Differences In Calcium Content Research Articles

Comparison of Calcium Content between Blossom-End Rot and Healthy Fruits in Red Pepper (Capsicum annuum L.) Grown in Open Field

The causes of blossom-end rot (BER) are still not understood clearly, although there was a lot of research on the development of BER over 100 years. This experiment was conducted to investigate the influence of Ca content of fruit on the incidence of BER in red pepper. BER affected fruits and healthy fruits had been harvested and the samples were divided into two parts, the upper and the lower, for chemical analysis to determine the content of mineral nutrients in the tissue. The content of nitrogen (N), phosphorus (P), potassium (K) and magnesium (Mg) was higher in the BER affected fruits and the difference of each nutrient content was revealed much larger in the lower part of the fruit but there were no significant differences in calcium content between the BER and the healthy fruits of red pepper, in this experiment. The results indicate that it is difficult to find a certain relation between the calcium content of the fruit and the occurrence of blossom-end rot.

In Vitro assessment of dentin erosion after immersion in acidic beverages: surface profile analysis and energy-dispersive X-ray fluorescence spectrometry study

The aim of this study was to investigate the effects of some acidic drinks on dentin erosion, using methods of surface profile (SP) analysis and energy-dispersive X-ray fluorescence spectrometry (EDXRF). One hundred standardized dentin slabs obtained from bovine incisor roots were used. Dentin slabs measuring 5x5 mm were ground flat, polished and half of each specimen surface was protected with nail polish. For 60 min, the dentin surfaces were immersed in 50 mL of 5 different drinks (Gatorade®, Del Valle Mais orange juice®, Coca-Cola®, Red Bull® and white wine), 20 blocks in each drink. The pH of each beverage was measured. After the erosive challenge, the nail polish was removed and SP was analyzed. The mineral concentration of dentin surfaces was determined by means of EDXRF. Data were analyzed statistically by ANOVA and Tukey's test (α=0.05). SP analysis showed that Red-Bull had the highest erosive potential (p<0.05). EDXRF results exhibited a decrease in phosphate in the groups immersed in Red-Bull, orange juice and white wine (p<0.05), and no significant difference in calcium content between the reference surface and eroded surface. In conclusion, this study demonstrated that all studied beverages promoted erosion on root dentin and Red Bull had the highest erosive potential. There was no correlation between pH of beverages and their erosive potential and only the P content changed after erosive challenge.

Impact of Season and Sex on Calcium and Phosphorus Content in the Meat of Roach (Rutilus rutilus L.) from the Brda River (Poland, Bydgoszcz)

The aim of this work was to compare the concentrations of calcium and phosphorus and Ca/P ratio in the meat of females and males of the roach (Rutilus rutilus L.) caught from the Brda River. The study involved 40 roach individuals caught in fall and spring (10 females and 10 males from each season). The muscle samples for analyses were taken from the large side muscle of the fish body above the lateral line. Ca concentrations were determined by atomic absorption spectrophotometry; P content was analyzed by the colorimetric method. Calcium concentration in the meat of analysed roach was higher in samples collected from fish caught in spring and equaled 1.82 g kg(-1) in females and 1.93 g kg(-1) in males. Values for individuals from autumn amounted to 0.83 and 1.10 g kg(-1), respectively. Statistically significant differences in calcium content were detected between individuals caught in different seasons, but samples taken from females and males caught within one season did not differ substantially. The mean value of P in the meat of analysed roach caught in spring was higher than in fish from autumn, and it was respectively 2.24 g kg(-1) in females and 2.30 g kg(-1) in males from spring, and 1.89 g kg(-1) in the tissue of females and 2.01 g kg(-1) in males in fish from autumn. The ratio of calcium to phosphorus in the meat of analysed wild roach ranged from 0.43:1 to 0.82:1. A negative and statistically significant correlation between Ca and P concentrations was found in the meat and the body length of analysed roach from the Brda River.

