Journal of Physiology & Pathology in Korean Medicine
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v.21
no.5
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pp.1278-1284
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2007
Solanum Iyratum Thunb (Solanaceae) has multiple applications in korean traditional medicine because of its cytotoxic, immunological and anti-inflammatory activities. However, no study on the anti-arthritic activity of Solanum Iyratum Thunb has been reported in vivo. Rheumatoid arthritis (RA) is a systemic autoimmune disease with chronic inflammation characterized by hyperplasia of synovial cells in affected joints, which ultimately leads to the destruction of cartilage and bone. Cytokine production were assessed during CIA(collagen-induced arthritis) model mice in lymph node (LN), in knee joint and spleen, using ELISA. DBA/1j mice were immunized with bovine type II collagen. After a second collagen immunization, mice were treated with Solanum Iyratum Thunb (SLT) orally at 400, 200 mg/kg once a day for 4 weeks. The severity of arthritis within the knee joints was evaluated by histological assessment of cartilage destruction and pannus formation. SLT significantly suppressed the progression of CIA and inhibited the production of TNF-alpha and IFN-gamma in serum and spleen cell culture supernatant. The erosion of cartilage was dramatically reduced in mouse knees after treatment with SLT. In conclusion, our results demonstrates that SLT significantly suppressed the progression of CIA. This action was characterized by suppression of arthritis index, cartilage erosion and synovial cell infiltration and the decreased production of $TNF-{\alpha}$, $IFN-{\gamma}$, CD4+, CD19+, CD3+/CD69+ cells (in lymph node), CD11b+/Gr-1 + (in knee joint).
Bone remodeling results from the combined process of bone resorption and new bone formation which is regulated in part by some of the polypeptide growth factors such as platelet derived growth factor(PDGF), which has been known to be an important local regulator of bone cell activity and participate in normal bone remodeling. This process includes strictly regulated gene expression of several bone matrix proteins such as type I collagen and osteopontin, a 44 kDa phosphorylated glycoprotein, which has important roles in bone formation. The purpose of this study is to evaluate the effecs of PDGF-BB on the mRNA expression of bone matrix protein, type I collagen and osteopontin, in MC3T3- E1 cell culture. Cells were seeded at $5{\times}10^5$ cells in 10 ml of minimum essential medium alpha(${\alpha}-MEM$) containig 10% fetal bovine serum, 10 mM beta glycerophosphate. 0.1, 1, 10 ng/ml PDGF-BB were added to the cells for the day 3, 7, 14, 21, 28 and cultured for 24 hours. Type I collagen cDNA, Hf677, and osteopontin cDNA were used as probes for northern blot analysis. Total cellular RNA was purified at indicated day and northern blot analysis was performed. The results were as follows : Type I collagen mRNA expressions were higher at the day 3 and 7, and lower in the day 14, 21 in the control groups. In the experimental groups, mRNA expressions were increased when 0.1 ng/ml PDGF-BB were added on the day 3, 7, 21, and decreased in dose-dependent manner on the day 14, decreased at all added dose on the day 28. Osteopontin mRNA expressions were highest in the day 21 groups and lowest in the day 14 groups in the control groups. Interesting results were shown in the day 14 and 21 groups. We found that osteopontin mRNA level was increased in dose dependent manner in the day 14 groups, and decreased dose dependent manner in the day 21 groups. In conclusion, PDGF-BB may have various control effects on type I mRNA expression in the growth and differentiation process of MC3T3-E1 cells and may have contrary regulatory effects on osteopontin mRNA expression. For examples, when the baseline level of osteopontin mRNA was low, as in the day 14, PDGF-BB up-regulated osteopontin mRNA expression in dose dependent manner, and when the baseline level was high as in the day 21, PDGF-BB down-regulated dose dependent manner. Thus, it may be useful for clinical application in periodontal regeneration procedure if further study were performed.
