• Title, Summary, Keyword: Dermal matrix

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Periodontal tissue engineering by hPDLF seeding on scaffold (Scaffold상에 식립한 사람치주인대섬유모세포를 통한 치주조직공학)

  • Kim, Seong Sin;Kim, Byung-Ock;Park, Joo-Cheol;Jang, Hyun-Seon
    • Journal of Periodontal and Implant Science
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    • v.36 no.3
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    • pp.757-765
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    • 2006
  • Human periodontal ligament fibroblasts (hPDLF) are very important for curing the periodontal tissue because they can be differentiated into various cells. A tissue engineering approach using a cell-scaffold is essential for comprehending today's periodontal tissue regeneration procedure. This study examined the possibility of using an acellular dermal matrix as a scaffold for human periodontalligament fibroblast (hPDLF). The hPDLF was isolated from the middle third of the root of periodontally healthy teeth extracted for orthodontic reasons. The cells were cultured in a medium containing Dulbecco's modified Eagle medium supplemented with 10% fetal bovine serum at $37^{\circ}C$ in humidified air with 5% $CO_2$. The acellular dermal matrix(ADM) was provided by the US tissue banks(USA). Second passage cells were used in this study. The hPDLF cells were cultured with the acellular dermal matrix for 2 days, and the dermal matrix cultured by the hPDLF was transferred to a new petri dish and used as the experimental group. The control group was cultured without the acellular dermal matrix, The control and experimental cells were cultured for six weeks. The hPDLF cultured on the acellular dermal matrix was observed by Transmission Electron microscopy (TEM). Electron micrography shows that the hPDLF was proliferated on the acellular dermal matrix. This study suggests that the acellular dermal matrix can be used as a scaffold for hPDLF.

HEALING PATTERNS OF THE ACELLULAR DERMAL MATRIX DEPEND ON GRAFT METHOD IN THE RABBIT EARS (가토의 귀에서 무세포성 진피 기질의 이식 방법에 따른 치유 양상)

  • Ryu, Jae-Young;Ryu, Sun-Youl
    • Maxillofacial Plastic and Reconstructive Surgery
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    • v.31 no.3
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    • pp.216-221
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    • 2009
  • Purpose: The retention of the basement membrane complex, which was the unique feature of the acellular dermal matrix ($AlloDerm^{(R)}$), plays an important role in the normal process of wound healing. The present study was aimed to compare the healing of the acellular dermal matrix according to the graft method in the rabbit ear. Materials and methods: Six mature rabbits weighing about 3.0 kg were used, $10\;{\times}\;5\;mm$ sized subcutaneous pockets were created between the ear skin and the underlying perichondrium. In the control group, the acellular dermal matrix was grafted with the basement membrane facing toward the perichondrium. On the contrary, the acellular dermal matrix was grafted with the basement membrane facing toward the skin side in the experimental group I. In the experimental group II, the acellular dermal matrix was grafted like rolled configuration with basement membrane side in. The grafted site was picked at 3, 7, and 21 days after the graft. Serial sections were processed by H-E stain and examined under light microscopy to assess the healing patterns. Results: There was no distinct volume loss in the gross examination, but resorption was observed from the edge of the acellular dermal matrix in the histological examination. The space of resorption was replaced by the newly formed fibrous tissues and vessels. The inflammatory cells were more increased at 7 days after the graft than the early days. However, inflammation was decreased at 21 days after the graft. Regardless of the graft direction, no differences were observed between the control and the experimental group I in the healing patterns. Conclusion: These results suggest that the acellular dermal matrix can be used simply and effectively without regard to the graft direction as a substitute of autogenous material for repairing soft tissue defect.

