复合富血小板血浆的酶处理异种骨修复兔桡骨缺损

目的:自源性的富血小板血浆可促进骨组织及软组织的修复,又不存在疾病传播及免疫排斥的可能。实验拟验证应用复合富血小板血浆的酶处理异种骨修复节段性骨缺损的可行性。方法:实验于2006-07/12在解放军昆明总医院实验动物中心完成。①实验分组:选用新西兰大耳白兔20只,体质量2.0～2.5kg,共40只前肢,随机分为复合酶组,单纯酶组,脱蛋白骨组,空白组。每组共10个标本。②实验方法:制作富血小板血浆及酶处理的异种骨并制备桡骨中段15mm的节段性骨缺损模型,将上述骨材料植入兔桡骨缺损处,其中复合酶组:植入复合富血小板血浆的酶处理异种骨;单纯酶组:植入酶处理异种骨;脱蛋白骨组:植入部分脱蛋白骨;空白组:不植入任何材料。③实验评估:分别于术后4,8,12周取材,X线片及苏木精-伊红染色分别观察骨缺损区的新骨形成情况。同时对各组骨密度进行测定。结果:纳入新西兰大耳白兔20只,均进入结果分析,所有手术无术后感染,无动物死亡,植入骨无脱落。①所有动物麻醉苏醒后均恢复进食,2周伤口愈合,未出现感染及渗液等。富血小板血浆中的血小板含量均为全血的4倍以上。②在同一时间点,酶处理后异种骨与部分脱蛋白骨在成骨能力方面差异无显著性意义,而复合富血小板血浆的酶处理异种骨在8周时骨缺损部分修复;12周时完全修复。空白组骨缺损未修复。③8,12周时复合酶组骨密度较单纯酶组、脱蛋白骨组高,差异有显著性意义(P<0.05);单纯酶组与脱蛋白骨组比较无明显差异(P>0.05)。结论:经酶处理后异种骨可用于节段性骨缺损的修复,复合富血小板血浆后有明显加速骨愈合的作用。     

同种异体脱蛋白骨复合纤维蛋白胶体内构建组织工程骨

背景：已有实验证明复合纤维蛋白胶的异体骨在体外培养中更适合种子细胞生长和增殖。目的：观察复合纤维蛋白胶的脱蛋白骨与单纯脱蛋白骨作为骨髓间充质干细胞支架材料的差异。方法：将28只新西兰大白兔随机分为3组,实验组在兔背部植入复合纤维蛋白胶的脱蛋白骨与自体骨髓间充质干细胞,对照组在兔背部植入单纯脱蛋白骨与自体骨髓间充质干细胞,空白组在兔背部植入单纯脱蛋白骨。结果与结论：随着植入时间的延长,各组植入物碱性磷酸酶活性逐渐增强,但实验组强于对照组与空白组（P〈0．01）。实验组与对照组术后4,8周均有新骨形成,且实验组成骨量高于对照组（P〈0．01）,空白组未见新骨形成。表明复合纤维蛋白胶的脱蛋白骨新骨形成能力优于单纯脱蛋白骨,可作为骨髓间充质干细胞的复合型载体构建于组织工程骨。     

