• Title/Summary/Keyword: Wound culture

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PRODUCTION OF TRANSFORMING GROWTH FACTOR-${\beta}_1$ IN HUMAN FIBROBLASTS INDUCED WITH BACTERIAL TOXINS (세균 독소를 작용시킨 섬유아 세포에서 Transforming Growth Factor-${\beta}_1$의 생성)

  • Lee, Seong-Geun;Kim, Kwang-Hyuk
    • Journal of the Korean Association of Oral and Maxillofacial Surgeons
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    • v.26 no.4
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    • pp.345-354
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    • 2000
  • TGF-${\beta}_1$ is a potent chemotactic factor for inflammatory cells and fibroblasts. It also stimulates the celluar source and components of extracellular matrix and the production of proteinase inhibitors. Collectively, these biologic activities lead to the accumulation and stabilization of the nascent matrix, which is vital to wound healing. The objective of this study is to investigate production of TGF-${\beta}_1$ in vitro fibroblast culture in the presence of Staphylococcus enterotoxin B(SEB) and/or lipopolysaccharide(LPS) and to elucidate the role of TGF-${\beta}_1$ which may be responsible for wound healing. The fibroblasts were originated from facial dermis and hypertrophic scar in 26 year-old male patient. In the presence of LPS($0.01{\mu}g$, $0.1{\mu}g$, $1.0{\mu}g$), SEB($0.01{\mu}g$, $0.1{\mu}g$, $1.0{\mu}g$) respectively, cells($5{\times}10^3ml$) were cultivated in vitro. At 1, 3, and 5 days after incubation, cells were counted. Also, cells($2.5{\times}10^5ml$) were cultivated in EMEM with LPS(0.01, 0.1 and $1.0{\mu}g$), SEB(0.01, 0.1 and $1.0{\mu}g$) respectively and LPS($0.1{\mu}g$) and SEB($0.1{\mu}g$) in combination for 24, 48, and 72 hours respectively. Culture supernatants were harvested at 1, 2, and 3 days after incubation period and triplicate culture supernatants were pooled and TGF-${\beta}_1$ was assayed in duplicate. The results were as follows. 1. In facial dermal fibroblast induced with SEB and LPS respectively or in combination, the suppression of cell proliferation occurred very significantly at 1 day after incubation, compared with the control. In SEB exposure, the production of TGF-${\beta}_1$ was decreased very significantly at 1 day after incubation, compared with the control. However, in LPS, SEB and LPS exposure, the production of TGF-${\beta}_1$ was increased very significantly at 1 day after incubation, compared with the control. 2. In hypertrophic scar fibroblast induced with SEB and LPS respectively or in combination, the suppression of cell proliferation did not occur at 1 day after incubation, compared with the control. In SEB and LPS exposure in combination, the production of TGF-${\beta}_1$ was increased very significantly at 1 day after incubation, compared with the control. However, the production of TGF-${\beta}_1$ did not occur in SEB and LPS exposure respectively. In conclusion, the concentration of bacterial toxins and the incubation period correlated with cell proliferation and production of TGF-${\beta}_1$ very significantly and both fibroblasts have different phenotype each other in this regard. This data suggest that the significant production of TGF-${\beta}_1$ may develope abnormal wound healing associated with tissue fibroproliferative disorder, such as hypertrophic scar and keloid formation.

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Complicated Wound Infection Following Transvenous Endocardial Pacemaker (경정맥(經靜脈) 내적(內的) Pacemaker 이식후(移植後) 발생(發生)한 복잡한 감염합병증(感染合?症)의 1치험례(治驗例))

