Heparin이 망내계세포에 미치는 영향에 관한 초미형태학적 연구

이동철,김중길,임정교,손윤경,서인수,손태중 慶北大學校 醫科大學 1991 慶北醫大誌 Vol.32 No.3

저자들은 heparin 투여후 세망내피계의 초미형태학적 변화를 관찰하기 위해 Sprague-Dawley계 숫쥐에 heparin과 endotoxin을 투여한 후 비장을 채취하여 실험에 이용하였다. 실험군은 heparin 전처치군과 heparin 후처치군으로 나누었으며 비교군으로 heparin 단독투여군과 endotoxin 단독투여군을 설정하여 각각을 투약후 비장을 채취하여 광학현미경 및 전자현미경적 검색을 실시하였다. 실험결과를 요약하면 heparin 단독투여군은 대식세포, 형질세포양 림프아구 및 다핵백혈구가 증가되었고, 대식세포의 탐식능도 항진되었다. 그러나 대식세포자체의 소기관 발달은 현저하지 않았다. Endotoxin 단독투여군은 시간이 갈수록 대식세포의 탐식능이 저하되고, 파괴가 증가되어 수가 감소하였다. 미소혈전이 자주 관찰되었으며 기타세포들의 파괴가 관찰되었다. Heparin 전처치 및 후처치군은 공히 endotoxin군에 비해 대식세포 탐식능이 증가되었다. 이상의 성적으로 보아 heparin의 망내세포에 대한 영향은 주로 대식세포의 탐식능과 수의 증가로 생각되며 이는 정상 및 병적상태 모두에서 일어난다고 생각된다. 또한 ET 투여 전후 heparin을 투여한 실험군에서 공히 대식세포 탐식능이 증가되었으므로 이는 ET shock 후 조직손상으로 분비되는 세망내피계 저하물질에 대해 heparin이 길항작용을 할 것으로 생각된다. 또한 heparin의 대식세포 탐식능항진에 대한 효과는 세포의 직접적인 자극이라기 보다는 탐식과정중 특히 oposonization에 효과를 미칠 것으로 생각된다. The authors studied an ultrastructural changes of reticuloendothelial system after administration of heparin. Male Sprague-Dawley rats were examined after administration of heparin and endotoxin. The experimental groups were divided into heparin pretreated groups and heparin posttreated groups. The control groups were divided into heparin-only groups and endotoxin-only groups. Each animals were sacrificed and spleens were extirpated and examined by light and electron microscopy. The results obtained were summarized as follows: Heparin-only groups showed proliferation of macrophages, lymphocytes and neutrophils. The phagocytic activity of marcophage is enhanced. The cytoplamic organellar change is not remarkable except increase of secondary lysosomes. Endotoxin-only groups showed decrease of phagocytic cells. The phagocytic activity is also depressed. Destruction of macrophages and other cells are noted. Microthrombi are frequently seen. Both heparin pretreated and heparin posttreated groups revealed relative enhancement of phagocytic activity compaired to endotoxin-only groups. From the result of the experimental study, it appears that the effect of heparin on the reticuloendothelial system is enhancement of phagocytic activity of macrophages both in the coditions of normal and pathologic one. And it also suggests that heparin may act as an antagonizing factor to the reticuloendothelial depressing agent that may be derived from injured organ of endotoxin shock. And the effect of heparin to the phagocytic activity of macrophage may be related to the process of phagocytosis such as opsonization or to the direct cellular stimulation.

Comparison of Two Methods for Heparin Sensitivity; Activated Partial Thromboplastin Time Assay using in vitro Heparin-spiked Sample and Anti-Xa Assay using in vivo Heparin-treated Sample

( Bon-kyung Koo ),( Eui-hoon Kwon ),( Kwang-hyun Ryu ),( Jae-won Yun ),( Hee-jin Kim ) 대한임상검사과학회 2011 대한임상검사과학회지(KJCLS) Vol.43 No.4