多孔羟基磷灰石与富血小板血浆和纤维蛋白胶复合修复骨缺损

背景：自体骨移植是治疗骨缺损的最理想方法,但来源有限,供区有一定的并发症,所以寻找自体骨的替代材料一直是骨科学领域的研究方向。目的：观察珊瑚多孔羟基磷灰石、富血小板血浆和纤维蛋白胶复合物修复骨缺损的效果。方法：在新西兰大白兔双侧前臂桡骨中段截骨1．5cm制成骨缺损模型,随机分为3组,实验组植入珊瑚多孔羟基磷灰石、富血小板血浆和纤维蛋白胶复合物,对照组植入自体骨,空白对照组未植入任何物质。结果与结论：①X射线：实验组术后12周时骨缺损基本修复,塑性完全,愈合过程与对照组同步：空白对照组骨缺损无明显变化。②组织病理学：实验组与对照组术后12周时骨缺损基本修复,出现成熟板层骨及哈佛氏管;空白对照组仅见大量成纤维细胞增生,未见骨质形成。③生物力学：术后2周时实验组最大扭矩和抗扭刚度优于对照组（P〈0．05）,术后12周时两组最大扭矩和抗扭刚度差异无显著性意义。表明珊瑚多孔羟基磷灰石、富血小板血浆和纤维蛋白胶复合物具有促骨质愈合的作用,甚至在术后早期修复骨缺损的效果优于自体骨。     

骨形态发生蛋白2复合异种脱蛋白骨支架材料修复山羊大段骨缺损

背景：单纯骨形态发生蛋白2修复骨缺损只有骨诱导性，单纯异种脱蛋白骨只有骨传导性。目的：评估异种脱蛋白骨复合骨形态发生蛋白2修复山羊大段长骨缺损的效果。设计、时间及地点：随机分组设计、动物对照观察实验，于2005—03／2007—02在解放军第三军医大学创伤、烧伤与复合伤国家重点实验室完成。材料：山羊24只用于制备大段长骨缺损模型。市售猪股骨用于制备脱蛋白骨。重组骨形态发生蛋白2为美国Biosource公司产品。方法：山羊右侧胫骨中下段截除胫骨总长度20％制备节段性骨缺损模型。24只山羊按植入材料的不同分为3组，单纯异种脱蛋白骨组、自体骨组和异种脱蛋白骨＋重组骨形态发生蛋白2组，每组8只。主要观察指标：植入后4，8，12，16，20，24周X射线评估骨缺损情况。植入后24周取新生骨组织进行双能X射线、组织学、生物力学检测修复效果。结果：植入后12，24周自体骨组和异种脱蛋白骨＋重组骨形态发生蛋白2组X射线评分均高于单纯异种脱蛋白骨组，差异有显著性意义（P〈0．05）。植入后24周自体骨组和异种脱蛋白骨＋重组骨形态发生蛋白2组骨密度和骨矿含量均高于单纯异种脱蛋白骨组，差异有显著性意义（P（0．05）。自体骨组和异种脱蛋白骨＋重组骨形态发生蛋白2组之间比较差异均无显著性意义（P〉0．05）。植入后24周生物力学测试表明，自体骨组〉异种脱蛋白骨＋重组骨形态发生蛋白2组〉单纯异种脱蛋白骨组。植入后24周，自体骨组、异种脱蛋白骨＋重组骨形态发生蛋白2组，新生骨与最初两断端连成一体，新生骨骨小梁排列整齐有序。单纯异种脱蛋白骨组，新生骨与最初两断端部分相连。结论：重组骨形态发生蛋白2复合异种脱蛋白骨修复山羊胫骨大段缺损成骨能力与自体骨相当。     

自体浓缩骨髓移植促进腱骨愈合

背景：前交叉韧带断裂后应用自体腘绳肌腱移植进行重建应用较多,腱-骨愈合的时间较长,限制了患者交叉韧带重建后早期的功能活动。目的：用浓缩自体骨髓植于前交叉韧带重建腱-骨界面,观察其界面愈合情况。方法：32只新西兰大白兔行双侧膝关节半腱肌肌腱前交叉韧带重建,随机选取一侧膝关节作为实验组,胫骨侧腱骨界面植入自体浓缩骨髓,对侧不植入作为对照组。结果与结论：组织学显示,前交叉韧带重建后2周对照组移植肌腱与骨隧道的界面由肉芽组织填充,连接组织松散,有成骨活动;实验组移植肌腱与骨隧道界面肉芽组织连接亦不紧密,骨隧道侧成骨活动较活跃。前交叉韧带重建后12周对照组腱骨结合部组织界面变模糊,可见胶原纤维-纤维软骨-钙化的软骨组织-骨组织的移行带改变;实验组显示明显的胶原纤维-纤维软骨-钙化的软骨组织-骨组织的移行带改变,潮线清晰。前交叉韧带重建后4,8,12周,实验组最大抗拉脱强度数值显著大于对照组（P〈0.05）。提示自体浓缩骨髓可以增强腱骨界面的抗拉伸强度,有利于腱-骨界的组织愈合。     