The Durability of Elastin-Incorporated Collagen Matrix for Dermal Substitute in Vitro Condition (In vitro 환경에서 엘라스틴을 혼합한 콜라겐 진피 지지체의 내구성)

  • Lew, Dae Hyun;Hong, Jong Won;Tark, Kwan Chul
    • Archives of Plastic Surgery
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    • v.35 no.1
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    • pp.7-12
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    • 2008
  • Purpose: Since the report of artificial dermis manufacturing method using collagen by Yannas in 1980, collagen has been effectively used as dermal substitute with its merits such as, lower antigeneicity, controllable biodegradation rate, and minimal inflammatory cytotoxic properties in the dermal tissue engineering field. However, weak mechanical durability was the main drawback of collagen dermal substitute. To improve its stability, mechanical or chemical cross-linking was used. Despite of such process, its clinical use was restricted due to weak durability. To improve the durability of collagen matrix, we designed elastin-incorporated collagen matrix and compared its durability with conventional collagen matrix. Methods: 15mm diameter with 4mm thick collagen dermal matrix was made according to Yannas protocol by mixing 0.5% bovine collagen and chondroitin-6-sulfate followed by degassing, freeze drying, dehydrodermal cross-linking and chemical cross-linking procedure. In elastin incorporated collagen matrix, same procedure was performed by mixing elastin to previous collagen matrix in 4:1 ratio(collagen 80% elastin 20%). In comparison of the two dermal matrix in vitro tests, matrix contracture rate, strain, tensile strength, was measured and stiffness was calculated from comparative analysis. Results: In terms of matrix contracture, the elastin-incorperated added collagen dermis matrix showed 1.2 times more contraction compared to conventional collagen matrix. However, tensile strength showed 1.6 times and stiffness showed 1.6 times increase in elastin-incorporated matrix. Conclusion: Elastin incorperated collagen matrix manufactured by our team showed increased durability due to improvement in tensile strength and stiffness compared to previous collagen matrix($Integra^{(R)}$).

Enhancing Dermal Matrix Regeneration and Biomechanical Properties of $2^{nd}$ Degree-Burn Wounds by EGF-Impregnated Collagen Sponge Dressing

  • Cho Lee Ae-Ri
    • Archives of Pharmacal Research
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    • v.28 no.11
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    • pp.1311-1316
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    • 2005
  • To better define the relationship between dermal regeneration and wound contraction and scar formation, the effects of epidermal growth factor (EGF) loaded in collagen sponge matrix on the fibroblast cell proliferation rate and the dermal mechanical strength were investigated. Collagen sponges with acid-soluble fraction of pig skin were prepared and incorporated with EGF at 0, 4, and 8 $\mu$g/1.7 $cm^{2}$. Dermal fibroblasts were cultured to 80$\%$ confluence using DMEM, treated with the samples submerged, and the cell viability was estimated using MTT assay. A deep, $2^{nd}$ degree- burn of diameter 1 cm was prepared on the rabbit ear and the tested dressings were applied twice during the 15-day, post burn period. The processes of re-epithelialization and dermal regeneration were investigated until the complete wound closure day and histological analysis was performed with H-E staining. EGF increased the fibroblast cell proliferation rate. The histology showed well developed, weave-like collagen bundles and fibroblasts in EGF-treated wounds while open wounds showed irregular collagen bundles and impaired fibroblast growth. The breaking strength (944.1 $\pm$ 35.6 vs. 411.5 $\pm$ 57.0 Fmax, $gmm^{-2}$) and skin resilience (11.3 $\pm$ 1.4 vs. 6.5 $\pm$ 0.6 mJ/$mm^{2}$) were significantly increased with EGF­treated wounds as compared with open wounds, suggesting that EGF enhanced the dermal matrix formation and improved the wound mechanical strength. In conclusion, EGF-improved dermal matrix formation is related with a lower wound contraction rate. The impaired dermal regeneration observed in the open wounds could contribute to the formation of wound contraction and scar tissue development. An extraneous supply of EGF in the collagen dressing on deep, $2^{nd}$ degree-burns enhanced the dermal matrix formation.