Carotenoid and mineral content of different morphotypes of Centella asiatica L. (Gotukola)

Six morphotypes of Centella asiatica designated G1, G2, G3, G7, G8 and G12 were analyzed for carotenoid content, proximate components and selected dietary minerals. Potassium was found to be the major element present in C. asiatica and the content in G1 and G3 (6,165.0 ± 125.8 mg/100 g dry weight) was significantly (P < 0.05) higher than all other morphotypes. C. asiatica is more popular for its higher contents of iron and calcium. However, no significant (P>0.05) differences in calcium content among the morphotypes were observed. The imported variety G7 had significantly higher (P < 0.05) mean content of iron (74.3 ± 34.1 mg/100 g dry weight). The morphotypes G8 and G12 contained significantly higher (P < 0.05) content of β-carotene (77.2 ± 5.1 μg/g fresh weight) and lutein (133.5 ± 10.1 μg/g, fresh weight) respectively. Lutein and β-carotene levels of these two types were also significantly higher (P < 0.05) than those in the other varieties. The study did not provide any evidence to believe that a given morphotype is superior to another in its overall nutritional value.

Interspecies differences in calcium content and requirement in four freshwater cladocerans explained by biokinetic parameters

We tested the hypothesis that cladocerans with higher calcium (Ca) content are more susceptible to Ca limitation by conducting life table experiments using four cladoceran species with contrasting Ca contents (0.06–2.24% of dry weight). Populations of Daphnia carinata and Daphnia galeata with high Ca content might collapse when the ambient Ca concentration is &lt; 0.5 mg L−1, whereas Ceriodaphnia dubia, with intermediate Ca content, and Moina macrocopa, with low Ca content, are well adapted to that low Ca level. However, Ca content is not a good proxy of the susceptibilities to Ca limitation within the genus of Daphnia. We propose an index, which considers both the Ca demand (i.e., Ca content under Ca‐sufficient conditions) and the ability of cladocerans to extract and retain Ca in low‐Ca environments, to explain the differences in cladoceran tolerances to Ca deficiency. We also used physiologically based biokinetic parameters, including the influx rate and efflux rate constant of Ca, to predict the interspecies differences of specific Ca content. Low‐Ca species had a lower influx rate and a higher efflux rate constant of Ca than the high‐Ca species. A Ca concentration of 2 mg L−1 was sufficient to keep Daphnia spp. from extinction because of Ca limitation.

Accumulation of calcium in the centre of leaves of coriander (Coriandrum sativum L.) is due to an uncoupling of water and ion transport

The aim of this study is to understand the parameters regulating calcium ion distribution in leaves. Accumulation of ions in leaf tissue is in part dependent on import from the xylem. This import via the transpiration stream is more important for ions such as calcium that are xylem but not phloem mobile and cannot therefore be retranslocated. Accumulation of calcium was measured on bulk coriander leaf tissue (Coriandrum sativum L. cv. Lemon) using ion chromatography and calcium uptake was visualized using phosphor-images of 45Ca2+. Leaves of plants grown in hydroponics had elevated calcium in the centre of the leaf compared with the leaf margin, while K+ was distributed homogeneously over the leaf. This calcium was shown to be localised to the mesophyll vacuoles using EDAX. Stomatal density and evapotranspiration (water loss per unit area of leaf) were equal at inner and outer sections of the leaf. Unequal ion distribution but uniformity of water loss suggested that there was a difference in the extent of uncoupling of calcium and water transport between the inner and outer leaf. Since isolated tissue from the inner and outer leaf were able to accumulate similar amounts of calcium, it is proposed that the spatial variation of leaf calcium concentration is due to differential ion delivery to the two regions rather than tissue/cell-specific differences in ion uptake capacity. There was a positive correlation between whole leaf calcium concentration and the difference in calcium concentration between inner and outer leaf tissue. Exposing the plants to increased humidity reduced transpiration and calcium delivery to the leaf and abolished this spatial variation of calcium concentration. Mechanisms of calcium delivery to leaves are discussed. An understanding of calcium delivery and distribution within coriander will inform strategies to reduce the incidence of calcium-related syndromes such as tip-burn and provides a robust model for the transport of ions and other substances in the leaf xylem.