  • Lee, D.Y.;Yoon, Y.J.;Cho, B.K.;Hong, S.N.
    • Journal of Chest Surgery
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    • v.9 no.2
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    • pp.265-270
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    • 1976
  • Implantation of a permanent pacemaker is a widely accepted procedure for the patient with complete heart block.As a result of these device, the prognosis for patients with Adams-Stokes syndrome caused by complete A-V block and other cardiac arrhythmia have become much more optimistic. Permanent pacemaker implantation by means of a transvenous approach has made the operative risk much less and the procedure simpler. However, a number of complications have been reported in the literature regarding transvenous endocardial pacemaker implantation during the last a decade. The patient presented in this paper is a 26-year old girl who was implanted with a permanent pacemaker at 14 years of age because of a congenital A-V block. Following first exchange of pulse generator, the electrode (lead) was fractured, so that by the pulse generator, a change to the transvenous technique of implantation was made, After this, there were episodes of recurrent wound infection on three occasions, even though the site of pulse generator implantation was exchanged to the contralateral side of chest wall, massive doses of antibiotics were administered and sensitivity tests for coagulase positive staphylococcal infection were performed. Though there was no definite evidence of blood stream infection by blood culture, we decided not to use the transvenous technique and not to implant the pulse generator in the chest wall because the venous system and the entire anterior chest wall appeared to be diseased or contaminated by virulent pyogenic organisms. Finally this intractable systemic and local wound infection was successfully controlled by myocardial lead implantation via a subxiphoid approach and implantation of the pulse generator far down in the abdominal wall. The causes and routes of recurrent wound infection and possible blood born infection in this particular patient are still obscure. We strongly believe that myocardial pacemaker implantation is much safer than transvenous endocardial pacemaker implantation & myocardial pacemaker implantation is a definite method for controlling such an intractable wound infection. following transvenous pacemaker implantation.

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THE INDUCTIVE CAPACITY OF PRIMARY CULTURED ORAL MUCOSAL KERATINOCYTES IN SKIN WOUND HEALING OF ATHYMIC NUDE MICE (배양된 구강점막 각화상피세포가 누드마우스 피부 창상 치유에 미치는 효과)

  • Kim, Hyun-Sil;Kim, Nam-Hee;Kim, Jin;Cha, In-Ho
    • Journal of the Korean Association of Oral and Maxillofacial Surgeons
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    • v.30 no.4
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    • pp.308-315
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    • 2004
  • Aim: The aim of this study was to investigate the mechanism of promoted skin wound healing in skin defects with primary cultured oral mucosal keratinocytes. Materials and methods: Thirty adult female nude mice weighing $20{\pm}2g$ were used for the experiment. Primary cultured and suspended oral mucosal keratinocytes, labeled with BrdU, were scattered onto $1.5cm{\times}1.5cm$ sized full thickness skin defects in the experimental group(N=15), and no grafts were placed the control group(N=15). They were sacrificed at 3 days, 1 week and 2 weeks after the treatment respectively. Histological examination of each wounds were performed to review the healing progress on measuring the length from the wound margin to regenerating epithelial front. The role of keratinocytes were assessed by double immunohistochemical staining with Anti-BrdU and Anti-cytokeratin AE1/3. Results: In the experimental group the wound was completely covered with regenerating epithelia in 2 weeks, but partially regenerated in the control group. The immunohistochemical studies unexpectedly reveal that most of regenerating epithelial cells were induced from marginal epithelium of the margin, not from the scattered keratinocytes. Conclusion: We could successfully confirm that graft of primary cultured oral mucosal keratinocytes promotes the regeneration of skin defects.

Treatment for Hydrofluoric Acid Chemical Burn Using Acticoat® (Acticoat®를 이용한 불산에 의한 화학 화상의 치료)

