• Biomedical Science Letters
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• v.6 no.2
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• pp.119-129
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• 2000

The skin is an organ that has many important roles, including protection against infection, regulation of temperature and fluid loss, and sensory function. Injury to the skin, wound repair normally involves: (1) balanced activity of inflammation, (2) the re-epithelial phase and (3) the matrix formation of remodeling phase. Thus, skin wound healing is a finely controlled biological process involving a series of complex cellular interactions. Laser therapy is being implemented with increasing frequency in medicine. Low intensity laser is one that is capable of producing an energy density so low that any biologic alterations are the result of direct irradiation effect, not thermal events. This study was designed to evaluate the efficacy of low intensity laser therapy on skin wound healing in rabbits. A total of 10 male rabbits (New Zealand White Rabbit), age 8 weeks were used. Skin wound were surgically created dorso-lateral on the flank of 10 rabbits (2$\times$2 cm/damage areas). The experimental animals were treated with 5Hz (830 nm wave length) low-intensity laser (MILTA-01 Model) daily for 10 min (1.6 J/$cm^2$) for 12 days. Control animals were sham treated with the laser head. Laser irradiation animals showed a complete remodeling of the epithelial layer, a positive repair of connective tissues, and enhanced the wound closure rate over time as compared to the control animals. Especially, laser irradiation groups improved fibroblast activity, cellular content, granulation tissue formation, and collagen deposition which is resulted in improving the tensile strength of the wound. These findings suggest that laser photostimulation could accelerate healing of open wound in rabbits, and may be benefit in the treatment of open wound, including decubitis ulcers.

• Archives of Plastic Surgery
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• v.34 no.3
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• pp.291-297
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• 2007

Purpose: Platelet-rich plasma(PRP) contains protein growth factors, which are actively secreted by platelets to promote wound healing. However, it is not clear whether the injection of PRP into the autologous fat grafts increases the survival rate and the degree of angiogenesis. Methods: New Zealand White rabbit ears were injected fat with PRP, saline, insulin or isoproterenol (n=8/each group) for observation of the survival and degree of angiogenesis of the injected fat. The volume of the harvested fat and the degree of angiogenesis from dorsum of rabbit ears were evaluated 4, 8, and 12 weeks after the autologous fat graft. The degree of angiogenesis was measured with microvascular density (MVD) counts. Results: The volume of harvested fat decreased in a time-dependent manner after autologous fat grafts, but the decrease rate in volume of harvested fat was slower in PRP-injected group compared to that of other control groups. The difference in the volume of the harvested fat between PRP-injected group and other control groups became significant from 4 weeks after the autologous fat graft, and was maintained up to 12 weeks. However, there was no significant difference between PRP-injected group and insulin-injected group 8 and 12 weeks after the autologous fat graft. On the contrary, MVD counts increased in a time-dependent manner after autologous fat grafts. The MVD counts were significantly higher in PRP-and insulin-injected groups than in other control groups from 4 weeks after the autologous fat graft, and these differences were maintained up to 12 weeks. There was no correlation between mean platelet numbers and the volume of harvested fat. Conclusion: The present study demonstrates that PRP-injection into autologous fat grafts increases the survival rate and the degree of angiogenesis. Thus, PRP injection with autologous fat grafts would be a promising tool for maintaining the volume of the grafted fat.

• Applied Microscopy
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• v.27 no.2
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• pp.121-130
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• 1997

It has been demonstrated that majority of cells in the mammalian body such as myocytes and epithelial cells of skin and intestine respond to mechanical force or environmental factors and exhibit partial disruption of cell membrane, i. e., cell wounding, even in a physiological condition. Myocardial cells are rather apt to be wounded than other cells since they are definitely exposed to mechanical stress by contraction-relaxation and blood flow. However, the mechanism how myocardial cells protect themselves against cell wounding is not yet clarified. On this background, the present study was performed to elucidate whether albumin leakage is related to cell wounding and to assess whether diltiazem, a potent calcium channel blocker, is beneficial in isoproterenol-induced cell wounding in the heart. Hearts isolated from New Zealand White rabbits ($1.5\sim2.0kg$ body weight, n=20) were perfused with Tyrode solution by Langendorff technique. After stabilization of baseline hemodynamics, the hearts were subjected to bolus administration of isoproterenol and diltiazem as following order: $1.6{\mu}M$ isoproterenol at zero min (the beginning point): $16{\mu}M$ diltiazem at 20min; $1.6{\mu}M$ isoproterenol at 25min; $16{\mu}M$ isoproterenol at 45 min; $160{\mu}M$ diltiazem at 65 min; $16{\mu}M$ isoproterenol at 70 min. During all experiments, the left ventricular function was recorded, albumin leakage in the coronary effluents was analyzed by electrophoresis and Western blot, and myocardial cell membranes were examined by conventional transmission electron microscopy. Data were analyzed by t-test and linear regression test. Isoproterenol significantly increased the inotropic and chronotropic contractions, coronary flow, and frequency of arrhythmia, however, diltiazem did not influence on hemodynamics except decrease in the frequency of arrhythmia and a slight decrease in contractility. Isoproterenol also resulted partial disruption of myocardial cell membrane and inclose in albumin leakage, while diltiazem pretreatment showed number of electron-dense plaques in the cell membrane and a tendency of decrease in albumin leakage. These results indicate that albumin leakage may be an indirect index of cell wounding in the heart and diltiazem nay be beneficial to protect myocardial cells against isoproterenol-induced cell wounding. It is likely that diltiazem promotes resealing process of the cell membrane.