Autotransplantation using the acellular dermal matrix seeded by periodontal ligament fibroblasts in minipig: histological evaluation as potential periodontal ligament substitutes (미니돼지에서 Acellular dermal matrix에 배양된 치주인대섬유모세포을 이용한 자가치아이식술: 치주인대로써의 잠재력에 대한 조직학적 평가)

  • Yu, Sang-Joun;Kim, Byung-Ock;Park, Joo-Cheol;Jang, Hyun-Seon
    • Journal of Periodontal and Implant Science
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    • v.37 no.1
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    • pp.53-64
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    • 2007
  • The aim of this study was to examine the possibility of periodontal ligament regeneration when autotransplantation was used by the periodontal ligament fibroblasts cultured on the acellular dermal matrix in teeth without a periodontal ligament. One minipig was used in this study. The mandibular and maxillary permanent incisors were ex-tracted for the culture of the periodontal ligament cells. The roots of the unextracted teeth were classified into a positive control group, in which the normal periodontal ligament was preserved. The roots of the extracted teeth were divided into the following two groups: The negative control group, in which the periodontal ligament had been removed and the acellular dermal matrix was not applied; and an experimental group, in which the periodontal ligament had been removed and periodontal ligament fibroblast cultured on an acellular dermal matrix was applied. The prepared teeth were transplanted, and completely submerged using physical barrier membranes. The animal was sacrificed 4 weeks after the autotransplant. The transplanted teeth were examined histologically. In this study, the periodontal ligament was normal in the positive control group, and ankylosis was discovered on the denuded root surface in the negative control group. Periodontal ligament-like connective tissue was found adjacent to the denuded root and the new cementum-like layer of hard tissue was formed in the experimental group. These results suggest that the periodontal ligament fibroblasts cultured on the acellular dermal matrix may play a role in regenerating the periodontal ligament-like tissue with new cememtum-like tissue formation.

Treatment of Aplasia Cuti Congenita Using Allogenic Dermal matrix and Cultured Epithelial Autograft: A Case Report (동종사체진피와 배양한 자가상피세포를 이용한 선천성 피부 형성 부전증 환자의 치험례)

  • Lee, Jin Hwa;Kim, Yong Kyu;Lee, Sang Joon
    • Archives of Plastic Surgery
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    • v.33 no.5
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    • pp.672-675
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    • 2006
  • Purpose: The purpose of this study is to document the surgical methods used in infants with aplasia cutis congenita treated with allogenic dermal matrix and cultured epithelial autografts. Methods: The large defects in both lower legs were replaced with allogenic dermal matrix to avoid the postoperative hypertrophic scar contracture and a full-thickness skin biopsy was taken from right groin area simultaneously. We sent the specimen to a commercial laboratory for culture and obtained cultured epithelial autografts($Holoderm^{(R)}$) after 2 weeks, placed it over the allogenic dermal matrix. Results: The skin-defected area were nearly epithelialized after 2 weeks and there were no significant problem on during 6 months follow-up. Conclusion: The surgical method using allogenic dermal matrix and cultured epithelial autograft provided an excellent coverage of large skin defects of infant with aplasia cutis congenita.

Peptide Hydrolysates from Astragalus membranaceus Bunge Inhibit the Expression of Matrix Metalloproteinases in Human Dermal Fibroblasts

  • Park, Sun Ki;Van Hien, Pham;Van Luong, Hoang;Yan, Shao-Wei;Byun, Sang Yo
    • KSBB Journal
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    • v.29 no.5
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    • pp.380-384
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    • 2014
  • Inhibition effects of peptide hydrolysates from Astragalus membranaceus Bunge. on the expression of the matrix metalloproteinases (MMPs) in human dermal fibroblasts were evaluated in vitro. Crude peptides were obtained by the hydrolysis of proteins extracted from A. membranaceus. Peptides were purified partially by the basis on the molecular weight using 40% polyacrylamide gel electrophoresis before treatment with human dermal fibroblasts. Basis on the doseeffect experiments, expressions of MMPs including MMP-1, MMP-3, MMP-8, MMP-13 in human dermal fibroblasts were evaluated. Expressions of MMP-1, MMP-3, MMP-8 and MMP-13 were reduced in 43%, 5%, 22% and 57% respectively. The mass spectrometric analysis of partially purified peptides from A. membranaceus, which strongly inhibit expressions of MMPs, indicated that the peptides were composed of molecules below 1500 Da.

