转化生长因子β对退变颈椎间盘成纤维细胞的成骨诱导作用

目的 探讨转化生长因子β（Transforming Growth Factor β,TGF-β）对退变的颈椎间盘纤维环中成纤维细胞成骨诱导的作用,以进一步揭示颈椎病的发病机理。方法 利用体外细胞培养技术,建立颈椎病患者的退变颈椎间盘纤维环中成纤维细胞的培养体系,观察在TGF-β条件培养液诱导下的成骨表现。结果 非条件培养组和条件培养组在传代的颈椎病患者纤维环成纤维细胞培养的各时间点的细胞增殖活力（MTT）测定无明显差异（P〉0．05）,TGF-β条件培养组的ALP活力经测定与空白对照组ALP活力有显著性差异（P〈0．05）,TGF-β组的成纤维细胞所形成的骨钙素分泌量均与空白对照组有显著性差异（P〈0．05）。结论 TGF-β对纤维环成纤维细胞体外有明确的诱导成骨作用,退变的颈椎间盘纤维环成纤维细胞存在成骨潜能。     

山羊颈椎间盘纤维环成纤维细胞成骨潜能的体外观察

目的 诱导培养椎间盘纤维环成纤维细胞，探讨其成骨潜能，比较重组人骨形成蛋白2（recombinant human bone morphogenetic protein2，rhBMP-2）和肿瘤坏死因子α（tumor necrosis factor α，TNF—α）对其成骨的不同影响。方法利用体外细胞培养技术，建立实验山羊颈椎间盘纤维环成纤维细胞培养体系，根据培养液的不同，分为空白对照组、TNF-α组（加入50U／ml TNF-α）、rhBMP-2组（加入0．1μg／ml rhBMP-2）和TNF—α＋rhBMP-2组（加入50U／ml TNF-α＋0．1μg／ml rhBMP-2），培养3周观察纤维环成纤维细胞在条件培养液诱导下的成骨表型变化、碱性磷酸酶（alkaline phosphatase，ALP）和骨钙素（osteocalcin，OC）等成骨指标变化。结果 各实验组培养液对细胞增殖无明显影响。rhBMP-2组和TNF-α＋rhBMP-2组细胞趋化性生长明显，矿化结节出现，茜素红染色呈深红色钙阳性反应。各实验组ALP活力与空白对照组比较差异均有统计学意义（P〈0．05）。rhBMP-2组与TNF—α＋rhBMP-2组成纤维细胞的OC分泌量与空白对照组比较差异均有统计学意义（P〈0．05），TNF—α组与空白对照组比较差异无统计学意义（P〉0．05）。结论 体外培养的椎问盘纤维环成纤维细胞有成骨潜能，rhBMP-2对纤维环成纤维细胞有明确的诱导成骨作用。     

肿瘤坏死因子-α和骨形态发生蛋白-2诱导成纤维细胞表达成骨表型的协同作用机制

目的:探讨肿瘤坏死因子-α(TNF-α)和骨形态发生蛋白-2(BMP-2)诱导成纤维细胞表达成骨表型的可能机制。方法:分离、纯化人真皮成纤维细胞,使其生长在分别含一定浓度的TNF-α、BMP-2和TNF-α与BMP-2联合培养液的干预条件下,采用MTT和RT-PCR技术,检测成纤维细胞增殖和C-myc、BMP-2、BMP-4mRNA表达状况的变化。结果:单独应用TNF-α和联合应用TNF-α与BMP-2可刺激成纤维细胞增殖;TNF-α诱导成纤维细胞表达C-myc mRNA和BMP-2 mRNA;TNF-α与BMP-2联合应用可诱导成纤维细胞表达BMP-4 mRNA。结论:TNF-α可诱导成纤维细胞转化,并赋予其新的生物特征;外源性BMP-2和内源性BMP-2、BMP-4的协同效应是成纤维细胞表达成骨表型的机制之一。     

