• Biotechnology and Bioprocess Engineering:BBE
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• v.7 no.5
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• pp.303-310
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• 2002

Cell therapy applied to wound healing or tissue regeneration presents a revolutionary realm to which principles of gene engineering and delivery may be applied. One promising application is the transplantation of cells into the wounded tissue to help the tissue repair. However, when cells are transplanted from in vitro to in vivo, immune rejection occurs due to the immune response triggered by the activation of T-cell, and the transplanted cells are destroyed by the attack of activated T-cell and lose their function. Immune suppressant such as FK506 is commonly used to suppress immune rejection during transplantation. However, such kind of immune suppressants not only suppresses immune rejection in the periphery of transplanted cells but also suppresses whole immune response system against pathogenic infection. In order to solve this problem, we developed a method to protect the desired cells from immune rejection without impairing whole immune system during cell transplantation. Previously, we reported the success of constructing glomerular epithelial cells for removal of immune complex, in which complement receptor of type 1 (CR1) was over-expressed on the membrane of renal glomerular epithelial cells and could bind immune complex of DNA/anti-DNA-antibody to remove immune complex through phagocy-tosis [1]. Attempting to apply the CR1-expressing cells to cell therapy and evade immune rejection during cell transplantation, we constructed three plasmids containing genes encoding a soluble fusion protein of cytolytic T lymphocyte associated antigen-4 (CTLA4Ig) and an enhanced green fluorescent protein (EGFP). The plasmids were transfected to the above-mentioned glomerular epithelial cells to express both genes simultaneously. Using the clone cells for cell transplantation showed that mice with autoimmune disease prolonged their life significantly as compared with the control mice, and two injections of the cells at the beginning of two weeks resulted in remarkable survivability, whereas it requires half a year and 50 administrations of proteins purified from the same amount of cells to achieve the same effect.

• The Plant Pathology Journal
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• v.24 no.2
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• pp.118-124
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• 2008

Skin sooty dapple disease, a fungal disease that lowers Asian pear fruit quality, has emerged recently in Korea but has not yet been thoroughly characterized. This disease affects the surface of fruit, leaves, and young shoots of the Asian pear, typically appearing as a dark or pale black dapple on the fruit surface. The disease initiates on the fruit with small circular lesions that become bigger, eventually spreading to form large circular or indefinite lesions. Sparse dark or flourishing white-greyish aerial mycelia and appearance of a dark or pale black dapple on the fruit surface are typical signs of this disease. The disease was severe during cold storage of the Niitaka and Chuhwangbae varieties, but more limited on the Gamcheonbae and Hwangkeumbae varieties. To identify causal pathogens, 123 fungal isolates were obtained from lesions. The fungi that caused typical skin sooty dapple disease symptoms in our bioassay were identified. Based on their morphological characteristics, 74% of the isolates were Cladosporium sp. and 5-7 % of the isolates were Leptosphaerulina sp., Tripospermum sp., or Tilletiopsis sp. None of the isolates caused severe soft rot by injection to a wound plug, but some of the Cladosporium sp. isolates caused mild maceration. Therefore this microbiol complex cannot account for the soft rot also observed in stored fruits. The high frequency of isolation of Cladosporium sp. from disease tissues and bioassay on pear fruit surface suggest that Cladosporium sp. could be a major pathogen in the microbial complex associated with skin sooty dapple disease of the Asian pear in Korea.

• Journal of the Korean Association of Oral and Maxillofacial Surgeons
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• v.33 no.4
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• pp.322-330
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• 2007

Backgrounds: To overcome limited amount of autogenous mucosa for the reconstruction of various mucosal defect including oral mucosal defect, tissue engineered mucosa has been recently introduced. However, introduced conventional technique of tissue engineered mucosa still have serious pitfalls such as long fabrication time, fragility of the reconstructed mucosa, and complexity of the technique. Aim of the study: To examine whether the complex of preconfluent autologous keratinocytes and autologous PRP(Platelet rich plasma) can reconstruct oral mucosa on the muscular flap with easier and faster way compared to conventional mucosal tissue engineering technique. Materials and methods: One day before the operation, oral mucosa(3mm in diameter) were taken and treated for extraction of oral keratinocytes according to the routine manner. The day of operation, oral keratinocytes were prepared in the laboratory and then moved to the operating theater. Autologous PRP was also prepared and then mixed with oral keratinocytes just before grafting on the prepared muscular flap. After keratinocyte-PRP complex was seated, then a sterilized rubber sheet was placed on the graft and the elevated skin flap was replaced and sutured. Biopsies were proceeded at 3, 5, 7, 14 and 21 days. Tissue samples were evaluated clinically, histologically, and immunohistochemically. Results: All of the oral keratinocyte-PRP complexes were successfully grafted on the recipient sites(100%). On 3 days after the operation, 1-2 continuous epithelial layer and many inflammatory cells were observed. On 5 days after the operation, increase of layers of keratinocyte was observed with less inflammatory response. Thickness of the layers was gradually increased from 7 to 21 days after the operation. Cytokeratin confirms epithelium in every specimen. Conclusions: Preconfluent graft of autogenous oral keratinocytes mixed with autogenous PRP have successfully reconstructed myo-mucosal flap. This technique could be a useful alternative for oral mucosal reconstruction in the near future.

