贫血小板血浆和根面脱矿对人牙周膜成纤维细胞在病变牙根面附着和增殖的影响

目的：观察贫血小板血浆和根面脱矿单独及联合应用对人牙周膜成纤维细胞在病变牙根面的附着及增殖的影响,探讨PPP和根面脱矿处理在促牙周组织再生中的可能作用。方法：实验分4组,经PPP、EDTA和两者联合处理的病变根片的3组作为实验组,未处理的病变根片作对照组。分别通过细胞计数法、四唑盐比色法[MTT]和扫描电镜检测人牙周膜成纤维细胞在不同处理病变牙根表面的附着、增殖和形态。结果：与未处理组相比,PPP组及根面脱矿组均能促进人牙周膜成纤维细胞对病变根面的附着（P〈0.05）,二者联合处理促细胞附着效果明显增强（P〈0.01）;未处理组中人牙周膜成纤维细胞体外培养48h与培养24h相比,细胞在病变根片上的无增殖（P〈0.05）;单独PPP或脱矿处理组中人牙周膜成纤维细胞体外培养48h与培养24h相比,细胞在病变根片上的附着及增殖增加,但无统计学意义,PPP和根面脱矿联合处理组中细胞在病变根片上的附着及增殖明显增加（P〈0.05）。结论：PPP能牢固地附着于病变根面,PPP和根面脱矿单独和联合处理病变根片能明显加强人牙周膜成纤维细胞的附着和增殖,而PPP和根面脱矿联合处理还有显著的促细胞增值效应。     

富血小板血浆和贫血小板血浆处理的生物降解膜对牙周韧带成纤维细胞影响的扫描电镜观察

目的 :探索富血小板血浆 (platelet -richplasma ,PRP)和贫血小板血浆 (platelet -poorplasma,PPP)处理后的生物降解膜对附着的人牙周韧带成纤维细胞形态的影响。方法 :体外培养人牙周韧带成纤维细胞 ,应用梯度密度离心 ,从人新鲜全血中获取PRP和PPP ,并分别预处理两种生物降解膜。体外细胞培养 2 4h ,扫描电镜观察PRP和PPP对两种生物降解膜的附着效果 ,以及附着于膜上人牙周韧带成纤维细胞的超微结构。结果 :PRP和PPP能有效地贴附于生物降解膜 ,在未处理和处理后的生物降解膜上均可见附着的呈梭形或长扁形的人牙周韧带成纤维细胞 ,但在处理膜上人牙周韧带成纤维细胞明显地被包裹于PRP和PPP中 ,仅见增大的胞体、胞浆突起的轮廓 ,细胞彼此重叠或融合成片状。结论 :有效地贴附于生物降解膜的PRP和PPP能将人牙周韧带成纤维细胞网织在一起 ,并促进细胞的增殖。提示经PRP和PPP处理的生物降解膜可能更利于引导牙周组织再生。     

牙周膜成纤维细胞与三种可吸收引导组织再生膜生物相容性实验研究

目的：探讨牙周膜成纤维细胞与可吸收引导组织再生膜生物相容性，为牙周组织工程中支架材料选择提供依据。方法：将人牙周膜成纤维细胞与三种商品化引导组织再生膜：BME-10XR(中国医学科学院生物医学工程研究所)、BioMeshR(韩国Samyang公司)及Bio-GideR(美国Osteohealth公司)体外复合培养，进行形态学观察、细胞附着及增殖的检测。结果：人牙周膜成纤维细胞可在三种可吸收膜上附着、增殖，并复层生长。细胞在三种膜上的附着无显著性差异(P＞0．05)；在BMK-10XR及Bio-GideR上生长、繁殖明显好于BioMesh R(P＜0．05)。结论：BME-10X R及Bio-Gide R具有良好的生物相容性，有望成为牙周膜成纤维细胞的载体材料应用于牙周组织工程研究.     