The monitoring of heparin therapy is using almost aPTT assay. This study is compare to estimating aPTT therapeutic range using in vitro heparin-spiked sample and aPTT therapeutic range using in vivo heparin-treated sample. Normal pooled plasma was collected from 20 healthy representative individuals. 11 concentration of heparinized plasmas from 0 U/mL to 1.0 U/mL at intervals of 0.1 U/mL made by addition of heparin to normal pooled plasma were measured aPTT. The aPTT therapeutic range was performed through correlation analysis between heparin level 0.2 to 0.4 U/mL and aPTT. 30 plasmas from patients on heparin therapy were measured aPTT and anti-Xa activity. The aPTT therapeutic range was performed through correlation analysis between anti-Xa activity 0.3 to 0.7 U/mL and aPTT. The aPTT therapeutic range corresponded by heparin level-vs-aPTT value regression analysis was 60.7 to 102.4 seconds. The aPTT therapeutic range corresponded by anti-Xa activity-vs-aPTT value regression analysis was 85.3 to 147.5 seconds. The validation of heparin sensitivity using in-vitro heparin sample was not considered. The establishing aPTT therapeutic range is recommended anti-Xa activity using in-vivo sample.

Aprotinin을 투여한 개심술 환자에서 Kaolin과 Celite Activator를 이용한 Activated Coagulation Time(ACT) 측정의 비교

김정택,선경,이춘수,백완기,조상록,김현태,김혜숙,박현희,김광호,Kim, Joung-Taek,Sun, Kyung,Lee, Choon-Soo,Baik, Wan-Ki,Cho, Sang-Rock,Kim, Hyun-Tae,Kim, Hea-Sook,Park, Hyun-Hee,Kim, Kwang-Ho 대한흉부심장혈관외과학회 1998 Journal of Chest Surgery (J Chest Surg) Vol.31 No.9

개심수술에서 Aprotinin에 의한 ACT가 연장되는가를 알아보기 위해 서로 다른 표면 촉매제인 kaolin (K-ACT)과 celite(C-ACT)를 이용하여 동시에 측정 비교하였다. 개심수술을 받은 22명의 성인을 대상으로 하여 Hemocron 8000 system을 이용하여 동시에 ACT를 측정 하였는데 aprotinin과 heparin 투여 전(Phase I), Aprotinin투여 후 heparin 투여 전(Phase II), heparin투여 5분 후(Phase III), haparin투여 30분 후(Phase IV), heparin투여 60분 후(Phase V), heparin투여 90분 후(Phase VI), protamin투여 30분 후(Phase VII)에 각각 측정하였다. Phase I, II, III에 두 군간에 차이가 없었으나 heparin투여 30분 후에는 C-ACT가 928$\pm$400초 K-ACT가 572$\pm$159초였고 60분 후에는 C-ACT가 888$\pm$254초 K-ACT가 535$\pm$186초 90분 후에는 C-ACT가 686$\pm$141초 K-ACT가 484$\pm$54초로 K-ACT에 비해 C-ACT가 통계학적으로 의의있게 증가하였다. 그러나 protamin투여 후에는 C-ACT가 137$\pm$26초 K-ACT가 139$\pm$28초로 두군간에 차이가 없었다. 이상의 결과에서 aprotinin투여 후 ACT는 연장이 되는 것이 아니라 activator로 celite를 사용했기 때문인 것으로 생각된다. 결론적으로 aprotinin을 투여한 개심수술에서 정확한 ACT수준을 측정하기 위하여 celite activator보다 kaolin activator를 사용해야 하며 heparin은 보통용량을 투입하여야 할 것으로 생각된다. Background: High-dose aprotinin has been reported to enhance the anticoagulant effects of heparin during cardiopulmonary bypass ; hence, som authors have advocated reducing the dose of heparin in patients treated with aprotinin. Material and Method: The ACT was measured before, during and after cardiopulmonary bypass, with Hemochron 801 system using two activators of celite(C-ACT) and kaolin(K- ACT) as surface activator. From June, 1996 to February, 1997, 22 adult patients who were scheduled for elective operation were enrolled in this study. Result: The ACT without heparin did not differ between C-ACT and K-ACT. At 30 minutes after anticoagulation with heparin and cardiopulmonary bypass, the average C-ACT was 928${\pm}$400 s; K-ACT was 572${\pm}$159s(p<0.05). After administration of protamine, C-ACT was 137${\pm}$26 s; K-ACT was 139${\pm}$28s, which were not statistically significant. Conclusion: Our results showed that the significant increase in the ACT during heparin- induced anticoagulation in the presence of aprotinin was due to the use of celite as surface activator, rather than due to enhanced anticoagulation of heparin by aprotinin. We conclude that the ACT measured with kaolin provides better monitoring of cardiac surgical patients treated with high dose aprotinin than does the ACT measured with celite. The patients treated with aprotinin should receive the usual doses of heparin.

