Secondary metabolites from the fruit body of Phellinus linteus were evaluated for their proliferative effect on human osteoblast-like cells. 3-[4,5-Dimethylthiazole-2-y1]-2,5-diphenyl-tetraxolium bromide (MTT) assay and alkaline phosphatase (ALP) activity assay were used to assess the effect those isolates on the human osteoblast-like cell line (Saos-2). Activity-guided fractionation led to the isolation of ALP-activating phenolic compounds through the extraction of P. linteus, solvent partitioning, and repeated silica gel and octadecyl silica gel (ODS) column chromatographic separations. From the result of spectroscopic data including nuclear magnetic resonance (NMR), mass spectrometry (MS), and infrared spectroscopy (IR), the chemical structures of the compounds were determined as 4-(4-hydroxyphenyl)-3-buten-2-one(1), 2-(3',4'-dihydroxyphenyl)-1,3-benzodioxole-5-aldehyde (2), 4-(3,4-dihydroxyphenyl)-3-buten-2-one (3), 3,4-dihydroxybenzaldehyde (4), and protocatechuic acid methyl ester (5), respectively. This study reports the first isolation of compounds 1-3 and 5 from P. linteus. In addition, all phenolic compounds stimulated proliferation of the osteoblast-like cells and increased their ALP activity in a dose-dependent manner ($10^{-8}$ to $10^{-1}\;mg/mL$). The present data demonstrate that phenolic compounds in P. linteus stimulated mineralization in bone formation caused by osteoporosis. The bone-formation effect of P. linteus seems to be mediated, at least partly, by the stimulating effect of the phenolic compounds on the growth of osteoblasts.
Kim, Byoung-Kook;Kim, Yong-Gun;Lee, Ho-Jae;Choi, Dae-Sung;Dan, Jinmyoung
Journal of the Korean Orthopaedic Association
/
v.53
no.6
/
pp.552-557
/
2018
Vitamin D plays a major role in bone metabolism, and its deficiency has an impact on fracture risk and healing. Low vitamin D levels are a cause of poor bone mineralization and have been associated with a significantly higher risk of physeal injury in children. This paper presents a case of a 13-year-old boy with a vitamin D deficiency, who sustained multiple sequential epiphyseal injuries at various areas. This report suggests that vitamin D deficiency is not only a significant cause of the clinical disease itself, but also an important factor affecting the successful recovery of injuries.
Hyun-Su Baek;Se-Jin Park;Eun-Gyung Lee;Yong-Il Kim;In-Ryoung Kim
The Korean Journal of Physiology and Pharmacology
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v.28
no.5
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pp.423-433
/
2024
Dental pulp stem cells (DPSCs) are a type of adult stem cell present in the dental pulp tissue. They possess a higher proliferative capacity than bone marrow mesenchymal stem cells. Their ease of collection from patients makes them well-suited for tissue engineering applications, such as tooth and nerve regeneration. Chios gum mastic (CGM), a resin extracted from the stems and leaves of Pistacia lentiscus var. Chia, has garnered attention for its potential in tissue regeneration. This study aims to confirm alterations in cell proliferation rates and induce differentiation in human DPSCs (hDPSCs) through CGM treatment, a substance known for effectively promoting odontogenic differentiation. Administration of CGM to hDPSC cells was followed by an assessment of cell survival, proliferation, and odontogenic differentiation through protein and gene analysis. The study revealed that hDPSCs exhibited low sensitivity to CGM toxicity. CGM treatment induced cell proliferation by activating cell-cycle proteins through the Wnt/β-catenin pathway. Additionally, the study demonstrated that CGM enhances alkaline phosphatase activation by upregulating the expression of collagen type I, a representative matrix protein of dentin. This activation of markers associated with odontogenic and bone differentiation ultimately facilitated the mineralization of hDPSCs. This study concludes that CGM, as a natural substance, fosters the cell cycle and cell proliferation in hDPSCs. Furthermore, it triggers the transcription of odontogenic and osteogenic markers, thereby facilitating odontogenic differentiation.