• Archives of Plastic Surgery
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• v.42 no.4
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• pp.446-452
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• 2015

Background Various techniques are used for performing breast reduction. Wise-pattern and vertical scar techniques are the most commonly employed approaches. However, a vertical scar in the mid-lower breast is prominent and aesthetically less pleasant. In contrast, a semicircular horizontal approach does not leave a vertical scar in the mid breast and transverse scars can be hidden in the inframammary fold. In this paper, we describe the experiences and results of semicircular horizontal breast reductions performed by a single surgeon. Methods Between September 1996 and October 2013, our senior author used this technique in 38 cases in the US and at our institution. We used a superiorly based semicircular incision, where the upper skin paddle was pulled down to the inframammary fold with the nipple-areola complex pulled through the keyhole. Results The average total reduction per breast was 584 g, ranging from 286 to 794 g. The inferior longitudinal pedicle was used in all the cases. The average reduction of the distance from the sternal notch to the nipple was 13 cm (range, 11-15 cm). The mean decrease in the bra cup size was 1.7 cup sizes (range, a decrease of 1 to 3). We obtained very satisfactory results with a less noticeable scar, no complication such as necrosis of the nipple or the skin flap, wound infection, aseptic necrosis of the breast tissue, or wound dehiscence. One patient had a small hematoma that resolved spontaneously. Conclusions This technique is straightforward and easy to learn, and offers a safe, effective, and predictable way for treating mammary hypertrophy.

• Journal of the korean academy of Pediatric Dentistry
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• v.24 no.1
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• pp.41-57
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• 1997

The present study investigated the effects of hyperbaric oxygen therapy on periodontal wound healing of replanted rat tooth. 80 rats (Sprague-Dawley strain) weighting $130{\pm}5gm$ were selected and divided into experimental and control group, each group consisting of 40 rats. Rats were administered 0.4% ${\beta}$-aminoproprionitrile for 5 days to achieve gentle tooth extraction. The maxillary first molars were extracted under anesthesia with pentobarbital, washed in sterile distilled water, treated with bacterial collagenase to remove collagen fibers on the root surfaces. After washing in water overnight, the mesial root surface were demineralized by application of citric acid, washed, dried and stored at $4^{\circ}C$. Immediately after tooth extraction and bleeding control, the treated molars extracted previously from other rats were replanted. The experimental group was exposed to hyperbaric oxygen at 2.5 atm. for 2 hrs. a day during experimental period. Eight animals of each group were sacrificed 1, 3, 6, 8, 10 days after reimplantation of teeth by intracardiac perfusion with 4% paraformaldehyde. The replanted molars and surrounding tissues were cut, demineralized, dehydrated and embedded in paraffin. Sections were stained with azan, toluidine blue and hematoxylin. Some other sections were stained by means of immunostaining achieved by the avidinbiotin complex method. The results as follows; 1. Experimental group showed fast healing of gingival epithelium. 2. Macrophage and newly formed blood vessels appeared early in the gingival connective tissue of experimental group. 3. Experimental group showed fast, abundant fibroblast proliferation and regularity of collagen fiber. 4. In both group, collagen was distributed along the collagen fiber. The distribution was strong and regular in the experimental group. 5. In the regenerated periodontal ligament of experimental group, fibers showed regular arrangement and invaded root surface fast.