With the development of cardiac surgical technique, we need more prosthetic materials for repairing the intra- and extracardiac defects. Although bovine pericardial bioprosthesis treated with glutaraldehyde (GA) solution is one of the most popular materials, it has a drawback of later calcific degeneration. The purpose of this study is to investigate the effectiveness of several materials and methods in reducing the calcific degeneration of bovine pericardium. Material and Method: Forty square-shaped pieces of bovine pericardia were fixed in 0.625% GA solution with 4 g/L MgCl$_2$ㆍ6$H_2O$ as a control group (group 1). Other 40 pieces pre-treated with 1 % SDS(group 2) and 40 pieces post-treated with 8% glutamate (group 3) and 2% chitosan (group 4) were also fixed in the same GA solution. Other 40 pieces pre-treated with 1% SDS and post-treated with 8% glutamate and 40 pieces post-treated with 2% chitosan were also fixed in the same GA solution (group 5, 6). The pericardial pieces were implanted into the belly of 40 Fisher 344 rats subdermally and were extracted 1 month, 2 months, 3 months, and 6 months after the implantation. With an atomic absorption spectrophotometry, we measured the calcium amount deposited and examined the tissue with microscope. Result: The calcium deposition in 1 month was less in group 2, 5, 6 than that in group 1 (p<0.05). It was most prominent in group 5 (p<0.01). This finding continued in 2 month. In 3 month, the calcium deposition was less in group 3 and 4 as well as group 2, 5, and 6 than in group 1. In 6 month, the calcium deposition in group 2, 3, 4, 5, and 6 was less than that in group 1 and the difference was more than that of 1, 2, and 6 month. The microscopic calcium deposition was also less in group 2 and 5. Calcium deposition developed in the whole layer of pericardium, beginning with the surrounding the collagen fiber and progressing inwardly. Conclusion: Pre-treatment with SDS, post-treatment with glutamate or chitosan, and SDS pre-treatment and post-treatment with glutamate or chitosan were effective in reducing the calcium deposition in bovine pericardium. Moreover, the combined method of SDS pre-treatment and glutamate post-treatment was more effective than other methods.
Kim, Jin-Young;Park, Se-A;Kang, Hyun-Mi;Kim, Eun-Su;Kim, Hae-Kwon
Development and Reproduction
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v.11
no.2
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pp.85-96
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2007
Mesenchymal stem cells (MSC) are promising candidates for cell-based therapies. One major obstacle for their clinical use is the unsafety of fetal bovine serum (FBS), which is a crucial part of all media currently used for the culture of MSC. We investigated the effect of human cord serum (HCS) on the growth response, mRNA and protein expressions of human amnion-derived stem cells (HAM). HAM were isolated from the amnion after a Caesarean section and cultured in DMEM supplemented with 10% FBS, 5% HCS or 10% HCS. During culture, their biological characteristics at earlier and later passages were analyzed using RT-PCR and immunocytochemistry. Regardless of serum sources, HAM showed the prominent expression of Oct-4, Rex-1, SCF, FGF-5, BMP-4, nestin, GATA-4, NCAM and HLA ABC genes. The expression profile was observed even at later passages. Similarly, HAM cultured in either FBS or HCS exhibited the distinct protein expression of collagen I, II, III and XII, fibronectin, $\alpha$-smooth muscle actin, vimentin, CK18, CD54, FSP, TRA-1-60, SSEA-3, -4 and HLA ABC. However, desmin expression was only observed in HAM cultured in medium supplemented with FBS and vWF expression was only found in HAM cultured in medium supplemented with HCS. Overall pattern of gene and protein expression of HAM was typical of known adult stem cells such as bone marrow-derived MSC. In conclusion, HCS could be as effective as FBS for the culture of HAM.
In the study, the protease activity of kiwifruit (Actinidia deliciosa Planch) cultivated in Korea was estimated, with specific examination of proteolytic effects on myofibrilar protein. The crude protease extract of kiwifruit was prepared in two ways; one in which the kiwifruit was homogenized with buffer followed by centrifugation, and the other were the supernatant was precipitated by saturated ammonium sulfate followed by dialysis. The former had 21.23 mM/mL of protease activity, which corresponded to 112.28 mM/g kiwifruit utilized, and the latter had 11.58 mM/mL and 45.80 mM/g of kiwifruit. The crude protease extract of the kiwifruit showed high specificity for casein substrate followed by bovine serum albumin, egg white, collagen, and elastin, in order. The enzyme lost proteolytic activity in acidic conditions such as pH 2-3, and at high temperatures over $60^{\circ}C$. It showed optimal activity in both pH 3.0 and pH 7.5 as well as at $40^{\circ}C$ for casein substrate and at $50^{\circ}C$ for myofibrilar protein substrate. The proteolytic activity toward casein was high with up to 0.5M salt, followed by a sharp decrease beyond this concentration. On the other hand the proteolytic activity for myofibrilar protein decreased steadily with increasing of salt concentration. Kiwifruit has been used as a for meat tenderizer for in home cooking and these results support the its tenderizing effectiveness of kiwifruit especially for Korean style marinating of meat for cooking.