Computational modelling identifies the impact of subtle anatomical variations between amphibian and mammalian skeletal muscle on spatiotemporal calcium dynamics

The physical sites of calcium entry and exit in the skeletal muscle cell are distinct and highly organised in space. It was investigated whether the highly structured spatial organisation of sites of Ca(2+) release, uptake and action in skeletal muscle cells substantially impacts the dynamics of cytosolic Ca(2+) handling and thereby the physiology of the cell. Hereto, the spatiotemporal dynamics of the free calcium distribution in a fast-twitch (FT) muscle sarcomere was studied using a reaction-diffusion computational model for two genotypes with known anatomical differences. A computational model of a murine FT muscle sarcomere is developed, de novo including a closed calcium mass balance to simulate spatiotemporal high stimulation frequency calcium dynamics at 35 degrees C. Literature data on high-frequency calcium dye measurements were used as a first step towards model validation. The murine and amphibian sarcomere models were phenotypically distinct to capture known differences in positions of troponin C, actin-myosin overlap and calcium release within the sarcomere between frog and mouse. The models predicted large calcium gradients throughout the myoplasm as well as differences in calcium concentrations near the mitochondria of frog and mouse. Furthermore, the predicted Ca(2+) concentration was high at positions where Ca(2+) has a regulatory function, close to the mitochondria and troponin C.

O.612 BMP-7 effects on osteogenic differentiation of stem cells

The objective of this study was to examine the osteoinductive capacity of different concentrations of BMP-2 on bone marrow stromal cells in vitro. Further, we intended to determine whether titanium provided with an increased surface roughness is more efficient in osteoblast differentiation than machined titanium. Therefore, 20,000 cells/ml were seeded and cultured on machined and grit-blasted titanium discs for 4, 8 and 16 days. Different concentrations of rhBMP-2 (0, 10, 100, 1000 ng/ml) were supplemented to the medium for 8 days of culturing. To evaluate cellular proliferation and differentiation, specimens were examined for DNA, alkaline phosphatase activity, and calcium content. Morphological appearance of the specimens at 8 and 16 days of incubation was evaluated using scanning electron microscopy. Two separate experimental runs were performed.Evaluation of the DNA and alkaline phosphatase data revealed that a significant difference existed for these data between both experimental runs. Further analysis of the DNA figures learned that roughening of the titanium surface and addition of BMP-2 had no effect on cell proliferation. The alkaline phosphatase analysis and calcium measurements revealed that BMP-2 stimulated the early differentiation of osteogenic cells on machined titanium substrates in a dose-dependent manner. After 16 days of culture, no significant differences in calcium content could be observed anymore between machined and roughened titanium surfaces. Further, the data revealed that the machined surfaces showed a significant increase in calcium deposition when 100 and 1000 ng/ml BMP-2 were supplemented to the medium. However, the roughened surfaces showed this significant enhancement in calcium content only with 1000 ng/ml BMP-2. In addition, SEM evaluation revealed a dose-dependent response to BMP-2. Increasing BMP-2 concentrations resulted in more calcified globular accretions on bone surfaces than when no BMP-2 was added.On the basis of our results, we conclude that (1) due to the heterogeneous nature of bone marrow, experimental results with primary rat bone marrow cells are difficult to reproduce from one experiment to the other, and (2) addition of rhBMP-2 in the medium stimulates the early differentiation and matrix mineralization of osteogenic cells on machined titanium surfaces in a dose-responsive manner. Further, we concluded that our roughened titanium surfaces had no effect on proliferation and differentiation of primary derived rate bone marrow cells.

