骨基质明胶治疗肿瘤性骨缺损

基质明胶（ＢＭＧ）具有较强的诱导成骨作用已被实验研究证实，但临床应用报道不多。我院临床应用ＢＭＧ治疗肿瘤性骨缺损９例，治疗效果表明ＢＭＧ植入后４周，腔洞内已出现新骨生成，８周以后成骨作用显著，密度均匀，相互连成一片，密度接近正常骨组织。术后观察植入ＢＭＧ后组织相容性良好，具有使用方便，病人无痛苦，成骨作用快的优点，是治疗肿瘤性骨缺损理想的植入材料。     

骨诱导性载体复合骨形态发生蛋白的双重成骨作用

本文用猪骨基质明胶（ｐＢＭＧ）复合猪骨形态发生蛋白（ｐＢＭＰ）植入小鼠肌肉内，以期达到ｐＢＭＧ和ｐＢＭＰ双重诱导成骨的叠加作用，并与单独植入的明胶海绵、ｐＢＭＧ、ｐＢＭＰ和无机骨－ｐＢＭＰ（ＡＯＢ－ｐＢＭＰ）的成骨效应相比较。结果显示，含有ｐＢＭＰ的各组术后４天有明显的间充质细胞化，新骨在１０天左右开始形成。ＡＬＰ的活性高峰出现在术后２１～２８天，组间钙、磷含量差异具有高度显著性（Ｐ＜０．０１），明胶海绵组最低，而ｐＢＭＧ－ｐＢＭＰ组含量最高。各组的钙、磷相关系数为高度正相关。作者讨论了ｐＢＭＧ复合ｐＢＭＰ后的双重成骨作用及其优越性，认为ＡＬＰ主要由诱导性新骨成熟期的成骨细胞和骨细胞所产生，新骨中的钙、磷含量因呈高度正相关，两者均可作为新骨定量的指标，但以钙含量为主。     

骨形态蛋白与骨基质明胶复合修复骨缺损的实验研究

通过骨形态蛋白（ＢＭＰ）与其前身物质骨基质明胶（ＢＭＧ）复合，修复家兔桡骨缺损，同时以ＢＭＰ与羟基磷灰石（ＨＡ）复合作实验对照。结果发现：ＢＭＰ／ＢＭＧ组第４周即在骨缺损区出现纤维性骨痂，第８周可见大量的骨小梁形成，第１２周ＢＭＰ完全被吸收，为新生的骨组织所代替，第１６周出现髓腔再通现象。而ＢＭＰ／ＨＡ复合组，虽然在第４、８周也有纤维性骨痂形成，但ＨＡ不被吸收；第１２、１６周与第８周相似，无髓腔再     

用同种异体骨基质明胶修复颅骨大面积缺损

本文报道同种异体骨基质明胶促进颅骨大面积缺损愈合的实验结果。以家兔颅顶骨直径1.0mm圆形骨缺损作为颅骨缺损模型,分别植入同种异体骨基质明胶、脱钙骨基质和不脱钙骨。植入后4周、8周、12周进行X线摄片和组织学位检查,结果表明同种骨基质明胶没有免疫排斥反应,并具有良好的骨诱导作用。术后8周已有大量新骨出现于骨缺损中央部,12周已使骨缺损完全愈合。说明同种骨基质明胶是修复儿童颅骨缺损较理想的植骨材料。     

聚乳酸作为骨形态发生蛋白载体修复骨缺损的实验研究

目的 探讨聚乳酸（ｐｏｌｙｌｄａｃ ａｃｉｄ,ＰＬＡ）作为骨形态发生蛋白（ｂｏｎ ｅ ｍｏｒｐｈｏｇｅｎｅｔｉｃ ｐｒｏ－ｔｅｉｎ,ＢＭＰ）载体的可行性及观察其诱导成骨能力。方法 手术造成日本大耳白兔左尺骨中上段１２ｍｍ骨缺损实验模型。随机分为实验。对照及空白组,实验组植入以ＰＬＡ为载体的ＢＭＰ１０ｍｇ、对照组植入以牛松质骨基质为载体的ＢＭＰ１０ｍｇ、空白组不做任何处理,术后摄Ｘ线片观察各组不同时相骨缺损修复情况,并于术后第４、８、１２周观察各组缺损内组织学变化。图像分析骨小梁的生成量。结果 实验修复情况优于对照组,无论是骨连接发生时间还是骨成熟时间,实验组均较对照组提前２周左右,同期骨生成量也明显多于对照组,而空白组缺损内主要形成纤维组织。结论 ＰＬＡ可以作为ＢＭＰ的载体修复骨缺损,它比异种松质骨基质载体的成骨效     

