• Journal of the korean academy of Pediatric Dentistry
• /
• v.25 no.3
• /
• pp.635-648
• /
• 1998

The clinical use of fluoride with a well known osteogenic action in osteoporotic patients is rational, because this condition is characterized by impaired bone formation. However, its anabolic effect has not been demonstrated well in vitro. The purpose of this study was to investigate the effects of sodium fluoride on the physiological role of osteoblastic cell. Osteoblastic cells were isolated from fetal rat calvaria. The results were as follows : 1. Mineralized nodules were shown in osteoblastic cell cultures, which had been maintained in the presence of ascorbic acid and ${\beta}-glycerophosphate$ up to 21 days. When cultures were treated with pulses of 48 hr duration before apparent mineralization was occurring, 2-fold increased in their number was detected. 2. Alkaline phosphatase activity of osteoblastic cells was inhibited by sodium fluoride in dose dependent manner. 3. The effect of sodium fluoride on the osteoblastic cell proliferation was measured by the incorporation of $[^3H]$-thymidine into DNA. As a result, sodium fluoride at $1{\sim}100{\mu}M$ increased the $[^3H]$-thymidine incorporation into DNA in a dose dependent manner. 4. The signaling mechanism activated by sodium fluoride dose-dependently enhanced the tyrosine phosphorylation of the adaptor molecule $Shc^{p66}$ and their association with Grb2, one of earlier events in a MAP kinase activation pathway cascade used by a significant subset of G protein-coupled receptors. 5. The phosphorylation of CREB(cAMP response element binding protein)was inhibited by the sodium fluoride in MC3T3E1 cells. In conclusion, the results of this study suggested that the mitogenic effect of the sodium fluoride in MC3T3E1 cell was stimulated in a dose-dependent manner and suggested "an important role for the interaction between She and Grb2" in controlling the proliferation of osteoblasts.

• Nuclear Engineering and Technology
• /
• v.52 no.10
• /
• pp.2320-2327
• /
• 2020

The extreme hydrophobicity of expanded polytetrafluoroethylene (ePTFE) hinders bone-tissue integration, thus limiting the use of ePTFE in medical implant applications. To improve the potential of ePTFE as a biomaterial, 2-hydroxyethyl methacrylate (HEMA) was grafted onto the ePTFE surface using the gamma irradiation technique. The characteristics of the grafted ePTFE were successfully evaluated using attenuated total reflectance Fourier transform infrared (ATR-FTIR), field-emission scanning electron microscopy (FESEM)/energy dispersive X-ray (EDX), and X-ray photoelectron spectroscopy (XPS). Under the tensile test, the modified ePTFE was found to be more brittle and rigid than the untreated sample. In addition, the grafted ePTFE was less hydrophobic with a higher percentage of water uptake compared to the untreated ePTFE. The protein adsorption test showed that grafted ePTFE could adsorb protein, which was denoted by the presence of N peaks in the XPS analysis. Moreover, the formation of the globular mineral on the grafted ePTFE surface was successfully visualized using the FESEM analysis, with a ratio of 1.94 for Ca:P minerals by the EDX. To summarize, the capability of the modified ePTFE to show protein adsorption and mineralization indicates the improvement of the polymer properties, and it can potentially be used as a biomaterial for implant application.

• The korean journal of orthodontics
• /
• v.49 no.4
• /
• pp.205-213
• /
• 2019

Objective: The aim of this study was to evaluate the amount of tooth movement and histologic changes with different corticotomy designs and micro-osteoperforation in rabbits. Methods: The sample consisted of 24 rabbits divided into three experimental groups (triangular corticotomy [TC] and indentation corticotomy [IC] with flap, and flapless micro-osteoperforations [MP]) and a control. A traction force of 100 cN was applied by connecting the first premolars to the incisors. The amount of tooth movement was measured. Kruskal-Wallis test was used to assess differences in tooth movement between the groups. Micro-computed tomography, hematoxylin and eosin staining, and tartrate-resistant acidic phosphatase (TRAP) analysis were performed. Analysis of variance was applied to assess differences in TRAP-positive osteoclast count between the groups. Results: The amount of tooth movement increased by 46.5% and 32.0% in the IC and MP groups, respectively, while the bone fraction analysis showed 69.7% and 8.5% less mineralization compared to the control. There were no significant intergroup differences in the number of TRAP-positive osteoclasts. Conclusions: The micro-osteoperforation group showed no significant differences in the amount of tooth movement compared to the corticotomy groups, nor in the TRAP-positive osteoclast count compared to both corticotomy groups and control.

