• Journal of Korean Dental Science
• /
• 제14권1호
• /
• pp.12-25
• /
• 2021

Purpose: (i) To evaluate the biologic properties of a bi-layered 1-ethyl-3-(3-dimethylaminopropyl)carbodiimide hydrochloride-cross-linked collagen membrane (CCM) in vitro. (ii) To assess the efficacy of CCM for localized bone regeneration in vivo. Materials and Methods: Biodegradation of CCM compared to a native collagen membrane (NCM) was assessed in vitro. In vivo, twelve male New Zealand White rabbits were used. Four calvarial, circular defects (diameter 8 mm) were created in each animal. The sites were randomly allocated to i) CCM+biphasic calcium phosphate (BCP) (CCM-BCP group), ii) CCM alone (CCM), iii) BCP alone (BCP) and, iv) negative control (control). Animals were sacrificed at 2 (n=6) and 8 weeks (n=6). Outcome measures included: micro-computed tomography (μCT) analysis (total augmented volume [TAV], new bone volume) and histomorphometry (total augmented area [TAA], newly formed bone, remaining membrane thickness [RMT]). Result: CCM was more resistant to degradation than NCM. μCT analysis showed CCM-BCP (196.43±25.30 mm³) and BCP (206.23±39.13 mm³) groups had significantly (P<0.01) larger TAV than the control (149.72±12.28 mm³) after 8 weeks. Histomorphometrically, CCM-BCP group (17.75±5.97 mm²) had significantly (P<0.01) greater TAA compared to the CCM group (7.74±2.25 mm²) and the control (8.13±1.81 mm²) after 8 weeks. After 8 weeks, RMT was reduced by 67%. Conclusion: CCM can be a favorable choice of barrier membrane when performing guided bone regeneration (GBR) in localized bone defects. CCM has better resistance to degradation than the natural collagen membrane, in vitro. In vivo, CCM provides an advantageous integration of prolonged barrier function and biocompatibility for GBR.

• Journal of Chest Surgery
• /
• 제46권1호
• /
• pp.1-13
• /
• 2013

Background: Glutaraldehyde (GA) is a widely used cross-linking agent for improving mechanical properties and resistance to enzymatic degradation of collagenous tissue, but it has several drawbacks such as calcification and cytotoxicity. The aim of this study was to find the alternative effective cross-linking methods to GA. Materials and Methods: Bovine pericardium was processed with GA with ethanol+octanol and glycine detoxification, and polyethylene glycol (PG) space filler, dimethyl 3,3'-dithiobispropionimidate (DTBP), 1-ethyl-3-(3-dimethylaminopropyl) carbodiimide (EDC) treatment, and the physical fixation of ultraviolet irradiation were done. The biologic material properties of variously treated pericardial tissues were assessed by biochemical, mechanical and histological tests. Treated pericardial tissues were also implanted subcutaneously or intramuscularly into the rabbit for 10 weeks to assess the xenoreactive antibody response of immunoglobulin G and M, their anti-calcification effect. Results: The biochemical and mechanical properties of EDC fixed pericardial tissues were comparable to the GA fixed tissue. The cytotoxicity was lowest in space filler treated GA fixed group. In rabbit subcutaneous or intramuscular implantation models, decellularization, space filler, EDC treatment group showed significantly lower calcium content than GA only and DTBP treatment group (p<0.05, analysis of variance). The titer of anti $Gal{\alpha}1-3Gal{\beta}1$-4GlcNAc-R antibodies did not change in the postimplantation serial enzyme-linked immunosorbent assay. Hematoxylin and eosin and von Kossa staining showed that decellularization, space filler, EDC, and ultraviolet treatment had less inflammatory cell infiltration and calcium deposits. Conclusion: The decellularization process, PG filler, and EDC treatments are good alternative cross-linking methods compared to GA only fixation and primary amine of DTBP treatment for cardiovascular xenograft preservation in terms of the collagen cross-linking stability and in vivo anti-calcification effects.