自体浓缩骨髓移植促进腱骨愈合

背景:前交叉韧带断裂后应用自体腘绳肌腱移植进行重建应用较多,腱-骨愈合的时间较长,限制了患者交叉韧带重建后早期的功能活动。目的:用浓缩自体骨髓植于前交叉韧带重建腱-骨界面,观察其界面愈合情况。方法:32只新西兰大白兔行双侧膝关节半腱肌肌腱前交叉韧带重建,随机选取一侧膝关节作为实验组,胫骨侧腱骨界面植入自体浓缩骨髓,对侧不植入作为对照组。结果与结论:组织学显示,前交叉韧带重建后2周对照组移植肌腱与骨隧道的界面由肉芽组织填充,连接组织松散,有成骨活动;实验组移植肌腱与骨隧道界面肉芽组织连接亦不紧密,骨隧道侧成骨活动较活跃。前交叉韧带重建后12周对照组腱骨结合部组织界面变模糊,可见胶原纤维-纤维软骨-钙化的软骨组织-骨组织的移行带改变;实验组显示明显的胶原纤维-纤维软骨-钙化的软骨组织-骨组织的移行带改变,潮线清晰。前交叉韧带重建后4,8,12周,实验组最大抗拉脱强度数值显著大于对照组(P<0.05)。提示自体浓缩骨髓可以增强腱骨界面的抗拉伸强度,有利于腱-骨界的组织愈合。     

海螵蛸富血小板血浆液复合物修复兔软骨损伤的可行性

背景:富血小板血浆是自体血小板的浓集物,富含大量生长因子,可以促进组织中细胞和基质的再生,促进组织修复,但其容易流动,不能有效固定于损伤部位。目的:以海螵蛸生物支架固定富血小板血浆,观察海螵蛸富血小板血浆复合物修复兔软骨损伤的效果。方法:将21只大耳白兔随机均分为3组,在兔右膝关节负重髁软骨面制作类似Hulth骨软骨损伤模型,实验组于骨损伤处植入海螵蛸支架-自体富血小板血浆复合物,对照组于骨缺损处植入明胶海绵-富血小板血浆复合物,空白对照组不植入任何材料。术后12周取材,进行大体观察与组织学观察。结果与结论:术后12周,实验组软骨缺损区域明显缩小,组织学观察缺损区域出现类软骨细胞,排列整齐,成柱状分布,非常接近正常软骨组织;对照组软骨缺损区域有轻度缩小,组织学观察骨基质表面可见类软骨细胞基质组织,但排列较为紊乱;空白对照组软骨缺损区域有轻度缩小,组织学观察到只有骨细胞及骨基质,血管组织存在,未见到软骨及类软骨组织。实验组缺损直径及深度均低于其他两组(P<0.05)。结果表明海螵蛸支架-富血小板血浆复合物可促进软骨缺损愈合。     