• Maxillofacial Plastic and Reconstructive Surgery
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• v.15 no.3
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• pp.217-228
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• 1993

The purpose or this study was to observe the histological alteration of Langerhans cells on wound healing process in applying low energy laser irradiation. For this study, 50 Spraque-Dewly rats, weighing 150Gm or more were devided into control, experimental control group(0), 47.5Hz(1), 190Hz(3), 380Hz(5), 760Hz(7), lased group. All the experimental animals were made excision wound on buccal mucosa, 2mm depth, and lased with stoma laser (904nm, semconductor type ASGaAI, Sedalac France) 47.5Hz, 380Hz, 960Hz, 3minutes one time respectively except experimental control group. After the experiment, experimental animals were sacrificed after 24hours, 48hours, 72hours on each. Taken specimens were embedden in paraffin, sectioned 6-8u in thickness. And the langerhans cell were detected using ant S-100 protein antibody, and histochemically processed with Avidin Biotin complex method. All the Langerhan cells were calculated under light microspe in 400 multiplication field and standard deviation, probability test between each group were evaluated using statistical analysis system(S.A.S)program. Following results were obtained. 1. Langerhan cells were increased in experimental control group compared to that in control group(P<0.01). 2. 24hour after experiments, Langerhans cell were decreased compare to that in control group and control experimental group 5, 1, 3. Probability test shows significance between control experimental and 5, 1, 3 group on a =0.05 range. 3. 48our after experiment, Langerhans cells were decreased compare to that on experimental control group, and probability test shows significance between control experimental and 3, 7, 5 group an a=0.05 range. 4. 72hour after experiments, Langerhans cells were decreased compare to that on experimental control group and probability test on group comparison shows significance between control experimental and 1, 5 and 1 between 3, 7 between 3, and 5, between 7, respeilively on a=0.05 range. 5. Langerhans cells number in experimental group were decreased compare to that on experimental control group in applying laser irradiation.

• Molecules and Cells
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• v.39 no.4
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• pp.345-351
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• 2016

Wound healing is a complex physiological process necessitating the coordinated action of various cell types, signals and microRNAs (miRNAs). However, little is known regarding the role of miRNAs in mediating this process. In the present study, we show that let-7c miRNA is decreased in heat-denatured fibroblasts and that inhibiting let-7c expression leads to the increased proliferation and migration of dermal fibroblasts, whereas the overexpression of let-7c exerts an opposite effect. Further investigation has identified heat shock protein 70 as a direct target of let-7c and has demonstrated that the expression of HSP70 in fibroblasts is negatively correlated with let-7c levels. Moreover, down-regulation of let-7c expression is accompanied by up-regulation of Bcl-2 expression and down-regulation of Bax expression, both of which are the downstream genes of HSP70. Notably, the knockdown of HSP70 by HSP70 siRNA apparently abrogates the stimulatory effect of let-7c inhibitor on heat-denatured fibroblasts proliferation and migration. Overall, we have identified let-7c as a key regulator that inhibits fibroblasts proliferation and migration during wound healing.

• Archives of Plastic Surgery
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• v.32 no.5
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• pp.607-612
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• 2005

A traditional tie-over dressing may be applied to support the take of a skin graft. Although there are many advantage of this method, it has significant disadvantages, including time-consuming application. Furthermore, when the dressing is changed, the gauze becomes hard and can be stuck to the graft, causing damage and pain upon removal. The purpose of our study is to evaluate the effect of semi-occlusive dressing using polyurethane foam and film dressing($Allevyn^{(R)}$, $Opsite^{(R)}$) after full thickness skin graft. The authors treated 45 cases including burn scar contracture(n=38), syndactyly (n=1), absence of nipple-areolar complex(n=4), traumatic skin defect(n=1) and contact burn(n=1) with authors' method and 39 patients including burn scar contracture (n=39) with the tie-over dressing between 2000 and 2004. The patients in polyurethane foam and film dressing group ranged from 1 to 62 years of age (mean age, 15.1 years) and the patients in tie-over dressing group ranged from 2 to 60 years of age(mean age, 21.3 years). The postoperative results were analyzed according to the following measures: (1) the duration of graft-taking, (2) the admission period, (3) complications. Compared with the traditional tie-over dressing, polyurethane foam and film dressing was shown to be more successful in a reduced duration of graft-taking, in which was similar to the former in the rate of graft-taking, a reduced admission period and patient's discomfort. We concluded that semi-occlusive dressing using $Allevyn^{(R)}$ and $Opsite^{(R)}$ was an effective method after full thickness skin graft, which was easy to shape to difficult body locations, such as web spaces, fingers and maintains a moist environment for wound healing and does not stick to the wound.