Spraying calcium is not an effective way to increase structural calcium in litchi pericarp

Calcium in different forms was compared in the pericarp of cracked and healthy fruit in the cracking-susceptible litchi ( Litchi chinensis Sonn.) cultivar ‘Nuomici’. Incorporation of 45Ca into the cell walls of the pericarp and effects of sprays of calcium chloride (0.2% or 0.5%) on endogenous calcium and cracking incidence were examined. The results showed that calcium concentration in cracked fruit was significantly lower than in healthy fruit. Structural calcium as part of cell wall components contributed to the majority of the difference in calcium concentration between cracked fruit and healthy fruit. Surface spray of calcium chloride, either once or three times at different stages of fruit development exhibited no significant effect on fruit cracking and hardly increased calcium, especially structural calcium in the pericarp. X-ray microanalysis revealed that the applied calcium remained on pericarp surface. Radioisotope test showed that less than 0.1% of 45Ca applied on fruit surface was incorporated into the cell walls. It is suggested that spraying calcium on fruit surface is not an effective way to prevent litchi fruit cracking due to the difficulty in uptake of the calcium sprayed on fruit surface.

Effects of Nanocalcium Supplemented Milk on Bone Calcium Metabolism in Ovariectomized Rats

This study examined effects of calcium supplemented milk on bone loss in ovariectomized rats. Twenty four Sprague-Dawley female rats, 7 weeks-old, were divided into 4 groups, ovariectomized and fed diets containing: 1) control, no Ca supplemented milk, 2) ovx 1, Ca carbonate supplemented milk, 3) ovx 2, ionized Ca supplemented milk, and 4) ovx 3, nano Ca supplemented milk. All rats were fed 1 ml of milk containing 20 mg supplemented Ca. After 18 wk feeding, body weight gain and food efficiency ratio were significantly different between ovx 1 and ovx 3. Serum concentration of calcium and phosphorus were not different among groups. However, there was a significant difference in calcium content of dry femoral weight in ovx 3 compared with the control and ovx 2. In addition, femoral bone mineral density (g/cm 2 ) was significantly greater in ovx 3 than in other groups (p<0.05). The ovx 3 group showed the highest stiffness (N/mm), maximum energy (N) in femur and trabecular bone area (%). The present study indicated that nano Ca supplementation in milk may be an effective way to enhance bone calcium metabolism for ovariectomized rats.

Changes in selected physiological and laboratory measurements in elite horses competing in a 160 km endurance ride

Limited information exists about the physiological changes and clinical problems that occur in elite horses competing in high-speed 160 km endurance races. To provide initial data describing changes in physiological and laboratory measurements in horses competing in a high-speed, 160 km endurance race under temperate conditions and to compare data between horses that successfully completed the race and those that failed to finish. Body mass (BM) was measured, blood samples were collected, and veterinary examinations performed on horses before, during, and at the finish of a CEI*** 160 km endurance race. Of 36 horses participating in the study, 22 (61%) completed the race. Twelve horses were eliminated for lameness and 2 for persistent heart rate elevation. Mean speed of finishers was 15.2 km/h. Mean +/- s.d. BM loss of finishers at the end of the race (5.7 +/- 2.6%) was not different (P = 0.58) from BM loss of nonfinishers at elimination (6.7 +/- 34%). Similarly, there were no significant differences in heart rate or veterinary assessment of hydration at the race end for finishers as compared to the elimination point for nonfinishers. PCV increased while sodium, chloride and potassium concentrations decreased with exercise but differences between finishers and nonfinishers were not detected. In contrast, both total and ionised calcium concentrations decreased in successful horses but remained unchanged in nonfinishers. Elite endurance horses are more likely to be eliminated from competition for lameness than metabolic problems; however, it remains unclear whether these conditions are entirely distinct. The magnitude of the decrease in sodium concentration in both finishers and nonfinishers was greater than in previous reports of 160 km rides. These data should be of use for both organisers and participants in elite 160 km endurance races. The tendency toward hyponatraemia as well as the difference in calcium concentrations between finishers and nonfinishers warrant further study.

