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PLACON 방법으로 제조된 혈소판 풍부혈장(PRP)의 활성후 시간에 따른 성장인자의 정량적 변화

Title
PLACON 방법으로 제조된 혈소판 풍부혈장(PRP)의 활성후 시간에 따른 성장인자의 정량적 변화
Other Titles
Growth Factor Levels in PRP produced by PLACON and Their Changes by Time after Activation
Authors
김옥진
Issue Date
2008
Department/Major
임상치의학대학원 임상치의학과임플란트치의학전공
Publisher
이화여자대학교 임상치의학대학원
Degree
Master
Abstract
혈소판풍부 혈장(PRP)은 혈소판에 있는 알파입자의 탈과립에 의한 성장인자의 분비로 좀더 빠르고 효과적인 골 재생을 얻기 위한 상태를 제공한다. 혈소판의 활성으로 유리된 성장인자는 상처치유를 개시하고 조절한다. 그러나 혈소판과 성장인자의 수는 제조방법에 따라 매우 다르게 나타나며 PRP 활성후 시간경과에 따라 가변적이다. 본연구의 목적은 PLACON (Platelet Concentrator, Oscotec Inc., Korea)에 의해 제조된 PRP 내의 혈소판 농축 정도와 활성화 이후 경과된 시간에 따른 성장인자가 정량적으로 어떻게 변화하는지를 규명하고자 한다. 연구재료와 방법은 4 명의 건강한 성인(25-30세)으로부터 혈액이 채혈되어 PLACON 제조사의 지시대로 혈소판 농축혈장이 제조되었다. 전혈과 PRP의 혈소판 수치 측정을 위해 혈액세포 분석을 시행하고 전혈과PRP활성 후 즉시, 30분, 1시간 2시간에 PRP내에 유리된 PDGF-AB, PDGF-BB, TGF-β 의 양을 ELISA(Enzyme Linked immunosorbent Assay)방법으로 측정하였고, 다음과 같은 결과를 얻었다. 1) PLACON으로 제조된 PRP의 혈소판 수는 905±148 x10³ platelets /μL 로서 전혈의 혈소판 수 207±10 x10³platelets /μL 의 436% 였다. (P <0.05) 2) PLACON으로 제조된 PRP의 활성화 후 PDGF-AB는 직 후에 25,890±15,500pg/mL, 30분에 47,960±15,590 pg/mL, 1시간에 51,310±13,910pg/mL , 2시간에 49, 910±15,740pg/ml를 보였다. (P <0.05) 3) PDGF-BB 성장인자는 활성화 직 후에12,460 ± 7,149pg/mL, 30분에 32,180±9,036pg/mL, 1시간에 35,340 ±10,190pg/mL, 2시간 후에 32,000±8,362pg/mL를 보였다. (P <0.05) 4) 성장인자 TGF-β 는 직 후에 58,560 ± 14,330pg/mL, 30분 후에 113,800±20,640pg/mL, 1시간에114,900±18,320 pg/mL, 2시간에 105,000±23,540pg/ mL를 유지하였다. (P <0.05) 연구결과 다음과 같은 결론을 얻었다. 1. PLACON 혈소판 농축 원심분리기에 의해 효과적인 혈소판의 농축을 얻었다. 2. 제조된 PRP내에 유리된 성장인자의 양은 활성 후 즉시 증가하여 2시간 동안 유지되었다.;Platelet-rich plasma (PRP) works via the degranulation of a- granules in the platelets and secretion of growth factors provides conditions for obtaining more rapid and effective bone regeneration. Growth factors released from activated platelets initiate and modulate wound healing, but the concentration of platelets and growth factors can vary widely depending on the preparation methods and it can be changeable after activation. The aim of this study was to evaluate the platelet concentration efficacy of PLACON (Platelet Concentrator, Oscotec Inc. Korea) and quantitate the growth factor level changes after platelet activation in PRP produced by PLACON protocol. Whole blood was drawn from 4 healthy donors and PRP was prepared by PLACON as manufactures directions. Complete cell count on the whole blood samples and PRP were determined. Platelet-derived growth factor (PDGF)-AB, PDGF-BB, and Transforming growth factor (TGF)- β were measured using the enzyme-linked immunosorbent assay(ELISA) method at time 0, 1/2hr, 1hr and 2hrs after platelet activation. Results are as follows; 1. The concentration of platelet of PRP Produce by PLACON was 905 ±148 x10³ /mL, 436% of whole blood 207.5±10x10³ /mL (P<0.05) 2. PDGF-AB growth factors were released as 25,890±15,560 pg/ml at the time of activation, 47,960±15,590pg/ml after 30 minutes, 51,310±13,910 pg/ml after 1 hour, 49,910±15,740 pg/ml in 2 hours (P<0.05) 3. PDGF-BB growth factors were released as high as 12,460±7,149 pg/ml right after activation, 32,180±9,036 pg/ml after 30 minutes, 35,340±10,190 pg/ml after 1 hour, 32,000±8,362 pg/ml in 2 hours after platelet activation. (P<0.05) 4. Growth factors TGF-β were measured to 58,560±14,330 pg/ml soon after activation, 113,800 ±20,640 after 30 minutes, and 114,900±18,320pg/ml after 1 hour, and 105,000±23,540pg/ml in 2 hours after platelet activation. (P<0.05) Therefore, it can be concluded that the platelet in the PRP produced by PLACON was concentrated as high as 4.36 times than whole blood. Growth factor levels released in PRP increased immediately after platelet activation and were maintained for 2 hours.
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