This study was performed to evaluate the effect of Carthami tinctorius(HH) on osteoblast function and gene expression. The osteoblasts separated from the rat calvariae were cultivated to evaluate the cell function, and MG-63 cell was also cultivated for the test of mRNA synthesis. In this experiments, cell proliferation of rat calvarial cells was increased by HH. PKC activity, intracellular free calcium level and collgen synthesis from calvarial cells were increased by HH, but not PKA activity. And the mRNA of $PLA_2$, COX-2, and $PGE_2$ synthase from MG-63 were decreased by HH, but the mRNA of prostacyclin synthase was increased. It is concluded that HH might increase the proliferation of calvarial cell resulted from augumentation of osteoblast activity and its mRNA synthesis.
Recent in vitro and in vivo animal studies have reported that statin, a cholesterol-lowering drug, stimulate osteogenic differentiation. In the present study, we investigated the effect of simvastatin on osteogenic and adipogenic differentiation in primarily cultured human adipose-derived stem cells (hADSCs). The simvastatin treatment significantly increased the positive cell numbers in alkaline phosphatase and von Kossa staining, and enhanced the expression levels of bone morphogenic protein (BMP)-2, core binding factor alpha 1 (cbfa1), collgen type I and osteonectin mRNAs. Lastly, hADSCs were cultured in the adipogenic media with or without simvastatin to examine the effect of simvastatin on adipogenic differentiation. In the RT-PCR analysis, there were notable decreases in mRNA expression of aP1, C/EBP-$\alpha$ and PPAR-$\gamma$ in hADSCs cultivated in simvastatin-added medium, compared to those in simvastatin-free medium. It suggests that the adipogenic differentiation was significantly inhibited by simvastatin treatment. These observations indicate that simvastatin induces osteogenic differentiation and suppresses adipogenic differentiation in hADSCs.
Journal of Physiology & Pathology in Korean Medicine
/
v.23
no.5
/
pp.1095-1105
/
2009
This study was carried out to know the immunological effect of GDGT on CIA(collagen induced arthritis) mice, a model of rheumatoid arthritis. For this purpose, GDGT was orally administerd to mice with arthritis induced by collagen II and then value of cytotoxicity on hFLSs and liver, the arthritis index, immunocyte in paw joint and DNL, rheumatoid factor (IgG and IgM), collagen II specific antibody in the serum were measured. The cytotoxicity were not shown on hFLSs and liver. The arthritis index decreased significantly after 3 week. In total cell counts of DLN and paw joint, there was a significant increase in DLN and significant decrease in paw joint. In DNL, $CD19^+$, $CD3^+$, $CD4^+$, $CD3^+/CD69^+$, $CD8^+$, $CD4^+/CD25^+$, $ CD3^+/CD49b^+$ cells increased significantly. In Paw joints, $CD3^+$, $ CD4^+$, $CD4^+/CD25^+$ cells decreased significantly. The level of serum IgG and IgM decreased significantly. The level of collagen II in the serum was decreased significantly. Marginal erosion, necrotic chodrocytes, cartilage and bone degradation were improved in histological section of paw joints. The results present significant immunological effect of GDGT on rats with arthritis induced by collgen II. So we expect that GDGT should be used as a effective drugs for not only rheumatoid arthritis but also another auto-immune disease.
Journal of the Korean Association of Oral and Maxillofacial Surgeons
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v.35
no.2
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pp.120-124
/
2009
The concept of biostimulation of wounds by low-level laser therapy(LLLT) is attracting considerable attention. Although its effect on whole tissues has been studied quite extensively, the biological and cellular mechanisms underlying LLLT have not been clarified. In an experimental radius fracture in rabbits, Tang and Chai reported that LLLT enhanced the activity of red blood cells, macrophages, fibroblasts, chondrocytes, and osteoclasts within the fracture area. The purpose of the present study was to evaluate the effect of LLLT with a GaAlAs diode laser device on bone healing in rabbit mandibular fractures. We use 12 rabbits for this study. All rabbits were fractured mandible angle area using saw in anesthetic condition. In control group(n=6), none treatment was performed at fracture site. In experimental group(n=6), LLLT with a GaAlAs diode laser was radiated at fracture site daily for 7 days. All rabbits were sacrificed at 6 weeks later from performed fracture day. We studied the immunohistochemical staining of CD34 and Vimentin and the histochemical analysis for calcium and phosphorus content. The results were as follows. 1. In the histological and immunohistological staining, after 6week, fibroblasts, osteogenic cells and collgen fibers were observed more in experimental group than in control group. 2. In the histochemical analysis, the amount of calcium and phosphorus contents of the experimental group were more than the control group. From the results obtained, we suggest that the bone healing is stimulated by low-level laser irradiation in bone fractures.