富血小板凝胶与拔牙窝新骨形成和骨量保持

背景：富血小板血浆能否促进骨组织的修复再生存在一定的争议。目的：研究富血小板血浆/凝胶对拔牙窝骨愈合过程中新骨形成和骨量保持的可能调节作用,探讨富血小板血浆/凝胶与骨愈合的相互关系。方法：通过拔牙建立Beagle犬拔牙窝骨缺损模型,同期在拔牙窝导入富血小板血浆或复血小板凝胶,并设计对照组。术后2,4,8,12周分别进行大体观察、放射影像学检查、三维CT平扫＋重建、组织学检查拔牙窝颊舌侧牙槽嵴高度差、CT值以及新生骨面积。结果与结论：与富血小板血浆组和对照组比较,影像学结果表明富血小板凝胶组在第2,4,8周新骨形成的面积最大（P〈0.01）;组织学结果表明富血小板凝胶组在第2,4周新骨形成面积最大（P〈0.05）;在所有时间点上富血小板凝胶组颊舌侧牙槽嵴高度差值最小（P〈0.05）。在第12周,富血小板凝胶组颊舌侧牙槽嵴高度仍有2mm差值。提示,富血小板凝胶具有促进牙槽窝早期愈合的能力,但其单独使用时促进牙槽窝骨量保持的效能有限。     

富血小板纤维蛋白/异种冻干骨复合物修复骨缺损及骨整合的作用

背景：异种冻干骨具有很好的骨引导性及低抗原性,但骨诱导性相对较差;自体富血小板纤维蛋白具有很好的诱导成骨能力。目的：观察富血小板纤维蛋白/异种冻干骨复合物在修复种植体周骨缺损及骨整合过程中的作用。方法：取新西兰大白兔12只,于双侧下颌无牙区造极限骨缺损模型。在两侧缺损区的近中、远中端骨壁各植入1枚钛螺纹钉,使种植体临近缺损侧无骨支持,右侧缺损区填压入自体富血小板纤维蛋白/异种冻干骨复合物,作为实验组;左侧缺损区填入异种冻干骨,作为对照组。植入4,8,12周后取完整下颌骨标本进行大体形态观察、植体扭矩测试及组织学观察。结果与结论：植入4周时,实验组可见新生血管,成骨细胞数量多,呈单层紧密排列在新生骨小梁的表面;对照组可见炎性细胞浸润,纤维组织增多,新生血管稀疏,成骨细胞及新生骨数量较实验组少。植入8周时,实验组见大量新生血管,骨小梁密集,呈网状连接,部分已逐渐融合成岛状;对照组新生血管较实验组少,骨小梁纤细,散乱。植入12周时,实验组新生骨组织成熟,骨小梁钙化程度高,可见形成板状新骨;对照组骨小梁融合成片,趋向成熟,但成熟度不及实验组。实验组不同时间点种植体-骨结合强度高于对照组（P 〈0.05）。表明富血小板纤维蛋白/异种冻干骨复合物具有很好的骨修复与骨整合能力。     

透明质酸钠复合重组人骨形态发生蛋白2凝胶对自体半腱肌重建兔前交叉韧带腱-骨愈合的生物力学分析

背景：在前交叉韧带重建的早期,腱-骨连接处是整个移植物-骨隧道复合体的薄弱环节,所以腱-骨愈合的情况是关系韧带移植重建成败的关键。目的：观察透明质酸钠复合重组人骨形态发生蛋白2凝胶对自体半腱肌重建兔前交叉韧带后腱-骨愈合的影响。设计、时间及地点：随机对照动物实验,于2007-09/2008-06在潍坊医学院动物实验中心实验室完成。材料：将重组人骨形态发生蛋白2和透明质酸钠混合制成混悬凝胶注射剂。48只兔随机分成2组,每组24只48膝。方法：实验组随机选取一侧于胫骨隧道入口处注入重组人骨形态发生蛋白2与透明质酸钠凝胶（透明质酸钠-重组人骨形态发生蛋白2亚组）,另一侧仅做重建后骨道不予处理（未处理亚组）;对照组随机选取一侧于胫骨骨道入口处仅注入重组人骨形态发生蛋白2（重组人骨形态发生蛋白2亚组）,另一侧仅注入透明质酸钠（透明质酸钠亚组）。主要观察指标：术后2,4,8,12周大体观察肌腱移植物生长情况及在胫骨隧道内的愈合情况,生物力学拉伸试验测定其生物力学的性能。结果：透明质酸钠-重组人骨形态发生蛋白2亚组在术后2周隧道内见稀疏软骨样组织,4～8周时隧道内见沿骨隧道排列的软骨样组织,双股肌腱之间形成稳定组织连接。l2周时出现部分骨样组织包裹肌腱。其他亚组术后2周时隧道内仅为瘢痕组织,到术后12周隧道内肌腱与骨道壁之间仍充满瘢痕组织,未见稳定组织连接形成。4,8和12周透明质酸钠-重组人骨形态发生蛋白2亚组和重组人骨形态发生蛋白2亚组肌腱移植物部分断裂和完全断裂平均载荷高于未处理亚组与透明质酸钠亚组（P〈0.05）,同时,透明质酸钠-重组人骨形态发生蛋白2亚组高于重组人骨形态发生蛋白2亚组（P〈0.05）。结论：注入透明质酸钠-重组人骨形态发生蛋白2的肌腱移植物有良好的生物力学特性,能促进肌腱移植物在骨隧道内的早期腱-骨愈合。     

