Purpose: It has been reported that low-level semiconductor diode lasers could enhance the wound healing process. The periodontal ligament is crucial for maintaining the tooth and surrounding tissues in periodontal wound healing. While low-level semiconductor diode lasers have been used in low-level laser therapy, there have been few reports on their effects on periodontal ligament fibroblasts (PDLFs). We performed this study to investigate the biological effects of semiconductor diode lasers on human PDLFs. Methods: Human PDLFs were cultured and irradiated with a gallium-aluminum-arsenate (GaAlAs) semiconductor diode laser of which the wavelength was 810 nm. The power output was fixed at 500 mW in the continuous wave mode with various energy fluencies, which were 1.97, 3.94, and 5.91 $J/cm^2$. A culture of PDLFs without laser irradiation was regarded as a control. Then, cells were additionally incubated in 72 hours for MTS assay and an alkaline phosphatase (ALPase) activity test. At 48 hours post-laser irradiation, western blot analysis was performed to determine extracellular signal-regulated kinase (ERK) activity. ANOVA was used to assess the significance level of the differences among groups (P<0.05). Results: At all energy fluencies of laser irradiation, PDLFs proliferation gradually increased for 72 hours without any significant differences compared with the control over the entire period taken together. However, an increment of cell proliferation significantly greater than in the control occurred between 24 and 48 hours at laser irradiation settings of 1.97 and 3.94 $J/cm^2$ (P<0.05). The highest ALPase activity was found at 48 and 72 hours post-laser irradiation with 3.94 $J/cm^2$ energy fluency (P<0.05). The phosphorylated ERK level was more prominent at 3.94 $J/cm^2$ energy fluency than in the control. Conclusions: The present study demonstrated that the GaAlAs semiconductor diode laser promoted proliferation and differentiation of human PDLFs.
The purpose of this study was to evaluate histologically the effect of LiF-maleic acid added calcium aluminate(LM-CA) bone cement & CA-PMMA composite bone cement on the healing of calvarial defect in Sprague-Dawley rats. The critical size defects were surgically produced in the calvarial bone using the 8mm trephine bur. The rats were divided in three groups : In the control group, nothing was applied into the defect of each rat. LM-CA bone cement was implanted in the experimental group 1 and CA-PMMA composite bone cement was implanted in the experimental group 2. Rats were sacrificed at 2, 8 weeks after surgical procedure. The specimens were examined by histologic analysis, especially about the bone-cement interface and the response of surrounding tissue. The results are as follows; 1. In the control group, inflammatory infiltration was observed at 2 weeks. At 8 weeks, periosteum and duramater were continuously joined together in the defect area. But the center of defect area was filled up with the loose connective tissue. 2. In the experimental group 1, the bonding between implanted bone cement and the existing bone was seen, which more increased in 8 weeks than 2 weeks. Inflammatory infiltration and the dispersion of implanted bone cement particles were seen in both 2 weeks and 8 weeks. 3. In the experimental group 2, implanted bone itself had a dimensional stability and no bonding between implanted bone cement and the existing bone was seen in both 2 weeks and 8 weeks. Implanted bone cement was encapsulated by fibrous connective tissue. In addition, inflammatory infiltration was seen around implanted bone cement. On the basis of these results, when LM-CA bone cement or CA-PMMA composite bone cement was implanted in rat calvarial defect, LM-CA bone cement can be used as a bioactive bone graft material due to ability of bonding to the existing bone and CA-PMMA can be used as a graft material for augmentation of bone-volume due to dimensional stability.
Purpose: The carrier used as delivery agent for bone morphogenetic proteins(BMPs) should also act as a scaffold for new bone formation. Moreover, bone formation should be predictable in terms of the volume and shape. This study examined the osteogenic effect of macroporous biphasic calcium phosphate (MBCP) block combined with ePTFE membrane as a carrier for recombinant human bone morphogenetic proteins (rhBMP-2). In addition, the additive effect of ePTFE membrane on bone formation was evaluated. Materials and Methods: Eight-millimeter critical sized calvarial defects were created surgically in 28 male Sprague-Dawley rats. The animals were divided into 2 groups containing 14 animals each. The defects were treated with either rhBMP-2/MBCP block (rhBMP-2/MBCP group) or rhBMP-2/MBCP block/ePTFE membrane (rhBMP-2/MBCP/ePTFE group). A disc-shaped MBCP block (3 mm height and 8 mm diameter) was used as the carrier for the rhBMP-2 and ePTFE membrane was used to cover the rhBMP-2/MBCP block. The histologic and histometric parameters were used to evaluate the defects after 2- or 8-week healing period (7 animals/group/healing interval). Results: The level of bone formation in the defects of both groups was significantly higher at 8 weeks than that at 2 weeks (P < 0.05). The ePTFE membrane has no additional effect compared with the rhBMP-2/MBCP block only. However, at 8 weeks, rhBMP-2/MBCP/ePTFE group showed more even bone formation on the top of the MBCP block than the rhBMP-2/MBCP group. Conclusion: These results suggest that the ePTFE membrane has no additive effect on bone formation when a MBCP block is used as a carrier for rhBMP-2.
