In order to evaluate the effect of platelet-derived growth factor(PDGF-BB) and guided tissue regeneration(GTR) technique on the regeneration of destructed periodontal tissue,intentional through-and-through furcation defects(4mm in height) were made on both mandibular 2nd and 4th premolars of 8 adult male dogs(30-40lb). Experimental group 1 was composed of the premolars that were treated by only topical application of PDGF-BB with 0.05M acetic acid without any barrier membrane. Experimental group 2 was composed of the premolars that were treated by GTR with expanded polytetrafluoroethylene membrane(ePTFE : Gore-tex periodontal material, USA). Experimental group 3 was composed of the premolars that were treated by GTR with ePTFE after topical application of PDGFBE. Control group was composed of the premolars that were treated by coronally positioned flap operation only without use of PDGF-BB and ePTFE membrane. All ePTFE membranes were carefully removed 4 weeks after regenerative surgery, and all experimental animals were sacrificed 8 weeks after regenerative surgery. The light microscopic findings were as follows ; (1) In experimental group 1, rapid new bone formation along the-root surface with multiple ankylosis and root resorption by multinucleated giant cells, and dense connective tissue in the central portion of the furcation defects were observed. (2) In experimental group 2, it was observed that the furcation defects were filled with newly formed bone, Sharpey's fibers were embedded into new cementum on root dentin of furcation fornix area, but the central portion and the area under furcation fornix were still filled with dense connective tissue. (3) In experimental group 3, the furcation defects were regenerated with newly formed dense bone and regular periodontal ligament with Sharpey's fibers embedded into newly formed cementum and bone underneath fornix area. (4) In control group, unoccupied space, apical migration of epithelium, dense infiltration of inflammatory cells in subepithelial connective tissue in relation to heavy plaque accumulation, and root resorption by inflammatory reaction were shown, but any new cementum formation on resorbed dentin surface could not be observed. The present study demonstrated that the combined therapy of PDGF-BB and GTR could enhance the regeneration of destructed periodontal tissue.
Leukemia stem cells (LSCs) play important roles in leukemia initiation, progression and relapse, and thus represent a critical target for therapeutic intervention. Hence, it is extremely urgent to explore new therapeutic strategies directly targeting LSCs for acute myelogenous leukemia (AML) therapy. We show here that Angelica sinensis polysaccharide (ASP), a major active component in Dong quai (Chinese Angelica sinensis), effectively inhibited human AML $CD34^+CD38^-$ cell proliferation in vitro culture in a dose-dependent manner while sparing normal hematopoietic stem and progenitor cells at physiologically achievable concentrations. Furthermore, ASP exerted cytotoxic effects on AML K562 cells, especially LSC-enriched $CD34^+CD38^-$ cells. Colony formation assays further showed that ASP significantly suppressed the formation of colonies derived from AML $CD34^+CD38^-$ cells but not those from normal $CD34^+CD38^-$ cells. Examination of the underlying mechanisms revealed that ASP induced $CD34^+CD38^-$ cell senescence, which was strongly associated with a series of characteristic events, including up-regulation of p53, p16, p21, and Rb genes and changes of related cell cycle regulation proteins P16, P21, cyclin E and CDK4, telomere end attrition as well as repression of telomerase activity. On the basis of these findings, we propose that ASP represents a potentially important agent for leukemia stem cell-targeted therapy.
Lee, Won Jai;Lee, Dong Won;Hur, Jae Young;Lee, Young Dae;Park, Beyoung Yun;Rah, Dong Kyun
Archives of Plastic Surgery
/
v.35
no.2
/
pp.127-133
/
2008
Purpose: The purpose of this study is to evaluate the remodeling process of the various skin substitutes in 4th and 6th weeks following the transplantation when transplanted onto nude mice. Methods: Three types of artificial skin substitutes, such as PLGA scaffold with keratinocyte sheets(group 1), acellular human dermis($Surederm^{(TM)}$) and keratinocyte sheet(group 2), bioengineered skin($Neoderm^{(TM)}$)(group 3), were applied to the wound on nude mice. All mice were killed in 2, 4 weeks and/or 6 weeks after grafting and tissue samples were harvested from the back of mice. The changes in wound size, degree of angiogenesis, formation of basement membrane and epidermis, density of collagen fibers and neural restoration were examined. Results: There was no significant changes in wound size among the three groups. However, the size of wound decreased in the non-substituted group due to contracture. Degree of angiogenesis and systhesis of collagen or neurofilaments were mostly increased in bioengineered skin($Neoderm^{(TM)}$)(group 3), followed by acellular human dermis($Surederm^{(TM)}$) and keratinocyte sheet(group 2), PLGA scaffold with keratinocyte sheets (group 1). However, group 3 and group 2 showed similar thickness of basement membrane and epidermis. Conclusion: We found that degree of angiogenesis, formation of basement membrane and skin appendages, density of collagen fibers and neurofilaments can be the categories to evaluate the success of artificial skin substitution in early stages.