• Macromolecular Research
• /
• 제11권5호
• /
• pp.368-374
• /
• 2003

Porous scaffolds composed of gelatin and polysaccharides such as hyaluronic acid and $\beta$-glucan were prepared by using the freeze-drying method after cross-linking with l-ethyl-(3-3-dimethylaminopropyl) carbodiimide hydrochloride (EDC). The scaffold had an inter-connected pore structure with the sufficient pore size for use as a support for the growth of fibroblasts. Results for the contact angle and cell attachment confirmed that high gelatin content in a mixture was suitable for cellular attachment and distribution in two- or three-dimensional fibroblast cultures. However, the addition of polysaccharides aroused the synergistic effects of morphologic and mechanical property of gelatin-based scaffolds. To prepare the artificial dermis for the wound dressing to mimic the normal human dermal skin, fibroblasts were isolated from a child's foreskin, and cultured in gelatin-based scaffolds. An in vivo study showed that the artificial dermis containing the fibroblasts enhanced the wound healing rate and re-epithelialization of a full-thickness skin defect rather than the acellular scaffold after one week.

• 한국식품위생안전성학회지
• /
• 제9권3호
• /
• pp.123-131
• /
• 1994

An enzyme-linked immunosorbent assay(ELISA) was developed for the detection of residual gentamicin(GM) in edible animal products. The immunogen(GM-KLH conjugate) and coating antigen(GM-BSA conjugate) were prepared by coupling GM sulfate to keyhole limpet hemocyanin(KLH) and bovine serum albumin(BSA) in the presence of 1-ethyl-3-(3-dimethylaminopropyl)carbodiimide hydrochloride, respectively. Polyclonal antibody to GM was produced in rabbits(New Zealand White, female) by using the immunogen and the antibody titer was measured by indirect ELISA. A competitive ELISA was developed using GM-bovine serum albumin conjugate as a coating antigen, GM(as standards or sample), polyclonal antibody to GM, secondary antibody conjugated with horseradish peroxidase as an enzyme, and H2O2 and o-phenylenediamine dihydrochloride as a substrate and a chromophore, respectively. The detection limit of GM was 10 ng/ml and the standard curve of GM(n=26) was linear up to 10 $\mu\textrm{g}$/ml in this competitive ELISA system. There were no cross-reactivities of the partially purified antibody between GM and the various antibiotice such as amikacin, benzyl-penicillin, chloramphenicol, erythromycin, furazlidone, kanamycin, neomycin, oleandomycin, streptomycin, sulfathiazole and thiamphenicol(CR50<0.05%)

• Journal of Microbiology and Biotechnology
• /
• 제27권8호
• /
• pp.1483-1490
• /
• 2017

In this study, silver nanoparticles (AgNPs) were synthesized by the citrate reduction process and, with the assistance of n-hydroxysuccinimide and 1-ethyl-3-(3-dimethylaminopropyl) carbodiimide, were successfully loaded with the macromolecular drug vancomycin (VAM) to form AgNP-VAM bioconjugates. The synthesized AgNPs, VAM, and AgNP-VAM conjugate were characterized by UV-visible spectroscopy, zeta potential analysis, confocal microscopy, and transmission electron microscopy. The effect of loading VAM onto AgNPs was investigated by testing the internalization of the bioconjugate into Mycobacterium smegmatis. After treatment with the AgNP-VAM conjugate, the bacterial cells showed a significant decrease in UV absorption, indicating that loading of the VAM on AgNPs had vastly improved the drug's internalization compared with that of AgNPs. All the experimental assessments showed that, compared with free AgNPs and VAM, enhanced internalization had been successfully achieved with the AgNP-VAM conjugate, thus leading to significantly better delivery of the macromolecular drug into the M. smegmatis cell. The current research provides a new potential drug delivery system for the treatment of mycobacterial infections.