• Journal of the Korean Association of Oral and Maxillofacial Surgeons
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• v.33 no.5
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• pp.405-418
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• 2007

Mesenchymal stem cells(MSCs) have been though to be multipotent cells that can replicate that have the potential to differentiate into lineages of mesenchymal tissue including the bone, cartilage, fat, tendon, muscle, and marrow stroma. Especially, scaffolds to support cell-based tissue engineering are critical determinants of clinical efforts to regenerate and repair the body. Selection of a matrix carrier imvolves consideration of the matrix's role as a scaffold for physical support and host tissue integration as well as its ability to support of synergize the osteoinductive program of the implanted mesenchymal stem cell. The aim of this study is to evaluate the effect of autobone and Bio-$Oss^{(R)}$ to adherent mesenchymal stem cells as scaffolds on sinus augmentation with fibrin glue mixture in a rabbit model. 16 New Zealand White rabbits were divided randomly into 4 groups based on their time of sacrifice(1, 2, 4 and 8 weeks). First, mesenchymal stem cells were isolated from iliac crest marrow of rabbits and expanded in vitro. Cell culture was performed in accordance with the technique described by Tsutsumi et al. In the present study, the animals were sacrificed at 1, 2, 4 and 8 weeks after transplantation, and the bone formation ability of each sides was evaluated clinically, radiologically, histologically and histomorphologically. According to the histological observations, autobone scaffolds group showed integrated graft bone with host bone from sinus wall. At 2 and 4 weeks, it showed active newly formed bone and neovascularization. At 8 weeks, lamellae bone was observed in sinus graft material area. Radiologically, autobone with stem cell showed more radiopaque than Bio-$Oss^{(R)}$ scaffolds group. there were significant differences in bone volume between 4 and 8 weeks(p<0.05).

• Archives of Plastic Surgery
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• v.37 no.6
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• pp.721-725
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• 2010

Purpose: The survival of composite graft is dependent on three steps, (1) plasmatic imbibitions, (2) inosculation, and (3) neovascularization. Among the many trials to increase the survival rate of composite graft, prostaglandin E1 (PGE1) has beneficial effects on the microcirculatory level with vasodilating, antithrombotic, anti-inflammatory and neoangiogenic properties. Lipo-PGE1 which is lipid microspheres containing PGE1 had developed to compensate the systemic and local side effects of PGE1. This study was proposed to determine whether Lipo-PGE1 administration enhanced the survival of composite graft through neovascularization quantitatively in a rabbit ear model. Methods: Fourteen New Zealand White Rabbits each weighing 3~4 kg were divided in two groups: (1) intravenous Lipo-PGE1 injection group and (2) control group. A $2{\times}1\;cm$ sized, full-thickness rectangular composite graft was harvested in each auricle. Then, the graft was reaaproximated in situ using a 5-0 nylon suture. For the experimental group, $3{\mu}g$/kg/day of Lipo-PGE1 ($5{\mu}g$/mL) was administered intravenously through the marginal vein of the ear for 14 days. The control group was received no pharmacologic treatment. On the 14th postoperative day, composite graft of the ear was harvested and immunochemistry staining used Monoclonal mouse anti-CD 31 antibody was performed. Neoangiogenesis was quantified by counting the vessels that showed luminal structures surrounded by the brown color-stained epithelium and counted from 10 random high-power fields (400x) by independent blinded observer. Statistical analysis (Wilcoxon Signed Ranks test for nonparametric data) was performed using SPSS v12.0, with values of p<0.05 considered significant. Results: The mean number of the microvessels was $15.48{\pm}8.65$ in the experimental group and $9.82{\pm}7.25$ in the control group (p=0.028). Conclusion: The use of Lipo-PGE1 facilitated the neoangiogenesis, resulted in the improvement of the survival rate of graft. On the basis of this results, we could support wider application of Lipo-PGE1 for more effective therapeutic angiogenesis and successful survival in various cases of composite graft in the human.

