Fabrication and Characterization of Three-Dimensional Silk Fibroin Scaffolds Using a Mixture of Salt/Sucrose

문보미,박찬흠,김동규,박현정,주형우,이옥주,김정호,이정민,이중섭 한국고분자학회 2014 Macromolecular Research Vol.22 No.12

This study described the preparation of three-dimensional (3-D) silk fibroin (SF) scaffolds, which derivedfrom a mixed salt/sucrose leaching without the process of lyophilization. Compared with the previous method, thisfabrication of 3-D SF scaffold is simple and can have more diverse morphological appearances of pores dependingon the different salt/sucrose mixing concentrations. The correlation between pore shape and septum of SF scaffoldsor salt concentration was examined by SEM. Additionally, the effects of salt or glucose concentrations on mechanicalproperties, such as compressive modulus in dry and wet states, were studied. For the materials with rangedbetween 69-88%, water binding capacity decreased with increasing salt or decreasing sucrose. After cell seeding andculture, SEM images of scaffolds showed that cell infiltrations were increased along with increased sucrose concentration. Analysis of Masson’s trichrome staining showed positive findings for collagen deposits in scaffolds of the100% sucrose type. Thus, our new method can fabricate 3-D SF scaffolds with different characteristics, and may beused for various tissue engineering uses.

도축장 출하돈의 돼지써코바이러스 발생율과 유전형 분석

문보미 ( Bo-mi Moon ),추금숙 ( Keum-sook Chu ),김승채 ( Seung-chai Kim ),김환주 ( Hwan-ju Kim ),김다정 ( Da-jeong Kim ),김원일 ( Won-il Kim ) 한국동물위생학회 2023 韓國家畜衛生學會誌 Vol.46 No.4

This study was carried out to investigate the genotypic diversity of PCV2 and co-infection of PCV3 in the hilar lymph nodes of 700 randomly-selected slaughter pigs. Fourteen samples per each farm were obtained from 50 farms between February and August in 2022. Of the 50 farms, 44 farms that had been positive for PCV2 by RT-PCR were genotyped. As a result of PCV2 genotyping, positive rate of PCV2 DNA was 62.3% (436/700). Among the PCV2 DNA-positive samples, positive rate of a single PCV2 genotype was 79.1% (345/436), while multiple PCV2 genotypes were only detected in 20.9% (91/436). Of the 436 single infection cases, PCV2d genotype was most prevalent. Positive rates of PCV2 and PCV3 were 53.6% and 26.0% at the sample level, 5.1% and 8.0% at the farm level, respectively. And the co-positive rate of two viruses was 8.7% (61/700) at the sample level, 62.0% (31/50) at the farm level. These results demonstrate that PCV2 prevalence in slaughter pigs is very high and co-infection between different PCV2 genotypes and between PCV2 and PCV3 is relatively common. Therefore, genetic diversity and co-infection between other porcine circoviruses should be consistently monitored in the future.

조선시대 관문서 관(關)의 기원과 수용 -행이체계를 중심으로-

문보미 ( Bo Mi Moon ) 한국고문서학회 2010 古文書硏究 Vol.37 No.-

실크 피브로인을 함유한 비출혈용 복합 지혈제 개발에 대한 연구

문보미 ( Bo Mi Moon ),주형우 ( Hyung Woo Ju ),박현정 ( Hyun Jung Park ),이정민 ( Jung Min Lee ),이옥주 ( Ok Joo Lee ),박찬흠 ( Chan Hum Park ) 한국조직공학·재생의학회 2013 조직공학과 재생의학 Vol.10 No.2s