In order to investigate experimentally that the effect of aqua-acupuncture solution of Paeonia lactiflora on arthritis of mice induced by collagenII, the author performed several experimental items : that is increase, paw thickness, DTH, weight of spleen, hematological change, expression of $CD4^+$, $CD8^+$, $CD19^+$, gene expression of IL-1, IL-6, IL-12, IFN-${\gamma}$, TNF, proliferation of synovial cells and cytotoxicity. The results were obtained as follows; 1. Inhibitory effects of aqua-acupuncture solution of Paeonia lactiflora on arthritis induced by collgenII. 1) In incidence, paw edema, AI and DTH were inhibited as compared with control group. 2) The splenic weight was increased and the number of leukocytes was decreased as compared with control group. 3) The number of $CD4^+$, $CD8^+$ activated cells and surface-receptor expression were increased as compared with control group. 4) In hematological change, total protein, creatinine and LDH were decreased significantiy as compared with control group. 2. FACS analysis on normal BABL/c of spenic cells treated with aqua-acupuncture solution of Paeonia lactiflora. 1) Aqua-acupuncture solution of Paeonia lactiflora activated adhesive splenic cells of mice morphologically in vitro. 2) Aqua-acupuncture solution of Paeonia dactiflora enhanced the gene expression of IL-12 and also enhanced that of interferon-${\gamma}$ remarkably. 3) Aqua-acupuncture solution of Paeonia lactiflora reduced the number of $CD4^+$, $CD8^+$, $CD19^+$ activated cells and their surface-receptor expression as compared with control group. 3. Effects of aqua-acupuncture solution of Paeonia lactiflora on human synovial cells. 1) In cytotoxicity against synovial cells, aqua-acupuncture solution of Paeonia lactiflora didn't show cytotoxicity at concentration of $10-100{\mu}g/m{\ell}$ but showed significantly at concentration of $200-400{\mu}g/m{\ell}$ as compared with control group. 2) Aqua-acupuncture solution of Paeonia lactiflora reduced the gene expression of IL-6, IL-$1{\beta}$ and TNF-${\alpha}$. 3) Aqua-acupuncture solution of Paeonia lactiflora inhibited proliferation of synovial cells at concentration of 100 and $200{\mu}g/m{\ell}$.
The effect of collagen dissolution in acid conditioned dentin was morphologically examined by both scanning and transmission electron microscopy. 18 freshly extracted human molars and dentin bonding systems of All Bond 2, Scotchbond Multipurpose, Superbond D-Liner were used in this study. For SEM preparation, each 3 of ~ exposed dentin surfaces were acid conditioned by using various acids within the above three bonding systems respectively. After acid conditioning of the other 3 exposed dentin surfaces as above, they were treated with 1.7% NaOCl for 2 minutes. The remaining 3 dentin surfaces were acid conditioned and treated with 3.3 % NaOCl for 2 minutes. All of the specimens were then fixed in 4 % glutaraldehyde for 12 h at $4^{\circ}C$ and dehydrated in ethanols grades from 50 % to 100 %, then surface changes of the specimens were observed by using SEM. For TEM preparation, exposed dentin surfaces were acid conditioned with the same acid as SEM specimens and treated with 1.7%, 3.3 % NaOCl respectively, then applied with corresponding bonding agents. After the procedures were finished, composite resin were applied on the dentin surfaces and light cured. Small, rectangular sticks with end dimensions of approximately 1 by 1 mm were sectioned and further sample preparative techniques for transmission electron microscopy were performed in accordance with the procedures used for ultrastructural TEM observations of calcified tissues. The results were as follows : 1. In the 1.7 % NaOCl retreated specimens after acid conditioning, the porous dentin surface of intertubular dentin and wide opening of dentinal tubules were appeared. And there were fine irregularities on the intertubular dentin, indicating a clear difference as compared with the acid conditioned specimens. 2. In the 3.3% NaOCl retreated specimens after acid conditioning, the intertubular dentin was further eroded causing a more porous and wider opening of dentinal tubules. Moreover, sharp irregularities on the intertubular dentin were more evident than those of acid conditioned and 1.7% NaOCl retreated specimens. 3. In all of the acid conditioned specimens, the resin-dentin hybrid layer of approximately 3.5mm thickness was formed and the collapsed collagen layer was observed on the uppermost part of hybrid layer in the specimens applied with All Bond 2. The collgen fibrils of intertubular dentin in specimens applied with Scotchbond Multipurpose were running perpendicular to the interface, and electron dense black layer demarcated from the deep unaltered dentin was more evident in the specimen applied with Superbond D-Liner than any other specimens. 4. In the 1.7 % NaOCl retreated specimens after acid conditioning, the resin-dentin hybrid layer of approximately 2.5-3.0mm thickness was formed and the collapsed collagen layer and longitudinally running collagen fibrils as shown in the acid conditioned specimens were observed in the specimens applied with All Bond 2 and Superbond D-Liner. 5. In all of the 3.3% NaOCl retreated specimens after acid conditioning, the evidence of resin-dentin hybrid layer was not identified ; nevertheless, the longitudinally running collagen fibrils remained slightly in the specimens applied with All Bond 2.