Purpose: Deproteinized bovine bone or synthetic hydroxyapatite are 2 prevalent bone grafting materials used in the clinical treatment of peri-implant bone defects. However, the differences in bone formation among these materials remain unclear. This study evaluated osteogenesis kinetics in peri-implant defects using 2 types of deproteinized bovine bone (Bio-Oss® and Bio-Oss/Collagen®) and 2 types of synthetic hydroxyapatite (Apaceram-AX® and Refit®). We considered factors including newly generated bone volume; bone, osteoid, and material occupancy; and bone-to-implant contact. Methods: A beagle model with a mandibular defect was created by extracting the bilateral mandibular third and fourth premolars. Simultaneously, an implant was inserted into the defect, and the space between the implant and the surrounding bone walls was filled with Bio-Oss, Bio-Oss/Collagen, Apaceram-AX, Refit, or autologous bone. Micro-computed tomography and histological analyses were conducted at 3 and 6 months postoperatively (Refit and autologous bone were not included at the 6-month time point due to their rapid absorption). Results: All materials demonstrated excellent biocompatibility and osteoconductivity. At 3 months, Bio-Oss and Apaceram-AX exhibited significantly greater volumes of formation than the other materials, with Bio-Oss having a marginally higher amount. However, this outcome was reversed at 6 months, with no significant difference between the 2 materials at either time point. Apaceram-AX displayed notably slower bioresorption and the largest quantity of residual material at both time points. In contrast, Refit had significantly greater bioresorption, with complete resorption and rapid maturation involving cortical bone formation at the crest at 3 months, Refit demonstrated the highest mineralized tissue and osteoid occupancy after 3 months, albeit without statistical significance. Conclusions: Overall, the materials demonstrated varying post-implantation behaviors in vivo. Thus, in a clinical setting, both the properties of these materials and the specific conditions of the defects needing reinforcement should be considered to identify the most suitable material.
Purpose: The aim of the present study was to evaluate the effectiveness of using a mineralized bone cortical allograft (MBCA), with or without a resorbable collagenous membrane derived from bovine pericardium, on alveolar bone remodeling after immediate implant placement in a dog model. Methods: Six mongrel dogs were included. The test and control sites were randomly selected. Four biradicular premolars were extracted from the mandible. In control sites, implants without an allograft or membrane were placed immediately in the fresh extraction sockets. In the test sites, an MBCA was placed to fill the gap between the bone socket wall and implant, with or without a resorbable collagenous membrane. Specimens were collected after 1 and 3 months. The amount of residual particles and new bone quality were evaluated by histomorphometry. Results: Few residual graft particles were observed to be closely embedded in the new bone without any contact with the implant surface. The allograft combined with a resorbable collagen membrane limited the resorption of the buccal wall in height and width. The histological quality of the new bone was equivalent to that of the original bone. The MBCA improved the quality of new bone formation, with few residual particles observed at 3 months. Conclusions: The preliminary results of this animal study indicate a real benefit in obtaining new bone as well as in enhancing osseointegration due to the high resorbability of cortical allograft particles, in comparison to the results of xenografts or other biomaterials (mineralized or demineralized cancellous allografts) that have been presented in the literature. Furthermore, the use of an MBCA combined with a collagen membrane in extraction and immediate implant placement limited the extent of post-extraction resorption.