Water chemistry of mires in relation to the poor-rich vegetation gradient and contrasting geochemical zones of the north-eastern fennoscandian Shield

The relationship of the poor-rich vegetation gradient with water chemistry and geochemical bedrock zones was studied with a data set from 36 pristine mire sites in the middle boreal zone, Finland. The bipartition of bedrock zones into a granitoid zone and three other, more alkaline bedrock zones was clearly connected to the poor-rich gradient in the study area. Mires in the granitoid bedrock zone were predominantly bogs and poor fens, while most of the rich fen sites were located in other bedrock zones. In fens with strongly minerogenous hydrology, the geochemically distinct bedrock zones showed significant differences in concentrations of calcium, manganese, phosphorus, as well as in the total equivalent charge of base cations and the ratio of calcium and magnesium. In light of the indicator species baseda priori poor-rich classification anda posteriori correlations with vegetation ordinations and a poor-rich indicator cover index, pH showed the strongest relationship of the water chemical variables with the poor-rich gradient. High calcium concentrations only separated extremely rich fens from other categories. The poor-rich gradient of mires is considered as a primarily pH-related vegetation continuum. A more general use of pH-classes along with hydrological and vegetation based classification is suggested for mire classification instead of the “poor-rich” terminology.

GRIT-SITE SELECTION OF BLACK BRANT: PARTICLE SIZE OR CALCIUM CONTENT?

We examined selection of grit-ingestion sites by Black Brant ( Branta bernicla nigricans )o n South Humboldt Bay, California in relation to particle size and calcium content. We hypothesized that Brant site selection was dependent primarily upon calcium content and secondarily upon distribution of substrate particle size. We (1) mapped grit-ingestion sites, (2) ranked their importance by Brant abundance and individual movement probabilities between sites, (3) characterized Brant gizzard grit and compared it with grit available at ingestion sites, and (4) compared calcium content and particle-size distribution between ingestion sites and unused sites, and between primary and secondary ingestion sites. Brant repeatedly congregated at specific, discrete sites during the 2 years of observation. The distribution of gizzard-grit particle size was right-skewed toward larger particles ( .0.5 mm) relative to the proportional availability of particle sizes in the substrate. We found no significant differences in calcium content or particle size between sites where grit was ingested and unused sites. Within used sites, the calcium content of substrates at the primary ingestion site was significantly higher than at the secondary ingestion sites, as ranked by Brant abundance and between-site movement proba- bilities. Our findings from the field corroborate previous laboratory results, and confirm that calcium is a sig- nificant ecological factor for this species. Received 14 May 2004, accepted 19 October 2004. Preferred sites for gizzard-grit ingestion may be used faithfully by wild bird popula- tions for many decades (McIlhenny 1932), but site selection of this resource is understudied. It has long been accepted that gizzard grit (hereafter grit) is an essential aid for grinding food (Leopold 1931), and captive birds de- prived of grit experience elevated mortality (McCann 1939). In some waterfowl, amount of grit in the gizzard and size of grit particles are related to diet, with more grit and smaller particles in the gizzards of herbivores com- pared to omnivores and carnivores (Thomas et al. 1977, Skead and Mitchell 1983). Leo- pold (1933) proposed that grit could be a source of mineral calcium for birds. This is well-documented only for Ring-necked Pheas- ant (Phasianus colchicus), a species whose grit consumption is driven primarily by the need for calcium and only secondarily as a grinding material (McCann 1939)—and whose distribution and abundance is influ- enced by the availability of calciferous grit (Leopold 1931; McCann 1939, 1961; Dale

A fase estrogênica altera a resposta do osso e do metabolismo mineral de ratas com hipertireoidismo?