Kim, Jin-Soo;Kim, Jeong-Gyun;Cho, Soon-Yeong;Ha, Jin-Hwan;Lee, Eung-Ho
Applied Biological Chemistry
/
v.36
no.4
/
pp.290-295
/
1993
In order to utilize effectively fish skin from fish processing, characteristics of the yellowfin sole and dover sole skins were investigated. In the yellowfin sole, the crude protein content and yield of fish skin used for the preparation of gelatin were 22.3% and 11.3%, respectively and in the dover sole, 17.2% and 8.9%, respectively. In the yellowfin sole skin, the soluble and insoluble collagen occupied 66.1% and 33.9%, respectively and in the dover sole skin, 78.8% and 21.1%, respectively. No difference in the amino acid composition between soluble and insoluble collagen was detected. The sum of proline and hydroxyproline content in the collagen extracted from fish skin was lower than that of those from pork skin or bone. The molecular weight of the two major subunits from the soluble collagen in the yellowfin sole skin were found to be 143 KDa and 202 KDa. Those in the dover sole skin were 142 KDa and 207 KDa. The physico-chemical properties such as the melting point and gelling point of yellowfin sole skin gelatin were superior to those of dover sole skin gelatin.
Purpose: Fibrillar collagens like type I collagen, are the major constituent of the extracellular matrix and structural protein of bone. Also, it can be a scaffold for osteoblast migration. The purpose of this study is to estimate the effects of absorbable atelo-collagen sponge (Teruplug$^{(R)}$, Terumo biomaterials Co., Tokyo, Japan) insertion in tooth extraction sites on periodontal healing of the second molar, healing of the fractured mandibular bone and new bone formation of third molar socket after the extraction of the impacted third molar with mandibular angle fracture. Methods: In our study of six cases of mandibular angle fractures, all of them underwent the extraction of the third molar tooth & absorbable atelo-collagen sponge insertion in tooth extraction site. Three of them had a intraoral infection & oral opening to fracture site, two of the six had dental caries, and only one had reduction problem due to third molar position. Six consecutive patients with noncomminuted fractures of the mandibular angle were treated by open reduction and internal fixation using one noncompression miniplates and screws placed through a transoral incision. Results: All of the patients have showed good postoperative functions and have not experienced complications requiring second surgical intervention. There was well healing of the mandibular bone and the most new bone formation of third molar socket after the extraction of the impacted third molar with mandibular angle fracture. Conclusion: The results of this study suggest that absorbable atelo-collagen sponge is relatively favorable bone void filler with prevention of tissue collapse, food packing, and enhance periodontal healing. Thus, the use of atelo-collagen sponge and one noncompression miniplate seems to be relatively easy, safe, and effective for the treatment of fractures of the mandibular angle and third molar extraction.
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 purpose of this study is to investigate the effects of the low power Helium Neon-Infra Red(HeNe-IR) laser and the special electromagnetic therapeutic apparatus stimulation, which is usually designated as TDP by using the initial of Tending Diancibo Pu which is the Chinese inscribed with English, on wound healing in rat. The seventy-five Sprague-Dawley adult female and male rats were assigned to the experimental and control groups. Each rat was anesthetized with pentobarbital sodium, and three full-thickness incisions with 12mm length wert made on the back of the half cf the rats and three deep second degree burns were made on the back of the remainder of the rats. From 34 hours after being injured, the rats of the experimental laser group were irradiated with the 157mW electric power HeNe-IR laser for 2 minutes every day and the rats of the experimental TDP group were stimulated with TDP irradiation with the 4km spot distance for 20 minutes every day during the 17 days. The rats were sacrificed and the wound parts of its were incised on the first day, 4th day, 7th day, 10th day and 17th day respectively after the beginning of wound treatment with laser and TDP irradiation. The incised wound parts were processed appropriately for the light microscopic and electron microscopic examination. The length of incised wound was measured with microcaliper before the wound part was incised. There was a significant decrease in the length of the incised wound of the experimental laser and TDP group, compared with that of control group at 4th day, 7th day and 10th day(p<0.01) after surgery. Through the histological examination of the wound site, the more rapid epithelialization and collagen formation in experimental groups were showed, compared with control group. The histological results were analyzed and summarized as the follows; The epidermis begins to be regenerated and the granulation tissue begins to be changed to the mature pattern in the H-E stained incised skin of the laser and TDP treatment group on the 4th day. The epidermis shows the complete regeneration and the granulation tissue in the dermis in mostly to be matured in the laser and TDP treatment group on the 7th day, compared with control group. The chronic inflammatory cells are oberved and the necrosis of the collagen fibers are partially observed in control group on the 10th day. The dermis of the laser and TOP treatment group reveals relatively compactly arranged collgen bundles with the mature collagen fibers on the 10th day. The epidermis and dennis of the laser and TDP group are repaired normally and the hair follicles are well regenerated on the 17th day. The mild edema and the granulation tissue is observed in the dermis of the control groups and the delayed treatment process is observed on the 17th day. The Most of proliferated collagen fibrils are found to be compact and regular in electron micrograph of burn skin of the laser treatment group on the 10th day hut the interstitial eadema and some inflammatory cells are found in the control group. The above results suggest that through the visual and histological examination the epithelized epithelium and the proliferation of the collagen liters in the dermis occur very effectively with the low power laser treatment and the TDP treatment in the incised wound healing and the burn wound healing.
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