• 멤브레인
• /
• 제15권3호
• /
• pp.213-223
• /
• 2005

본 연구에서는 히알루론산나트륨/카르복시메틸셀룰로오스나트륨(HA/CMC)으로 구성된 유착방지 막을 제조하였고, 쥐에서 유착방지용에 대한 효과를 평가하였다. 유착방지 막은 HA/CMC 용액을 동결건조한 후 1-에틸-3-(3-디메틸아미노프로필)카보디이미드 (EDAC)로 가교하여 제조하였다. Sprague-Dawley쥐의 맹장/복벽 찰과상 모델에서 복부 중앙선 절개 후 장막과 복벽을 bone burr로 $1\times2\;(cm^2)$ 크기로 찰과상을 만들고, 상처 면 주위의 3곳을 봉합사로 고정시켰다. 상처난 장을 실험군의 경우 HA/CMC 막으로 덮어주었으며, 대조군은 아무처치도 하지 않은 상태로 복벽을 마주보게 하였다. 대조군의 대부분은 수술 후 7, 14, 21, 28일에 유착정도 3 이상을 나타내었고, 반면 실험군의 $60\~70\%$는 수술 후 14, 21, 28일에 유착정도 2 이하를 나타내었다. 이는 유착세기에서도 유사하였다. 일반적으로 유착정도와 유착세기는 수술 후 14일까지 점차 증가하였고, 수술 후 21일에는 거의 동일하거나 약간 증가하였으며, 28일째에는 감소하였다. 대조군은 유착정도와 유착세기, 유착면적에서 높은 수치를 나타내었다. 이 유착방지막은 수술 후 유착방지에서 좋은 임상결과를 나타낼 수 있을 것으로 기대된다.

• Journal of Chest Surgery
• /
• 제43권6호
• /
• pp.576-587
• /
• 2010

배경: 이종조직판막이나 조직이 임상적으로 이용되기 위해서는 조직의 장기간의 내구성이 동반된 효과적인 보존과 적절한 고정과정이 필요하다. 본 연구에서는 소심낭을 sodium dodecyl sulfate (SDS)와 N-lauroylsarcosinate를 이용하여 무세포화하고, 알파-갈락토시다아제를 처리한 뒤, 이를 다양한 방법으로 고정함으로써 조직손상과 면역반응을 최소화할 수 있는 효과적인 우심낭 보존 방법에 대해 알아보고자 하였다. 대상 및 방법: 우심낭으로부터 이종 이식 보철편을 채취한 뒤 SDS와 N-lauroylsarcosinate를 이용하여 무세포화 하였고, 알파-갈락토시다아제를 처리하였다. 이 두 가지 군에 대하여 각기 다른 조건 즉 glutaraldehyde (GA), l-ethyl-3-(3-dimethylaminopropyl)-carbodiimide (EDC)/Nhydroxysuccinamide (NHS)의 처리를 달리하여 고정하였다. 이후 각 군들의 물리적 성상, 인장강도, 열 안정성 검사, 세포독성 검사, 프로나아제 저항 시험과 Pronase-Ninhydrin 시험, 투과도와 유순도 검사, purpald 검사 등을 시행하였으며, H&E 염색과 DNA 정량, 알파-갈 항원결정인자 염색 등도 함께 시행하였다. 결과: GA 단독 고정군은 EDC/NHS 고정군에 비해 물리적인 성상과 열안정성에서 더 우수하였고, EDC/NHS와 GA의 이중 고정군은 EDC/NHS 고정군에 비해 물리적인 성상과 열안정성에서 더 우수하였으며, GA단독 고정군과 비교해서 열안정성에서 더 우수하였다. 프로나아제 저항 시험과 Pronase-Ninhydrin 시험을 통해 알아본 crosslinking 정도는, GA 고정 군과 EDC/NHS의 이중 고정군이 EDC/NHS 고정군에 비해 높은 것으로 관찰되었다. EDC/NHS와 GA의 이중 고정군은 GA 고정군에 비해 투과도와 유순도가 다소 증가하였으나, GA 고정군에 비해 인장강도가 높고 세포 독성이 감소하는 것으로 관찰되었다. 결론: GA와 EDC/NHS의 이중고정을 통해서, GA단독 고정의 독성을 줄이면서 효과적인 crosslinking을 시행할 수 있음을 확인하였다. 향후 생체내 시험을 통해서 GA와 EDC/NHS의 이중고정이 석회화의 감소와 조직부전에 어떠한 영향을 미치는지 추가적인 연구와 검증이 필요하다.