自体富含血小板血浆对兔骨折愈合的影响

背景:研究表明富含血小板血浆常通过同源异体制备,可应用于促进扁骨和松质骨的骨折愈合。目的:用Landsbergs法制备自体富含血小板血浆,观察其对兔桡骨骨折愈合影响。方法:将新西兰兔随机分成富含血小板血浆组和对照组,分别建立桡骨中下1/3处骨折模型,富含血小板血浆组中加入制备的自体富含血小板血浆凝胶,对照组建立骨折模型后直接缝合。采用Landsbergs法制备自体富含血小板血浆,并植入富含血小板血浆组的骨折端,并行骨折断端石膏外固定。结果与结论:兔骨折标本在建模后1,2,4,6周均有不同程度骨痂形成。富含血小板血浆组在建模后第4周X射线进行影像学评分明显高于对照组(P<0.05)。免疫组织化学检测显示:富含血小板血浆组Ⅰ型胶原量明显的高于对照组(P<0.05),可持续4周左右。结果证实,富含血小板血浆可通过增加骨痂中骨岛量和加速Ⅰ型胶原合成而明显促进长骨骨折愈合。     

In vivo evaluation of bioactive glass foams associated with platelet-rich plasma in bone defects

The objective of this study was to evaluate the use of bioactive glass foams produced by the sol-gel process, associated or not with platelet-rich plasma (PRP), in the regeneration of bone defects. Mongrel dogs (n = 14) were divided into two groups after having their superior first premolar removed. A small piece of vestibular bone from the alveolus was intentionally removed. The area was filled with bioactive glass foam produced by the sol-gel method. Two groups were tested: group A was the glass foam; group B was the same material associated with PRP, prepared from each animal. The other side of alveolar bone was used as a control group, in which the bone defect did not receive any biomaterial. The thickness of the bone area was measured before and after the intervention. After a period of 60 days implantation, the right and left bones were measured again, and a bone biopsy on both regions was conducted for histological analysis. The findings show an increase of bone thickness for both materials implanted compared to the control group. Group B, implanted with bioactive glass foam associated with PRP, showed a thicker bone area compared to Group A. Histological results indicate bone formation for both materials used. However, the bioactive glass associated with PRP gave rise to a more mature bone formation. These results show that bioactive glass foams processed by a sol-gel method is effective in maintaining the thickness of the alveolar ridge, and the use of PRP associated with the foams improve bone formation.     