Xiang Jin;Jin-Young Park;Jung-Seok Lee;Ui-Won Jung;Seong-Ho Choi;Jae-Kook Cha
Journal of Periodontal and Implant Science
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v.53
no.3
/
pp.207-217
/
2023
Purpose: Non-crosslinked and crosslinked collagen membranes are known to exhibit distinct degradation characteristics, resulting in contrasting orientations of the adjacent tissues and different biological processes. The aim of this study was to conduct a histomorphometric assessment of non-crosslinked and crosslinked collagen membranes regarding neovascularization, tissue integration, tissue encapsulation, and biodegradation. Methods: Guided bone regeneration was performed using either a non-crosslinked (BG) or a crosslinked collagen membrane (CM) in 15 beagle dogs, which were euthanized at 4, 8, and 16 weeks (n=5 each) for histomorphometric analysis. The samples were assessed regarding neovascularization, tissue integration, encapsulation, the remaining membrane area, and pseudoperiosteum formation. The BG and CM groups were compared at different time periods using nonparametric statistical methods. Results: The remaining membrane area of CM was significantly greater than that of BG at 16 weeks; however, there were no significant differences at 4 and 8 weeks. Conversely, the neovascularization score for CM was significantly less than that for BG at 16 weeks. BG exhibited significantly greater tissue integration and encapsulation scores than CM at all time periods, apart from encapsulation at 16 weeks. Pseudoperiosteum formation was observed in the BG group at 16 weeks. Conclusions: Although BG membranes were more rapidly biodegraded than CM membranes, they were gradually replaced by connective tissue with complete integration and maturation of the surrounding tissues to form dense periosteum-like connective tissue. Further studies need to be performed to validate the barrier effect of the pseudoperiosteum.
Composites of hydroxyapatite (HAp) and chitosan (CS) have been successfully used in bone healing in humans and animals. However, the characteristics of HAp and CS are different. Therefore, the effects of HAp/CS composites on canine bone formation could differ according to their ratio. This study investigated the therapeutic effects of different contents ratios (100, 80:20, 60:40 wt%) on bone defects in a canine model. Thirty intrabony cylindrical defects were created in the humeruses and femurs of 5 beagle dogs, and then the defects were implanted with different composites. The evaluations were performed using radiographs obtained at 10 weeks post-surgery and by histological findings. In radiographic evaluation including the grades of bone filling, periosteal and endosteal reactions, pure hydroxyapatite composite had a significant effect on bone filling, and chitosan containing the composites showed vigorous responses at the periosteum and endosteum. In histological findings, the defect implanted with pure hydroxyapatite had healed completely into mature bony tissue with an obvious osteon structure, and the defect implanted with chitosan containing the composites had the amount of fibrous connective tissue increased significantly within the cortical bone tissue. The results indicate that hydroxyapatite/chitosan composites are therapeutically useful, promoting effective bone healing in defects when the ratio of hydroxyapatite is high and enhanced fibrous connective tissue formation at the periosteum as more chitosan is added.
The demonstrations of collagenous fiber in mandibul ar bone formation were analized histochemically, using Carson's Mallary-Heidenhein stain, McManus' PAS reaction and H-E stain The stain ablity of osteoid increased with formation of bone tissue. In the surrounding areas of osteoblast and osteocyte were showed slight activity.
The purpose of this study was to evaluate new bone formation following guided bone regeneration by resorbable and nonresorbable membrane. Six adult mongrel dogs were used. The first, second, third, fourth premolars in the mandible of each dog were extracted. Two months after tooth extraction, a buccal dehiscence defect was surgically created on each edentulous area. The experimental sites were divided into three groups according to the treatment modalities ; Group I-a: surgical treatment only ; Group I -b: allogenic decalcified freezed dried bone grafting ; Group II-a : e- PTFE membrane placement only ; Group II-b : allogenic decalcified freezed dried bone grafting and e-PTFE membrane placement ; Group III-a : Vicryl(R) mesh placement only ; Group III-b : allogenic decalcified freezed dried bone grafting and Vicryl(R) mesh placement . The animals were sacrificed at 8 weeks after operation and the specimens were prepared for histologic and histometric examination. The results were as follows : Clinically, all defect sites were healed without exposure of barrier membrane after the eight weeks. In Group I-a, dense connective tissues were impinged in the bony defect area. Well vascularized and fibrous bone marrow indicated that bone formation was still taking place was found. In Group I-b, in areas closer to the periphery, lamellation of the newly formed bone would found. In Group II-a, beneath the e-PTFE membrane a dense layer of connective tissue covering the most external portions of the regenerated tissue was seen. The new bone surfaces were lined with osteoid and osteoblast. In Group II-b, a dense layer of connective tissue covering the most external portions of the regenerated tissue was observed beneath the e-PTFE membrane. A notable amount of alveolar ridge regeneration was seen with new rigdes with well-contoured form. In Group III-a, the new bone surface were lined with osteoid and osteoblast, indicating active bone formation. A clear demarcation could not be noted between the host bone and new bone. In Group III-b, a notable amount of alveolar ridge regeneration was seen with new ridges assuming wellcontoured form. In areas closer to the periphery, lamellation of the newly formed bone would found. As histometric examination, the amount of bone formation was gained from $12.8mm^2$ to $26.3mm^2$. It was significantly greater in group II-b and group III-b compared to other groups(p<0.05) . These results suggest that Vicryl(R) mesh after DFDB grafting used in guided bone regeneration could create and sustain sufficient space for new bone formation.
