This study was performed to examine the cytotoxic effects of nicotine on the seminiferous tubules and Leydig cells in mice testis. A different amount of nicotine (2~15 mg/kg, for two weeks, one dose of 100 mg/kg) were administered to four-month male mice, and then the optical microscopic findings of its effect on testis of the mice are as follows: 1. The group that 2 mg/kg of nicotine was administered showed normal findings that nucleus and cytoplasm of Leydig cells are distinct, while the other group that 5 mg/kg of nicotine was given to showed nucleus and cytoplasm are swollen and thickened a little, and slightly dyed. 2. The group that 10 mg/kg of nicotine was given had irregular arrangement of spermatogenesis inside seminiferous tubules so it was impossible to distinguish phages of seminiferous tubules. It was also impossible to observe cells due to fusion of their nucleuses, and distinct cytoplasm. 3. The group that 15 mg/kg of nicotine was administered showed destruction of nucleuses and cytoplasm of spermatocytes and sperms and a fill of fibered connective tissues so that it is impossible to observe rumens of seminiferous tubules.
The ultimate objective of periodontal treatment is to stop disease progression and to regenerate destroyed periodontal tissues and thereby regain normal function. Growth factors are naturally found polypetides which stimulate many cellular activities pertaining to wound healing by acting as signal molecule in controlling cell movement, proliferation, and matrix production. Platelet derived growth factor (PDGF) is 28,000-35,000 Da molecular weight dimeric protein with 2 long positively charged polypeptide chains connected by sulfide bonds. The purpose of this study is to evaluate histologically the initial guided tissue regeneration in a periodontal defect f a beagle dog treated with a biodegradable membrane formed with polylactic acid (poly-L-lactic acid) and polyglycolic acid loaded with 200ng/$cm^2$ platelet derived growth factor. 2 beagle dogs were used in he experiment. $5mm{\times}6mm$ alveolar bone defect was formed in upper and lower canines and third premolars and a reference notch was placed. PDGF-BB non-containing membrane was used as control. Each defect was randomly assigned to the test roup or the control group. The dogs were sacrificed 3 weeks after membrane placement. Toluidine blue and multiple staining was done for histological analysis. In the 3 week specimen in the control group, no new one formation could be seen. Small amount f bone resorption below the notch could be seen. In the notch, loose connective tissue with infiltration of inflammatory cells could be seen. Also thin discontinuous new cementum could be seen and the membrane still retained its structure. Where PDGF-BB containing membrane was used, new bone formation could be seen in the notch at weeks and also continuous thin cementum could be seen. PDL cells were observed between new bone and new cementum and some were attached to bone and cementum. These results suggest that new bone and cementum formation seen when PDGF-BB loaded membrane was used was due to inhibition of downgrowth of epithelial cells and also due to continuous release of the growth factor. Further study on the resorption characteristics of the membrane nd the release characteristics of the PDGF-BB is necessary. Also, development of a membrane easier to use clinically is necessary.
Among objectives of periodontal therapy. the principal one is the morphological and functional reconstruction of lost periodontal supporting tissues. This includes de novo formation of connective tissue attachment and the regrowth of alveolar bone. The use of enamel matrix derivative(EMD) may be a suitable means of regeneration new periodontal attachment in the infrabony defects. Implant used to replace lost tooth but, implantitis occurred after installation. The purpose of this study was to investigate the effects of EMD on differentiation and growth of osteoblast in titanium disc. Twentyfive millimeter diameter and 1mm thick Ti disc which was coated 25, 50, 100, 200${\mu}g$/ml of EMD(Emdogain(R)) used as experimental group, 25, 50, 100, 200ng/d of rhBMP-2 as positive control group, and no coat as negative control group. A human osteosarcoma cell line Saos-2 was cultured in Ti disc and cell proliferation and Alkaline phosphatase (ALP) activity were measured at 1 and 6 days. PCR was performed at 2 and 8 hours. Semi-quantitative RT-PCR for mRNA expressions of various osteoblastic differentiation markers -type I collagen, ALP, osteopontin, and bone sialoprotein - were performed at appropriate concentrations based upon the results of MTT and ALP assay. Cultured cell-disc complexes were prepared for scanning electron microscopy (SEM) at 2 hour. Data were analyzed using Mann-Whitney and repeated- measures 1-way analysis of variance(SPSS software version 10,SPSS. Chicago. IL). After culture, there was more osteoblast in EMD100${\mu}g$/ml than in EMD50, 200${\mu}g$/ml on day 6. There was significant difference in experimental and positive control group compared control group, as times go by(1 and 6 days). Alkaline phosphatase activity was different significantly in EMD100, 200${\mu}g$/ml and BMP100, 200${\mu}g$/ml on day 6. The results of reverse transcriptase-polymerase chain reaction (RT-PCR) showed that expression of mRNA for ALPase, collagen type I, osteopontin. hone sialoprotein and BMP-2 was detected at 2 hour and 8 hour in EMI 200${\mu}g$/ml subgroup and BMP100ng/ml subgroup. The results of this study suggest that application of enamel matrix derivative on osteoblast attached to titanium surface facilitate the expression of bone specific protein and the differentiation and growth of osteoblast.