경험적 항응고요법과 측정에 따른 개별적 항응고요법이 혈액투석시 항응고 및 투석에 미치는 효과

허영숙 ( Heo Yeong Sug ),장인선 ( Jang In Seon ),이명선 ( Lee Myeong Seon ),이유나 ( Lee Yu Na ),이형주 ( Lee Hyeong Ju ),송지숙 ( Song Ji Sug ),권정아 ( Kwon Jeong A ),이갑노 ( Lee Gab No ),신진호 ( Sin Jin Ho ),권영주 ( Kwon Yeo 대한신장학회 2004 Kidney Research and Clinical Practice Vol.23 No.2

목 적 : 혈액투석 중의 항응고요법은 체외순환회로에 혈액응고가 생기지 않으며 출혈을 일으키지 않을 만큼의 항응고 효과가 있어야 하지만, 대다수 투석실에서는 경험적으로 일정량의 헤파린을 균등하게 사용하여 과하거나 부족한 항응고 작용을 보이게 된다. 적절한 헤파린 투여를 위해서는, 그에 대한 개개인의 반응이 다양하므로 혈중 헤파린 농도를 측정하여 용량을 정하는 것이 필요하다. 이에 환자마다 필요한 헤파린 용량을 적용하는 항응고요법이 혈액투석 중의 항응고 및 투석에 미치는 효과를 파악하고자 하였다. 방 법 : 3개월 이상 유지혈액투석을 받고 있는 환자 24명을 대상으로 경험적으로 규정된 사용량인 헤파린 부하량과 유지량을 주고, 유지량은 투석 30분 후부터 종료 30분 전까지 지속적으로 주입하는 경험적 항응고요법 (empirical heparinization, EH)과 개개인의 activated clotting time (ACT) 추적으로 결정된 헤파린 부하량과 유지량을 주고, 유지량은 투석 개시부터 종료 1시간 전까지 지속적으로 주입하는 개별적 항응고요법 (individualized heparinization, IH)을 각각 임의 적용 후 교차 측정하였며, 2주 후 방법을 바꾸어 동일대상자 반복 측정 조사를 하였다. 결 과 : 분석된 대상자는 남자 8명, 여자 14명이였고, 당뇨 10명, 비당뇨 12명, 흡연자 3명, 비흡연자 19명이었다. 평균 연령 50.36±13.89세, 건체중 55.51±7.84㎏, 혈액투석 기간 54.48±66.09개월이었다. 투석 전후 헤모글로빈, 헤마토크리트, 혈소판수, 투석 전 알부민과 헤라핀 부하량은 두 방법이 차이가 없었으나, 헤파린 유지량은 개별적 항응고요법이 유의하게 많았다 (p=0.000). ACT의 적절한 연장과 유지 (p=0.000), 투석기 혈액 구획 용적 손실 (p=0.035), 혈액응고 (p=0.000)는 개별적 항응고요법이 항응고 효과가 좋았고, 출혈은 모두 없었다. 투석 전후, 다음 투석 전 요소질소와 투석 전 크레아티닌, 요소감소율과 투석적절도는 두 방법이 투석의 효과에 차이가 없었다. 결 론 : 이상의 결과로 ACT 측정에 따른 개별적 항응고요법이 경험적 항응고요법 보다 혈액 투석시 그에 따른 문제없이 투석의 효과를 유지하며 항응고 효과가 우월하였다. Background : Most chronic hemodialysis units select heparin doses on an empirical basis. Too little heparin causes clotting in the extracorporeal circuit and too much heparin may lead to excessive bleeding. We conducted a prospective, randomized, repeated cross over study to evaluate the effect of two different heparin regimens. The empirical standard dose regimen (empirical heparinization, EH) was used for all patients, and the individualized dose regimen (individualized heparinization, IH) determined by measuring the activated clotting time (ACT) was performed for more adequate heparinization during hemodialysis. Methods : Twenty-four outpatients with systemic heparinization who had been on hemodialysis for more than 3 months were enrolled. In both methods, anticoagulation was achieved with a loading dose and a continuous infusion of heparin. Each regimens were prescribed alternately, and repeated after 2 weeks later. The study evaluated pre-post dialytic Hgb, Hct, Platelet and predialytic albumin, heparin loading dose and infusion rate, ACT, total blood compartment volume (TBCV), visible blood clots, bleeding, pre-post dialytic and next predialytic BUN, predialytic Cr, URR, Kt/V_(urea). Results : Twenty-two patients were analyzed in this study. Pre-post dialytic Hgb, Hct, Platelet and predialytic albumin, heparin loading dose were not significantly different between two methods. But heparin infusion rate were significantly increased in individualized heparinization than in empirical heparinization. Activated clotting times were prolonged and maintained adequately in individualized heparinization during hemodialysis. The loss of TBCV and visible blood clots were significantly decreased in individualized heparinization than in empirical heparinization. There was no bleeding complication in two methods. Pre-post and next predialytic BUN, predialytic Cr, URR, Kt/V_(urea) were not significantly different between two methods. Conclusion : We concluded that the individualized heparinization can maintain adequate anticoagulation than the empirical heparinization without any other problems and compromising the delivery dose of dialysis. (Korean J Nephrol 2004;23(2):300-308)