• Animal Bioscience
• /
• v.36 no.2
• /
• pp.167-174
• /
• 2023

Phosphorus (P) is a macro mineral needed for bone mineralization and cell membrane structure and P is also involved in several fundamental pathways of metabolism in the body. Because of the low concentration and digestibility of P in plant ingredients that are the main components of diets for poultry and pigs, feed phosphates are usually included in diets in addition to the P contributed by plant ingredients. The most widely used feed phosphates in poultry and swine diets are dicalcium phosphate (DCP) and monocalcium phosphate (MCP), but tricalcium phosphate (TCP), monosodium phosphate (MSP), and magnesium phosphate (MgP) may be used as well. Because feed phosphates are mostly produced from rock phosphate, feed phosphates have impurities that contain minerals other than P. Concentrations of P in feed phosphates range from 14.8% (MgP) to 25.7% (MSP). The standardized total tract digestibility (STTD) of P in pigs ranges from 71% (TCP) to 95% (MSP). The STTD of Ca and the standardized ileal digestibility (SID) of P and Ca in feed phosphates fed to pigs and poultry have been determined only in a few experiments. Available data indicate that the STTD of Ca and SID of P in MCP are greater than in DCP in both poultry and pigs, but the SID of Ca is similar between DCP and MCP fed to broilers. Information on mineral concentrations and digestibility values in feed phosphates is needed in diet formulation for pigs and poultry, but if diets are formulated to contain equal concentrations of digestible P and Ca, it is unlikely that animal performance will be impacted by the source of feed phosphates used in the diet.

• Journal of Veterinary Science
• /
• v.24 no.5
• /
• pp.69.1-69.11
• /
• 2023

Background: Kalkitoxin (KT) is an active lipopeptide isolated from the cyanobacterium Lyngbya majuscula found in the bed of the coral reef. Although KT suppresses cell division and inflammation, KT's mechanism of action in vascular smooth muscle cells (VSMCs) is unidentified. Therefore, our main aim was to investigate the impact of KT on vascular calcification for the treatment of cardiovascular disease. Objectives: Using diverse calcification media, we studied the effect of KT on VSMC calcification and the underlying mechanism of this effect. Methods: VSMC was isolated from the 6 weeks ICR mice. Then VSMCs were treated with different concentrations of KT to check the cell viability. Alizarin red and von Kossa staining were carried out to examine the calcium deposition on VSMC. Thoracic aorta of 6 weeks mice were taken and treated with different concentrations of KT, and H and E staining was performed. Real-time polymerase chain reaction and western blot were performed to examine KT's effect on VSMC mineralization. Calcium deposition on VSMC was examined with a calcium deposition quantification kit. Results: Calcium deposition, Alizarin red, and von Kossa staining revealed that KT reduced inorganic phosphate-induced calcification phenotypes. KT also reduced Ca⁺⁺-induced calcification by inhibiting genes that regulate osteoblast differentiation, such as runtrelated transcription factor 2 (RUNX-2), SMAD family member 4, osterix, collagen 1α, and osteopontin. Also, KT repressed Ca²⁺-induced bone morphogenetic protein 2, RUNX-2, collagen 1α, osteoprotegerin, and smooth muscle actin protein expression. Likewise, Alizarin red and von Kossa staining showed that KT markedly decreased the calcification of ex vivo ring formation in the mouse thoracic aorta. Conclusions: This experiment demonstrated that KT decreases vascular calcification and may be developed as a new therapeutic treatment for vascular calcification and arteriosclerosis.

• Journal of the korean academy of Pediatric Dentistry
• /
• v.30 no.3
• /
• pp.391-405
• /
• 2003

