The aim of our study is to achieve complete periodontal tissue regeneration by the application of BMP and resorbable membrane. Three beagle dogs aged over one and half years and weighed 14 to 16 kg were used in this study. Mandibular 1st, 2nd premolars were extracted bilaterally. Horizontal furcation defects were induced around 3rd, 4th premolars bilaterally. BMP-4 were applied in the right side with resorbable membranes and only resorbable membranes were applied in the left side respectively. Each animal was sacrificed at 2, 4, and 8weeks, after regenerative surgery. Specimens were prepared with Hematoxylin-Eosin stain and Goldner's modified Masson Trichrome stain for light microscopic evaluation. The results were as follows: 1. At 2 weeks after regenerative surgery, downgrowth of junctional epithelium was observed both in the membraneapplied site and BMP-4-and-membrane-applied site. 2. At 4 weeks after regenerative surgery, resorbable membranes were completely resolved, therefore would not prevent downgrowth of junctional epithelium. New bone formation, new cementum formation and Sharpey's fiber were observed in BMP-4-andmembrane-applied site. 3. At 8 weeks after regenerative surgery, downgrowth of junctional epithelium was observed in the membrane-applied site. But, new cementum formation was observed in the same site. The extensive regeneration of new bone, new cementum and remarkable formation of Shapey's fiber were showed in BMP-4-and-membrane-applied site. 4. Resorbable membranes were resolved via the cell-mediated processes. 5. Periodontal tissue regeneration were better achieved in the BMP-4-andmembrane-applied site than in the membrane-applied site. Within the above results, BMP-4 may have the strong capability to form the new bone and resorbable membrane may be able to prevent the bony ankylosis. However, resolution rate of resorbable membrane may not be enough to protect rapid epithelial downgrowth for ideal periodontal regeneration. In conclusion, I suggest BMP-4 may have the strong possibility to be utilized in the clinical periodontal treat-
Objectives : This study investigated anti-inflammatory effects of Nardotidis seu Sulculii Concha water extract (NSCE) against restraint-induced stress. Methods : In vivo, NSCE was orally administered to male white mice at concentrations of 250 mg/kg and 500 mg/kg for 3 days, and then restraint-induced stress was induced for 6 hours. The level of liver damage was measured by serum aspartate aminotransferase (AST), alanine aminotransferase (ALT) and lactate dehydrogenase (LDH). The stress-related hormones such as cortisol and corticosterone were measured by ELISA assay. Also, western blot analysis was performed to detect expression of mitogen-activated protein kinase (MAPK) and cyclooxygenase-2 (COX-2) proteins. Pathological changes were observed by hematoxylin and eosin (H&E) staining of the liver tissue, and Immunohistochemical (IHC) staining was performed to examine liver inflammation through macrophage infiltration. Results : The AST, ALT, LDH and the stress related hormones such as cortisol and corticosterone were significantly decreased in the NSCE treated group compared with stress group. In histological analysis, H&E staining of liver tissues did not detect the hepatic injury or damage in all groups. As a result of IHC staining, it was confirmed that infiltration of macrophages was increased in the stress-induced group, but decreased in the group treated with NSCE. The COX-2 and MAPK proteins expression was significantly increased by restraint-induced stress, but these proteins were decreased in the NSCE treated group. Conclusions : These results suggest that NSCE has the anti-inflammatory activity in restraint-induced stress model, and it is believed that NSCE can be used for the prevention of liver inflammation.
Objective : This study was undertaken to investigate effects of Opuntia ficus-indica (OP)extract on an asthma murine model. Methods : The total number of cells and eosinophils in BALF and the infiltration of inflammatory cells into lung tissues were determined by hematoxylin and eosin staining. Results : The number of OVA-induced total cells and eosinophils, a phenomenon of asthma, were decreased by treatment of the animals with OP extract (200 mg/kg) (respectively, p < 0.001 and p<0.01). Furthermore, we showed that OP extract reduced the increased immune cell infiltration induced by ovalbumin (p < 0.01). The levels of interleukin (IL)-4 in BAL fluid were measured by enzyme-linked immunosorbent assay, because the eosinophilia is associated with a T helper (Th) 2 response including IL-4. The level of OVA-induced IL-4 was decreased by OP extract in BAL fluid (p < 0.05). We investigated whether OP extract reduced nitrite (NO) production on lipopolysaccharide (LPS)-stimulated RAW 264.7 sells, because asthma is an inflammatory disease. The level of LPS-induced NO production was decreased by OP extract (50, 100 and 200 mg/ml) on RAW 264.7 cells (respectively, p < 0.05, p<0.01 and p<0.05). Conclusions : Our results indicate that OP extract has an inhibitory effect on lung eosinophilia of asthma and suggest that OP extract is a therapeutic candidate in the treatment of inflammatory disease, including asthma.