Epistaxis is defined as active bleeding arising from the nasal mucosa. It constitutes one of the most common otolaryngology emergencies and can be severe or even fatal. Hemostasis is a process which causes bleeding to stop, meaning to keep blood within a damaged blood vessel which involves differential sequential events. This system in humans has been the most extensively researched. However, lot of research work is going on and lot more needs to be done to overcome the problem associated with this serious issue. This study was conducted to develop a new sponge type of biomaterial to be used for ideal hemostatic agent. The combination of three biomaterials; Silk fibroin (SF), Hyaluronic acid (HA), Gelatin (Gel) and Polyvinyl alcohol (PVA) as proposed hemostatic agent were tested against commercially available hemostatic pad (ChitoClot®). A comparative study of these silk based hemostasis agents designed in combination of SF/HA/PVA and SF/Gel/PVA sponges were tested on injured model of rats. Each sponge to be tested was made same size of (1×1 cm) then applied on top of injury site while applying a pressure of 5 N for 30 seconds. The In vivo experiments were carried by creating a proximal arterial injury in unilateral femoral arteries of 12 anesthetized SD rats. In conclusion, this study suggest a new developed silk fibroin-based sponges as hemostatic materials, which induced durable hemostasis and an increased blood clotting in animal studies. These initial studies explores the loopholes present in the existing ChitoClot® pad compared with that of fabricated silkbased sponges.

Induction of Systemic and Mucosal Immune Responses in Mice Orally Administered with Recombinant Attenuated Salmonella Expressing Subunits of P Fimbriae of Avian Pathogenic Escherichia coli

오인경,문보미,이존화,허진 한국임상수의학회 2011 한국임상수의학회지 Vol.28 No.3

Avian pathogenic Escherichia coli (APEC) causes a number of extraintestinal diseases in poultry. A virulence factor, P-fimbriae is firmly associated with the diseases. In this study, to develop an effective vaccine for the prevention of APEC, recombinant attenuatted Salmonella Typhimurium vaccines expressing PapA and PapG of P-fimbriae were evaluated whether these induced protective immune responses in murine models. Female BALB/c mice were primed and boosted orally at 7 and 10 weeks of age. In all immunized mice, the antigen-specific serum IgG levels were remained higher than those in the control mice from the fourth week post inoculation till the end of this study. In addition, antigen-specific serum IgG levels in the prime-booster immunized mice were enhanced as compared to the single immunized mice among each immunized group. The antigen-specific mucosal IgA levels in the mice immunized with each strain also induced higher than those in control mice. In addition, serum IgG and fecal IgA levels in mice administered with the combination of both strains were highly induced compared to those in mice immunized with each strain alone. These results indicated that PapA and PapG worked together for inducing high immune responses. To partly discern the nature of immunity induced by the strains, we quantified serum IgG subtypes IgG1 and IgG2a specific to antigens. The PapA and PapG strains biased the immunity to the Th1-type, as determined by the IgG2a/IgG1 ratio. On the other hand, the immunization with the both strains in combination produced mixed Th1- and Th2-type immune responses. These indicated that immunization with the combination of PapA and PapG could elicit both humoral and cell-mediated immunities. 조류병원성 대장균 (APEC)은 가금에서 많은 장관외 질병을 야기한다. 병원성 인자 중 하나인 P-fimbriae 또한 질병과 밀접하게 관련되어 있다. 이번 연구에서 마우스에 P-fimbriae의 subunit 들인 PapA와 PapG를 발현하는 재조합 약독화 Salmonella Typhimurium 백신균주 접종 후 APEC 방어에 필요한 면역반응 유도 여부를 알아보기 위한실험을 수행하였다. 각 백신 균주로 경구 접종 후 각 항원에 대한 serum IgG 항체 역가는 접종 후 4주째부터 대조군에 비해 항체 역가가 높게 유도되기 시작하여 실험이 끝날 때까지 높은 항체 역가가 계속 유지 되었다. 한번 접종 된경우보다 추가 접종 된 군에서 보다 높은 항체 역가가 관찰되었다. 더불어 각 항원에 대한 mucosal IgA 역가 또한 높게 유도되었다. 두 균주를 혼합하여 접종 하였을 경우에는 각 백신 균주를 접종하였을 경우보다 높은 serum IgG와mucosal IgA의 역가가 관찰되었다. 이 결과는 이들 백신 균주가 면역원성이 있음을 확인하는 결과였다. 또한 helper T cells type을 알아보기 위해 IgG1과 IgG2a의 항체 역가를 측정하여 본 결과 각 백신 균주를 접종하였을 경우에는IgG2a의 항체 역가가 IgG1의 항체 역가 보다 월등히 높은 이들 백신 균주를 접종 하였을 경우에는 Th1-type의 면역반응이 유도되지만 두 백신 균주를 혼합 접종하였을 경우에는 Th1-과 Th2-type 모두를 유도하는 것으로 관찰되었다. 이 결과를 통해서 이들 두 백신 균주를 혼합 접종 하였을 경우에는 세포성 면역뿐만 아니라 체액성 면역 모두를 유도할 수 있음을 확인할 수 있었다.