Purpose: The present study investigated the impact of 2 different suture techniques, the conventional crossed mattress suture (X suture) and the novel hidden X suture, for alveolar ridge preservation (ARP) with an open healing approach. Methods: This study was a prospective randomized controlled clinical trial. Fourteen patients requiring extraction of the maxillary or mandibular posterior teeth were enrolled and allocated into 2 groups. After extraction, demineralized bovine bone matrix mixed with 10% collagen (DBBM-C) was grafted and the socket was covered by porcine collagen membrane in a double-layer fashion. No attempt to obtain primary closure was made. The hidden X suture and conventional X suture techniques were performed in the test and control groups, respectively. Cone-beam computed tomographic (CBCT) images were taken immediately after the graft procedure and before implant surgery 4 months later. Additionally, the change in the mucogingival junction (MGJ) position was measured and was compared after extraction, after suturing, and 4 months after the operation. Results: All sites healed without any complications. Clinical evaluations showed that the MGJ line shifted to the lingual side immediately after the application of the X suture by $1.56{\pm}0.90mm$ in the control group, while the application of the hidden X suture rather pushed the MGJ line slightly to the buccal side by $0.25{\pm}0.66mm$. It was demonstrated that the amount of keratinized tissue (KT) preserved on the buccal side was significantly greater in the hidden X suture group 4 months after the procedure (P<0.05). Radiographic analysis showed that the hidden X suture had a significant effect in preserving horizontal width and minimizing vertical reduction in comparison to X suture (P<0.05). Conclusions: Our study provided clinical and radiographic verification of the efficacy of the hidden X suture in preserving the width of KT and the dimensions of the alveolar ridge after ARP.
The fruit of Terminalia chebula Retzius has been used as a panacea in India and Southeast Asia but its biological activities have not been fully elucidated. Here we report anti-arthritic and analgesic effect of NDI10218, a standardized ethanol extract of Terminalia chebula, on collagen-induced arthritis and acetic acid-induced writhing model, respectively. Arthritis was induced in DBA/1J mice by immunizing bovine type II collagen and mice were treated with NDI10218 daily for 5 weeks after the onset of the disease. NDI10218 reduced the arthritis index and blocked the synovial hyperplasia in a dose-dependent manner. The serum levels of pro-inflammatory cytokines TNF-${\alpha}$, IL-6, and IL-$1{\beta}$ were significantly reduced in mice treated with NDI10218. Production of the inflammatory IL-17, but not immunosuppressive IL-10, was also inhibited in splenocytes isolated from NDI10218-treated arthritis mice. Administration of NDI10218 markedly decreased the number of T cell subpopulations in the regional lymph nodes of the arthritis mice. Finally, NDI10218 reduced the number of abdominal contractions in acetic acid-induced writhing model, suggesting an analgesic effect of this extract. Taken together, these results suggest that NDI10218 can be a new therapeutic candidate for the treatment of rheumatoid arthritis.
Journal of the Korean Society of Laryngology, Phoniatrics and Logopedics
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v.16
no.2
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pp.129-134
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2005
Background and Objectives : Artecoll(R) is an injectable soft tissue filler, which is a suspension of polymethylmethacrylate microspheres in $3.5\%$ bovine collagen solution. The authors aimed to determine the clinical of Artecoll of Artecoll(R) as an injection material into the vocal fold to correct the glottal insufficiency caused by unilateral vocal cord paralysis. Materials and Methods : Forty-one consecutive patients with unilateral vocal cord paralysis received percutaneous Artecoll injections under local anesthesia. Acoustic, aerodynamic and stroboscopic analyses were prospectively provided before, 1 week and 3 months after injection. Perceptual GRBAS grading by speech language pathologists and subjective ratings of the hoarseness and aspiration by the patients were also obtained. Results : Aerodynamic parameter(maximal phonation time) were significantly improved after the injection (p<0.05). Acoustic parameters (jitter and shimmer) were improved at the 3rd month follow-up. GRBAS uading and patients own subjective scaling of hoarseness and aspiration also showed significant improvement (p<0.05). Early or delayed significant side effects were not observed. Conclusion : Vocal fold injection with Artecoll is a convenient, safe and useful method of temporarily correcting the glottal insufficiency. Further long-term follow-up studies will answer the usefulness and safety of the Artecoll injection laryngoplasty.
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