Objectives: Cornu cervi pantotrichum (CCP) has been widely used in Korean and China, as an anti-fatigue, anti-aging, and tonic agent to enhance the functions of the reproductive and the immune systems. Because CCP has various growth factors that play important roles in the development of hair follicles, we examined whether CCP pharmacopuncture solution (CCPPS) was capable of promoting hair growth in an animal model. Methods: One day after hair depilation, CCPPS were topically applied to the dorsal skin of C57BL/6 mice once a day for 15 days. Hair growth activity was evaluated by using macro- and microscopic observations. Dorsal skin tissues were stained with hematoxylin and eosin. Expressions of bromodeoxyuridine (BrdU), proliferating cell nuclear antigen (PCNA), and fibroblast growth factor (FGF)-7 were examined by using immunohistochemical staining. A reverse transcription polymerase chain reaction (RT-PCR) analysis was also conducted to measure the messenger RNA (mRNA) expression of FGF-7. Results: CCPPS induced more active hair growth than normal saline. Histologic analysis showed enlargement of the dermal papilla, elongation of the hair shaft, and expansion of hair thickness in CCPPS treated mice, indicating that CCPPS effectively induced the development of anagen. CCPPS treatment markedly increased the expressions of BrdU and PCNA in the hair follicles of C57BL/6 mice. In addition, CCPPS up regulated the expression of FGF-7, which plays an important role in the development of hair follicles. Conclusion: These results reveal that CCPPS facilitates hair re-growth by proliferation of hair follicular cells and up-regulation of FGF-7 and suggest that CCPPS can potentially be applied as an alternative treatment for patients with alopecia.
We elucidated the distribution of interstitial cells of Cajal (ICC) in human stomach, using cryosection and $c-Kit$ immunohistochemistry to identify $c-Kit$ positive ICC. Before $c-Kit$ staining, we routinely used hematoxylin and eosin (HE) staining to identify every structure of human stomach, from mucosa to longitudinal muscle. HE staining revealed that the fundus greater curvature (GC) had prominent oblique muscle layer, and $c-Kit$ immunostaining $c-Kit$ positive ICC cells were found to have typical morphology of dense fusiform cell body with multiple processes protruding from the central cell body. In particular, we could observe dense processes and ramifications of ICC in myenteric area and longitudinal muscle layer of corpus GC. Interestingly, $c-Kit$ positive ICC-like cells which had morphology very similar to ICC were found in gastric mucosa. We could not find any significant difference in the distribution of ICC between fundus and corpus, except for submucosa where the density of ICC was much higher in gastric fundus than corpus. Furthermore, there was no significant difference in the density of ICC between each area of fundus and corpus, except for muscularis mucosa. Finally, we also found similar distribution of ICC in normal and cancerous tissue obtained from a patient who underwent pancreotomy and gastrectomy. In conclusion, ICC was found ubiquitously in human stomach and the density of ICC was significantly lower in the muscularis mucosa of both fundus/corpus and higher in the submucosa of gastric fundus than corpus.