Histomorphometric Analysis of Heparin Effects on the rhBMP-2

정명호,이종헌 대한구강악안면병리학회 2016 대한구강악안면병리학회지 Vol.40 No.2

Although recombinant human Bone Morphogenetic Proteins-2 (rhBMP-2) is clinically useful for bone regeneration, induction of new bone formation requires a large amount of rhBMP-2 in humans. Many investigators have been concentrated efforts on searching materials which enhance the effect of rhBMP-2, and then heparin was found as a potentiating material to rhBMP-2. The purpose of this study were histomorphometrically to analyze the enhancing effect of heparin to rhBMP-2 and to study the mode of actions of heparin to rhBMP-2. Stem cells obtained from rabbit adipose tissue were divided into 4 groups according to heparin concentrations(0, 0.25, 2.5, and 25 μg/ml) with a constant rhBMP-2 concentration(150 ng/ml) and cultured for 2, 4, and 8 days. Naphtol AS phosphate-fast blue BB staining for alkaline phosphatase content and Alizarin red staining for calcium content were performed as time schedules and the morphology and osteoblastic activity were observed closely. 5 μg/ml rhBMP-2 was mixed with the following doses of heparin: 0, 0.25, 2.5, and 25 μg/ml. Each mixture was blotted into 0.5 g of multiporous anorganic bovine bone and was inserted into the critical sized calvarial defects(diameter of 0.8-mm) of rabbits. After 1, 3, and 6 weeks, the harvested tissues were processed and stained using H&E and Masson’s trichrome methods. And the areas of newly formed bone in the grafted material were measured and statistically analyzed. During culture experiment of adipose stem cells with rhBMP-2 and heparin, the degree of osteoblastic differentiation was increased with increasing heparin concentration, but the cellular degeneration was accelerated at higher concentration of heparin as time passed. Consequently the osteoblastic differentiation of progenitor cells were accelerated as the concentration of heparin increased. In addition, the progenitor cells exhibited full differentiations early showing fast degeneration. The higher the concentration of heparin, larger newly formed bone in grafted materials was obtained in initial period. However, the increased amount of the newly formed bone in grafted materials was progressively decreased at the higher concentration of heparin as time passed. In conclusion, the heparin has influence on the osteoinductive effect of rhBMP-2 in the initial stage of bone formation. The use of heparin with rhBMP-2 could offer cheaper, safer, and improve clinical results in grafting procedures.

Highly sensitive ratiometric detection of heparin and its oversulfated chondroitin sulfate contaminant by fluorescent peptidyl probe

Mehta, Pramod Kumar,Lee, Hyeri,Lee, Keun-Hyeung Elsevier 2017 Biosensors & Bioelectronics Vol.91 No.-