Craniosynostosis, known as a premature fusion of cranial sutures, is a developmental disorder characterized by precocious differentiation and mineralization of osteoblasts in the calvarial sutures. Recent genetic studies have demonstrated that mutation in the homeobox gene Msx2 causes Boston-type human craniosynostosis. Additionally, the phenotype of Dlx5 homozygote mutant mouse presents craniofacial abnormalities including a delayed ossification of calvarial bone. Furthermore transcription of osteocalcin, a mature osteoblast marker, is reciprocally regulated by the homeodomain proteins Msx2 and Dlx5. These facts suggest important roles of osteocalcin, Msx2 and Dlx5 genes in the calvarial bone growth and suture morphogenesis. To elucidate the function of these molecules in the early morphogenesis of mouse cranial sutures, we have first analyzed by in situ hybridization the expression of osteocalcin, Msx2 and Dlx5 genes in the developing parietal bone and sagittal suture of mouse calvaria during the embryonic (E15-E18) stage. Osteocalcin mRNA was found in the periosteum of parietal bones from E15, and gradually more highly expressed with aging. Msx2 mRNA was intensely expressed in the sutural mesenchyme, osteogenic fronts and mildly expressed in the dura mater during the embryonic stage. Dlx5 mRNA was intensely expressed osteogenic fronts and the periostem of parietal bones. To further examine the upstream signaling molecules of transcription factor Msx2 and Dlx5, we have done in vitro experiments in E15.5 mouse calvarial explants. Interestingly, implantation of BMP2-, BMP4-soaked beads onto the osteogenic fronts after 48 hours organ culture induced etopic expressions of Msx2 and Dlx5 genes. On the other hand, overexpression of $TGF{\beta}1$, GDF-6, -7, FGF-2, -4 and Shh did not induce the expression of Msx2 and Dlx5. Taken together. these data indicate that transcription factor Msx2 and Dlx5 play critical roles in the calvarial bone and suture development, and that BMP siganling is involved in the osteogenesis of calvarial bones and the maintenance of cranial sutures through regulating these two transcriotpn factors. Furthermore, different expression patterns between Msx2 and Dlx5 suggest their specific functions in the osteoblast differentiation.

• Journal of the korean academy of Pediatric Dentistry
• /
• v.27 no.2
• /
• pp.309-317
• /
• 2000

Bisphosphonates inhibit bone resorption in vivo and in vitro. Currently proposed mechanism of action of bisphosphonates involves both direct effect on osteoclasts and indirect effect through the mediation of osteoblasts. Recent understanding of molecular mechanism of osteoclastogenesis indicates that osteoclast differentiation is quite tightly regulated by signaling molecules from differentiating osteoblasts. Therefore this investigation was designed to elucidate the effect of bisphosphonate on osteoblast differentation. For this purpose, in vitro effects of etidronate and alendronate on the expression of Cbfa1 a master control gene of osteoblast differentiation, several bone marker genes, and formation of calcified nodules were evaluated. To evaluate the effect of bisphosphonate on calcified nodule formation, osteoblasts isolated from rat calvaria were cultured in a-MEM containing $10^{-4},\;10^{-5},\;10^{-6}M$ of etidronate or $10^{-6},\;10^{-7},\;10^{-8}M$ of alendronate for 15 days, and then stained by alizarin red to determine mineralization. To evaluate the effect of bisphosphonate on osteoblast differentiation, osteoblast cells were cultured in a-MEM containing $10^{-4},\;10^{-5},\;10^{-6}M$ of etidronate or $10^{-6}$ M of alendronate for 8 days. And then total RNA was extracted and northern blot analysis was done to examine the expression of Cbfa1, type I collagen, alkaline phosphatase, osteopontin and osteocalcin. The results were as follows: 1. Etidronate suppressed the calcification of bone nodule in dose dependent manner, while alendronate didn't. 2. The expression of Cbfa1 was decreased dose dependently by etidronate, but increased by alendronate. 3. Etidronate suppressed the expression of type I collagen, osteopontin and osteocalcin in dose dependent manner however alendronate promote the expression of osteoblast marker gene. 4. The expression of alkaline phosphatase was not affected either etidronate nor alendronate. These results suggest that etidronate suppressed the expression of Cbfa1 in dose dependent manner, and consequently the expression of osteoblast marker genes, such as type I collagen, osteopontin and osteocalcin were also suppressed in similar manner. And finally this decreased expression of osteoblastic marker gene prevent calcined bone nodule formation.

• Asian-Australasian Journal of Animal Sciences
• /
• v.16 no.11
• /
• pp.1642-1649
• /
• 2003