Kartogenin with PRP promotes the formation of fibrocartilage zone in the tendon–bone interface

Treatment of tendon–bone junction injuries is a challenge because tendon–bone interface often heals poorly and the fibrocartilage zone, which reduces stress concentration, at the interface is not formed. In this study, we used a compound called kartogenin (KGN) with platelet‐rich plasma (PRP) to induce the formation of fibrocartilage zone in a rat tendon graft–bone tunnel model. The experimental rats received KGN‐PRP or PRP injections in the tendon graft–bone tunnel interface. The control group received saline. After 4, 8 and 12 weeks, Safranin O staining of the tendon graft–bone tunnels revealed abundant proteoglycans in the KGN‐PRP group indicating the formation of cartilage‐like transition zone. Immunohistochemical and immuno‐fluorescence staining revealed collagen types I (Col‐I) and II (Col‐II) in the newly formed fibrocartilage zone. Both fibrocartilage zone formation and maturation were healing time dependent. In contrast, the PRP and saline control groups had no cartilage‐like tissues and minimal Col‐I and Col‐II staining. Some gaps were also present in the saline control group. Finally, pull‐out strength in the KGN‐PRP‐treated group at 8 weeks was 1.4‐fold higher than the PRP‐treated group and 1.6‐fold higher than the saline control group. These findings indicate that KGN, with PRP as a carrier, promotes the formation of fibrocartilage zone between the tendon graft and bone interface. Thus, KGN‐PRP may be used as a convenient cell‐free therapy in clinics to promote fibrocartilage zone formation in rotator calf repair and anterior cruciate ligament reconstruction, thereby enhancing the mechanical strength of the tendon–bone interface and hence the clinical outcome of these procedures. Copyright © 2017 John Wiley & Sons, Ltd.     

Bone regeneration in minipigs via calcium phosphate cement scaffold delivering autologous bone marrow mesenchymal stem cells and platelet‐rich plasma

Macroporous calcium phosphate cement (CPC) with stem cell seeding is promising for bone regeneration. The objective of this study was to investigate the effects of co‐delivering autologous bone marrow mesenchymal stem cells (BMSCs) and autologous platelet‐rich plasma (PRP) in CPC scaffold for bone regeneration in minipigs for the first time. Twelve female adult Tibet minipigs (12–18 months old) were used. A cylindrical defect with 10 mm height and 8 mm diameter was prepared at the femoral condyle. Two bone defects were created in each minipig, one at each side of the femoral condyle. Three constructs were tested: (1) CPC scaffold (CPC control); (2) CPC seeded with BMSCs (CPC‐BMSC); (3) CPC seeded with BMSCs and PRP (CPC‐BMSC‐PRP). Two time points were tested: 6 and 12 weeks (n = 4). Good integration of implant with surrounding tissues was observed in all groups. At 12 weeks, the CPC‐BMSC‐PRP group had significantly less residual CPC remaining in the defect than the CPC‐BMSC group and the CPC control (p < 0.05). The residual CPC volume for the CPC‐BMSC‐PRP group was half that of the CPC control. New bone formation for CPC‐BMSC‐PRP was more than two‐fold that of the CPC control (p < 0.05). CPC‐BMSC‐PRP had new blood vessel density that was nearly two‐fold that of the CPC control (p < 0.05). In conclusion, CPC scaffold with autologous BMSC‐PRP doubled the new bone regeneration and blood vessel density in minipigs compared with the CPC control. In the present study, the new macroporous CPC system with co‐delivered BMSC‐PRP has been shown to promote scaffold resorption and bone regeneration in large defects. Copyright © 2017 John Wiley & Sons, Ltd.     

自体富含血小板血浆对兔骨折愈合的影响

背景：研究表明富含血小板血浆常通过同源异体制备,可应用于促进扁骨和松质骨的骨折愈合。目的：用Landsbergs法制备自体富含血小板血浆,观察其对兔桡骨骨折愈合影响。方法：将新西兰兔随机分成富含血小板血浆组和对照组,分别建立桡骨中下1/3处骨折模型,富含血小板血浆组中加入制备的自体富含血小板血浆凝胶,对照组建立骨折模型后直接缝合。采用Landsbergs法制备自体富含血小板血浆,并植入富含血小板血浆组的骨折端,并行骨折断端石膏外固定。结果与结论：兔骨折标本在建模后1,2,4,6周均有不同程度骨痂形成。富含血小板血浆组在建模后第4周X射线进行影像学评分明显高于对照组（P〈0.05）。免疫组织化学检测显示：富含血小板血浆组Ⅰ型胶原量明显的高于对照组（P〈0.05）,可持续4周左右。结果证实,富含血小板血浆可通过增加骨痂中骨岛量和加速Ⅰ型胶原合成而明显促进长骨骨折愈合。     