  • Choi, Hwan Jun;Wee, Syeo Young;Choi, Chang Yong
    • Archives of Plastic Surgery
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    • v.35 no.4
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    • pp.407-412
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    • 2008
  • Purpose: Hydrofluoric acid(HF) is one of the most dangerous mineral acids with the dissociated fluoride ions. The initial corrosive burn is caused by free hydrogen ion, and the second and more severe burn is caused by penetration of fluoride ions into subcutaneous tissues. Silver is a cation producing dressing, an effective antimicrobial agent, but older silver-containing formulations are rapidly inactivated by wound environment, requiring frequent replenishment. But, $Acticoat^{(R)}$ is a relatively new form of silver dressing which helps avoid the problems of earlier agents. The aim of this study is to evaluate effects of $Acticoat^{(R)}$, silver-containing dressing on the treatment for HF injury wound. Methods: From september 2006 to september 2007, the study was carried out with 10 patients who had HF partial thickness burns. $Acticoat^{(R)}$ dressing and 10% calcium gluconate wet gauze dressings in 10 cases. As a principle, in the emergency treatment, partial or complete removal of the nail and early bullectomy along with copious washing with normal saline was done, depending on the degree of HF invasion of the wound. Wound was dressed with $Acticoat^{(R)}$ and 10% calcium gluconate solution. The effect of dressing was investgated by serial bacterial culture and wound exudates assessment. Results: We therefore reviewed 10 cases of HF-induced chemical burns and treatment principle. The 10 cases who came to the hospital nearly immediately after the injury healed completely without sequelae. Conclusion: As the industrial sector develops, the use of HF is increasing more and more, leading to increased incidences of HF-induced chemical burns. The education of patients regarding this subject should be empathized accordingly. In conclusion, $Acticoat^{(R)}$ dressing is a better choice for HF partial thickness burn injuries because of shorter healing time, less pain and more comfortable dressing.

Recombinant human granulocyte macrophage colony-stimulating factor (rhGM-CSF) could accelerate burn wound healing in hamster skin

  • Heo, Si-Hyun;Han, Kyu-Boem;Lee, Young-Jun;Kim, Ji-Hyun;Yoon, Kwang-Ho;Han, Man-Deuk;Shin, Kil-Sang;Kim, Wan-Jong
    • Animal cells and systems
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    • v.16 no.3
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    • pp.207-214
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    • 2012
  • Burns are one of the most devastating forms of trauma and wound healing is a complex and multicellular process, which is executed and regulated by signaling networks involving numerous growth factors, cytokines, and chemokines. Recombinant human granulocyte macrophage colony-stimulating factor (rhGM-CSF) was specifically produced from rice cell culture through use of a recombinant technique in our laboratory. The effect of rhGM-CSF on promotion of deep second-degree burn wound healing on the back skin of a hamster model was evaluated through a randomized and double-blind trial. As macroscopic results, hamster skins of the experimental groups showed earlier recovery by new epidermis than the control groups. Immunohistochemical reactions of proliferating cell nuclear antigen and transforming growth factor-b1, which are indicators of cell proliferation, were more active in the experimental group, compared with the control group. On electron microscopy, basal cells in the epidermis of the experimental group showed oval nuclei, prominent nucleoli, numerous mitochondria and abundant free ribosomes. In addition, fibroblasts contained well-developed rough endoplasmic reticulum with dilated cisternae. Bundles of collagen fibrils filled the extracellular spaces. Particularly, ultrastructural features indicating active metabolism for regeneration of injured skin at 15 days after burn injury, including abundant euchromatin, plentiful free ribosomes, and numerous mitochondria, were observed. These findings suggest that use of rhGM-CSF could result in accelerated deep second-degree burn wound healing in animal models.