Purpose: In this study, we investigated the effects of Cirsium setidens ethanolic extract (CS) on the development of alcoholic fatty liver and associated injury. Methods: Sprague-Dawley male rats were fed either Lieber-DeCarli control (C) or ethanol (35.5% of total calories) liquid diet with 0 (E), 100 mg/kgBW CS (E+LCS), or 500 mg/kgBW CS (E+HCS) for 8 weeks. Serum alanine aminotransferase (ALT) and aspartate aminotransferase (AST) activities as well as TG and cholesterol concentrations in the serum and liver tissues were measured by colorimetric assays. Liver histopathology was examined by Hematoxylin-eosin staining of the fixed liver tissues. Protein levels of phosphorylated-AMP activated protein kinase (p-AMPK), phosphorylated-acetyl CoA carboxylase (p-ACC), phosphorylated-nuclear factor kappa B (p-$NF{\kappa}B$), and $TNF{\alpha}$ were measured by Western blot analyses. Results: Both doses of CS markedly suppressed alcohol-induced lipid droplets accumulation in the liver tissues and significantly inhibited alcohol-induced increases in activities of serum ALT and serum AST. Similarly, CS significantly reduced hepatic and serum TG concentrations. Compared to groups fed alcohol only, CS supplementation strongly increased hepatic levels of p-AMPK and p-ACC. Further, CS significantly inhibited alcohol-induced phosphorylation of $NF{\kappa}B$, which was associated with reduced hepatic protein levels of $TNF{\alpha}$. Conclusion: Our data demonstrated that CS has a protective effect against alcoholic liver injury, which was associated with activation of AMPK and inhibition of $NF{\kappa}B$.
On the basis of the evidence that electrical stimulation could promote healing and regeneration of bone, this study was performed to investigate the effects of electrical stimulation on rat extraction socket, and to evaluate the potential of clinical application of electrical stimulation. Forty rats were used and divided into control groups(l0)and the experimental groups(30) in this study. The maxillary 1st molar were extracted in both groups. In experimental group, electrical stimulation was given at the current intensity of lmA(Test-1), l0mA(Test-2), 25mA(Test-3) each day. At 1,3,5,7 days after the tooth extraction, rats in both groups were serially sacrificed. And the specimens were prepared with Hematoxylin-Eosin stain for the light microscopic evaluation. The results of this study were as follows; 1. At 1 day after the extraction, the periodontal ligament was found in the extraction socket wall. The formation of blood clot with dense infiltration of inflammatory cells in control group and there were less inflammatory cells in test group. 2. At 3 day after the extraction, the cells and collagen of the periodontal ligament were so actively proliferated and synthesized that invaded into the connective tissue of the extraction sockets in the control group. There were the formation of new bone in the basal & lateral portion of socket wall in test -2 and -3. 3. At 5 days after the extraction, there were no formation of new bone in control group. But the more electrical stimulation was applied, the more formation of new bone in test group. 4. At 7 days after the extraction, the extraction sockets were almost filled with trabecular bone in each group. Bone maturarity was remarkable in test-3. 5. The electrical stimulation at l0mA and 25mA was more effective in the bone formation at 5 and 7 days after the extraction. From the above results, electrical stimulation could promote the extraction socket wound healing, and be utilized in the clinical application of the residual ridge expansion.
Endothelial cells are a vital constituent of most mammalian organs and are required to maintain the integrity of these tissues. These cells also play a major role in angiogenesis, inflammatory reactions, and in the regulation of thrombosis. Angiogenesis facilitates pulp formation and produces the vessels which are essential for the maintenance of tooth homeostasis. These vessels can also be used in bone and tissue regeneration, and in surgical procedures to place implants or to remove cancerous tissue. Furthermore, endothelial cell regeneration is the most critical component of the tooth generation process. The aim of the present study was to stimulate endothelial regeneration at a site of acute cyclophosphamide (CP)-induced endothelial injury by treatment with human umbilical cord-derived endothelial/mesenchymal stem cells (hEPCs). We randomly assigned 16 to 20-week-old female NOD/SCID mice into three separate groups, a hEPC ($1{\times}10^5$ cells) transplanted, 300mg/kg CP treated and saline (control) group. The mice were sacrificed on days 5 and 10 and blood was collected via the abdominal aorta for analysis. The alanine transaminase (ALT), aspartate aminotransferase (AST), serum alkaline phosphatase (s-ALP), and albumin (ALB) levels were then evaluated. Tissue sections from the livers and kidneys were stained with hematoxylin and eosin (HE) for microscopic analysis and were subjected to immunohistochemistry to evaluate any changes in the endothelial layer. CP treatment caused a weight reduction after one day. The kidney/body weight ratio increased in the hEPC treated animals compared with the CP only group at 10 days. Moreover, hEPC treatment resulted in reduced s-ALP, AST, ALT levels compared with the CP only group at 10 days. The CP only animals further showed endothelial injuries at five days which were recovered by hEPC treatment at 10 days. The number of CD31-positive cells was increased by hEPC treatment at both 5 and 10 days. In conclusion, the CP-induced disruption of endothelial cells is recovered by hEPC treatment, indicating that hEPC transplantation has potential benefits in the treatment of endothelial damage.