The effect of the estrogenic phase in the bone and in the mineral metabolism was studied in Wistar adult female rats kept under euthyroidism or hyperthyroidism for 60 days. The rats were divided, according to the stage of the estrous cycle, into four groups: 1) euthyroid (proestrus-estrus), 2) euthyroid (metaestrus-diestrus), 3) hyperthyroid (proestrus-estrus), and 4) hyperthyroid (metaestrus-diestrus). After 60 days the blood plasma was collected and the concentrations of free T4, estradiol, progesterone, calcium, phosphorus, and of alkaline phosphatase were determined. The bones (femur and tibia) were analysed microscopically. Despite of the functional state of the thyroid, the levels of estrogen were significantly higher in the proestrus-estrus. The estrogenic phase increased the plasmatic concentration of calcium significantly in the euthyroid rats but it did not alter the levels of phosphorus and alkaline phosphatase. In the hyperthyroid state no significant differences in the plasmatic concentrations of calcium, phosphorus and alkaline phosphatase throughout the cycle were found. The phases of the cycle did not also influence the bone morphology in the euthyroid and hyperthyroid states. It was concluded that the estrogenic phase increases the plasmatic concentration of calcium, even without altering the bone morphology of the euthyroid rats. In addition the estrogenic phase does not increase the plasmatic calcium and it does not modify the response of the bone as well as of the mineral metabolism under effect of the hyperthyroidism.

Effect of fibronectin- and collagen I-coated titanium fiber mesh on proliferation and differentiation of osteogenic cells.

The objective of this study was to evaluate the effects of fibronectin and collagen I coatings on titanium fiber mesh on the proliferation and osteogenic differentiation of rat bone marrow cells. Three main treatment groups were investigated in addition to uncoated titanium fiber meshes: meshes coated with fibronectin, meshes coated with collagen I, and meshes coated first with collagen I and then subsequently with fibronectin. Rat bone marrow cells were cultured for 1, 4, 8, and 16 days in plain and coated titanium fiber meshes. In addition, a portion of each of these coating treatment groups was cultured in the presence of antibodies against fibronectin and collagen I integrins. To evaluate cellular proliferation and differentiation, constructs were examined for DNA, osteocalcin, and calcium content and alkaline phosphatase activity. There were no significant effects of the coatings on cellular proliferation as indicated by the DNA quantification analysis. When antibodies against fibronectin and collagen I integrins were used, a significant reduction (p < 0.05) in cell proliferation was observed for the uncoated titanium meshes, meshes coated with collagen, and meshes coated with collagen and fibronectin. The different coatings also did not affect the alkaline phosphatase activity of the cells seeded on the coated meshes. However, the presence of antibodies against fibronectin or collagen I integrins resulted in significantly delayed expression of alkaline phosphatase activity for uncoated titanium meshes, meshes coated with collagen, and meshes coated with collagen and fibronectin. Calcium measurements did not reveal a significant effect of fibronectin or collagen I coating on calcium deposition in the meshes. Also, no difference in calcium content was observed in the uncoated titanium meshes and meshes coated with fibronectin when antibodies against fibronectin or collagen I integrins were present. Meshes coated with both collagen I and fibronectin showed significantly higher calcium content when cultured in the presence of antibodies to collagen and fibronectin integrins. A similar phenomenon was also observed for collagen-coated meshes cultured in the presence of antibodies to fibronectin integrins. No significant differences in osteocalcin content were observed between the treatment groups. However, all groups exposed to antibodies against fibronectin integrins showed a significant decrease in osteocalcin content on day 16. These results show that a fibronectin or collagen I coating does not stimulate the differentiation of rat bone marrow cells seeded in a titanium fiber mesh.

CALCIUM ABSORPTION AND DISTRIBUTION IN MATURE KIWIFRUIT PLANTS

Actinidia deliciosa (C.F. Liang et A. R. Ferguson L.) mature plants trained to “Pergola” system were grown in a semiarid environment located in Southern Italy. During the growing season measurements were carried out on sap-flow of canes grown in different light exposure (exposed and shaded) using the heat-balance method. The exposed condition (> 40% daily available PPFD) was maintained through summer pruning, while the shaded one (< 20% daily available PPFD) was obtained using a shading-net; meteorological data were recorded, too. At the end of growing season, calcium and potassium accumulation in leaves, fruits and shoots of exposed and shaded canes were determined. Throughout the season the sap-flow of shaded canes was on average 45% of the exposed one. At the end of growing season differences in calcium content were found in different organs of exposed and shaded canes. The calcium concentration in exposed and shaded leaves was about 3 g m-2 and 1.5 g m-2 respectively, while in fruit the concentration was 0.2 % D.M and 0.11% D.M. for exposed and shaded fruits. The results obtained indicate that a high level of light availability determine an increase in xylem stream in canes which is responsible for the increase of calcium accumulation in different organs. Besides this increase of fruit transpiration rate can improve fruit storage quality.

