富血小板纤维蛋白凝胶三维结构及其对人牙髓细胞体外增殖的影响

上海口腔医学 ›› 2015, Vol. 24 ›› Issue (3): 263-268.

富血小板纤维蛋白凝胶三维结构及其对人牙髓细胞体外增殖的影响

何璇,韦维,陈文霞

广西医科大学附属口腔医院牙体牙髓病科,广西南宁 530021

收稿日期:2014-09-25 出版日期:2015-06-20 发布日期:2015-07-24
通讯作者: 陈文霞,Tel:0771-5358225,E-mail:angelaxiacw@163.com
作者简介:何璇（1989-）,女,硕士,E-mail:hexuan269@163.com
基金资助:
国家自然科学基金（81160133）

Three-dimensional structure of platelet-rich fibrin gel and its effect on proliferation of human dental pulp cells in vitro

HE Xuan,WEI Wei,CHEN Wen-xia

Department of Endodontics, College of Stomatology, Guangxi Medical University. Nanning 530021, Guangxi Zhuang Autonomous Region, China

Received:2014-09-25 Online:2015-06-20 Published:2015-07-24
Supported by:
National Natural Science Foundation of China (81160133)

摘要/Abstract

摘要： 目的:分析富血小板纤维蛋白（platelet-rich fibrin,PRF）凝胶的三维结构,并探索不同浓度PRF凝胶析出液对人牙髓细胞（human dental pulp cells,hDPCs）体外增殖的影响,评价PRF凝胶作为牙髓组织再生支架的可行性。方法:采用Choukroun一步离心法,制备PRF凝胶,分别行组织学和扫描电镜（SEM）观察。将新鲜制备的PRF凝胶浸泡于DMEM培养基中,于第7天取出析出液。分别用含PRF 凝胶析出液浓度（体积分数）为25%（1PRF组）和75%（3PRF组）的DMEM培养基培养hDPCs。采用细胞计数试剂盒 8（CCK-8）检测24、48、72、96、120、144、168 h各时间点的细胞增殖活性。采用SPSS16.0软件包对数据进行统计学处理。结果:光镜和SEM观察结果均表明,血小板和白细胞主要分布在PRF凝胶的白膜层,此层由粗大而密集的纤维蛋白条索构成。CCK-8结果显示,在24、48、72、96 h,1PRF组和3PRF组的光密吸光度（OD）值与对照组相比,差异无显著性（P>0.05）;在120、144、168 h,1PRF组和3PRF组的光密度值显著高于对照组(P<0.05);1PRF组的OD值和3PRF组相比,差异无显著性(P>0.05)。结论:PRF凝胶的三维结构由纤维蛋白网构成,其白膜层聚集了大量可释放生长因子的血小板与白细胞;PRF 凝胶析出液对hDPCs体外增殖的促进作用具有时间依赖性,提示PRF凝胶有望成为牙髓再生的良好支架材料。

关键词: 富血小板纤维蛋白凝胶, 三维结构, 人牙髓细胞

Abstract: PURPOSE: This study was designed to analyze the three-dimensional structure of platelet-rich fibrin (PRF) gel as scaffold for dental pulp regeneration, and to investigate the effect of PRF gel on the proliferation of human dental pulp cells (hDPCs) in vitro. METHODS: PRF gel prepared by Choukroun’s protocols was evaluated by light microscope and scanning electron microscope (SEM). The extract of PRF gel was collected at the time point of 7th day. The experimental group was divided into 2 subgroups (25% and 75%) according to the volume fraction of PRF gel extract. The 2 subgroups were named 1PRF and 3PRF, respectively. The hDPCs were isolated and cultured in DMEM with 1PRF and 3PRF, respectively. Cell proliferation was detected using cell counting kit-8 (CCK-8) at 24, 48, 72, 96, 120, 144 and 168 h, respectively. The data was analyzed with SPSS16.0 software package. RESULTS: Platelet and leukocyte were mainly distributed at the junction between the red corpuscles and the PRF gel, which was called "buffy coat". "Buffy coat" was constructed by large and dense fibrin clusters. At 24, 48, 72 and 96 h, there was no significant difference (OD) value of optical density(OD) between experimental group and control group(P>0.05). At 120, 144 and 168 h, the OD value in the experimental group was significant higher than that of the control group (P<0.05); there was no significant difference of OD value between group 1PRF and 3PRF (P>0.05). CONCLUSIONS: The results confirm that the three-dimensional structure of PRF gel is constituted by fibrin network. A large amount of platelet and leukocyte which can release high quantities of growth factors are concentrated in the PRF gel. The extract of PRF gel can accelerate the proliferation of hDPCs in time-dependent manner. PRF gel may be a proper scaffold for pulp regeneration.

Key words: Platelet-rich fibrin gel, Three-dimensional structure, Human dental pulp cells

中图分类号:

R781.3

何璇,韦维,陈文霞. 富血小板纤维蛋白凝胶三维结构及其对人牙髓细胞体外增殖的影响[J]. 上海口腔医学, 2015, 24(3): 263-268.

HE Xuan,WEI Wei,CHEN Wen-xia. Three-dimensional structure of platelet-rich fibrin gel and its effect on proliferation of human dental pulp cells in vitro[J]. Shanghai Journal of Stomatology, 2015, 24(3): 263-268.

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