Purpose : To investigate osseointergration of titanium implants into the tibia of rabbits, which were fed a low calcium diet and irradiated. Materials and Methods : To prepare the experimental model, control group was fed a normal diet and experimental group was fed a low calcium diet for 4 weeks. And then, titanium implants were inserted into the tibia of each rabbit. Experimental group was subdivided into two groups; low calcium diet/non-irradiation group and low calcium diet/irradiation group. The low calcium diet/irradiation group was irradiated with a single absorbed dose of 15 Gy at the 5th postoperative day. At 12, 19, 33, 47, and 61 days after implantation (1, 2, 4, 6, and 8 weeks after irradiation), the bone formation in the bone-implant interface area was examined by light microscopy and fluorescent microscopy. Results and Conclusions: 1. In the control group, there began to form woven bone in the bone-implant interface area at 12 days after implantation. As the experimental time was going on, the amount of bone which was in contact with the implant was increased. 2. In the low calcium diet/non-irradiation group, there began to form woven bone in the bone-implant interface area at 19 days after implantation. Although the amount of bone which was in contact with the implant was increased as the experimental time was going on, the extent of increased bone was weak as compared with control group. 3. In the low calcium diet/irradiation group, there began to form woven bone incompletely in the bone-implant interface area at 19 days after implantation, but there were vascular connective tissues in the bone-implant interface area over the entire experimental period. 4. In the control group and low calcium diet/non-irradiation group, bone labeling bands were observed at 33 days after implantation, which suggests that the bone formation and remodeling was in process, but interstitial bone remodeling was not observed in the low calcium diet/irradiation group.
An ultimate goal of periodontal therapy is to stop the disease process and to regenerate a functionally-oriented periodontium destroyed as a result of periodontal disease. The purpose of this study was to observe the effect of grafting granulation tissue obtained from extraction socket on the regeneration of horizontal furcation defect. Six dogs were used in this study. All mandibular first and third premolars were extracted. At 2, 3, and 5 days after extraction, tissues were obtained from extraction socket of 1 mongrel dog and examined by light microscope. Granulation tissue obtained at 5 days after extraction was chosen as the graft material. Five days later, horizontal furcation defects were created surgically at mandibular second and fourth premolars in the right and left side of the 5 beagle dogs. The entrance area of the artificially prepared "key hole" defects were about $3\;4mm^2$. By random selections, 2 exposed furcation defects were grafted with granulation tissue obtained from extraction socket as experimental group and 1 furcation defect was as control. The flaps were replaced to their original position and sutured with 4-0 chromic cat-gut. Three dogs were sacrificed 4 weeks and two dogs 8 weeks after surgery, and the prepared specimens were examined by light microscope. At 4 weeks, furcations were filled with epithelial lining and fibrous connective tissue infiltrated with chronic inflammatory cells. New bone formation was observed in all groups. Only experimental group showed new cementum formation. At 8 weeks, new cementum, functional arrangement of new PDL fiber, root resorption, and some ankylotic union of newly formed alveolar bone and root surface were observed in all groups. Experimental group showed that epithelial downgrowth was inhibited and new bone formation was more active compared to control. The success rate of the furcation defect healing was higher in experimental group than control. These results suggested that grafting of granulation tissue obtained from extraction socket which combined with reconstructive periodontal flap surgery may promote periodontal regeneration of horizontal furcation defect.