Observations on the effect of BMP-2 on rat bone marrow cells cultured on titanium substrates of different roughness

The objective of this study was to examine the osteoinductive capacity of different concentrations of BMP-2 on bone marrow stromal cells in vitro. Further, we intended to determine whether titanium provided with an increased surface roughness is more efficient in osteoblast differentiation than machined titanium. Therefore, 20,000 cells/ml were seeded and cultured on machined and grit-blasted titanium discs for 4, 8 and 16 days. Different concentrations of rhBMP-2 (0, 10, 100, 1000 ng/ml) were supplemented to the medium for 8 days of culturing. To evaluate cellular proliferation and differentiation, specimens were examined for DNA, alkaline phosphatase activity, and calcium content. Morphological appearance of the specimens at 8 and 16 days of incubation was evaluated using scanning electron microscopy. Two separate experimental runs were performed. Evaluation of the DNA and alkaline phosphatase data revealed that a significant difference existed for these data between both experimental runs. Further analysis of the DNA figures learned that roughening of the titanium surface and addition of BMP-2 had no effect on cell proliferation. The alkaline phosphatase analysis and calcium measurements revealed that BMP-2 stimulated the early differentiation of osteogenic cells on machined titanium substrates in a dose-dependent manner. After 16 days of culture, no significant differences in calcium content could be observed anymore between machined and roughened titanium surfaces. Further, the data revealed that the machined surfaces showed a significant increase in calcium deposition when 100 and 1000 ng/ml BMP-2 were supplemented to the medium. However, the roughened surfaces showed this significant enhancement in calcium content only with 1000 ng/ml BMP-2. In addition, SEM evaluation revealed a dose-dependent response to BMP-2. Increasing BMP-2 concentrations resulted in more calcified globular accretions on bone surfaces than when no BMP-2 was added. On the basis of our results, we conclude that (1) due to the heterogeneous nature of bone marrow, experimental results with primary rat bone marrow cells are difficult to reproduce from one experiment to the other, and (2) addition of rhBMP-2 in the medium stimulates the early differentiation and matrix mineralization of osteogenic cells on machined titanium surfaces in a dose-responsive manner. Further, we concluded that our roughened titanium surfaces had no effect on proliferation and differentiation of primary derived rate bone marrow cells.

Bone formation by rat bone marrow cells cultured on titanium fiber mesh: effect of in vitro culture time.

The objective of this study was to examine the effect of cell culture time on bone formation by rat bone marrow cells seeded in titanium fiber mesh. As a seeding technique, a high cell suspension was used (3 x 10(6) cells/mL). Therefore, 30 meshes were repeatedly rotated in a 10 mL tube (containing 30 x 10(6) cells) on a rotation plate (2 rpm) for 3 h. Osteogenic cells were cultured for 1, 4, and 8 days on titanium fiber mesh and finally implanted subcutaneously in rats. Meshes without cells were also implanted subcutaneously in rats. DNA and scanning electron microscopy (SEM) analyses and calcium measurements determined cellular proliferation and differentiation during the in vitro incubation period of the mesh implants. Four weeks after implant insertion, the animals were sacrificed. The implants, with their surrounding tissue, were retrieved and prepared for histologic evaluation and calcium measurements. DNA analysis of the in vitro experiment showed a lag phase from day 1 through day 4, but a 42% increase in DNA between days 4 and 8. SEM and calcium measurements indicated an increase in calcium from day 1 to day 4, yet only a small but significant increase from days 4 to 8. Histologic analysis demonstrated that bone was formed in all day 1 and day 4 implants, and that the bone-like tissue was present uniformly through the meshes. The bony tissue was morphologically characterized by osteocytes embedded in a mineralized matrix, with a layer of osteoid and osteoblasts at the surface. The day 8 implants showed only calcium phosphate deposition in the titanium fiber mesh. Calcium measurements of the implants revealed that calcification in day 1 implants was significantly higher (p < 0.05) compared to day 4 and day 8 implants. No significant difference in calcium content existed between day 4 and day 8 implants. On the basis of our results, we conclude that 1) bone formation was generated more effectively in osteogenic cells by a short culture time after seeding in titanium fiber mesh; 2) dynamic cell seeding is probably more effective than static cell seeding; and 3) selection of the right cells from the heterogenous bone marrow population remains a problem.