Development of Hair Keratin Protein to Accelerate Oral Mucosal Regeneration

  • So-Yeon Kim
    • Journal of dental hygiene science
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    • v.23 no.4
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    • pp.369-377
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    • 2023
  • Background: In this study, we investigated the potential use of keratin for oral tissue regeneration. Keratin is well-known for its effectiveness in skin regeneration by promoting keratinization and enhancing the elasticity and activity of fibroblasts. Because of its structural stability, high storability, biocompatibility, and safety in humans, existing research has predominantly focused on its role in skin wound healing. Herein, we propose using keratin proteins as biocompatible materials for dental applications. Methods: To assess the suitability of alpha-keratin protein as a substrate for cell culture, keratin was extracted from human hair via PEGylation. Viabilities of primary human gingival fibroblasts (HGFs) and human oral keratinocytes (HOKs) were assessed. Fluorescence immunostaining and migration assays were conducted using a fluorescence microscope and confocal laser scanning microscope. Wound healing and migration assays were performed using automated software to analyze the experimental readout and gap closure rate. Results: We confirmed the extraction of alpha-keratin and formation of the PEG-g-keratin complex. Treatment of HGFs with keratin protein at a concentration of 5 mg/ml promoted proliferation and maintained cell viability in the test group compared to the control group. HOKs treated with 5 mg/ml keratin exhibited a slight decrease in cell proliferation and activity after 48 hours compared to the untreated group, followed by an increase after 72 hours. Wound healing and migration assays revealed rapid closure of the area covered by HOKs over time following keratin treatment. Additionally, HOKs exhibited changes in cell morphology and increased the expression of the mesenchymal marker vimentin. Conclusion: Our study demonstrated the potential of hair keratin for soft tissue regeneration, with potential future applications in clinical settings for wound healing.

Effectiveness of Nanocrystalline Silver(Acticoat®) Dressing at Wound Infected by Multidrug Resistant Bacteria (다제내성(多劑耐性)을 지닌 균주에 감염된 상처에서 Nanocrystalline Silver(Acticoat®) 드레싱의 효용)

  • Jeong, Tae Kwang;Yang, Ho Jik
    • Archives of Plastic Surgery
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    • v.34 no.6
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    • pp.691-696
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    • 2007
  • Purpose: The emergency of multi-drug resistant stains of bacteria represents a challenge in the field of plastic surgery. Especially, MRSA(methycillin-resistant Staphylococcus aureus) and Pseudomonas aeruginosa have strong pathogenicity as well as multi-drug resistance so that they have become a lot more problematic strains. This study has been planned to reduce the bacterial burden by applying $Acticoat^{(R)}$(Smith & Nephew Healthcare, Hull, England)dressing into the chronic wounds infected by multi-drug resistant strains and to facilitate their healing. Methods: Nanocrystalline silver dressings($Acticoat^{(R)}$) were applied to chronic wound infected by MRSA or Pseudomonas aeruginosa. Multi-drug resistant bacteria were smeared over a slide glass using sterilized cotton swabs and gram stains were performed directly before and after applying $Acticoat^{(R)}$ dressings at 1, 24, 48 and 72 hours. The gram-stained slides were observed using an optical microscope magnified 1000 times(${\times}1000$). The bacterial counts of the control group(0 hour) were compared to those of the experimental groups(1, 24, 48, and 72 hour). Paired T-test was used to assess a statistical significance. MRSA was cultured in two BAPs(blood agar plate) and two MacConkey plates with streak plate method. None were interventions on one culture plate, while on the other culture plate, $Acticoat^{(R)}$ was placed in a square shape and cultured for 72 hours at $37^{\circ}C$, then plates were examined. Pseudomonas aeruginosa was cultured in the same manner as MRSA. Results: There are the large amount of declination of bacterial counts with statistical significance after $Acticoat^{(R)}$ dressing. The bacteria grew in culture plate without specific intervention, but no bacteria grew in culture plate with applying of $Acticoat^{(R)}$ dressing. Conclusion: We believe that $Acticoat^{(R)}$ dressing could be used as an effective method of treating chronic wounds which are infected by multi-drug resistant organisms.

The healing effect of rhGM-CSF on uninfected wounds (rhGM-CSF(Leucogen)의 비감염성 상처 치유 효과에 관한 연구)