陶瓷样异种骨复合骨髓异位成骨的实验研究

目的了解陶瓷样异种骨（ＣＸＢ）与骨髓（ＢＭ）复合移植的成骨作用。方法将ＣＸＢ与ＢＭ复合及单纯ＣＸＢ植入兔的骶棘肌内,术后２、４、８、１２、１６及２４周取出植入材料作组织学及组织化学检查,观察植入材料的成骨作用。结果ＣＸＢ与ＢＭ复合植入后２周开始有软骨和新骨生成,以后软骨逐渐钙化成骨,８周时出现髓腔,并随植入时间的延长,新骨生成增多,成骨更为典型。而单纯ＣＸＢ植入后无新骨或软骨形成。结论ＣＸＢ与ＢＭ复合移植通过骨传导、骨诱导及提供成骨细胞或成软骨细胞而成骨,是一种理想的骨移植替代材料,可望在临床上用于骨缺损的修复。     

同种异体骨在斜外侧椎间植骨融合中的应用

目的观察同种异体骨在斜外侧椎间融合(OLIF)术中的应用效果。方法采用OLIF同种异体骨椎间植骨融合治疗93例腰椎退行性疾病患者。比较手术前后疼痛VAS评分、ODI评分及椎间隙高度,观察术后椎间植骨融合情况。结果 93例患者均获得随访,时间12～24个月。术后1个月VAS评分、ODI评分均较术前明显下降(P 0.05)。术后2周、12个月椎间隙高度均较术前明显改善(P 0.05)。末次随访91例(97.85%)植骨融合。无植骨吸收、脱落、骨不连、假关节形成及螺钉松动、断裂等现象。结论 OLIF术中植入同种异体骨可以获得良好的近期疗效。     

同种皮质骨融合支架的研制及临床应用

目的研究同种皮质骨融合支架行后路椎间融合术治疗腰椎滑脱的临床疗效。方法对12例腰椎滑脱患者,用同种皮质骨融合支架行后路椎体间融合加内固定术。根据手术前后的椎间隙高度的变化和JOA评分,观察临床疗效。结果12例均获得随访,时间3-6个月,术后椎间隙高度明显恢复,无神经系统并发症,无融合支架脱出,切口无感染,JOA评分术后改善率为93.5%。结论该手术保留的骨性终板有利于防止植入物陷入椎体,能有效地维持椎间隙高度。术中无须取自体骨,且植骨面积最大化,有利于融合,植骨内固定一次完成。同种皮质骨融合支架置入牢固,具有骨传导和骨诱导作用,手术操作简单,是一种良好的椎间融合材料。     

内源性骨形态发生蛋白在骨修复中的作用

目的：研究内源性骨形态发生蛋白（ＢＭＰ）对骨修复的作用。方法：３６只家兔,环形切除双侧胫骨中段骨膜后剥脱部分胫侧骨皮质,右侧钻孔达骨髓腔,左侧不钻孔作为对照。术后３ｄ,１、２、４、６、８周分批处死取材,用石蜡切片做ＢＭＰ免疫组化染色。结果：ＢＭＰ免疫组化染色显示,对照染色阴性,始终无骨痂形成。实验侧术后３ｄ多形间充质细胞染色呈阳性,术后１周内源性ＢＭＰ浓度达高峰,成骨能力增强。术后６周阳性细胞消失,但随着骨重建过程增强,骨基质染色变深。结论：骨损伤后内源性ＢＭＰ呈阶段性分泌,且对骨损伤的修复起重要作用     

可降解多孔β—TCP／rhBMP—2人工骨的骨诱导活性研究

目的：检测可降解多孔β－ＴＣＰ／ｒｈＢＭＰ－２人工骨的诱导成骨能力。方法：自制要降解多孔β－ＴＣＰ与ｒｈＢＭＰ－２复合后植入４５只小鼠股后肌袋作为实验组，单纯β－ＴＣＰ植入对侧作对照，术后２４，７２小时，１，２，４，８周分别取材做组织学检查，２，４周取材的部分标本做扫描电镜观察：１，２，４周取材的部分标本做碱性磷酸酶（ＡＬＰ）活性测定。结果：实验组有大量新生软骨和骨形成，随着时间的推移，生成量逐渐     