Silk Fibroin based Hydrogel for Regeneration of Burn Induced Wounds

주형우,이옥주,문보미,Faheem A. Sheikh,이정민,김정호,박현정,김동욱,이민채,김수현,박찬흠,이혜란 한국조직공학과 재생의학회 2014 조직공학과 재생의학 Vol.11 No.3

The present study deals with fabrication of hydrogel composed of silk fibroin, calcium alginate and car-boxymethyl cellulose. A simple mixing of calcium alginate and carboxymethyl cellulose and silk fibroin wasadopted to fabricate new silk fibroin based hydrogel (SFH) type. The adhesive strength, water content and cytotox-icity of fabricated hydrogel was comparatively investigated against standard control as medical gauze (C) andPurilon Gel®(PG). The results from the adhesive strength indicated that presented hydrogels had least adhesivestrength but had good cell viability than the C and PG. The cell toxicity results after 1 day of culturing NIH 3T3fibroblasts indicated that prepared hydrogels comparably promoted cellular growth. The in vivoexperiments usingSprague-Dawley were done by creating a second-degree burn on the back of rats to study the effect of healing pro-cess. The gross examination investigations at 1, 3, 7, 14 and 21 days indicated a progressive effect induced by theSFH. The histological examinations using Hematoxylin and eosin (H&E) and Masson’s trichrome (MT) stainingafter 3, 7, 14 and 21 days indicated the complete collagen deposition and dermic formation using the presented SFH. Our results suggest that SFH can be considered as promising new candidate matrices for burn wound regeneration.

Fabrication and Characterization of Hydrocolloid Dressing with Silk Fibroin Nanoparticles for Wound Healing

이옥주,김정호,문보미,Janet Ren Chao,윤재호,주형우,이정민,박현정,김동욱,김승주,박혜상,박찬흠 한국조직공학과 재생의학회 2016 조직공학과 재생의학 Vol.13 No.3

Hydrocolloid dressings have been developed for many types of wound healing. In particular, dressing is a critical component in the successful recover of burn injuries, which causes a great number of people to not only suffer from physical but also psychological and economic anguish each year. Additionally, silk fibroin is the safest material for tissue engineering due to biocompatibility. In this study, we fabricated hydrocolloid dressings incorporating silk fibroin nanoparticles to enhance the efficacy of hydrocolloid dressing and then use this silk fibroin nanoparticle hydrocolloid dressing (SFNHD) in animal models to treat burn wounds. The structures and properties of SFNHD were characterized using tensile strength and Cell Counting Kit-8 assay. The results indicated the structural stability and the cellular biocompatibility of the hydrocolloid dressing suggesting that SFNHD can be applied to the treatment of wounds. To demonstrate the capacity of a silk fibroin hydrocolloid dressing to treat burn wounds, we compared SFNHD to gauze and Neoderm®, a commercially available dressing. This study clearly demonstrated accelerated wound healing with greater wound structural integrity and minimal wound size after treatment with SFNHD. These observations indicate that SFNHD may be an improvement upon current standard dressings such as Gauze and Neoderm® for burn wounds.

Application of a Collagen Patch Derived from Duck Feet in Acute Tympanic Membrane Perforation

김수현,정주연,박현정,문보미,박예리,이옥주,Md. Tipu Sultan,김동규,박혜상,이준호,박찬흠 한국조직공학과 재생의학회 2017 조직공학과 재생의학 Vol.14 No.3