The purpose of this study was to investigate whether the ashed tooth powder is utilized as an alternative material of the implant to recovery the bony defect. For this purpose its biocompatibility was evaluated comparing to the synthetic calcium phosphate compounds, such as Syntograft and Calcitite, as well as the vacuum firing porcelain (Ceramco Inc.) which is anticipated to use as a matrix to aid sintering. Bony defects to exposure the bone marrow, $3{\times}5$ mm in size, were created in the right and left tibias of fifteen rabbits, and then the ashed tooth powder at $950^{\circ}C$, the porcelain powder, Syhtograft and Calcitite were inserted in the defects of twelve rabbits of the experimental group and the blood clot only was filled in the defects of three rabbits of the control group. The experimental and control rabbits were sacrificed at 1st, 2nd 3rd week after implantation and the histologic examination was performed. The ashed tooth powder in order to make the needed form of the implant was molded using the cylindrical mold 1 cm high, 1 cm in diameter under the pressure of $1000kg/cm^2$ and the ashed tooth powder was sintered at $1100^{\circ}C$ for 1 hour and the mixture of the porcelain powder and the ashed tooth powder at the weight ratio of 7:3, 6:4, 5:5, 4:6 were molded in the same manner and were sintered at $925^{\circ}C$. From this sintered material, square shaped implants were prepared in the dimension of $2{\times}4{\times}6mm$. The prepared implants were surgically placed in the subperiosteum of lateral surfaces of the right and left mandibular bodies. The dogs were sacrificed at 4 weeks, and then the specimens were examined using the light and scanning electron microscopes. The results of this study were obtained as follows: 1. Any inflammatory response was not noted after implanting of the ashed tooth powder, Syntograft, Calcitite and the porcelain powder during the whole experimental period after implantation. 2. Induction of the new bone formation was significantly shown in the ashed tooth powder, Syntograft and Calcitite. 3. The more the porcelain powder was contained in the implants, the more the porosity was and the bigger the pore size was under the scanning electron microscope. And there was ingrowing of the fibrous connective and the osteoid tissue. 4. The osteoid tissues were found to be directly fused to the implant of the ashed tooth powder, and the mixture implant of the porcelain powder and the ashed tooth powder at the weight ratio of 4:6 under the light and scanning electron microscopes.
Specific endometrial preparation should occur during periimplantation period. That is a progress of serial differentiation and is absolute in implantation of embryo and successful pregnancy. Remodeling of tissues shown during embryogenesis is regulated by various factors including extracellular matrix (ECM). Marked changes during pregnancy are including embryo migration, decidual response, and differentiation of placenta in placental animals including human. These changes to successful implantation in embryo and uterus have to prepare the competence for attachment of embryo and uterus, and invasion defense of uterus. During these changes, ECM dramatically changes for maintaining the uterine and embryonic functions. The major component of most connective tissue is collagens. It is very complex and hard to explore the mechanisms for ECM modulation. Recently using high throughput methodology, PCR-select cDNA subtraction method, microarray, many candidate genes have been identified. Steroid hormones have fundamental role in implantation and maintenance of pregnancy. Dermatopontin, a regulator of collagen accumulation, is regulated spatio-temporally in the uterus by primarily progesterone through progesterone receptors at the time of implantation. Modulation of extracellular matrix is critically regulated by cascade of gene net-works which are regulated by cascade of sex steroid hormones. Pathological regulation of uterine extracellular matrix reported in diabetic patients. To know the extracellular modulation is essential to understanding implantation, feto-placental development and overcome the paths involved in female reproduction. Though ECM composed with very various components and it is complex, the present review focused on the fate of collagens during periimplantation period.