富血小板血浆和乏血小板血浆包被的屏障膜对成骨细胞附着的影响

目的：观察富血小板血浆（plateletrich plasma，PRP）和乏血小板血浆（platelet poor plasma，PPP）包被的屏障膜对人牙槽骨成骨细胞附着的影响。方法取第4代人牙槽骨成骨细胞用于实验。健康成人的全血经过两次离心得到PRP和PPP。将A膜（GoreTex-ePTFE^TM膜）、B膜（GoreTex-Resolut^TM膜）和C膜（Inion-GTR^TM膜）冲切成直径为3mm的圆片并固定于24孔培养板底，用盖玻片作为阳性对照，将包被液分为PRP、PPP、磷酸盐缓冲液（phosphate buffer solution，PBS）3个组，分别包被屏障膜或玻片（仅用磷酸盐）2h，将成骨细胞以5×10^7个／L每孔接种于屏障膜或玻片上并孵育24h使细胞附着。苏木素染色，光镜下观察并计数，扫描电镜观察成骨细胞附着膜上的形态。结果：PRP组包被的A、B、C膜上的细胞数分别为23、35和41；PPP组包被的A、B、C膜上的细胞数分别15、12和22；PBS包被的A、B、C膜上的细胞数分别为3、4和6。成骨细胞在PRP、PPP组的附着数量明显高于磷酸盐组（P〈0．05）；PRP组较PPP组的附着数量多（P〈0．05）；盖玻片上的附着数量显著多于3种屏障膜。3种屏障膜相比，B膜和C膜的成骨细胞附着量高于A膜（P〈0．05）。扫描电镜结果显示，PRP组屏障膜上成骨细胞呈梭形，贴壁好，膜表面可见血小板、交织成网状的纤维蛋白，细胞呈现复层生长；PPP组或磷酸盐组成骨细胞贴壁不完全，呈圆形。结论：PRP和PPP能促进成骨细胞在屏障膜上的附着数量；PRP能改善成骨细胞在屏障膜上的附着方式。     

促进牙周膜成纤维细胞在病变牙根面的附着和增殖

富血小板血浆(platelet-rich plasma,PRP)含有丰富的生长因子和含量较高的TGF-β和PDGF,能促进体外培养的牙周成纤维细胞和成骨细胞的增殖和胶原的形成[1],Lekovic[2]将PRP、牛骨粉联合应用治疗慢性牙周炎骨缺损患者取得较好的效果,刘琪等[3]的研究也表明PRP处理的生物降解膜能     

碱性成纤维细胞生长因子对成骨细胞和牙周膜成纤维细胞迁移、增殖的影响

目的　明确碱性成纤维细胞生长因子(bFGF)对体外成骨细胞和牙周膜成纤维细胞迁移、增殖的影响,以 探讨在牙种植体组织界面局部应用bFGF诱导类牙周膜形成的可行性。方法　同一只SD大鼠来源的成骨细胞和 牙周膜成纤维细胞经传代培养至第4代,建立体外创伤模型,分别在普通培养基和含bFGF的培养基中培养,观察 细胞迁移情况,四唑盐比色实验(MTT)测定细胞的增殖速度。结果　普通培养基中,成骨细胞迁移速度快于成纤维 细胞。加bFGF培养基中牙周膜成纤维细胞迁移速度明显快于其他各组,同时MTT结果显示加入bFGF能明显促进 两种细胞的增殖。结论　bFGF能明显促进牙周膜成纤维细胞的增殖、移行。     

放线共生放线杆菌表面相关物质对人牙周膜成纤维细胞在牙根片上附着的影响

目的：观察放线共生放线杆菌表面相关物质对人牙周膜成纤维细胞在牙根片上附着的影响。方法：应用细胞计数法和扫描电镜观察法。结果：100mg/L组对人牙周膜成纤维细胞在根面附着有明显抑制作用,表现为正常牙根片上附着细胞数明显少于对照组（P＜0．05）;扫描电镜显示细胞生长数量减少,细胞突起伸展不充分。结论：此物质可抑制人牙周膜成纤维细胞在牙根面上的附着。     