Arterial thrombosis and its associated diseases are considered to constitute a major healthcare problem. Arterial thrombosis, defined as blood clot formation in an artery that interrupts blood circulation, is associated with many cardiovascular diseases. Oxidative stress is one of many important factors that aggravates the pathophysiological process of arterial thrombosis. Apurinic/apyrimidinic endonuclease 1/redox factor-1 (Ref-1) has a multifunctional role in cells that includes the regulation of oxidative stress and anti-inflammatory function. The aim of this study was to investigate the therapeutic effect of adenovirus-mediated Ref-1 overexpression on arterial thrombosis induced by 60% FeCl3 solution in rats. Blood flow was measured to detect the time to occlusion, thrombus formation was detected by hematoxylin and eosin staining, reactive oxygen species (ROS) levels were detected by high-performance liquid chromatography, and the expression of tissue factor and other proteins was detected by Western blot. FeCl3 aggravated thrombus formation in carotid arteries and reduced the time to artery occlusion. Ref-1 significantly delayed arterial obstruction via the inhibition of thrombus formation, especially by downregulating tissue factor expression through the Akt-GSK3β-NF-κB signaling pathway. Ref1 also reduced the expression of vascular inflammation markers ICAM-1 and VCAM-1, and reduced the level of ROS that contributed to thrombus formation. The results showed that adenovirus-mediated Ref-1 overexpression reduced thrombus formation in the rat carotid artery. In summary, Ref-1 overexpression had anti-thrombotic effects in a carotid artery thrombosis model and could be a target for the treatment of arterial thrombosis.
In this study, we intend to develop a granulation tissue formation model. As a pilot experiment, a contrast agent was injected into the pylorus in 3 rats, the normal pylorus lumen size was confirmed, and a stent was placed. Stent migration was confirmed in to the duodenum within 1 week. In this experiment, stent was sutured and fixed to induce granulation tissue formation after gastrostomy under a fluoroscopic guidance. Twenty rats were divided into Healthy Group / Gastrostomy Group. After anesthesia of the Gastrostomy Group, an abdominal incision was performed, and gastrostomy was performed under a fluoroscopic guidance, and a stent was placed into the pylorus. In order to prevent stent migration due to peristalsis, suture between the pylorus and the proximal end of the stent was performed. Postoperative behavior and weight changes were monitored daily. Four weeks after surgery, gastrointestinal fluoroscopy imaging was performed and rats were sacrifices. To evaluate the degree of granulation formation, the stent was sectioned transversely. Gastrostomy group was statistically significantly higher than Healthy Group in granulation area ratio (all p<.001). In conclusion, it is considered that the level of tissue overgrowth formation for preclinical evaluation of the pylorus stricture model through gastrostomy is appropriate as a research evaluation tool.
The present study was aimed to evaluate the effect of demineralized freeze-dried bone (DFDB) and resorbable hydroxyapatite (RHA) on bone formation in the extracted socket. The lower left and right 2nd and 3rd premolar were extracted in adult dogs. The one group was grafted with DFDB into the extracted socket, and the other group grafted with RHA. The extracted socket was sutured without any graft materials as control. The animals were killed on the 1st, 2nd, 4th, and 8th week after the graft for macroscopic and microscopic examination. Results obtained were as follows : 1. Macroscopically, nor infection of the graft site and dislodgement of the grafted material were noted in any animals used. 2. Young trabeculae of osteoid were formed in the socket wall in control group at 2 weeks after the graft. Osteoid tissue was formed in DFDB group at 1 week after graft, and a fine osteoid tissue was grown through the RHA particles in RHA group at 2 weeks graft. 3. The grafted groups showed more rapid bone formation than the control. Between the grafted groups, DFDB group showed more rapid formation than RHA group, DFDB group showed osteoinductive bone formation and RHA group showed osteoconductive bone formation. These results suggest that DFDB and RHA are useful to preserve the alveolar bone and to improve new bone formation by immediate grafting into the extraction sockets.
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