Dong-Hun Lee;Eun Chae Lee;Sang-Won Park;Ji young Lee;Kee-Pyo Kim;Jae Sang Oh
Journal of Korean Neurosurgical Society
/
v.67
no.3
/
pp.333-344
/
2024
Objective : Markers of neuroinflammation during ischemic stroke are well characterized, but additional markers of neural damage are lacking. The study identified associations of behavioral disorders after stroke with histologic neural damage and molecular biological change. Methods : Eight-week-old, 25 g male mice of the C57BL/6J strain were subjected to middle cerebral artery occlusion (MCAO) to induce ischemic stroke. The control group was a healthy wild type (WT), and the experimental group were designed as a low severity MCAO1 and a high severity MCAO2 based on post-stroke neurological scoring. All groups underwent behavioral tests, realtime polymerase chain reaction, triphenyltetrazolium chloride (TTC) staining and Hematoxylin and Eosin staining. One-way analysis of variance was used to analyze statistical significance between groups. Results : In TTC staining, MCAO1 showed 29.02% and MCAO2 showed 38.94% infarct volume (p<0.0001). The pro-inflammatory cytokine interleukin (IL)-1β was most highly expressed in MCAO2 (WT 0.44 vs. MCAO1 2.69 vs. MCAO2 5.02, p<0.0001). From the distance to target in the Barnes maze test, WT had a distance of 178 cm, MCAO1 had a distance of 276 cm, and MCAO2 had a distance of 1051 (p=0.0015). The latency to target was 13.3 seconds for WT, 27.9 seconds for MCAO1, and 87.9 seconds for MCAO2 (p=0.0007). Prospero homeobox 1 (Prox1) was most highly expressed in MCAO2 (p=0.0004). Doublecortin (Dcx) was most highly expressed in MCAO2 (p<0.0001). Conclusion : The study demonstrated that histological damage to neural cells and changes in brain mRNA expression were associated with behavioral impairment after ischemic stroke. Prox1 and Dcx may be biomarkers of neural damage associated with long-term cognitive decline, and increased expression at the mRNA level was consistent with neural damage and long-term cognitive dysfunction.
Objective: Orthodontic root resorption (ORR) due to orthodontic tooth movement is a difficult treatment-related adverse event. Caspases are important effector molecules for apoptosis. At present, little is known about the mechanisms underlying ORR and apoptosis in the cementum. The aim of the present in vivo study was to investigate the expression of tartrate-resistant acid phosphatase (TRAP), caspase 3, caspase 8, and receptor activator of nuclear factor kappa-B ligand (RANKL) in the cementum in response to a heavy or an optimum orthodontic force. Methods: The maxillary molars of male Wistar rats were subjected to an orthodontic force of 10 g or 50 g using a closed coil spring. The rats were sacrificed each experimental period on days 1, 3, 5, and 7 after orthodontic force application. And the rats were subjected to histopathological and immunohistochemical analyses. Results: On day 7 for the 50-g group, hematoxylin and eosin staining revealed numerous root resorption lacunae with odontoclasts on the root, while immunohistochemistry showed increased TRAP- and RANKL-positive cells. Caspase 3- and caspase 8-positive cells were increased on the cementum surfaces in the 50-g group on days 3 and 5. Moreover, the number of caspase 3- and caspase 8-positive cells and RANKL-positive cells was significantly higher in the 50-g group than in the 10-g group. Conclusions: In our rat model, ORR occurred after apoptosis was induced in the cementum by a heavy orthodontic force. These findings suggest that apoptosis of cementoblasts is involved in ORR.
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