An experiment was conducted to evaluate the effects of phytase supplementation on the growth performance, nutrients utilization and bone mineralization in broiler chickens. Day-old broiler chicks (n=480) were equally devided into eight treatment groups and fed maize or wheat based isocaloric, isonitrogenous and isocalcium diets having two non phytate phosphorus (NPP) concentrations (0.50% and 0.30%) and two phytase levels (0 and 500 phytase units/kg diet) in a 42 days growth trial. Maize based dietary treatments were MC (NPP 0.50%, MN (NPP 0.30%), MNP (MN+500 units of phytase) and MCP (MC+500 units of phytase), whereas wheat based experimental diets were WC (NPP 0.50%), WN (NPP 0.30%), WNP (WN+500 units of phytase) and WCP (WC+500 units of phytase). The NPP levels were maintained by dicalcium phosphate. Reduction in dietary NPP depressed live weight gain and feed intake and increased feed conversion ratio (FCR). Phytase supplementation to low NPP (0.30%) diets significantly (p<0.05) improved the growth performances of broilers. The supplementation to low NPP diets allowed complete, safe and economic replacement of dietary inorganic P (dicalcium phosphate) to reduce feed cost per kg live weight gain of broilers. Reduction in dietary NPP did not affect retention of nutrients except phosphorus (P) but had a significant (p<0.05) depression in tibia ash and minerals (Ca, P) concentration in serum and tibia ash. Phytase supplementation at low NPP level was effective (p<0.05) in improving the retention of dry matter, Ca and P and Ca and P concentration in serum and tibia ash. However, the supplementation was not effective at high level of NPP (0.50%). There were no significant (p>0.05) differences in carcass quality among dietary treatments. The response of phytase was greater in low NPP and maize based diets as compared with high NPP and wheat based diets, respectively. The results show that phytase supplementation to low NPP (0.30%) diets improved the growth performance, relative retention of nutrients (N, Ca and P) and minerals (Ca, P) status of blood and bone in broiler chickens, with a better efficacy in maize based diets.

• Asian-Australasian Journal of Animal Sciences
• /
• v.20 no.6
• /
• pp.933-938
• /
• 2007

An experiment was conducted to study the laying performance, shell quality, bone mineralization, hatchability of eggs and performance of progeny (weight at day one and 14 d of age, P content in day old chick, leg score and survivability of chicks) of synthetic broiler breeders fed different levels of non-phytate phosphorus (NPP). Six levels of NPP (1.2, 1.8, 2.4, 3.0, 3.6 and 4.2 g/kg diet) at a constant calcium (Ca) level (30 g/kg) in a maize-soya-deoiled rice bran based diet were tested. Levels of dicalcium phosphate, shell grit and deoiled rice bran were adjusted to achieve the desired levels of NPP and Ca. Each level of NPP was fed with a weighed quantity of feed (160 g/b/d) to 40 female broiler breeders from 25 to 40 weeks of age housed in individual cages. Each bird was considered as a replicate. Egg production, feed/egg mass, body weight, egg weight, shell weight, shell thickness, egg specific gravity, serum Ca content and tibia breaking strength were not influenced (p>0.05) by the variation in dietary NPP levels tested. Increasing the dietary levels of NPP did not influence the hatchability of eggs, phosphorus (P) contents both in egg yolk and day old chick, chick body weight at day one and 14 d of age, leg score and survivability of chicks up to 14 d of age. Maximum response ($p{\leq}0.01$) in shell breaking strength, tibia ash and serum inorganic P contents were observed at NPP levels of 2.09, 2.25 and 3.50 g per kg diet, respectively. The retention of Ca increased, while the P retention decreased ($p{\leq}0.01$) with increasing dietary levels of NPP. Though maximum responses in shell breaking strength, bone ash and serum inorganic P were observed at NPP higher than 1.2 g/kg diet, the broiler breeder performance in terms of egg production, shell quality, hatchability of eggs and progeny performance and their survivability was not influenced by dietary NPP concentrations. It is concluded that synthetic broiler breeders maintained in cages do not require more than 1.2 g NPP/kg diet with a daily intake of 192 mg NPP/b/d during 24 to 40 weeks of age.

• Journal of The Korean Society of Inherited Metabolic disease
• /
• v.16 no.3
• /
• pp.141-147
• /
• 2016

Hypophosphatasia is caused by the mutations in ALPL, which encodes tissue-nonspecific alkaline phosphatase (TNSALP). It can be inherited either in an autosomal dominant or recessive manner. Clinically, hypophophosphatasia is characterized by skeletal findings similar to those in rickets or osteomalacia, but serum alkaline phosphatase levels are decreased in the affected patients. Hypophosphatasia can be classified into six clinical forms according to age at diagnosis and severity of symptoms: perinatal lethal, infantile, childhood, adult, odontohypophosphatasia, and perinatal benign. As being a very rare disease, only one case has been reported in Korean population. Here we describe a case with perinatal benign hypophosphatasia with recessive ALPL mutations. Bowing of lower legs was detected in prenatal period and low serum alkaline phosphatase level was noted after birth. During the follow-up evaluation for 4.5 years, bone mineralization and legs bowing were improved but the growth retardation was persistent. As the recombinant bone-targeted human TNSALP became available, the clinical improvement of the affected patients is expected including the case described here with this treatment. More efforts are needed to identify the cases affected by hypophosphatasia.