<P><B>Abstract</B></P> <P>The selective and sensitive detection of heparin, an anticoagulant in clinics as well as its contaminant oversulfated chondroitin sulfate (OSCS) is of great importance. We first reported a ratiometric sensing method for heparin as well as OSCS contaminants in heparin using a fluorescent peptidyl probe (<B>Pep1</B>, pyrene-GSRKR) and heparin-digestive enzyme. <B>Pep1</B> exhibited a highly sensitive ratiometric response to nanomolar concentration of heparin in aqueous solution over a wide pH range (2~11) and showed highly selective ratiometric response to heparin among biological competitors such as hyaluronic acid and chondroitin sulfate. <B>Pep1</B> showed a linear ratiometric response to nanomolar concentrations of heparin in aqueous solutions and in human serum samples. The detection limit for heparin was calculated to be 2.46nM (R<SUP>2</SUP>=0.99) in aqueous solutions, 2.98nM (R<SUP>2</SUP>=0.98) in 1% serum samples, and 3.43nM (R<SUP>2</SUP>=0.99) in 5% serum samples. <B>Pep1</B> was applied to detect the contaminated OSCS in heparin with heparinase I, II, and III, respectively. The ratiometric sensing method using <B>Pep1</B> and heparinase II was highly sensitive, fast, and efficient for the detection of OSCS contaminant in heparin. <B>Pep1</B> with heparinase II could detect as low as 0.0001% (w/w) of OSCS in heparin by a ratiometric response.</P> <P><B>Highlights</B></P> <P> <UL> <LI> Highly sensitive and selective ratiometric detection of heparin among various biological heparin competitors </LI> <LI> The detection limit for heparin was measured to be 2.46nM </LI> <LI> Highly sensitive ratiometric detection of OSCS in heparin using only one heparin-digestive enzyme (heparinase II) </LI> <LI> Ratiometric detection of as low as 0.0001% (w/w) of OSCS in heparin </LI> </UL> </P>

Boronic acid-functionalized oligoamine colorimetric heparin receptor

Joseph, Vincent S.,Hong, Jong-Dal Elsevier 2018 Colloids and surfaces. A, Physicochemical and engi Vol.543 No.-

<P><B>Abstract</B></P> <P>Here we developed a synthetic colorimetric heparin receptor (denoted as OHR) bearing phenylboronic acid (PBA) and intermediary cationic ammonium groups on a geometrically flat biocompatible oligoamine framework. OHR showed remarkable affinity for heparin (at physiological conditions) with a binding constant comparable to the best such values reported previously. The high affinity of OHR for heparin was ascribed primarily to the flat molecular geometry of the receptor, which apparently allowed for effective interactions of the cationic ammonium and PBA groups of OHR with the functional groups along the bulky polymer chain of heparin. Moreover, this high affinity of OHR for heparin under physiological conditions as well as the high biocompatibility of OHR are features that make OHR a promising candidate for use in potential biomedical applications including the bedside detection of heparin levels in human blood during surgery, and the use of heparin reversal agents.</P> <P><B>Graphical abstract</B></P> <P>We developed a synthetic colorimetric heparin receptor OHR bearing phenylboronic acid and intermediary cationic ammonium groups on a geometrically flat biocompatible oligoamine framework. OHR showed remarkable affinity for heparin (at physiological conditions), which was ascribed primarily to the flat molecular geometry of the receptor, which apparently allowed for effective interactions of OHR with the functional groups along the bulky polymer chain of heparin.</P> <P>[DISPLAY OMISSION]</P>

헤파린민감도평가를 위한 두 가지 방법의 비교; 체외에서 헤파린 혼합 혈장을 사용한 aPTT 검사와 헤파린 치료중인 환자 혈장을 사용한 anti-Xa 검사

구본경,권의훈,유광현,윤재원,김희진 대한임상검사과학회 2011 대한임상검사과학회지(KJCLS) Vol.43 No.4

The monitoring of heparin therapy is using almost aPTT assay. This study is compare to estimating aPTT therapeutic range using in vitro heparin-spiked sample and aPTT therapeutic range using in vivo heparin-treated sample. Normal pooled plasma was collected from 20 healthy representative individuals. 11 concentration of heparinized plasmas from 0 U/mL to 1.0 U/mL at intervals of 0.1 U/mL made by addition of heparin to normal pooled plasma were measured aPTT. The aPTT therapeutic range was performed through correlation analysis between heparin level 0.2 to 0.4 U/mL and aPTT. 30 plasmas from patients on heparin therapy were measured aPTT and anti-Xa activity. The aPTT therapeutic range was performed through correlation analysis between anti-Xa activity 0.3 to 0.7 U/mL and aPTT. The aPTT therapeutic range corresponded by heparin level-vs-aPTT value regression analysis was 60.7 to 102.4 seconds. The aPTT therapeutic range corresponded by anti-Xa activity-vs-aPTT value regression analysis was 85.3 to 147.5 seconds. The validation of heparin sensitivity using in-vitro heparin sample was not considered. The establishing aPTT therapeutic range is recommended anti-Xa activity using in-vivo sample. .