We investigated the utility of the duck-feet collagen extraction patching procedure in the traumatic tympanic membrane (TM) perforation via a comparison with spontaneous healing or paper patch. Fifty-six ears of adult male Sprague-Dawley rats, each weighing in the range of 250 to 300 g, were used for the animal studies. Sixteen rats had oneside ear in the control group and the opposite-side ear in the treated groups. The remaining twelve rats had a one-side ear with the duck-feet collagen patch and the opposite-side ear with a paper patch. The proliferating cell nuclear antigen (PCNA) expression cells were calculated among the 200 basal cells, and the expression percentage was identified as the labeling index. The healing of the perforation in the duck-feet collagen patch group was confirmed to be more rapid compared to the healing of the other groups. PCNA staining was observed in the migrating portion of PCNA enhanced cell to collagen scaffold in Post operative day (POD) 7 of collagen patch group. Thus, the adhesive effect of the duck-feet collagen patch to perforated margin was better than that of the paper patch. After completing the healing process, the collagen patch shrank and detached from the tympanic membrane (POD 14). In this study, we confirmed that the use of a duck-feet collagen patch had the advantage of early healing, inducing natural TM contour, and disappearing ability after the patch effect is complete.

전북지역 돼지유행성설사 바이러스 Spike 유전자 염기서열 및 계통분석

박미연 ( Mi-yeon Park ),문보미 ( Bo-mi Moon ),강수진 ( Su-jin Gang ),이종하 ( Jong-ha Lee ),박진우 ( Jin-woo Park ),조성우 ( Sung-woo Cho ),허철호 ( Cheol-ho Her ) 한국동물위생학회(구 한국가축위생학회) 2021 韓國家畜衛生學會誌 Vol.44 No.2

Although many swine farms continuously vaccinated to sow to prevent Porcine epidemic diarrhea(PED), PED has occurred annually in swine herds in Jeonbuk province, Korea. In the present study, the small intestine and feces samples from 17 farms where severe watery diarrhea and death of newborn piglets occurred in 2019 were collected, amplified by RT-PCR and determined the complete nucleotide sequences of the spike (S) glycoprotein genes of nine Jeonbuk PEDV isolates. The spike (S) glycoprotein is an important determinant for molecular characterization and genetic relationship of PEDV. These nine complete S gene isolates were compared with other PEDV reference strains to identify the molecular diversity, phylogenetic relationships and antigenicity analysis. 9 field strains share 98.5∼100% homologies with each other at the nucleotide sequence level and 97.3∼100% homologies with each other at the amino acid level. The nine Jeonbuk PEDV isolates were classified into G2b group including a genetic specific signal, S-indels (insertion and deletion of S gene). In addition, comparisons the neutralizing epitopes of S gene between 9 field strains and domestic vaccine strains of Korea mutated 12-15 amino acids with SM-98-1 (G1a group) and mutated 0-3 amino acids with QIAP1401 (G2b group). Therefore, the development of G2b-based live vaccines will have to be expedited to ensure effective prevention of endemic PED in Korea. In addition, we will need to be prepared with periodic updates of preventive vaccines based on the PEDV variants for the re-emergence of a virulent strain.

Effect of Pore Sizes of Silk Scaffolds for Cartilage Tissue Engineering

한갑수,강길선,송정은,Nirmalya Tripathy,김형석,문보미,박찬흠 한국고분자학회 2015 Macromolecular Research Vol.23 No.12

The aim of this study was to investigate the effects of silk fibroin scaffold, a natural biodegradable polymer scaffold, on the adhesive and proliferative behaviors of chondrocytes. Various silk fibroin scaffolds were produced using the salt extraction method, and scaffolds with different pore sizes (90-180, 180-250, 250-355, and 355- 425 μm) were constructed based on the size of the salt particles. Chondrocytes were seeded on the scaffolds and incubated. The produced scaffolds were analyzed with Fourier transform-infrared spectroscopy and exhibited characteristics similar to those of natural silk in terms of chemical composition and structure. Moreover, we found that the mechanical strength decreased as the pore size increased. Scanning electron microscopy images confirmed the existence of pores in the silk fibroin scaffold. Additionally, scaffolds with smaller pore sizes facilitated improved cell adhesion. Using MTT analysis, we found that scaffold with pore sizes of 90-180 and 180-250 μm provided the best environment for cell proliferation. The amount levels of sulfated glycosaminoglycan (sGAG) and collagen were highest for scaffolds with a pore size of 90-180 μm. In gene expression analysis, scaffolds with pore sizes of 90-180 and 180-250 μm showed the highest expression of the chondrocytes marker aggrecan and type II collagen. Collectively, these data suggest that silk fibroin scaffolds with smaller pore sizes (90-250 μm) provide the best environment for adhesion and proliferation of chondrocytes.