 microg/mg). In binary mixtures, acid fractions exhibited stronger inhibitory effect in forming HAAs than base fractions. With the set of experimental data obtained from this work, no relationships between specific HAA formation potential and various organic fractions concentrations in binary mixtures could be formulated. Among the various individual HAA species obtained from the chlorination of each individual organic fraction, dichloroacetic acid (DCAA) was found to be predominant. On the other hand, the chlorination of binary organic fraction mixtures often led to the formation of monochloroacetic acid (MCAA) as the predominant HAA species.

Extracellular matrix and growth factors are the crucial factors that regulate healing and regenerating processes in human periodontal ligament cells. The purpose of this study was to examine the effects of type I collagen and insulin-like growth factor-I (IGF-I) on osteopontin (OPN) expression. The data showed that OPN expression was significantly decreased when cells were cultured on collagen-coated plates. Addition of IGF-I obviously induced OPN expression only in a collagen-coated condition, suggesting an attenuating effect of IGF-I on the decrease of OPN expression. Cells treated with a combination of inhibitory antibody to beta1 integrin and IGF-I showed the same level of OPN expression as those treated with either inhibitory antibody to beta1 integrin or IGF-I alone. These results indicate that IGF-I counteracts with the inhibitory signal from type I collagen through beta1 integrin receptor.

CM-chitin and CM-chitosan films were successfully crosslinked by microwave treatment. Crosslinking of the microwave-treated CM-chitin films involved mainly the carboxylate and the secondary alcohol groups, while crosslinking of microwave-treated CM-chitosan films involved the carboxylate and the amino groups. In addition, the crystallinity of CM-chitin increased with increasing microwave treatment time, whereas an increase in the crystallinity of the microwave-treated CM-chitosan films was not observed. At a similar percentage of weight loss, the crosslinking of either CM-chitin or CM-chitosan films by microwave treatment required much less stringent condition when compared with the crosslinking by autoclave treatment. Based on both direct and indirect cytotoxicity assays, the cytotoxicity of the microwave-treated CM-chitin films was negative, while that of the microwave-treated CM-chitosan films was positive. Human fibroblasts adhered on the surface of microwave-treated CM-chitosan films much better than on the surface of microwave-treated CM-chitin films.

Resin adsorption techniques using three types of resin (DAX-8, AG-MP-50, and WA-10) were employed to characterize the raw water (RW) from the major 3 million m3/day (793 million gal/day) drinking water treatment plant in Bangkok, Thailand. The dissolved organic carbon (DOC) mass distribution sequences of the six organic fractions in raw water, from high to low, were hydrophilic neutral (HPIN), hydrophobic acid (HPOA), hydrophilic acid (HPIA), hydrophobic neutral (HPON), hydrophilic base (HPIB), and hydrophobic base (HPOB). HPIN and HPOA were the two main precursors for trihalomethanes formation (THMFP) in this water source following chlorination. The chlorination of HPON and HPIN fractions only led to the formation of mostly chloroform, while other organic fractions formed both chloroform and bromodichloromethane. The linear dependency between each organic fraction concentration and THMFP indicated that the reactions of each organic fraction with chlorine were first-order.

The fractionation of raw water from Bangkhen water treatment plant, Bangkok, Thailand revealed that the mass distribution sequence of the six organic fractions from high to low was hydrophilic neutral (HPIN), hydrophobic acid (HPOA), hydrophilic acid, hydrophobic neutral, hydrophilic base, and hydrophobic base. The main organic matter components in raw water were HPIN and HPOA, which were also the two most important contributors of trihalomethane formation potential (THMFP). Linear dependencies between the level of each organic fraction and the formation potential of THM species were observed, which suggested the reactions between the organic fraction and chlorine during the chlorination were first order. The fractionation led to a deviation of bromide concentration in each organic fraction from the original concentration, and this affected the formation of brominated THM species. However, this effect was demonstrated to be within an acceptable range. The chlorination of an individual organic fraction resulted in a higher level of THMFP than that of the raw water and mixed fractions, indicating an inhibitory effect between the organic species.