Yoo, Han-Seok;Chung, Kang-Hyun;Lee, Kwon-Jai;Kim, Dong-Hee;An, Jeung Hee
Nutrition Research and Practice
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v.11
no.3
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pp.190-197
/
2017
BACKGROUND/OBJECTIVES: Gallus gallus domesticus (GD) is a natural mutant breed of chicken in Korea with an atypical characterization of melanin in its tissue. This study investigated the effects of melanin extracts of GD on osteoblast differentiation and inhibition of osteoclast formation. MATERIALS/METHODS: The effects of the melanin extract of GD on human osteoblast MG-63 cell differentiation were examined by evaluating cell viability, osteoblast differentiation, and expression of osteoblast-specific transcription factors such as bone morphogenetic protein 2 (BMP-2), small mothers against decapentaplegic homologs 5 (SMAD5), runt-related transcription factor 2 (RUNX2), osteocalcin and type 1 collagen (COL-1) by reverse transcription-polymerase chain reaction and western blotting analysis. We investigated the inhibitory effect of melanin on the osteoclasts formation through tartrate-resistant acid phosphatase (TRAP) activity and TRAP stains in Raw 264.7 cell. RESULTS: The melanin extract of GD was not cytotoxic to MG-63 cells at concentrations of $50-250{\mu}g/mL$. Alkaline phosphatase (ALP) activity and bone mineralization of melanin extract-treated cells increased in a dose-dependent manner from 50 to $250{\mu}g/mL$ and were 149% and 129% at $250{\mu}g/mL$ concentration, respectively (P < 0.05). The levels of BMP-2, osteocalcin, and COL-1 gene expression were significantly upregulated by 1.72-, 4.44-, and 2.12-fold in melanin-treated cells than in the control cells (P < 0.05). The levels of RUNX2 and SMAD5 proteins were higher in melanin-treated cells than in control vehicle-treated cells. The melanin extract attenuated the formation of receptor activator of nuclear factor kappa-B ligand-induced TRAP-positive multinucleated RAW 264.7 cells by 22%, and was 77% cytotoxic to RAW 264.7 macrophages at a concentration of $500{\mu}g/mL$. CONCLUSIONS: This study provides evidence that the melanin extract promoted osteoblast differentiation by activating BMP/SMADs/RUNX2 signaling and regulating transcription of osteogenic genes such as ALP, type I collagen, and osteocalcin. These results suggest that the effective osteoblastic differentiation induced by melanin extract from GD makes it potentially useful in maintaining bone health.
This study was carried out in order to study early histologic changes and repair reaction appling to extrusive force for 3rd premolar of adult dogs. After 1 week of extrusive force with elastic chain, one of dogs was sacrified and after 3 weeks retention period, another dog was sacrified. The paraffin sections of samples were stained with Hematoxylin - Eosin and Masson's Trichrome and were examed by light microscopy . The obtained results as follows 1. In Hematoxylin - Eosin and Masson Trichrome stain of control group , the periodontal ligament width was constant from apical third to cervical third of the root and periodontal fiber arrangement was horizontal or oblique in cervical third. oblique in middle third, oblique in apical third of root. in alveolar bone, smooth appearance was shown 2. In Group 1, all periodontal fiber arrangement was oblique toward tooth, and the periodontal ligament width increased Partially PDL was ruptured in apex. In MT stain, immature bone formation was seen at alveolar crest area. Active bone formation was observed along the one side of alveolus, and apical portion of pulp was involved with blood vessel rupture , vacuolization of pulp tissue and hyperemia 3. In Group 2, most periodontal ligament arrangement and PDL width was repaired and fiber density increased. In MT stain, mineralization of immature bone on the alveolar crest was progressed. In pulp, vacuole and hyperemia was diminished and fibrotic change was diminished 4. After 3 week periodontal ligament has more repair ability than pulp tissue. pulp was involved with vacuolization and fibrosis, so it takes more time for repair.
An experiment was conducted (28 to 44 weeks) to study the laying performance, shell quality, and nutrient retention of White Leghorn layers fed different levels of non-phytate phosphorus (NPP). Six levels of NPP (0.15, 0.18, 0.21, 0.24, 0.27 and 0.30%) at a constant calcium (Ca) level (3.5%) in maize-soya-deoiled rice bran based diets were formulated, and each experimental diet was offered ad libitum for 16 weeks to five replicates with five birds in each replicate. The body weight of WL layers fed diet containing 0.15% NPP was significantly (p<0.05) lower than those fed diet with 0.30% NPP, at 44 weeks of age. However, the hen day egg production, egg weight, daily feed intake and feed consumed per dozen eggs were not influenced by the variation in the NPP levels in the diet. The bone ash content was significantly (p<0.05) higher in the birds fed 0.30% NPP as compared with those fed diets up to 0.24% NPP. Bone ash content was intermediate in the birds fed diet containing 0.27% NPP. The tibia strength followed the same trend as that of bone ash. Dietary NPP content had no influence on serum Ca and protein concentration and activity of alkaline phosphatase. However, serum inorganic P concentration increased linearly with NPP content in the diet. The concentration of P was significantly (p<0.05) higher in the birds fed 0.27% NPP or higher as compared with those fed 0.15% NPP. Levels of dietary NPP had no influence on egg quality parameters like shell wt, shell thickness, shell strength and specific gravity. The retention of nutrients such as DM, N and Ca were comparable among the WL layers fed different levels of NPP. However, the retention of P decreased linearly with increase in the level of NPP in the diet. The retention of P in the birds fed diets up to 0.24% NPP in the diet was comparable, however further increasing the content of NPP (either 0.27% or 0.30%) reduced the retention of P. Based on the results of the present study, 0.15% NPP (180 mg/b/d) in the diets of WL layers is adequate for optimum production performance during 28 to 44 weeks of age, however, WL layers require 0.27% NPP (324 mg /b/d) in the diet for optimum production with better bone mineralization.