Repairing articular cartilage defects with tissueengineering cartilage in rabbits

Objective: To investigate the effect of cancellous bone matrix gelatin ( BMG ) engineered with allogeneic chondrocytes in repairing articular cartilage defects in rabbits. Methods: Chondrocytes were seeded onto three-dimensional cancellous BMG and cultured in vitro for 12 days to prepare BMG-chondrocyte complexes. Under anesthesia with 2.5% pentobarbital sodium (1ml/kg body weight), articular cartilage defects were made on the right knee joints of 38 healthy New Zealand white rabbits (regardless of sex, aged 4-5 months and weighing 2. 5-3 kg) and the defects were then treated with 2. 5% trypsin. Then BMG-chondrocyte complex ( Group A, n = 18 ), BMG (Group B, n = 10), and nothing (Group C, n = 10) were implanted into the cartilage defects, respectively. The repairing effects were assessed by macroscopic, histologic, transmission electron microscopic ( TEM ) observation, immunohistochemical examination and in situ hybridization detection, respectively, at 2, 4, 8, 12 and 24 weeks after operation. Results: Cancellous BMG was degraded within 8 weeks after operation. In Group A, lymphocyte infiltration was observed around the graft. At 24 weeks after operation, the cartilage defects were repaired by cartilage tissues and the articular cartilage and subchondral bone were soundly healed. Proteoglycan and type II collagen were detected in the matrix of the repaired tissues by Safranin-O staining and immunohistochemical staining, respectively. In situ hybridization proved gene expression of type II collagen in the cytoplasm of chondrocytes in the repaired tissues. TEM observation showed that chondrocytes and cartilage matrix in repaired tissues were almost same as those in the normal articular cartilage. In Group B, the defects were repaired by cartilage-fibrous tissues. In Group C, the defects were repaired only by fibrous tissues. Conclusions: Cancellous BMG can be regarded as the natural cell scaffolds for cartilage tissue engineering. Articular cartilage defects can be repaired by cancellous BMG engineered with allogeneic chondrocytes. The nature of repaired tissues is closest to the normal cartilage. Local administration of trypsin can promote the adherence of repaired tissues to host tissues. Transplantation of allogeneic chondrocytes has immunogenicity, but the immune reaction is weak.     

同种骨基质明胶加石膏修复骨缺损的实验研究

目的研究骨基质明胶加石膏复合物在骨缺损修复过程中的作用。方法新西兰兔１６只,桡骨干缺损为植骨区,实验组植入骨基质明胶加石膏复合物（１６侧）,对照组分别植入骨基质明胶（８侧）和酒精储存骨（８侧）,观察骨缺损的修复情况。结果实验组成骨活跃,１２周时骨缺损已基本修复。对照组也有成骨现象,但进展不如实验组。结论骨基质明胶加石膏复合物为一种良好的骨移植材料。     

颈椎椎间融合器融合效果的在体动物研究

目的在活体山羊颈椎上比较三面皮质自体骨、垂直圆柱体Harmscage及方盒形Carboncage的椎间融合效果。方法取24只成年雄性山羊,随机分为三组:自体骨组、Harmscage组及Car-boncage组。所有动物均行C3-4椎间盘切除术并分别植入以上内植物。于术前、术后即刻及术后1、2、4、8、12周摄颈椎正侧位X线片,于侧位X线片上测量平均椎间高度、椎间角及前凸角;术后12周处死动物,取C3,4节段标本进行组织学评估。结果术后1周Carboncage组的平均椎间高度、椎间角及前凸角大于Harmscage组和自体骨组(P<0.05);术后12周两个cage组的平均椎间高度、椎间角及前凸角均大于自体骨组,差异有统计学意义(P<0.05)。术后12周依据融合分级标准对三组进行影像学评估,和自体骨组比较,两个cage组的骨融合效果略好,但无统计学意义;组织形态学亦发现两个cage组的椎间融合效果较好,但与自体骨组比较差异无统计学意义。垂直圆柱体Harmscage中形成的新生骨量多于方盒形Carboncage。结论方盒形Carboncage具有良好的椎间支撑能力,而垂直圆柱体Harmscage的椎间融合效果更好。     