• Journal of Periodontal and Implant Science
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• v.39 no.sup2
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• pp.223-230
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• 2009

Purpose: Synthetic bone products such as biphasic calcium phosphate (BCP) are mixtures of hydroxyapatite (HA) and ${\beta}$-tricalcium phosphate (${\beta}$- TCP). In periodontal therapies and implant treatments, BCP provides to be a good bone reconstructive material since it has a similar chemical composition to biological bone apatites. The purpose of this study was to compare bone regeneration capacity of two commercially available BCP. Methods: Calvarial defects were prepared in sixteen 9-20 months old New Zealand White male rabbits. BCP with HA and ${\beta}$- TCP (70:30) and BCP with Silicon-substituted hydroxyapatite (Si-HA) and ${\beta}$-TCP (60:40) particles were filled in each defect. Control defects were filled with only blood clots. Animals were sacrificed at 4 and 8 week postoperatively. Histomorphometric analysis was performed. Results: BCP with HAand ${\beta}$- TCP 8 weeks group and BCP with Si-HA and ${\beta}$- TCP 4 and 8 weeks groups showed statistically significant in crease (P <0.05) in augmented area than control group. Newly formed bone area after 4 and 8 weeks was similar among all the groups. Residual materials were slightly more evident in BCP with HA and ${\beta}$- TCP 8 weeks group. Conclusions: Based on histological results, BCP with HA and ${\beta}$- TCP and BCP with Si-HA and ${\beta}$- TCP appears to demonstrate acceptable space maintaining capacity and elicit significant new bone formation when compared to natural bone healing in 4 and 8 week periods.

• Maxillofacial Plastic and Reconstructive Surgery
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• v.27 no.6
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• pp.510-522
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• 2005

To assess the clinical applicability of bio-artificial mucosa which was made with autologous oral keratinocytes and human acellular dermal matrix, the formation of basement membrane and stratification of oral keratinocytes were evaluated. Six New Zealand white rabbits (around 2kg in weight) were anesthetized and its buccal mucosa was harvested (1.0 $\times$ 0.5cm size). Oral keratinicytes were extracted and cultured primarily with the feeder layer of pretreated NIH J2 3T3 fibroblast. These confluent cells were innoculated on the human acellular dermal matrix and cultured in multiple layer by air-rafting method. After 3, 5, 7, 10, 14 days of culture, each cultured bio-artificial mucosa was investigated the number of epthelial layer of by H&E stain and toluidine blue stain. The immuhohistochemical methods were used to evaluate the cell division capacity, the formation of basement membrane, and it's property of specific cells (PCNA, cytokeratin 14, laminin). Transmission electromicroscopy was used for the attachment between cells and matrix with the number of hemidesmosome. In result, the numbers of layer of stratified growth of oral keratinocyte cultured on the human acellular dermal matrix and the number of hemidesomal attachment between epithelial cells and human acellular dermal matrix were similar to the layers of normal oral mucosa after 10 days of culture. The cell division rate, basement membrane formation and proliferation rate increased as culture period increased. With these results, bio-artificial mucosa with autologous oral epithelial cells cultured on the acellular dermal matrix had clinically adaptable properties after 10 days' culture and this new bio-artificial mucosa model with relatively short culture time can be expected clinical applicability.

• Archives of Plastic Surgery
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• v.36 no.5
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• pp.531-537
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• 2009