Effect of Avidin and Biotin in Attachment of Human Adipose Stem Cells to Micronized Acellular Dermal Matrix (지방줄기세포가 무세포 분쇄진피기질(Acellular micronized dermal matrix) 부착에 있어 Avidin과 Biotin의 효과)

  • You, Gyeol;Rhie, Jong Won;Lim, Jin Soo
    • Archives of Plastic Surgery
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    • v.35 no.1
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    • pp.1-6
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    • 2008
  • Purpose: In tissue engineering, it is important that the scaffolds have high affinity with cells for making efficient use of cells. The authors studied the binding affinity of human adipose stem cells(ASCs) to micronized acellular dermal matrix(alloderm) using biotin and avidin linkages.Methods: Human ASCs were harvested from adipose tissue obtained by abdominoplasty. ASCs($1{\times}10^4$, $5{\times}10^4$, $1{\times}10^5$, $5{\times}10^5$, $1{\times}10^6$, $5{\times}10^6$ cells) were attached to micronized alloderm(1mg) in three groups; 1) control group in which no ASCs and alloderm was treated; 2) serum group in which alloderm was exposed to fetal bovine serum; and 3) biotin group in which biotinylated cells were attached to biotinylated alloderm. The binding affinities were determined 1 day after making ASC-alloderm complexes. The proliferation rates were determined by XTT assays in 4, 7, 14, and 21 days and scanning electron microscopic examination was performed in 7 and 21 days after culture of ASC-alloderm complexes.Results: The binding affinities of the biotin group were significantly increased in all cell concentrations. Maximum binding affinity was observed at $5{\times}10^4/mg$ of micronized dermal matrix in biotin group. The viabilities were lowest in biotin group in contrast to binding affinity, but the difference was not significant. SEM showed well attachment of cells to micronized dermal matrix in all groups. Conclusion: The use of avidin/biotin facilitated human ASCs attaching to micronized acellular dermal matrix. This attachment would not disturb adipose stem cells viabilities. The present study suggests that avidin/ biotin can be used as making efficient use of cells in adipose tissue engineering.

Protective actions of Rubus coreanus ethanol extract on collagenous extracellular matrix in ultraviolet-B irradiation-induced human dermal fibroblasts

  • Bae, Ji-Young;Lim, Soon-Sung;Choi, Jung-Suk;Kang, Young-Hee
    • Nutrition Research and Practice
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    • v.1 no.4
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    • pp.279-284
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    • 2007
  • Solar ultraviolet (UV) irradiation leads to distinct changes in the skin connective tissues by degradation of collagen, which is a major structural component in the extracellular matrix. UV irradiation induces the production of matrix metalloproteinases (MMP) capable of attacking native fibrillar collagen and responsible for inhibiting the construction of collagenous extracellular matrix. In this study, we attempted to investigate the protective actions of Rubus coreanus ethanol extract (RCE) on the MMP production and the consequent procollagen/collagen degradation in UV-B-irradiated human dermal fibroblasts. The analytical data showed that Rubus coreanus ethanol extract was mostly comprised of cyanidin 3-rutinoside. Pre-treatment of fibroblasts with this extract inhibited UV-B-induced production of MMP-1, MMP-8 and MMP-13 in dose-dependent manners. In addition, Western blot analysis and immunocytochemical staining assay revealed that RCE markedly augmented the cellular levels of procollagen/collagen declined in UV-B-exposed dermal fibroblasts. These results demonstrate that RCE blocks UV-B-induced increase of the collagen degradation by inhibiting MMP production. Thus, RCE may act as an agent inhibiting excessive dermal collagen degradation leading to the skin photoaging.