Morphologic, histochemical, and functional analysis of platelet-rich plasma activity on skeletal cultured cells

BACKGROUND: Platelet-rich plasma (PRP) is a medium containing concentrated amounts of growth factors in a form that is easy to handle in regenerative sites. The aim of this study was to assess the effect of PRP on the differentiation of cultured skeletal cells and the capability of PRP to induce the production of some osteogenesis-related molecules and mineralization.
STUDY DESIGN AND METHODS: Flow cytometry (cellular antigens), real-time quantitative polymerase chain reaction (RT-qPCR; bone morphogenetic protein [BMP] messengers), alkaline phosphatase (ALP; osteogenic expression), and calcification analyses were performed on 24- and 48-hour human bone cells (HBCs) and 143B and SaOS-2 (osteosarcoma) cell cultures to study the effect of PRP on proliferation and differentiation of skeletal cultured cells. PRP was added using different protocols since no studies are available on bone cultures treated in the long term with PRP.
RESULTS: Flow cytometry showed PRP induction toward a nonhemopoietic lineage in HBCs; RT-qPCR showed enhanced mRNA encoding for BMP2 in HBCs, BMP6 and BMP7 in 143B cultures, and BMP2 and BMP7 in SaOS-2 cultures. Better ALP and calcification results were obtained in SaOS-2 cultures when PRP was added more frequently at shorter intervals while poor results were obtained after single PRP addition.
CONCLUSIONS: The results highlight induction of bone cell proliferation and differentiation by PRP. Since repeated administration of PRP is needed to achieve the best results, an almost continuous delivery system of PRP, or better a controlled release of growth and differentiation factors, using biomaterials might provide increased performance at bone regeneration sites.     

不同时期注射富血小板血浆对牵张成骨的影响

目的确定富血小板血浆(platelet-rich plasma,PRP)在牵张成骨中的最佳注射时期。方法将36只大白兔随机分为4组,每组9只。Ⅰ组为对照组不注射PRP,其余3组分别在延迟期、牵张期和固定期注射0.5ml PRP于牵张间隙中。牵张结束后2、4、8周每组各处死3只动物取材。双能量X线骨密度仪(dualenergy X-ray absorptionmetry,DEXA)测量新生骨骨密度,采用方差分析法对数据进行统计分析;HE染色光镜下观察新骨生成情况。结果实验组新骨生成速率高于对照组;牵张结束后2、4周时,牵张期注射PRP组骨密度测量和组织学观察与其余两实验组间差异有统计学意义(P<0.05)。结论PRP对牵张成骨具有促进作用,牵张期注射PRP可缩短牵张成骨过程。     

小型猪富血小板血浆对牙周膜干细胞的成骨诱导

背景:组织工程技术为牙周炎致骨组织缺损的修复提供了新的思路。目的:探寻富血小板血浆在小型猪牙周膜干细胞成骨诱导中的作用。方法:采集贵州小型猪静脉血,三次离心制备富血小板血浆。采用组织块法分离培养小型猪牙周膜干细胞,分别将含体积分数0.8%,1.0%,1.2%的富血小板血浆与牙周膜干细胞共同培养3,7,14,21d,以未添加富血小板血浆的牙周膜干细胞作为对照。结果与结论:体积分数0.8%,1.0%,1.2%的富血小板血浆成骨诱导第14天,碱性磷酸酶活性达到峰值,其中体积分数1.0%的富血小板血浆诱导的细胞碱性磷酸酶活性最高,第21天时碱性磷酸酶活性降低。茜素红染色显示富血小板血浆成骨诱导第21天细胞染色呈阳性。单克隆纯化后细胞的生长曲线从第3天起进入对数生长期,第8天细胞数量达到顶峰。说明富血小板血浆具有诱导牙周膜干细胞成骨的能力,以体积分数1.0%时诱导成骨效率最优。     