Purpose: Alveolar bone grafting has become an essential process in the treatmemt of alveolar cleft patient for stabilization of the maxillary arch, elimination of oronasal fistula, the reconstruction of the soft tissue nasal base support, and creation of bony support for tooth eruption for implant. The use of Autologous iliac cancellous bone is preferable because of the adequate quantity and high osteoinductive potential. However, even with iliac bone, insufficient osteoregeneration and absorption occur due to several factors such as the patient's age, cleft width, functional stress, and others. In order to increase osteoregeneration where the iliac bone is placed, the present study is associated with bone marrow aspirate (BMA). The experimental study evaluated the efficacy of osteoregeneration in normal cleft rabbits when alveolar bone grafting was performed with autologous iliac corticocancellous bone. Methods: Twenty - four New Zealand White rabbits were divided randomly into 2 groups (BMA, control). All animals underwent harvesting of corticocancellous bone graft from the right posterior iliac crest via standard surgical technique. $1m{\ell}$ of BMA were obtained by scraping the needle and aspirate with $10m{\ell}$ syringe from the contralateral iliac bone wall. The muco - periosteal flap on the palate was elevated. A mixture of Equal bone's volumes with BMA and saline as its control was inserted into the cleft. Animals were sacrificed at 2, 4, and 8 weeks and maxilla was harvested for dental peri - apical X-ray, bone matrix density (BMD),and histologic analysis. Result: BMD of regenerated bone to the cleft in the rabbits was higher than that of the control rabbits. X-ray, histologic analysis showed that increased osteoregeneration and low absorption rate were observed in the BMA group. Conclusion: Our experimental study showed BMA enhanced the osteoregeneration and survival rate of alveolar bone grafting. BMA is easy to extract & cost - time effective. So it can be an effective enhancers for bone grafting mixtures.

• The Korean Journal of Nuclear Medicine
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• v.24 no.1
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• pp.124-132
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• 1990

It has been reported that hematoma is one of the most crucial factors in fracture healing since callus formation is disturbed by washing out the hematoma near a fracture site. However, it is not clear why the hamatoma is important and how it plays a role during the fracture healing. In order to investigate the role of hematoma in the process of fracture healing, the osteogenic potential by subperiosteal transplantation have been studied. Experimental fractures by operation were made at the mid-shaft of the tibia in New Zealand white rabbits. Removal of hematoma at the fracture site was done after 2 and 3 days from experimental fracture, and the removed hematoma was transplanted into the subperiosteal area at the mid-shaft of the ulna of each rabbit. As control groups, we have performed 3 different procedures 1) the hematoma was transplanted into the muscular layers at the thigh and forearm; 2) autologous blood clots were transplanted into the subperiosteal area of the ulna; and 3) sham operation without a transplantation into the subperiosteal area. After transplantation, serial bone scintigraphy and simple radiography were performed at 4 days, 1 week, and 2 weeks to detect an abnormality. The results of bone scintigraphy were positive in 5 of 6 experimental group. However, all in three control groups were negative. Histological observation of the positive bone revealed new bone formation with trabeculation. These results suggest the hematoma in fracture site has osteogenic potential in the subperiosteal area which can be demonstrable by bone scintigraphy and histologic findings. Therefore, it is considered that hematoma of the fracture site plays an important role in the process of fracture healing. Further biochemical investigation using various experimental models is mandatory to apply this preliminary result to the treatment of clinical delayed union or nonunion.

• Journal of Ginseng Research
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• v.23 no.1 s.53
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• pp.13-20
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• 1999

We previously reported that Korean red ginseng (KRG) has a relaxation effect on the smooth muscles of corpus cavernosum via nitric oxide (NO) pathway and calcium and potassium channels. However, it is suggested that the active ingredients of KRG might be different depending on the sources of preparation, and there might be differences in actions for different compositions. We first investigated the composition of KRG saponins according to the extractions of the various sources of KRG, then with these extractions the relaxation effects were evaluated in vitro and hemodynamical in vivo using New Zealand white rabbit and rat corpus cavernosum. The total compositions of ginsenoside $(G-Rb_1,\;-Rb_2,\;-Rc,\;-Rd,\;G-Re,\;-Rf,\;-Rg_1)$ in fractionated KRG saponin designated as TS-1, TS-2, TS-3 were $41\%,\;40\%,\;and\;62\%,$ respectively, and the ratios of PD saponin and PT saponin (PD/PT) were 1,55, 1.72, 2.25, and 2.61, the values of which were statistically significant. In vitro studies using the rabbit corpus cavernosal muscle strips, the KRG saponin relaxed cavernosal strips in a dose-dependent manner, and same results were observed in in vivo studies, that KRG saponin increased the intracavernosal pressure in the rat. There was difference in the efficacy according to fractionation techniques. The differences in the total contents of ginsenosides did not affect relaxation, rather PT saponin content was statistically related to the degree of cavernosal relaxation, and this action presumed to be mediated by NO pathway and calcium and potassium channels. In conclusion, KRG exerts relaxation which is a key step in erection via combination of effects on NO system or calcium and potassium channels. The efficacy of this action is different to the sources of ginseng, which is affected by the different composition of ginsenosides $(G-Rb_1,\;-Rb_2,\;-Rc,\;-Rd,\;G-Re,\;-Rf,\;-Rg_1).$ Thus the further studies on the active ingredients such as minor ginsenosides and non-saponin components of red ginseng with maximum potency should be sought.