Ipriflavone (IP), a non-hormonal isoflavone derivative, has been shown to interfere with bone remodeling by inhibiting bone resorption and stimulating bone formation. IP consistently increased the amount of Ca incorporated into the cell layer by mesenchymal stem cells (MSCs). In this study, we developed the novel IP loaded poly(L-lactide-co-glycolide) (PLGA) scaffolds for the possibility of the application of the tissue engineered bone. IP/PLGA scaffo1ds were prepared by solvent casting/salt leaching method and were characterized by porosimeter, scanning electron microscopy, determination of residual salt amount, differential scanning calorimetry, and X-ray diffractometer, respectively. IP/PLGA scaffolds were implanted into the back of athymic nude mouse to observe the effect of IP on the osteoinduction compared with control PLGA scaffo1ds. Thin sections were cut from paraffin embedded tissues and histological sections were stained H&E, von Kossa, and immunohistochemical staining for Type I collagen and osteocalcin. It can be observed that the porosity was above 91.7% and the pore size was above 101 $\mu\textrm{m}$. Control scaffo1d and IP/PLGA scaffo1ds of 50% IP were implanted on the back of athymic nude mouse to observe the effect of IP on the induction of cells proliferation for 9 weeks. The evidence of calcification, osteoblast, and osteoid from the undifferentiated stem cells in the subcutaneous sites and other soft connective tissue sites having a preponderance of stem cells has been observed. From these results, it seems that IP plays an important role for bone induction in IP/PLCA scaffolds.
Journal of the Korean Association of Oral and Maxillofacial Surgeons
/
v.31
no.5
/
pp.379-389
/
2005
Obstructive adenitis of the salivary gland following salivary stone or infectious disease of the gland and surrounding tissues is a common disease. It is often difficult to decide whether to perform total excision of the gland or to consider conservative treatment. The present study was designed to investigate histological, histochemical changes of submandibular gland after ligating the excretory duct for identifying the results of gland duct obstruction. A group of 40 rat of Sprague-Dawley weighing about $200{\sim}220gm$ were used in the present study. 30 rats had ligation of the main excretory ducts of submandibular glands just at the exit from the glands. For controls, 10 rats had a sham operation without duct ligation. They were inducted into euthanagia state by intracardial Ketamine injection in 1, 2, 3, 5, 7, 14, 21, 28, 35 and 42 days after the ligation. In each ligation period, 3 animals were used for ligation and one animal was for control. The submandibular glands were dissected out at sacrifice and stained with H&E, PAS, mucicarmine stain and histological examination were carried out under the light microscope. After examination and comparison of all specimens, the results were as follows: 1. In the features of H&E stain, acini disappeared by degrees after the ligation of the excretary duct and interstitial cells were displaced into fibrous connective tissue. Salivary gland had been atrophied with enlarging ducts and proliferating ductal cells. 2. Through total experimental period, a lot of vessels were observed and the atrophy of serous gland was severer than that of mucous gland. 3. The deep portion of submandibular glands showed severe degeneration rather than superficial portion of them after the ligation. 4. The changes which had enlarged ducts and proliferating ductal cells were observed in entire gland and more prominent in serous gland than mucous gland after the ligation. 5. Although PAS and mucicarmine reactions were decreased gradually after the ligation with the lapse of time, since 2 to 3 weeks they were strong positive reactions on entire gland, especially on duct-like structure. So, we can suggest that salivary gland will be atrophied but, survived acini will be redistributed around the ducts.
We observed the salivary ducts of two species of snails, Achatina fulica and Incilaria fruhstoferi with an electron microscope, and obtained the following results. The intralobular and interlobular ducts of Achatina fulica assume the forms of round or ellipsoidal doughnuts. The boundaries between the endothelial cells are not clear. It is also found that the cytoplasm of the endothelial cells consists of the membrane infolded in interdigital form, and there are well -developed microvilli at the apical portion of the cytoplasm. On the other hand, the intralobular and interlobular ducts of Incilaria fruhtoferi consist of the irregular simple columnar epithelia. The high electron dense cytoplasm is filled with the irregular round granules. The microvilli at the apical portion of the cytoplasm are not so well-developed as those in Achatina fulica. In the salivary duct of Achatina fulica, the lumen has narrow and long tubular structure. The boundaries between the endothelial cells are not clear. The cytoplasm is full of many vacuoles and electron lucent granules. At the apical portion of the cytoplasm, lots of short and thin microvilli are found. The salivary duct of Incilaria fruhstorferi is wider ($65\times250{\mu}m$ in diameter) than that of Achatina fulica, and consists of endothelial cells of the same structures. At the apical portion of those endothelial cells, a lot of junction apparatus such as desmosomes are observed. The vessels in the salivary ducts of Achatina fulica and Incilaria fruhstoferi are observed mainly in the connective tissues between the salivary glands. The endothelial cell of the vessel has the irregular structure and looks dark due to the high electron density. These cells protrude their filopodia and phagocytosize foreign bodies.
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