帽式颈椎椎间融合器的活体实验研究

目的 本研究在活体山羊颈椎上从椎间支撑能力、二期生物力学稳定性及椎间融合组织学方面比较三面皮质骨、Harms椎间融合器（cage）、Carboncage及自行所研制的帽式颈椎椎间融合器（hat　type　cervical　intervertebral　fusion　cage,HCIFC）的术后效果。方法　取32头山羊随机平均分为4组：第1组自体三面皮质髂骨;第2组Harmscage;第3组Carboncage;第4组HCIFC。分别行C3,4椎间盘切除术并分别植入以上内植物。术后共观察12周并测量平均椎间高度（DSH）、椎间角（IVA）及前凸角（LA）,处死后进行影像学、生物力学及组织学评估。结果 术后1周方盒形HCIFC、Carboncage组的平均DSH、IVA及LA要显著大于垂直圆柱体Harmscage和三面皮质骨组（P〈0．05）;术后12周时3个cage组的平均DSH、IVA及LA显著高于三面皮质骨组（P〈0．05）。垂直圆柱体Harmscage轴向旋转和侧屈时平均刚度显著大于其他组（P〈0．001）,轴向旋转时的最大活动度要显著低于其他组（P〈0．05）。与三面皮质骨组相比,3个cage组的椎间融合效果略好。结论HCIFC的椎间支撑能力、生物力学性能及椎间融合效果均适合于临床使用。     

Bone matrix gelatin. Clinical application in 38 cases

Bone matrix gelatin (BMG) was prepared and used clinically as graft-stuff in 38 patients. It was used to fill the bone defects resulted from excision of benign bone tumors (22 patients) and debridement of chronic osteomyelitis (3), to treat delayed union of fractures (5), and to make posterior spinal fusion (8). Fourty-five days after operation, speckles or flakes of dense shadow appeared in the film at the original defects and fields. In 2 to 6 months, bony union took place in 36 patients (95%). Failure was noted in 2 patients with chronic osteomyelitis. Because of its excellent osteo-inductive property and nonantigenic treat, BMG has been proved to be an ideal substitute for bone graft. It is easy to prepare, safe for storage at room temperature and sound for transportation.     

切除卵巢对骨基质明胶修复大鼠颅骨缺损成骨效能的影响

目的　探讨实验性骨质疏松状态对骨基质明胶修复骨缺损成骨效能的影响。方法　将 6 8只雌性SD大鼠随机分为去卵巢组和对照组。 90d后两组各抽取 10只测定骨密度以确定骨质疏松模型的建立。其余 4 8只大鼠制备直径 8mm颅骨极量骨缺损并同期植入骨基质明胶颗粒。植入术后 2 1和 5 6d两组每次分别处死 12只大鼠 ,通过不脱钙组织磨片、四环素荧光示踪及骨组织计量学动态参数测定、扫描电镜和X射线显微分析技术观察骨缺损区修复情况。结果　骨密度测定指标显示 ,去卵巢 90d后骨质疏松状态建立。骨基质明胶植入术后 2 1d ,均有新骨在两实验组骨缺损内形成 ,去卵巢组新骨形成较少 ,四环素标记成骨区域较稀疏。植入术后 5 6d ,对照组骨缺损基本修复 ,去卵巢组骨缺损部分区域为纤维结缔组织充填。植入术后 2 1d、5 6d荧光双标间距和平均矿化沉积速率均较对照组低 ,差异有统计学意义 (P <0 0 1)。骨缺损区新生骨痂的钙磷比值定量分析 ,去卵巢组均低于对照组 ,差异有统计学意义 (2 1d :0 4 892± 0 0 342、0 5 4 75± 0 0 2 0 4 ,P <0 0 1;5 6d :1 0 95 0± 0 0 4 37、1 195 5± 0 0 4 6 3,P <0 0 1)。结论　实验性骨质疏松降低了骨基质明胶的成骨效能。雌激素可能在生物骨替代材料参与的骨重建中发挥重要     

Dose-dependent effects of combined IGF-I and TGF-β1 application in a sheep cervical spine fusion model