富血小板血浆促进骨修复的理论研究与临床实践

目的:归纳总结富血小板血浆的发展、作用机制及目前还存在的问题,为富血小板血浆的临床应用提供参考。资料来源:应用计算机检索Medline1995-01/2005-06关于富血小板血浆对骨修复的作用的相关文献。检索词“platelet-richplasma,bone,re-pair”,同时应用计算机检索万方数据库1998-01/2005-06期间的上述相关文章,限定文章语言种类为中文,检索词“富血小板血浆,骨,修复”。资料选择:选择有关富血小板血浆和骨修复的中外研究文献;未排除非随机对照的研究文献。资料提炼:49篇关于富血小板血浆与骨修复的文献,其中40篇符合标准。排除的9篇文献是因为重复的同一研究。对剩余的40篇文献进行分类整理,予以综述。资料综合:富血小板血浆在1998年首先被应用于临床修复下颌骨缺损,在颌面缺损处植入自体骨与富血小板血浆的复合物,骨密度影像测定发现富血小板血浆组与单用自体骨的对照组相比,其骨成熟度是对照组的1.62～2.61倍。在随后的这些年里,富血小板血浆已经在很多医学领域被用来促进组织修复,其特点:安全,简便,廉价。但目前仍有一些问题有待进一步研究和解决。结论:富血小板血浆包含多种生长因子,且已被证明可促进骨组织和软组织的修复。富血小板血浆经离心自体血而制成,制作简单,安全。由于是自源性的,根本上排除了疾病传播及免疫排斥的可能。     

Bone marrow mesenchymal stem cells,platelet‐rich plasma and nanohydroxyapatite–type I collagen beads were integral parts of biomimetic bone substitutes for bone regeneration

Platelet rich plasma (PRP), which includes many growth factors, can activate osteoid production, collagen synthesis and cell proliferation. Nanohydroxyapatite‐type I collagen beads (CIB), which mimetic natural bone components, are not only flexible fillers for bone defect but also encourage osteogenesis. Bone marrow mesenchymal stem cells (BMSCs) are often used as an abundant cell source for tissue engineering. We used a rabbit model to combine PRP, CIB and BMSCs (CIB+PRP+BMSC) into a bone‐like substitute to study its impact on bone regeneration, when compared to defect alone, PRP, CIB+PRP, and PRP+BMSC. CIB+PRP upregulated more alkaline phosphatase (ALP) activity in BMSCs than PRP alone at 4 weeks postoperation. CIB+PRP+BMSC and PRP+BMSC did not differ significantly in DNA content, total collagen content, and ALP activity at 8 weeks. In histological assay, both CIB+PRP+BMSC and PRP+BMSC showed more bone regeneration at 4 and 8 weeks. Higher trabecular bone volume in tissue volume (BV/TV) (31.15±2.67% and 36.93±1.01%), fractal dimension (FD) (2.30±0.18 and 2.65±0.02) and lower trabecular separation (Tb.Sp) (2.30±0.18 and 1.35±0.16) of CIB+PRP+BMSC than of other groups at 4 and 8 weeks, and approach to of bone tissue (BV/TV=24.35±2.13%; FD=2.65±0.06; Tb.Sp=4.19±0.95). CIB+PRP+BMSC significantly enhanced new bone formation at 4 week. Therefore, nanohydroxyapatite‐type I collagen beads combined with PRP and BMSCs produced a bone substitute with efficiently improved bone regeneration that shows promise to repair bone defects. Copyright © 2012 John Wiley & Sons, Ltd.