牙周膜成纤维细胞在三维静电纺丝纳米纤维支架上的培养

目的:研究人牙周膜成纤维细胞与静电纺丝聚乳酸/聚己内酯纳米纤维支架体外培养的生物相容性.方法:分离、培养人牙周膜成纤维细胞,接种在静电纺丝纳米纤维支架上,与常规培养条件下的细胞进行比较,观察生长形态、生长曲线、倍增时间及活性.结果:人牙周膜成纤维细胞生长情况与常规培养基本一致,2组间的倍增时间比较无统计学差异(P＞0.05),活细胞百分率与正常培养无明显统计学差异(P＞0.05).结论:人牙周膜成纤维细胞在三维静电纺丝纳米纤维上生长、增殖,该支架材料具有很好的生物相容性,可作为牙周膜组织工程候选支架材料.     

体外培养碱性成纤维细胞因子对人牙周膜成纤维细胞生物学特性的影响

目的：探讨体外培养过程中，碱性成纤维细胞生长因子（bFGF）对人牙周膜成纤维细胞（hPDLFs）生物学特性的影响。方法：体外分离培养人牙周膜成纤维细胞并鉴定，加入不同浓度bFGF（1ng／ml、10ng／ml、50ng／ml、100ng／ml）培养，MTT方法检测细胞增殖情况；并对细胞进行矿化诱导，检测细胞的碱性磷酸酶活性。结果：hPDLFs呈星形或长梭形，免疫组化波丝蛋白阳性，角蛋白阴性，证实该细胞来源可靠。bFGF在一定浓度范围内，与细胞增殖成正比，而在本实验培养条件下（10％FBS）bFGF最大效应浓度为10ng／ml。矿化诱导条件下，碱性磷酸酶活性明显增加，在联合应用bFGF的情况下，100ng／ml组碱性磷酸酶活性明显高于其他组。结论：不同浓度bFGF对人牙周膜成纤维细胞生物学行为的影响不同，在一定浓度条件下，低浓度bFGF促进人牙周膜成纤维细胞的增殖，而高浓度bFGF作用于人牙周膜成纤维细胞可能更易于分化。     

bFGF对人牙髓、牙周膜成纤维细胞生物学效应的研究

目的：了解碱性成纤维细胞生长因子（bFGF）对人牙髓、牙周膜成纤维细胞的生物学效应,为bFGF在牙髓、牙周病中的治疗研究提供新的实验依据。方法：利用3H-TdR掺入法观察在bFGF作用下人牙髓、牙周膜成纤维细胞DNA和胶原蛋白合成的情况。结果：20ng/mL～60ng/mLbFGF可明显促进人牙髓、牙周膜成纤维细胞DNA的合成（P〈0.01）,在40ng/mL浓度时牙髓,牙周膜成纤维细胞DNA合成最高,40ng/mLbFGF作用于牙髓,牙周膜成纤维细胞,24～48h可使细胞DAN合成显著增多,牙髓成纤维细胞在36h时DNA合成达最高峰,牙周膜成纤维细胞在24h时DNA合成达最高峰;bFGF对牙髓,牙周膜成纤维细胞胶原蛋白的合成无明显促进作用（P〉0.05）。结论：人牙髓、牙周膜成纤维细胞是bFGF的靶细胞,其胞膜上可能有bFGF特异性受体的存在,也表明bFGF在牙髓、牙周组织的创伤愈合中可能起重要作用。     