The major goals of periodontal therapy are the functional regeneration of periodontal supporting structures already destructed by periodontal disease as well as the reduction of signs and symptoms of progressive periodontal disease. There have been many efforts to develop materials and therapeutic methods to promote periodontal wound healing. There have been increasing interest on the chitosan made by chtin. Chitosan is a derivative of chitin made by deacetylation of side chains. Chitosan has been widely studied as bone substitution and membrane material in periodontology. Many experiments using chitosan in various animal models have proven its beneficial effects. Tetracycline has been considered for use in the treatment of chronic periodontal disease and gingivitis. The aim of this study is to evlauate the osteogenesis of tetracycline blended chitosan membranes on the calvarial critical size defect in Sprague Dawley rats. An 8mm surgical defect was produced with a trephine bur in the area of the midsagittal suture. The rats were divided into five groups: Untreated control group versus four experimental group. Four types of membranes were made and comparative study was been done. Two types of non-woven membranes were made by immersing non-woven chitosan into either the tetracycline solution or chitosan-tetracycline solution. Other two types of sponge membranes were fabricated by immersing chitosan sponge into the tetracycline solution, and subsequent freeze-drying. The animals were sacrificed at 2 and 8 weeks after surgical procedure. The specimens were examined by histologic analyses. The results are as follows: 1. Clinically the use of tetracycline blended chitosan membrane showed great healing capacity. 2. The new bone formations of all the experimental group, non-woven and sponge type membranes were greater than those of control group. But, there was no significant difference between the experimental groups. 3. Resorption of chitosan membranes were not shown in any groups at 2 weeks and 8 weeks. These results suggest that the use of tetracycline blended chitosan membrane on the calvarial defects in rats has significant effect on the regeneration of bone tissue in itself. And it implicate that tetracycline blended chitosan membrane might be useful for guided tissue regeneration.
Chlorhexidine and Listerine are widely used in dentistry due to its effectiveness on plaque control and bactericidal action. The effects of these agent on chronic gingivitis and wound healing following surgical periodontal therapy in human has been favorable. Understanding the effects of chlorhexidine and Listerine on human gingival fibroblast will provide the rationale for its use during the healing process of periodontal surgery. The purpose of this study was to compare the effects of chlorhexidine and Listerine on human gingival fibroblast. Human gingival fibroblasts were cultured from the healthy gingiva on the extracted premolar of orthodontic patients. Human gingival fibroblast were trypsinized and cultured in growth medium added range of 0.0012-0.12% chlorhexidine and 1-100% Listerine mouth wash solution. The cell used in this study were between fifth to eighth passage number. The cell morphology were examined by inverted microscope and the cell activity were measured by MIT assay. The Morphology of gingival fibroblast added Chlorhexidine and Listerine at the concentration of all range were became globular and lost their cytoplasmic process. Our results indicate that a 0.0012 concentration of chlorhexidine and 1% concentration of Listerine were shows minimal cytotoxicity, but above these concentraion, there was a significant difference between the cell activity in the experimental group and control group(p
Chitosan has been known as a wound healing agent. The purpose of this study was to evaluate the biocompatibility and guided bone regenerative effect of chitosan and chitosan-cellulose membranes. The effects of chitosan and chitosan-cellulose membranes on the growth and survival of human periodontal ligament cells were examined by rapid colorimetric MTT(tetrazolium) assay, and the tissue response and resorption pattern were observed by implanting the membranes into the subcutaneous tissue of the back of rats for 6 weeks. To evaluate the guided bone regenerative potential of membranes, the amount of newly formed bone in the rat calvarial defects(8mm in diameter) was measured by histomorphometry and radiomorphometry 1,2 and 4 weeks after implantation of membranes. Chitosan and chitosan-cellulose membranes showed no adverse effect on the growth and survival of human periodontal ligament cells. When membranes were subcutaneously implanted, inflammatory reaction was observed at 1 week and which gradually subsided 2weeks after implantation. Membranes remained intact throughout the experimental period of 6 weeks. Radiomorphometric analysis of the craniotomy sites revealed that chitosan and chitosan-cellulose membrane implanted sites showed increased radiopacity over control. Statistically significant differences with control were found in chitosan-cellulose membrane implanted group at 2 and 4 weeks, and chitosan membrane implanted group at 4 weeks(P<0.05). Histomorphometric data indicated a pattern of osseous healing similar to radiomorphometric analysis. There was a statistically significant difference between control and chitosan-cellulose membrane implanted group at 4 weeks(P<0.05). These results implicate that chitosan and chitosan-cellulose membrane might be useful for guided bone regeneration.