  • Han, Seung Kyu;Kim, Byung Soo;Kim, Aeree;Seo, Jae Hong;Choi, Chul Won;Shin, Sang Won;Kim, Yeul Hong;Kim, Woo Kyung;Kim, Jun Suk
    • IMMUNE NETWORK
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    • v.1 no.1
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    • pp.32-35
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    • 2001
  • Background: rhGM-CSF has been shown to enhance the migration and proliferation of endothelial cells and to promote keratinocyte growth. This study was tried to evaluate the effect of rhGM-CSF dressing on the uninfected wounds. Methods: Thirty Sprague-dawley white mice(250-300g) were selected in this study. The number of wound with the diameter of 5 mm, was 3 in left and 3 in right at the symmetric sites, respectively. The site of rhGM-CSF dressing was decided by a randomization. rhGM-CSF($Leucogen^{(R)}$) was diluted in the distilled water($5{\mu}g/mL$). The experimental wound group was dressed by l mL of distilled water mixed with rhGM-CSF and control wound group was dressed by l mL of distilled water. The dressing was done, every 24 hours. The criteria of comparison were the duration of wound healing duration, histologic findings and the bacterial culture of wound sites. Results: The duration of wound healing was $10.3{\pm}1.7days$ in experimental group and $10.2{\pm}2.8days$ in control group, without significant difference. There was no specific difference of histologic findings between both groups. The pathogen was not found, at all. Conclusion: It seems to be that rhGM-CSF has no prominent effect on the uninfected wound healing in the mice without immune suppression.

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Effect of Silver sulfadiazine on the Skin Cell Proliferation and Wound Healing Process in Hairless Mouse 2nd degree Burn Model (설파디아진은의 피부세포 증식 및 화상모델에 있어서의 상처치유과정에 미치는 영향)

  • Cho, Ae-Ri
    • Journal of Pharmaceutical Investigation
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    • v.32 no.2
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    • pp.113-117
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    • 2002
  • Cyto-toxic effect of silver sulfadiazine (Ag-SD) on keratinocytes and its implication on wound healing process were investigated in $2^{nd}$ degree bum hairless mouse model. As a dermal model, HaCat (immortalized keratinocytes) monolayer culture in DMEM with 10% FBS was used. Cyto-toxicity of Ag-SD was estimated by measuring the cell viability using neutral red assay after adding the drug. The $2^{nd}$ degree bum was prepared on hairless mouse back skin (1 cm diameter) and dressings with Ag-SD were applied for 96 hr. The process of re-epithelialization and the presence of inflammatory cells were investigated and histology with Hematoxylin-Eosin staining was performed. Ag-SD displayed highly cyto-toxic effect on cultured HaCat cells in a concentration dependent manner $(1-100\;{\mu}g/mL)$. Topical application of Ag-SD (2%) could control the infection: no inflammatory cells were observed in histology. However the cyto-toxic effect of Ag-SD on skin cells induced the impairment in epidermal regeneration.

Subeschar culture using a punch instrument in unstageable wounds

  • Jung, Han Byul;Lee, Yong Jig
    • Archives of Plastic Surgery
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    • v.47 no.3
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    • pp.228-234
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    • 2020
  • Background A patient's overall condition sometimes does not allow for the complete removal of a dead eschar or injured slough in cases involving a pressure-injury skin lesion. This frequently occurs in clinical practice, particularly in bedridden and older patients receiving home care or intensive care. Even after debridement, it is also difficult to manage open exudative wounds in these patients. Nevertheless, when a mature or immature eschar is treated without proper debridement, liquefaction necrosis underneath the eschar or slough tends to reveal a large, open wound with infectious exudates. We hypothesized that if the presence of any bacteria under the eschar can be evaluated and the progression of the presumed infection of the subeschar can be halted or delayed without creating an open wound, the final wound can be small, shallow, and uninfected. Methods Using a punch instrument, we performed 34 viable subeschar tissue cultures with a secure junction between the eschar and the normal skin. Results The bacterial study had 29 positive results. Based on these results and the patient's status, appropriate antibiotics could be selected and administered. The use of suitable antibiotics led to relatively shallow and small exposed wounds. Conclusions This procedure could be used to detect potentially pathogenic bacteria hidden under black or yellow eschars. Since subeschar infections are often accompanied by multidrug-resistant bacteria, the early detection of hidden infections and the use of appropriate antibiotics are expected to be helpful to patients.