自体来源富血小板血浆对大鼠牙髓细胞增殖作用的研究

目的:初步探讨富血小板血浆及之中生长因子对大鼠牙髓细胞增殖的作用.方法:采用Landesberg 法制备PRP,ELISA测定PRP中两种主要生长因子:转化生长因子(TGF - β1)和血小板源性生长因子(PDGF -AB)的浓度;CCK-8法观察5％、10％PRP在48h时对牙髓细胞的增殖作用;在5％PRP中加入TGF-β1抗体和PDGF-AB抗体分别拮抗TGF-β1和PDGF-AB的作用并观察对细胞增殖的影响.结果:所制备的PRP中血小板含量为1790.58±388.08×109个/L,ELISA的方法测定PRP中TGF-β1及PDGF-AB的浓度分别为3.080 ng/ml和10.706 ng/ml.CCK-8法测定5％和10％PRP对牙髓细胞均有增殖作用(P＜0.05,与对照组相比);屏蔽生长因子可以明显改变5％PRP对大鼠牙髓细胞的增殖作用(P＜0.05);拮抗PDGF-AB组比拮抗TGF - β1组降低增殖作用明显(P＜0.05).结论:本实验制备的PRP含有高浓度的TGF - β1及PDGF-AB,不同浓度的PRP均能有效促进牙髓细胞增殖;屏蔽PDGF-AB明显降低5％PRP对大鼠牙髓细胞的增殖作用.     

Effect of autologous and allogenic platelet-rich plasma on human gingival fibroblast function

Oral Diseases (2012) 18 , 494–500 Objective: Platelet‐rich plasma (PRP) has been proposed as a method of delivering growth factors to enhance regeneration. The aim of this study was to investigate the use of autogenous and allogenic PRP and platelet‐poor plasma (PPP) on migration and proliferation of human gingival fibroblasts in vitro. Methods: Various concentrations of PRP, as well as PPP, were prepared from autologous and allogenic sources and applied to primary gingival fibroblasts. Migration was determined by assessing the fibroblast response to a concentration gradient. ³H‐thymidine incorporation and crystal violet colorimetric assays were utilized to assess DNA synthesis and proliferation. Results: Platelet‐rich plasma provides a significant migratory stimulus to gingival fibroblasts. Furthermore, the various concentrations of PRP (50%, 20% and 10%) do not promote DNA synthesis in the short term (24 h), but over the longer term (5 days) they stimulate an increase in cell proliferation. Compared with PPP, PRP was superior in terms of encouraging migration, but was inferior in terms of promoting DNA synthesis and cell proliferation. No difference was noted between the autologous and allogenic PRP preparations on cell function. Conclusion: Both PPP and PRP promote gingival fibroblast migration and proliferation in vitro, without differences between preparations obtained from autologous and allogenic sources.     

Attachment of periodontal fibroblasts to barrier membranes coated with platelet-rich plasma

BACKGROUND: This study evaluated the attachment of human periodontal ligament (HPDL) and human gingival (HG) fibroblasts onto commercially available barrier membranes coated with platelet-rich plasma (PRP). METHODS: Gore-Tex Regenerative Membrane (GTN1), Inion GTR Biodegradable system (INION), and Gore-Resolut XT Regenerative membrane (GTRX) were studied. The membranes were pre-wetted with phosphate buffered saline before exposure to PRP for two hours. HPDL or gingival fibroblasts were then seeded onto the membranes and incubated in culture medium for 24 hours. The samples were then fixed and processed for light and scanning electron microscopy evaluation evaluation. RESULTS: PRP-coated membranes showed greater cell attachment compared to non-coated membranes. The INION membrane showed the greatest attachment followed by the GTN1 and GTRX membranes. The cells showed increased cytoplasmic extensions and appeared more intertwined around individual fibres of the membranes compared to uncoated control membranes. CONCLUSION: HPDL and HG fibroblasts have a distinct reaction with different membranes depending on both microstructure and composition of the membrane. Coating membranes with PRP had a significant impact on the way HPDL and HG fibroblasts attach to certain barrier membranes by increasing both the amount and the quality of attachment.     