The present study evaluates the effects of dura mater barrier membranes In class III furcation defects on the regeneration of periodontal tissues in dogs. Experimental class III furcation defects were created surgically by removing alveolar bone horizontally down to 4mm from CEJ in mandibular premolars of adult dogs. Dura mater barrier membranes were applied bucco-lingually in the test group, and flap surgery only with no membranes in the control group. The healing was evaluated clinically and histologically after 8weeks. Clinically, the test group showed slight exposures of the membranes, while the control group showed no furcation exposure, The test specimens showed new bone formation coronal to the notch, while the control specimens had new bone formation up to the level of the notch. New cementum was observed in both groups. The test specimens showed functional arrangements of connective tissue fibers between new bone and new cenentum, while irregular arrangements were observed in the controls. No root resorption or ankylosis were observed in either groups,These results suggest that dura mater resorbable barrier membranes on class III furcation defects may be effective in regeneration of alveolar bone and peridontal ligament.
The periodontal flap surgery is the most widely utilized surgical procedure to reduce the pocket depth and to access the subgingival root surfaces for scaling and root planing. The diagnosis of the periodontal lesion and the objective of the surgery will dictate the type of flap procedure which will be utilized to obtain the best result. The incisions, type of flap and the selection of suturing design must be planned and executed to fit the problem. Periodontal flaps are designed to preserve gingival integrity and to gain access to root surfaces for residual calculus removal and to thoroughly remove granulation tissue so bone defects can be visualized and treated. Gentle and efficient procedures result in optimum healing and minimal postoperative pain. When flaps need to be repositioned apically or less often, coronally, then the flaps must sit passively at the appropriate level before suturing. To ensure this, buccal and lingual flaps need to be elevated beyond the mucogingival junction so the elasticity of the mucosa allows for flap mobility. Sometimes it may be necessary to extend the flap elevation apically with a split incision approach to minimize the effect of the less elastic periosteum. Vertical incisions can aid in flap positioning by allow ing the clinician to suture the flap at a different level to the adjacent untreated gingiva. In osseous periodontal surgery, flaps are apically positioned to minimize postoperative pocket depth. In regenerative periodontal surgery including implant surgery, soft tissue cove rage of bony defects, graft materials, membranes, and bio logic agents is important so sulcular incisions and light suturing techniques are crucial.
Incipient changes of the periodontal tissue in the pressure zones of rat molar subjected to the experimental force were studied by the transmission electron microscope. Experimental animals were consisted in 3 control and 21 experimental rats, of which one maxillary first molar was moved buccally with a fixed appliance which were exerting the force of 15 gm. After experimental period of 1 hour, 3 hours, 6 hours, 24 hours, 2 days, 3 days and 7 days, the animal were sacrificed with cardiac perfusion of $2.5\%$ glutaraldehyde in the sodium cacodylate buffer and the experimental teeth with surrounding periodontal structures were processed for electron microscope. At the beginning of the tooth movement, periodontal ligaments of the pressure were compressed and collagenous fibers were arranged parallel to the root of the teeth and cell free zones in company with cell necrosis were followed. Cell free zones at the periodontal ligaments appeared in the 3 hour survival group, and getting severe with time lapse it became widespread in 2-3 day survival group and undermining bone resorption as a healing process was observed in 7 day survival group. Dilatation of mitochondria and swelling of the rER in the fibroblast and other connective tissue cells in the periodontal ligament were observed in the 3 hour survival group, which were characteristics of the incipient changes in the compressed periodontal ligament. Dilatation of nuclear membrane and pyknosis were followed by the destruction of the nucleus and cell membrane. There were no evidence in cell damage or necrosis of the alveolar bone adjacent to the hyalinized area of periodontal ligaments.
After periodontal surgery, the potential healing responses were occurred by interaction among junctional epithelium, gingival connective tissue, alveolar bone and periodontal ligament. The only cell that created periodontal regeneration was derived from periodontal ligament. The aim of the study was to evaluate the regenerative effects of the collagen membrane($collacote^{\circ}C$) and autogenous connective tissure graft with periosteum. Experimental periodontitis were created in furcation area of 4 adult dogs with bone removal and gutta percha packing. After 6 weeks later, the gutta percha was removed and experiment was performed divided by 3 groups. 1) Flap operation(control group). 2) Flap operation with collage membrane(Experimental group I). 3) Flap operation with autogenous connective tissue graft with periosteum (Experimental group II). After dogs were sacrificed after two and three weeks, specimens were prepared and stained with hematoxylin-eosin and masson-trichrome stain for light microscopic study. The results were as follows : 1. In all gruoups, connective tissue compartments were increased from two to three weeks especially in experimental group I. 2. Collagen membrane and connective tissue were increased collagen deposits of periodontal ligament. Therefore collagen fiber attached to tooth surface was seen. 3. In al experimental groups, newly forming alveolar bone was seen. 4. Collagen membrane and connective tissue were which prevented proliferation of epithelium, aided connective tissue new attachment and influenced periodontal regeneration.
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