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박민언,이광철,류강현,임원봉 한국생물공학회 2019 Biotechnology and Bioprocess Engineering Vol.24 No.6
The use of multi-porous scaffolds for bone tissue engineering has been shown to improve osteogenesis. Although the exact mechanisms by which these scaffolds promote new bone formation have not yet been recognized well, related hypotheses can be found in many previous studies. The aim of this study was to examine the influence of the modulation of the pore size of beta- TCP ceramics on osteogenic differentiation in rats with calvarial defects. Treatments with macro- and meso-sized particles of NaCl, which was used as a porogen, were carried out during scaffold manufacturing, and the pore sizes of the beta-TCP scaffolds were measured by SEM. New bone formation was evaluated by Micro-CT, H&E staining, and immunohistochemical analysis at week 4 after the operation. It was observed that the multi porosity of beta-TCP ceramics was controlled by treating the scaffolds with NaCl particles of two sizes (500–800 μm and 10–50 μm) during the firing process. Histological and immunohistochemical analysis of type 1 collagen and osteocalcin protein expression showed that the new bone formation and mineralization in case of the multi-porous beta-TCP ceramics comprising macroand meso-sized pores were significantly higher than those in case of single porous macro- or meso-sized scaffolds. In conclusion, the distribution and size of the pores, as well as the surface structure of the scaffolds, may play an important role for osteogenic differentiation in vivo.
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이광철,고영종,박민언,김보라,현훈,임원봉 한국생물공학회 2017 Biotechnology and Bioprocess Engineering Vol.22 No.6
Osteopetrosis belongs to a group of rare genetic diseases typically treated with bone marrow transplantation. This approach is not effective in a recently identified form of the disease caused by mutations in the receptor activator of NF-κB ligand (RANKL) gene. In these patients, replacement therapy and RANKL delivery may be a more valid approach than transplantation. Here, we describe the construction of a recombinant gene encoding regions of RANKL (rRANKL), including the biologically active regional loop sequence. We present detailed methods for the cloning, expression, and purification of the recombinant protein. The activity of rRANKL including the active region was assessed in vitro and mature osteoclast generation was evaluated in vivo using a mouse model. We provide a proof of concept for the therapeutic potential of full-length and selected active regions of rRANKL in the treatment of osteopetrosis, warranting clinical assessment.
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척수 손상 백서에서 경피신경전기자극을 통한 미세전류 치료 효과
손홍문,임원봉,김영욱,고영종,박민언,김보라 대한척추외과학회 2019 대한척추외과학회지 Vol.26 No.1
Study Design: Animal study. Objectives: To investigate the effects of microelectric treatment by transcutaneous electrical nerve stimulation (TENS) on functional recovery and histological changes in a rat model of spinal cord injury (SCI). Summary of Literature Review: The effects of TENS on spasticity and its underlying mechanisms remain unclear. Materials and Methods: SCI was induced by a 1.5-mm impactor with 200,000–260,000 dyne after laminectomy. Rats were divided into the following groups: group I (normal control), group II (microelectric treatment of 0 A), group III (microelectric treatment of 100 μA for 1 hr/day), group IV (microelectric treatment of 400 μA for 1 hr/day), and group V (microelectric treatment of 400 μA for 24 hr/day). After inducing SCI, rats were assessed by a sensory test with von Frey filaments and the locomotor recovery test (BBB rating scale) at 1, 4, 7, 14, 21, and 28 days. To evaluate spinal cord damage, histopathological studies were performed with hematoxylin and eosin. Brainderived neurotrophic factor (BDNF) and TrkB immunohistochemistry studies were performed at 28 days. Results: In groups IV and V, the BBB score had significantly improved on days 21 and 28 after SCI, and the TENS-treated groups showed significant neuronal recovery. After SCI, groups IV and V showed a significant recovery of locomotor function and the motor sensory response of the withdrawal threshold to 3.5 g. In addition, necrotic tissue and cystic spaces in the spinal cord were significantly reduced and BDNF/TrkB-positive cells were highly expressed in groups III, IV, and V. Conclusions: Microelectric treatment can play a role in facilitating the recovery of locomotion following SCI. 연구 계획: 동물 실험목적: 척수 손상 백서 동물 모델에서 경피신경자극을 통한 미세전류 치료가 기능적 회복과 조직학적 변화에 미치는 영향을 알아보고자 하였다. 선행 연구문헌의 요약: 경피신경자극의 척수 기능 향상에 대한 보고는 드물다. 대상 및 방법: 실험 쥐의 후궁 절제 후 1.5 mm impactor를 이용하여 200,000-260,000 dyne의 외력으로 척수 손상을 시켰다. 실험군은 그룹 I; 비수술군, 그룹 II; 미세전류치료(0 A) 군, 그룹 III; 미세전류치료(100 μA, 하루 한시간) 군, 그룹 IV; 미세전류치료(400 μA, 하루 한시간) 군, 그룹 V; 미세전류치료 (400 μA, 하루 24시간) 군으로 나누어 실험을 진행하였다. 척수 손상 후 1, 4, 7, 14, 21일과 28일에 BBB척도를 이용한 운동 기능 검사와 von-Frey monofilament를 이용한 감각 검사를 시행하였다. 또한 손상 부위의 조직학적 변화 관찰을 위한 H&E 염색 및 신경성장인자의 발현 분석을 위한 BDNF, Trk-b의 면역조직화학염색을 수상 후 28일에 시행하였다. 결과: 그룹 IV, V군에서 척수 손상 후, 21일과 28일에서 BBB 점수가 상당히 향상하였으며, 유의한 신경원 회복을 보였다. 또한 척수 손상 후 그룹 IV, V군에서 유의한 운동기능 향상과 감각기능 회복을 보였으며, 척수의 괴사조직이 상당히 감소됨을 확인할 수 있었다. 또한 그룹 III, IV, V군에서는 BDNF 및Trk-β의 발현이 증가하였다. 결론: 척수 손상 후 경피신경전기자극 치료는 운동 기능 회복의 향상에 도움이 되리라 사료된다. 약칭 제목: 급성 척수 손상의 미세전류 치료
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