富血小板血浆对牙髓成纤维细胞附着、增殖的影响

目的 :探讨富血小板血浆 (PRP)对牙髓成纤维细胞附着、增殖的影响。方法 :利用原代细胞体外培养技术、PRP提取技术及四唑盐比色法 (MTT)检测 5 %PRP对牙髓成纤维细胞附着的影响和不同浓度PRP(5 %、10 %、2 0 %、30 %、4 0 %、5 0 % )对牙髓成纤维细胞增殖的影响。结果 :在附着实验中 ,5 %PRP组所测得的平均光密度值 (OD值 )明显高于空白对照组 (P <0 .0 1)。在增殖实验中 ,各不同浓度PRP组所测得的平均OD值均明显高于对照组 (P <0 .0 1)。结论 :PRP可促进牙髓成纤维细胞的附着及增殖     

The effect of platelet-rich plasma on the cellular response of rat bone marrow cells in vitro

The aim of this study was to investigate the effect of platelet-rich plasma (PRP) on the proliferation and the differentiation of rat bone marrow cells (RBMCs). PRP, platelet-poor plasma (PPP), and bone marrow cells were derived from the rats (hearts and tibia) and the cells were cultured with or without PRP or PPP (0 [control]), 0.2 approximately 10 microL/mL). The proliferation of RBMCs was measured on days 2 and 4, and alkaline phosphatase (ALP) staining and activity measurement were evaluated to determine the effect of PRP on the differentiation on days 4 and 8. PRP enhanced the proliferation significantly compared to the control group (P < .05). These enhancements were greater than ones induced by the addition of PPP. ALP staining appeared to show that PRP decreased the number of ALP positive cells and ALP activity significantly (P < .05). Our results demonstrate that PRP stimulates the proliferation but suppresses the differentiation of RBMCs.     

血小板血浆对人牙髓细胞增殖的影响

目的：观察血小板血浆及其细化成分对人牙髓细胞增殖的影响。方法：使用因正畸而拔除的人牙的牙髓细胞，用细胞定量测定试剂盒测定细胞增殖情况。结果：5％的多血小板血浆（platelet-rich plasma，PRP）、5％和10％的洗净血小板（washed platelet，WPLT）均明显地促进了人牙髓细胞的增殖，而且WPLT的作用较PRP更显著。乏血小板血浆（platelet-poor plasma，PPP）呈浓度依赖性地抑制了人牙髓细胞增殖，5％WPLT促进人牙髓细胞增殖的作用。结论：去除血浆成分的WPLT对培养的人牙髓细胞增殖有明显的促进作用；血浆中可能存在对抗血小板生长因子作用的因子。     

The contribution of platelet-derived growth factor, transforming growth factor-beta1, and insulin-like growth factor-I in platelet-rich plasma to the proliferation of osteoblast-like cells

The aim of this study was to investigate the effect of platelet-rich plasma (PRP) on the proliferation of osteoblast-like cells in vitro. PRP was prepared using a centrifuge; the number of platelets (n = 32) and the levels of platelet-derived growth factor-AB (PDGF-AB), transforming growth factor-beta1 (TGF-beta1), and insulin-like growth factor-I (IGF-I) were measured (n = 16). For the proliferation assay, SaOS-2 was cultured in the presence of platelet-poor plasma (PPP), whole blood, or PRP. The cell number was counted after 36 and 72 hours. To investigate the effect of each growth factor, the cells were cultured with PRP in the absence or presence of neutralizing antibodies, and counted as described. The mean platelet count of PRP was 1546.36 +/- 382.25 x 10(3)/microL, and the mean levels of PDGF-AB, TGF-beta1 and IGF-I were 0.271 +/- 0.043, 0.190 +/- 0.039, and 0.110 +/- 0.039 ng/1500 x 10(3) platelets, respectively. Cell proliferation was enhanced in all PRP groups in a dose-dependent manner, and all neutralizing antibodies significantly suppressed proliferation compared with the PRP group, lacking antibody, at 36 hours. However, at 72 hours, the neutralizing antibodies of PDGF and TGF-beta1, but not IGF-I, significantly suppressed proliferation. These results show the beneficial abilities of PRP in the proliferation of osteoblast-like cells from the standpoint of growth factors, including the contribution of each factor.