Background: The lipopolysaccharide (LPS) of A. actinomycetemcomitans is one of the major pathogenic factors in periodontal disease. It induces secretion of proinflammatory cytokines and is involved in alveolar bone destruction. We hypothesized that the LPS of A. actinomycetemcomitans could affect the activation of matrix metalloproteinase (MMP)-2 and the expression of receptor activator of nuclear factor kappa B ligand (RANKL) and osteoprotegerin in human periodontal ligament (HPDL) cells leading to the destruction of periodontium., Methods: HPDL cells were cultured in serum-free medium with or without the LPS of A. actinomycetemcomitans for 36 hours. The activation of MMP-2 was analyzed by zymography. Changes of the expression of RANKL and osteoprotegerin (OPG) were examined by reverse transcription-polymerase chain reaction and supported by Western blot analysis., Results: The activation of MMP-2 could be induced by the LPS of A. actinomycetemcomitans in HPDL cells and could be inhibited by a serine protease inhibitor. This result suggested that the LPS might activate MMP-2 through a serine protease-dependent pathway. This activation was also blocked by NF-kappaB inhibitor, which indicated the involvement of NF-kappaB. The upregulation of RANKL but not OPG by the LPS was found in both transcription and translation and could be reduced by indomethacin. In addition, serine protease inhibitor also inhibited the upregulation of RANKL, suggesting the activity of serine protease., Conclusions: The effect of the LPS of A. actinomycetemcomitans on HPDL cells is serum-independent and the induction of the activation of MMP-2 and the expression of RANKL are serine protease-dependent pathways. The results suggest the role of HPDL cells in the pathogenesis of periodontitis.

Objective: Matrix metalloproteinase 2 (MMP-2) has been implicated to play a role in pathogenesis of periodontal disease. We recently reported that Porphyromonas gingivalis supernatant could activate MMP-2 in human periodontal ligament (HPDL) cells. In this study, activation of MMP-2 by Actinobacillus actinomycetemcomitans supernatant and the mechanism was investigated., Methods: HPDL cells were treated with either A. actinomycetemcomitans or P. gingivalis supernatant for 48 h. To verify the mechanism, pretreated inhibitors were used. Gelatin zymography, RT-PCR and Western blot analysis were used to detect the activation of MMP-2, expression of MT1-MMP and TIMP-2 mRNA and the proteins, respectively., Results: The supernatant from A. actinomycetemcomitans could activate MMP-2 in HPDL cells similar to that from P. gingivalis but by a different mechanism. Activation by A. actinomycetemcomitans supernatant was correlated with a reduction of TIMP-2 secretion without any alteration of MT1-MMP, while activation by P. gingivalis increased MT1-MMP but no change of TIMP-2 was found., Conclusion: The supernatant from A. actinomycetemcomitans and P. gingivalis could induce the activation of MMP-2 possibly through the imbalance of MT1-MMP and TIMP-2 in HPDL cells but by different mechanisms. The imbalance of MT1-MMP and TIMP-2 may be another factor that is involved in the severity of periodontal disease.

The objectives of this study were to evaluate the effect of heat application after silanation and the pH effect of silanated solution on flexural properties and hydrolytic durability of experimental composites. A spherical-shaped filler was silanated with 0.66 mass% of gamma-MPTS in 70 vol% ethanol solution. For the heating effect, the filler was heated after silanation for 3 hours at 50 degrees C, 80 degrees C and 110 degrees C. For the effect of pH, ethanol solution with HCl solution of pH 1, 2 and 4 were used. All fillers were mixed to make photopolymerized experimental composite resins. All specimens after immersion in 37 degrees water for 24 hours were additionally immersed in Soxhlet's extractor for another 1, 7 and 28 days. A three-point bending test was performed in 37 degrees C water. The heat application of 50 degrees C and 80 degrees C increased flexural strength while heat of 110 degrees C and pH showed no effects on flexural properties at p<0.05.

1,25(OH)2D3 and TGF-beta can influence the function and differentiation of dental pulp fibroblasts. In this study, we examined the effect of 1,25(OH)2D3 and TGF-beta on the synthesis of SPARC and ALP activity in human pulp fibroblasts. Two isoforms of SPARC, the 43 and 38 kDa, were detected in this cell type. TGF-beta increased the synthesis of SPARC about 2.5-fold after 3 days of treatment but had no effect on the ALP activity. On the contrary, 1,25(OH)2D3 increased ALP activity 2-fold but had no effect on SPARC. The combination of TGF-beta and 1,25(OH)2D3 significantly induced SPARC synthesis and ALP activity by 5 and 9 folds, respectively (P<0.05). This finding suggested the synergistic effect between TGF-beta and 1,25(OH)2D3 in dental pulp fibroblasts on the synthesis of SPARC and ALP activity. This interaction could influence the function and differentiation of dental pulp fibroblasts.