胰岛素样生长因子-1对成纤维细胞向成骨细胞转化的影响

目的 探讨胰岛素样生长因子-1对成纤维细胞(Fb)向成骨细胞转化的影响. 方法 Fb来源于成年新西兰大白兔,通过分离、纯化、培养而得到.实验分为3组:空白对照组、成骨诱导组和实验组.空白对照组:常规培养液培养,不加入任何诱导剂及干预因子;成骨诱导组:成骨诱导液为常规培养液中加入浓度为1×10-8 mol/L地塞米松、50 mg/L维生素C、10 mmol/L β-甘油磷酸钠;实验组:成骨诱导液中加入终浓度为50 ng/mL的IGF-1.采用四甲基偶氮唑蓝(MTT)比色法检测各组细胞增殖情况,检测各组细胞碱性磷酸酶(ALP)活性,测定各组细胞培养后6d后的骨钙素含量,采用钙钴染色评价培养后2周的成骨能力.结果 实验组细胞增殖较其他两组旺盛,差异有统计学意义(P＜0.05),而空白对照组和成骨诱导组差异无统计学意义(P＞ 0.05).ALP活性和骨钙素测定:实验组ALP表达明显高于其他两组,差异有统计学意义(P＜0.05);成骨诱导组ALP表达高于空白对照组,差异有统计学意义(P＜0.05).实验组钙化结节数量明显较其他两组多,成骨诱导组次之,空白对照组未见钙化结节.结论 胰岛素样生长因子-1可以促进Fb在成骨过程中的增殖以及增加其成骨能力.     

诱导立体网架上成纤维细胞表达成骨表型及其机制的研究

目的探讨诱导条件下的成纤维细胞在三维结构的聚乙醇酸（PGA）网架上表达成骨表型的可行性,以及肿瘤坏死因子α（TNF-α）对成纤维细胞骨形态发生蛋白（BMP）受体表型表达的影响。方法分离、纯化人皮肤成纤维细胞,实验用第2代细胞：（1）种植成纤维细胞于PGA网架上,进行体外旋转培养,并用含TNF-α（50U／ml）和BMP-2（0．1μg／ml）的条件培养液进行诱导。于1d,3、6周后利用倒置相差显微镜、扫描电镜、四环素荧光标记、茜素红染色方法观察细胞生长、骨样组织形成和矿化物沉积情况。上清液生化检测分析成骨性标志物分泌情况;（2）接种成纤维细胞于预置玻片或75cm。培养瓶中,用含TNF-α（50U／ml）的条件培养液进行一次性或连续性干预,采用逆转录聚合酶链反应和免疫组织化学技术,于2．4、6、8d后分别检测BMPⅠ型受体（BMPR-ⅠA和BMPR-ⅠB）mRNA表达及蛋白形成状况。结果诱导3周后三维网架上的成纤维细胞表达成骨表型,分泌成骨细胞特征性标志物：骨钙素（OCN）和骨特异性碱性磷酸酶（B-AKP）;分泌大量细胞外基质形成骨样组织;平面培养发现,TNF-α（50U／ml）连续干预8d可增高BMPR-ⅠB的mRNA表达和蛋白合成。结论三维立体网架上的成纤维细胞在诱导条件下能够向成骨型细胞转化,形成骨样组织,有希望作为一种新的成骨型细胞的种子细胞源;TNF-α为BMP-2的靶向作用提供条件,TNF-α和BMP-2联合应用是调节成纤维细胞表型转化的诱导条件之一。     

骨形态发生蛋白和碱性成纤维细胞生长因子激发纤维环细胞成骨的潜能

目的研究联合使用重组人骨形态发生蛋白（recombinant human bone morphogenetic protein-2,rhBMP-2）和碱性成纤维细胞生长因子（basic fibroblast growth factor,bFGF）椎间盘纤维环细胞成骨潜能的激发作用。方法向体外培养的纤维环细胞中分别及联合加入rhBMP-2和bFGF,观察纤维环细胞的表型表达特点。结果联合使用rh-BMP-2和bFGF能够明显促进椎间盘细胞增殖,提高细胞内碱性磷酸酶活力,增加I型胶原分泌,提高钙盐沉积程度,提高骨钙素的表达水平。结论联用rhBMP-2和bFGF能够诱导纤维环细胞向成骨细胞方向分化,分泌钙盐并形成钙结节。     

rhBMP-2对不同培养时间骨髓基质干细胞成骨能力的影响

目的：探讨不同培养阶段骨髓基质干细胞成骨能力的变化及BMP-2对其成骨能力的影响。方法：培养兔骨髓基质干细胞，测定第3代和第18代细胞碱性磷酸酶及骨钙素活性；测定BMP-2对不同培养时间骨髓基质干细胞成骨能力的影响；测定rhBMP-2对细胞增殖的影响。结果：细胞传至第18代后，分泌的骨钙素（OC）水平及ALP活力明显降低，与第3代细胞相比，差异显著（P〈0．05）。第3代细胞在rhBMP-2的诱导下，分泌的OC水平及ALP活力在原基础上进一步升高，与对照组相比，差异显著（P〈0.05）。结第18代细胞在rhBMP-2的诱导下，分泌的OC水平及ALP活力在原基础上升高，与对照组相比，差异不显著（P〉0．05）；随培养时间的延长，各组细胞数量均有所增加，rhBMP-2诱导组与对照组细胞无显著性差异（P〉0．05）。细胞的传代次数对细胞增殖无明显影响（P〉0．05）。结论：rhBMP-2对细胞增殖无影响。随着传代次数的增加，骨髓基质干细胞的成骨能力下降，rhBMP-2促进其成骨的能力亦下降。     