• Journal of the Korean Association of Oral and Maxillofacial Surgeons
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• v.32 no.3
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• pp.189-199
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• 2006

Purpose of study: Partial thickness skin graft is the golden standard regimen for full-thickness skin defect caused by burn or trauma. However, in case of extensive burns of more than 50% of total body surface area, the donor site is not sufficient to cover all defects. As a second choice, allograft, xenograft and synthetic materials have been used to treat skin defect. Among them the amniotic membrane(AM) was used as a biological dressing for centuries because of its potential for wound healing. In this study, quantification of EGF in AM and effect of AM-collagen complex on full thickness skin defects was examined. Materials & Methods: The concentration of EGF in fresh, deep frozen and freeze-dried AM was evaluated by ELISA. EGF-R immunostaining was performed in freeze-dried AM. SD rats weighing 250${\sim}$300g was used for wound healing experiment. Three full thickness skin defects(28mm diameter) were made on dorsal surface of SD rat. The control group was covered by Vaselin gauze and AM-collagen complex and $Terudermis^{(R)}$. was grafted in two other defects. Healing area, Cinamon's score were evaluated before biopsy. Grafted sites were retrieved at 3 days, 1 week, 2 weeks and 4 weeks after operation. H & E and Factor VIII immunohistochemical stain was performed to evaluate the microscopic adhesion and structural integrity and microvessel formation. Results: 1. EGF concentration of fresh, deep frozen and freeze-dried AM showed similar level and EGF-R was stained in epithelial layer of freeze-dried AM. 2. At 4 weeks after grafting, the healing area of AM-collagen and Terudermis group was 99.29${\pm}$0.71% and 99.19${\pm}$0.77 of original size. However, that of control group was 24.88${\pm}$2.90. 3. The Cinamon's score of AM-Collagen and $Terudermis^{(R)}$. group at 4 weeks was 15.6${\pm}$1.26 and 14.6${\pm}$3.13 and that of control group was 3.7${\pm}$0.95. Significant difference was observed among control and experimental groups(p<0.05). 4. Histologic examination revealed that AM protected leukocyte infiltration and epithelial migration was nearly completed at 4 weeks. $Terudermis^{(R)}$. group showed mild neutrophil infiltration until 2 weeks and completion of epithelization at 4 weeks. Control group showed massive leukocyte infiltration until 4 weeks. 5. Microvessels were increased sharply at 1 week and control group at 1 and 4 week showed significant differences with $Terudermis^{(R)}$. group of same interval(p<0.05) but no differences were found with AM group(p<0.05). Conclusion: EGF and EGF-R were well preserved in freeze-dried AM. AM attached to collagen acted as excellent biologic dressing which had similar effect with $Terudermis^{(R)}$. AM showed anti-inflammatory action and healing was completed at 4 weeks after full-thickness skin defect.

• Archives of Reconstructive Microsurgery
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• v.9 no.2
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• pp.114-119
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• 2000

The soft tissue defects including the Achilles tendon are complex and very difficult to reconstruct. Recently, several free composite flaps including the tendon have been used to reconstruct large defects in this area in an one-stage effort. Our case presents a patient reconstructed with free composite dorsalis pedis flap along with the extensor digitorum longus and superficial peroneal nerve for extensive defects of the Achilles tendon and surrounding soft tissue. A 36-year-old-man sustained an open injury to the Achilles tendon. He was referred to our department with gross infection of the wound and complete rupture of the tendon associated with loss of skin following reduction of distal tibial bone fracture. After extensive debridement, $6{\times}8cm$ of skin loss and 8cm of tendon defect was noted. Corresponding to the size of the defect, the composite dorsalis pedis flap was raised as a neurosensory unit including the extensor digitorum longus to provide tendon repair and sensate skin for an one-stage reconstruction. One tendon slip was sutured to the soleus musculotendinous portion, the other two were sutured to the gastrocnemius musculotendinous portion with 2-0 Prolene. The superficial peroneal nerve was then coaptated to the medial sural cutaneous nerve. The anterior tibial artery and vein were anastomosed to the posterior tibial artery and accompanying vein in an end to end fashion. After 12 months of follow-up, 5 degrees of dorsiflexion due to the checkrein deformity and 58 degrees of plantar flexion was achieved. The patient was able to walk without crutches. Twopoint discrimination and moving two-point discrimination were more than 1mm at the transferred flap site. The donor site healed uneventfully. Of the various free composite flaps for the Achilles tendon reconstruction when skin coverage is also needed, we recommand the composite dorsalis pedis flap. The advantages such as to control infection, adequate restoration of ankle contour for normal foot wear, transfer of the long tendinous portion, and protective sensation makes this flap our first choice for reconstruction of soft tissue defect including the Achilles tendon.