It has been reported that matrix metalloproteinase (MMP) produced by host cells plays a major role in periodontal tissue destruction. In addition, secreted virulence factors from Porphyromonas gingivalis can alter MMP secretion and cause activation in host cells that lead to the tissue degradation. In this study, we examine the effects of P. gingivalis supernatant on matrix metalloproteinase-2 (MMP-2) activation in human periodontal ligament (HPDL) cells. Cultures of HPDL cells were treated with P. gingivalis supernatant for 48 h and the level of MMP-2 activation was monitored by gelatin zymography. The profound activation of MMP-2 was seen only in the treated group. The activation of MMP-2 was inhibited by MMP inhibitors phenanthroline and EDTA, but not serine protease or cysteine protease inhibitors. To study the correlation between the expression of membrane-type-1 matrix metalloproteinase (MT1-MMP) and the activation of MMP-2, the level of MT1-MMP was analyzed. The results from reverse-transcription polymerase chain reaction (RT-PCR) and Western analysis indicated that P. gingivalis supernatant up-regulated the expression of MT1-MMP in both transcription and translation levels within 48 h. These results suggest that P. gingivalis supernatant can activate MMP-2 in HPDL cells and the mechanism of activation may involve the increased amount of MT1-MMP. It is possible that the activation of MMP-2 by P. gingivalis plays a role in the process of chronic periodontitis.

In a previous study, we presented evidence that the synthesis of hyaluronan by hypertrophic chondrocytes is one of the principal factors driving the interstitial expansion of the growth plate (Pavasant et al., J. Cell Sci. 109: 327-334, 1996). To test this possibility further, we used two different approaches to examine the effects of hormones on the production of hyaluronan in the growth plate. In the first approach, we examined the growth plate of the lit/lit mouse that lacks growth hormone and found that its hypertrophic lacunae were smaller and contained less hyaluronan than those of wild type mice. Moreover, the ratios of hyaluronan staining density to total area of the lacunae were similar for the lit/lit and the wt/wt mice, indicating that the amount of hyaluronan is directly related to lacuna size. In the second approach, we examined the effects of hormones on segments of the epiphysial growth plate placed in organ culture. Under normal culture conditions, a band of hyaluronan staining progressed across the length of the growth plate, reflecting the maturation of chondrocytes into the hypertrophic stage. When insulin-like growth factor-1, a factor known to promote chondrocyte maturation, was added to the culture medium, the production of hyaluronan and the enlargement of the lacunae were stimulated. In contrast, when either parathyroid hormone or transforming growth factor-beta 1, both of which inhibit chondrocyte differentiation, was added to the medium of cultured segments, new pericellular hyaluronan was not detected and the lacunae did not enlarge. Taken together, these results indicate that factors that either stimulate or inhibit the maturation of epiphysial chondrocytes have a corresponding effect on the production of hyaluronan. This, in turn, further supports the importance of hyaluronan in the process of lacuna enlargement.

In vitro analyses of basement membrane invasiveness employing Matrigel (a murine tumor extract rich in basement membrane components) have been performed on human breast cancer model systems. Constitutive invasiveness of different human breast cancer (HBC) cell lines has been examined as well as regulation by steroid hormones, growth factors, and oncogenes. Carcinoma cells exhibiting a mesenchymal-like phenotype (vimentin expression, lack of cell border associated uvomorulin) show dramatically increased motility, invasiveness, and metastatic potential in nude mice. These findings support the hypothesis that epithelial to mesenchymal transition (EMT)-like events may be instrumental in the metastatic progression of human breast cancer. The MCF-7 subline MCF-7ADR appears to have undergone such a transition. The importance of such a transition may be reflected in the emergence of vimentin expression as an indicator of poor prognosis in HBC. Matrix degradation and laminin recognition are highlighted as potential targets for antimetastatic therapy, and analyses of laminin attachment and the matrix metalloproteinase (MMP) family in HBC cell lines are summarized. Matrigel-based assays have proved useful in the study of the molecular mechanisms of basement membrane invasiveness, their regulation in HBC cells, and their potential as targets for antimetastatic therapy.