Comparison of Two Methods for Heparin Sensitivity; Activated Partial Thromboplastin Time Assay using in vitro Heparin-spiked Sample and Anti-Xa Assay using in vivo Heparin-treated Sample

Koo, Bon-Kyung,Kwon, Eui-Hoon,Ryu, Kwang-Hyun,Yun, Jae-Won,Kim, Hee-Jin 대한임상검사과학회 2011 대한임상검사과학회지(KJCLS) Vol.43 No.4

The monitoring of heparin therapy is using almost aPTT assay. This study is compare to estimating aPTT therapeutic range using in vitro heparin-spiked sample and aPTT therapeutic range using in vivo heparin-treated sample. Normal pooled plasma was collected from 20 healthy representative individuals. 11 concentration of heparinized plasmas from 0 U/mL to 1.0 U/mL at intervals of 0.1 U/mL made by addition of heparin to normal pooled plasma were measured aPTT. The aPTT therapeutic range was performed through correlation analysis between heparin level 0.2 to 0.4 U/mL and aPTT. 30 plasmas from patients on heparin therapy were measured aPTT and anti-Xa activity. The aPTT therapeutic range was performed through correlation analysis between anti-Xa activity 0.3 to 0.7 U/mL and aPTT. The aPTT therapeutic range corresponded by heparin level-vs-aPTT value regression analysis was 60.7 to 102.4 seconds. The aPTT therapeutic range corresponded by anti-Xa activity-vs-aPTT value regression analysis was 85.3 to 147.5 seconds. The validation of heparin sensitivity using in-vitro heparin sample was not considered. The establishing aPTT therapeutic range is recommended anti-Xa activity using in-vivo sample.

헤파린이 rhBMP-2에 미치는 영향에 관한 조직계측 분석

정명호,이종헌 대한구강악안면병리학회 2016 대한구강악안면병리학회지 Vol.40 No.2

Although recombinant human Bone Morphogenetic Proteins-2 (rhBMP-2) is clinically useful for bone regeneration, induction of new bone formation requires a large amount of rhBMP-2 in humans. Many investigators have been concentrated efforts on searching materials which enhance the effect of rhBMP-2, and then heparin was found as a potentiating material to rhBMP-2. The purpose of this study were histomorphometrically to analyze the enhancing effect of heparin to rhBMP-2 and to study the mode of actions of heparin to rhBMP-2. Stem cells obtained from rabbit adipose tissue were divided into 4 groups according to heparin concentrations(0, 0.25, 2.5, and 25 μg/ml) with a constant rhBMP-2 concentration(150 ng/ml) and cultured for 2, 4, and 8 days. Naphtol AS phosphate-fast blue BB staining for alkaline phosphatase content and Alizarin red staining for calcium content were performed as time schedules and the morphology and osteoblastic activity were observed closely. 5 μg/ml rhBMP-2 was mixed with the following doses of heparin: 0, 0.25, 2.5, and 25 μg/ml. Each mixture was blotted into 0.5 g of multiporous anorganic bovine bone and was inserted into the critical sized calvarial defects(diameter of 0.8-mm) of rabbits. After 1, 3, and 6 weeks, the harvested tissues were processed and stained using H&E and Masson’s trichrome methods. And the areas of newly formed bone in the grafted material were measured and statistically analyzed. During culture experiment of adipose stem cells with rhBMP-2 and heparin, the degree of osteoblastic differentiation was increased with increasing heparin concentration, but the cellular degeneration was accelerated at higher concentration of heparin as time passed. Consequently the osteoblastic differentiation of progenitor cells were accelerated as the concentration of heparin increased. In addition, the progenitor cells exhibited full differentiations early showing fast degeneration. The higher the concentration of heparin, larger newly formed bone in grafted materials was obtained in initial period. However, the increased amount of the newly formed bone in grafted materials was progressively decreased at the higher concentration of heparin as time passed. In conclusion, the heparin has influence on the osteoinductive effect of rhBMP-2 in the initial stage of bone formation. The use of heparin with rhBMP-2 could offer cheaper, safer, and improve clinical results in grafting procedures.