Naked DNA and standard vectors have been previously used for gene delivery. Among these, PEI can efficiently condense DNA and has high intrinsic endosomal activities. The aim of this study is to investigate whether the cationic polycation PEI could increase the transfection efficiency of BMP expressing DNA using a vector-loaded collagen sponge model. BMP-2/pcDNA3.1 plasmid was constructed by subcloning human BMP-2 cDNA into the pcDNA3.1 plasmid vector. PEI/DNA complexes were prepared by mixing PEI and BMP-2/pcDNA3.1 and the constructed complexes were loaded into the collagen sponges. In vitro studies, BMSCs were transfected with the PEI/BMP-2/pcDNA3.1 complexes from collgen sponge. The level of secreted BMP-2 and alkaline phosphatase activities of transfected BMSCs were significantly higher in PEI/BMP-2/pcDNA3.1 group than in BMP-2/pcDNA3.1 group (p<0.05). Transfected BMSCs were cultured and mineralization was observed only in cells treated with PEI/BMP-2/pcDNA3.1 complexes. In vivo studies, PEI/BMP-2/pcDNA3.1/collagen, BMP-2/pcDNA3.1/collagen and blank collagen were grafted in skeletal muscle of nude mice. Ectopic bone formation was shown in PEI/BMP-2/pcDNA3.1/collagen grafted mouse 4 weeks postimplantation, while not in BMP-2/pcDNA3.1 grafted tissue. This study suggests that PEI-condensed DNA encoding for BMP-2 is capable of inducing bone formation in ectopic site and might increase the transfection rate of BMP-2/pcDNA3.1. As a non-viral vector, PEI offers the potential in gene therapy for bone engineering.
The bones of the human body support the structures of the body and provide protection for a person's internal organs. Bone metabolic diseases are on the rise due to a significant increase in life expectancy over a short period of time. Therefore, we investigated the osteoblast differentiation promoting and osteoclastogenesis inhibitory activities of fermented Benincasa hispida cong. (HR1901-BS, HR1901-BSaf). We evaluated the alkaline phosphatase (ALP) activity of MC3T3-E1 mouse calvarial-derived osteoblasts. We also evaluated expression of ALP, osteocalcin (OCN), and runt-related transcription factor 2 (Runx2), which regulate osteoblast differentiation. To assess effects on osteoclast formation, tartrate-resistant acid phosphatase (TRAP) activity in RAW264.7 cells was analyzed. ALP activity increased by 121-136% and 140-156%, respectively in the presence of HR1901-BS and HR1901-BSaf. Expression of osteoblast differentiation factor also increased significantly. We also confirmed that HR1901-BS and HR1901-BSaf decreased TRAP activity in osteoclasts by 35-47% and 23-39%, respectively. Our results showed that fermented Benincasa hispida cong. (HR1901-BS, HR1901-BSaf) increase bone mineralization and osteoblast differentiation activity in MC3T3-E1 cells, and inhibit bone resorption activity in RAW264.7 cells. In conclusion, fermented Benincasa hispida cong. (HR1901-BS, HR1901-BSaf) can be used as an effective natural resource for preventing and treating bone-related diseases.
Self-potential and electrical resistivity methods were conducted for locating the auriferous mineralized zone, called Jija Vein, of Sambo mine, Limsu-ri, Haeje-myeon, Muahn-gun, Jeollanam-do. The host rocks of the mineralization include gneiss, rhyolite and felsic dyke. Ore vein is mainly fissured-filling type and sulfide minerals, such as pyrite, are disseminated in country rock. By the electrical responses from SP and surface resistivity methods., the mineralized zone is supposed to extend about 360 m directed N5W with the width of 20 m to 30 m. From resistivity tomograms using inclined borehole to surface, the ore body shape is interpreted as the width of 20 m in depth 40 m to 50 m.
Purpose: This review aims at introducing 3 modeling approaches classified into 3 categories based on the purpose (estimation or prediction), structure (linear or non-linear) and phase (steady-state or dynamic-state); 1) statistical approaches, 2) kinetic modeling and 3) mechanistic modeling. We hope that this review can be a useful guide in the model-based approach of calcium metabolism as well as illustrates an application of engineering tools in studying biosystems. Background: The meaning of biosystems has been expanded, including agricultural/food system as well as biological systems like genes, cells and metabolisms. This expansion has required a useful tool for assessing the biosystems and modeling has arisen as a method that satisfies the current inquiry. To suit for the flow of the era, examining the system which is a little bit far from the traditional biosystems may be interesting issue, which can enlarge our insights and provide new ideas for prospective biosystem-researches. Herein, calcium metabolic models reviewed as an example of application of modeling approaches into the biosystems. Review: Calcium is an essential nutrient widely involved in animal and human metabolism including bone mineralization and signaling pathways. For this reason, the calcium metabolic system has been studied in various research fields of academia and industries. To study calcium metabolism, model-based system analyses have been utilized according to the purpose, subject characteristics, metabolic sites of interest, and experimental design. Either individual metabolic pathways or a whole homeostasis has been modeled in a number of studies.
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