In vivo evaluation of cellular and acellular bovine pericardia fixed with a naturally occurring crosslinking agent (genipin)

A cell extraction process was employed in the study to remove the cellular components from bovine pericardium, leaving a framework of largely insoluble collagen and elastin. It was hypothesized in the literature that this process may decrease the antigenic load (or increase the biocompatibility) within the material. Additionally, acellular tissues may provide a natural microenvironment for host-cell migration to regenerate the tissue. The study was to evaluate the biocompatibility of cellular and acellular bovine pericardia fixed with a naturally occurring crosslinking agent (genipin) implanted subcutaneously in a growing rat model. Additionally, the tissue regeneration rate in the genipin-fixed acellular tissue was investigated. The glutaraldehyde-fixed counterparts were used as controls. The results indicated that the degrees in inflammatory reaction for the genipin-fixed cellular and acellular tissues were significantly less than their glutaraldehyde-fixed counterparts. Additionally, it was noted that the inflammatory reactions for the glutaraldehyde-fixed cellular and acellular tissues lasted much longer than their genipin-fixed counterparts. The tissue regeneration rate for the genipin-fixed acellular tissue was significantly faster than its glutaraldehyde-fixed counterpart. The calcium content of each studied group, analyzed by atomic absorption, did not change significantly until at the 52nd week, postoperatively. The differences in calcium content between the cellular and acellular tissues were insignificant for both the glutaraldehyde- and genipin-fixed groups throughout the entire course of the study. In summary, the biocompatibility of the genipin-fixed cellular and acellular tissues was superior to their glutaraldehyde-fixed counterparts. The genipin-fixed acellular tissue provided a better microenvironment for tissue regeneration than its glutaraldehyde-fixed counterpart, due to its low cytotoxicity. These results suggested that the genipin-fixed acellular tissue might be used as a tissue-engineering matrix in the clinical applications.

Calcium deficiency provoked by the application of different forms and concentrations of Ca 2+ to soil-less cultivated muskmelons

The influences of deficiency concentrations of calcium compounds other than the calcium nitrate usually used, on the development of Cucumis melo L. cv. Revigal cultivated in rockwool were studied under adverse calcium absorption conditions (low solar radiation, temperature and calcium and high sodium and magnesium levels in the nutrient solution). A standard Ca(NO 3) 2 concentration was used for comparison and two different concentrations of Ca-EDTA, CaCl 2 and CaSO 4 (equivalent to half and to one quarter of the calcium in the control) were used to create calcium deficiencies. The demand for calcium increased substantially during the period of the greatest vegetative growth. Ca-EDTA produced greater Ca deficiency symptoms than CaCl 2 and CaSO 4, and was not effective in alleviating calcium-associated problems. There were no significant differences in biomass production between the sulphate and chloride treatments with the same calcium levels, although there were great differences in biomass production in the different plant fractions between the two calcium levels applied. The biomass differences were greater half way through the cultivation cycle than at the end. The use of calcium concentrations below 3 mM in the nutrient solution, which represents a net daily calcium uptake of less than 1 mmol per plant throughout the cultivation cycle was insufficient and provoked deficiency symptoms in leaves and fruits in the conditions of this experiment. Calcium levels at the end of the growing cycle were significantly higher in the control than in other treatments in all plant fractions. There were very few differences in calcium content in fractions between the chloride and sulphate treatments, while the Ca-EDTA treatments led to noticeably lower calcium levels in the different fractions.