원저 : 백서에서 혈소판 풍부 혈장의 제작과 유효성에 대한 실험적 연구
Original Articles : Fabrication of Platelet-Rich Plasma in a Rat Model and the Efficacy Test in Vitro
이상훈, 이부규, 안강민, 조영욱, 지현숙,
소속 상세정보
이상훈 ( Lee Sang-Hun ) - 가톨릭대학교 의과대학 내과학교실
이부규 ( Lee Bu-Kyu ) - 울산대학교 의과대학 서울아산병원 치과학교실
안강민 ( Ahn Kang-Min ) - 울산대학교 의과대학 서울아산병원 구강악안면외과
조영욱 ( Cho Young-Uk ) - 울산대학교 의과대학 서울아산병원 진단검사의학과
지현숙 ( Chi Hyun-Sook ) - 울산대학교 의과대학 서울아산병원 진단검사의학과
KMID : 0360120070290020113
Abstract
Purpose: Platelet-rich plasma (PRP) is known to accelerate and/or enhance hard and soft tissue healing and regeneration. As such, PRP has been used in various clinical fields of surgery. Recently there have been several attempts to use PRP in the field of tissue engineering. However, some controversies still exist on exact mechanism and benefits of PRP. Therefore various animal experiments are necessary to reveal the effect of the PRP. However, even if animal experiment is performed, the efficacy of the experiment could not be validated due to absence of an animal PRP model. The purpose of this study is to establish rat PRP model by comparing several PRP fabricating methods, and to assay growth factor concentration in the PRP.
Materials and Methods: Rat blood samples were collected from nine SD rat (body weight: 600-800g). PRP was prepared using three different PRP fabricating methods according to previously reported literatures. (Method 1: 800 rpm, 15 minute, single centrifuge; Method 2: 1000 rpm, 10 minute, double centrifuge; Method 3: 3000 rpm, 4min and 2500 rpm, 8 min, double centrifuge). Platelet counts were evaluated in an automated machine before and after PRP fabrications. In terms of growth factor assay, prepared PRP were activated with 100 unit thrombin and 10% calcium chloride. Growth factor (PDGF-BB, VEGF) concentrations on incubation time were determined by sandwich-ELISA technique.
Results: An average of 3ml (via infraorbital venous plexus) to 15ml (via celiac axis) the rat blood could be collected. By using Method 3 (3000 rpm, 4 min and 2500 rpm, 8 min, double centrifugation), around 1.5ml of PRP could be prepared. This method allowed us to concentrate platelet 3.77-fold on average. PDGF-BB concentration (mean, 1942.10 pg/ml after 1 hour incubation) and VEGF concentration (mean, 952.71 pg/ml after 1 hour incubation) in activated PRP were higher than those in untreated blood. Also PDGF-BB showed constant concentration during 4-hour incubation, while VEGF concentration was decreased after 1 hour.
Conclusion: Total 11,000 g minute separation and condensation double centrifuge method can produce efficient platelet-rich plasma. Platelet-rich plasma activated with thrombin has showed higher concentrations of growth factors such as PDGF-BB and VEGF, compared to the control group. Platelet-rich plasma model in a rat model was confirmed in this study.
키워드
Tissue engineering;Platelet-rich plasma;Growth factor
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