Combined IGF-I and TGF-β1 application by a poly-(D,L-lactide) (PDLLA) coated interbody cage has proven to promote spine fusion. The purpose of this study was to determine whether there is a dose-dependent effect of combined IGF-I and TGF-β1 application on intervertebral bone matrix formation in a sheep cervical spine fusion model. Thirty-two sheep underwent C3/4 discectomy and fusion. Stabilisation was performed using a titanium cage coated with a PDLLA carrier including no growth factors in group 1 (n=8), 75 μg IGF-I plus 15 μg TGF-β1 in group 2 (n=8), 150 μg IGF-I plus 30 μg TGF-β1 in group 3 (n=8) and 300 μg IGF-I plus 60 μg TGF-β1 in group 4 (n=8). Blood samples, body weight and temperature were analysed. Radiographic scans were performed pre- and postoperatively and after 1, 2, 4, 8, and 12 weeks. At the same time points, disc space height and intervertebral angle were measured. After 12 weeks, the animals were killed and fusion sites were evaluated using quantitative computed tomographic (CT) scans to assess bone mineral density, bone mineral content and bony callus volume. Biomechanical testing was performed and range of motion, and neutral and elastic zones were determined. Histomorphological and histomorphometrical analysis were carried out and polychrome sequential labelling was used to determine the time frame of new bone formation. In comparison to the group without growth factors (group 1), the medium- and high-dose growth factor groups (groups 3 and 4) demonstrated a significantly higher bony callus volume on CT scans, a higher biomechanical stability, an advanced interbody bone matrix formation in histomorphometrical analysis, and an earlier bone matrix formation on fluorochrome sequence labelling. Additionally, the medium- and high-dose growth factor groups (groups 3 and 4) demonstrated a significantly higher bony callus volume, a higher biomechanical stability in rotation, and an advanced interbody bone matrix formation in comparison to the low-dose growth factor group (group 2). No significant difference could be determined between the medium- and the high-dose growth factor groups (groups 3 and 4, respectively). The local application of IGF-I and TGF-β1 by a PDLLA-coated cage significantly improved results of interbody bone matrix formation in a dose-dependent manner. The best dose-response relationship was achieved with the medium growth factor dose (150 μg IGF-I and 30 μg TGF-β1). With an increasing dose of these growth factors, no further stimulation of bone matrix formation was observed. Although these results are encouraging, safety issues of combined IGF-I and TGF-β1 application for spinal fusion still have to be addressed. This study was carried out with the support of the MBI – Max Biedermann Institut der Steinbeis Stiftung (RS/KF-82042).     

"双相"组织工程软骨修复兔关节骨软骨缺损

目的探讨“双相”异体骨基质明胶（bonematrixgelatin,BMG）作为组织工程软骨载体,与同体骨髓间充质干细胞（marrowmesenchymalstemcells,MSCs）结合,构建组织工程软骨修复兔关节骨软骨缺损的效果。方法4月龄新西兰兔32只,雌雄不限,体重2～3kg。①体外实验：取5只新西兰兔,处死后取髂骨和四肢骨,制备一侧松质骨,一侧皮质骨的“双相”异体BMG载体,扫描电镜观察。另取新西兰兔18只,抽取骨髓,分离MSCs并诱导成软骨分化;将诱导而来的软骨前体细胞与“双相”BMG载体复合构建组织工程软骨,分别于1、3和5周取材行Masson、PAS染色和扫描电镜观察。②体内实验：将抽取骨髓的18只及余下的9只新西兰兔制成双侧股骨内髁骨软骨缺损模型,将前期制备的组织工程软骨同体植入18只兔的右股骨内髁骨软骨缺损（A组）,左侧缺损移植异体BMG（B组）,其余9只双侧软骨缺损未予处理作为空白对照（C组）,分别于术后1、3和6个月取材,行大体、组织学和Ⅱ型胶原mRNA原位杂交观察,改良Wakitani法评分,比较各组修复效果差异。结果①体外实验：“双相”BMG松质骨面孔隙大小100-800μm,细胞于其中增生,形成富含细胞的软骨层;皮质骨面孔隙大小10～40pm,细胞层状覆盖于其表面,可作为起支撑作用的软骨下骨。②体内实验：A组术后1个月即可重建关节骨软骨缺损;修复软骨在观察期内逐渐变薄,但在6个月内始终保持关节面及软骨下骨结构完整。B、C组未能修复缺损,缺损周边软骨磨损加剧。改良Wakitani评分显示A组在3个时间点的各项评分结果,除6个月软骨厚度外,其它指标均优于B、C组,且差异有统计学意义（P〈0．01）。Ⅱ型胶原mRNA原位杂交显示,A组缺损区修复组织中细胞阳性染色率明显高于B、C组,且差异有统计学意义（P〈0．01）。结论“双相”异体BMG可作为组织工程软骨载体材料,其结合自体MSCs诱导的软骨前体细胞制备的组织工程软骨,可修复兔关节软骨和软骨下骨。     

自体外周血干细胞与骨基质明胶复合移植治疗骨缺损

目的观察自体外周血干细胞（autologous peripheral blood stem cells,APBSC）/表面脱钙骨基质明胶（bone matrix gelatin,BMG）共移植修复节段性骨缺损的疗效。方法 36例患者随机分为APBSC/BMG组（A组）及BMG组（B组）各18例,均行手术治疗,并行牢固内外固定。结果术后平均愈合时间A组为5个月,B组为8个月,两组间差异具有统计学意义（P〈0.05）;治疗期间患者未见明显不良反应。结论 APBSC/BMG复合移植较单纯BMG移植治疗骨缺损疗效好,无不良反应。     

Comparison of BMP-2 and combined IGF-I/TGF-ß1 application in a sheep cervical spine fusion model

Growth factors have proven to promote spine fusion. However, no comparative evaluation of growth factors in spinal fusion has yet been performed. The purpose of this study was to compare the efficacy and safety of combined IGF-I and TGF-ss1 application with BMP-2 application and autologous cancellous bone graft at an early time point in a sheep cervical spine fusion model. Thirty-two sheep underwent C3/4 discectomy and fusion. They were divided into four groups, according to their treatment: group 1, titanium cage ( n=8); group 2, titanium cage filled with autologous cancellous iliac crest bone grafts ( n=8); group 3, titanium cage coated with a poly-(D,L-lactide) (PDLLA) carrier including BMP-2 (5% w/w) ( n=8); group 4, titanium cage coated with a PDLLA carrier including IGF-I (5% w/w) and TGF-ss1 (1% w/w) ( n=8). Blood samples, body weight and temperature were analysed. Radiographic scans were performed pre- and postoperatively and after 1, 2, 4, 8 and 12 weeks. At the same time points, disc space height and intervertebral angle were measured. After 12 weeks, the animals were killed and fusion sites were evaluated using functional radiographic views in flexion and extension. Quantitative computed tomographic scans were performed to assess bone mineral density, bone mineral content and bony callus volume. Biomechanical testing was carried out and the values for range of motion, and neutral and elastic zone were determined. Histomorphological and histomorphometrical analysis were performed and polychrome sequential labelling was used to determine the time frame of new bone formation. The results showed that, in comparison to the group treated with the cage alone (group 1), the cage plus BMP-2 group (group 3) and the cage plus IGF-I and TGF-ss1 group (group 4) demonstrated a significantly higher fusion rate in radiographic findings, a higher biomechanical stability, a more advanced interbody fusion in histomorphometrical analysis, and an accelerated interbody fusion on fluorochrome sequence labelling. In comparison to the bone graft group (group 2), the BMP-2 (group 3) and IGF-I/TGF-ss1 group (group 4) showed significantly less residual motion on functional radiographic evaluation, higher bone mineral density of the callus and higher biomechanical stability in extension, rotation and bending. The BMP-2 group showed significantly less residual motion on functional radiographic evaluation and higher intervertebral bone matrix formation on fluorochrome sequence labelling at 9 weeks in comparison to the IGF-I/TGF-ss1 group. In contrast, the IGF-I/TGF-ss1 group showed a significantly higher bone mineral density of the callus than the BMP-2 group. In comparison to the autologous cancellous bone graft group, both growth factors (BMP-2 and combined IGF-I and TGF-ss1) significantly improved the biomechanical results of interbody fusion. No systemic side effects were observed for either growth factor. On the basis of these preliminary results, it would appear that combined IGF-I/TGF-ss1 application yields equivalent results to BMP-2 application at an early time point in anterior sheep cervical spine fusion.