中药增骨I、Ⅲ号调节rhBMP-2对成纤维细胞的影响

目的 观察中药增骨Ⅰ、Ⅲ号调节体外重组人骨形态发生蛋白－2（rhBMP－2）对体外成纤维细胞的影响。方法 体外分离培养人成纤维细胞。然后用中药增骨Ⅰ、Ⅲ号及增骨Ⅰ、Ⅲ 复合rhBMP-2分别作用于成纤维细胞,使用倒置相差显微镜观察其形态,噻唑蓝（MTT）比色分析其增殖、放免测定骨钙素（BGP）含量。结果 增骨Ⅰ、Ⅲ MTT比色结果与对照差异有显性（P＜0．05）,中药复合rhBMP-2组与对照组比色结果差异非常显（P＜0．01）。增骨Ⅰ、Ⅲ号及中药复合rhBMP-2都可促进成纤维细胞BGP的表达,与对照组相比结果差异分别为有显性（P＜0．05）和非常显性（P＜0．01）。结论 增骨Ⅰ、Ⅲ号能激活成纤维细胞向成骨细胞分化,能增强rhBMP-2诱导成骨,促进成纤维细胞成骨表型的表达。     

山羊颈椎间盘纤维环组织成骨潜能的体内观察

目的：观察山羊颈椎椎间融合器内填充松质骨、纤维环组织后在体内的组织学变化过程，以了解纤维环组织在骨融合过程中的成骨潜能。方法：实验山羊按常规颈椎前路减压、内同定术式施术，术中随机取C2～C6椎间隙中相邻的两个间隙．每个间隙各置入两枚钛合金颈椎窄心螺纹式柱状内同定器(CHTF)，分别填充单纯松质骨(A组)：松质骨 纤维环(B组)：纤维环(C组)及空白对照(D组)。术后应用X线片、颈椎CT扫描等影像学检查及组织切片观察植骨融合及局部组织反应情况。结果：X线片及CT示内置CHTF与椎体的骨一金属界面周围有骨组织生长．CHTF与椎体终板接触部位有成骨现象，骨桥形成。A组切片观察示新生软骨、骨小梁存在，原植入骨坏死：B组纤维组织有坏死．原骨小梁、纤维环周围新牛骨存在，新生软骨堆积；C组术后6周纤维组织内有纤维软骨存在．术后12周新生软骨存在；D组术后6周组织学观察无阳性染色结果，术后12周有少量新生软骨。结论：颈椎间盘纤维环组织有成骨潜能，成骨形式可能是成纤维细胞的软骨化骨。     

重建端粒酶活性的正常人成纤维细胞的成骨潜能研究

目的　通过重建端粒酶活性延长人成纤维细胞寿命 ,并对其成骨潜能进行研究 ,为解决组织工程骨修复种子细胞老化问题提供实验依据。方法　将人端粒酶催化亚基 (h TERT)基因用电穿孔法导入正常人原代成纤维细胞 ,用 TRAP- PCR检测细胞端粒酶活性 ,用β-半乳糖苷酶活性测定评价细胞衰老情况。在此基础上用骨形成蛋白(BMP- 2 )和肿瘤坏死因子 -α(TNF-α)联合诱导已重建端粒酶活性的成纤维细胞在体外培养条件下成骨 ,用四环素活体标记和茜素红染色显示钙盐结节形成。结果　转染 h TERT的人成纤维细胞能稳定表达端粒酶活性 ,培养超过 5 0代后细胞仍保持β-半乳糖苷酶阴性。经 BMP- 2和 TNF-α诱导后 ,转染后传 80代的人成纤维细胞仍可形成四环素标记和茜素红染色均为阳性的钙盐结节。结论　重建端粒酶活性、寿命延长的人成纤维细胞仍然维持了其本身所具有的成骨潜能     

颈椎不稳在交感型颈椎病发病中的作用

目的：研究交感型颈椎病的病理因素及治疗方法。方法：回顾分析了1988-2000年收治的20例手术治疗的交感型颈椎病患者。根据术前及术后颈椎伸屈侧位X光片判断有无颈椎不稳。结果：20例患者术前均有颈椎不稳,颈椎不稳主要发生在C3-C4和C4-C5,颈椎高位硬膜外封闭对大部分患者有短期效果。每例患者均于不稳节段行颈前路融合术,手术有效率为90％。结论：颈椎不稳是导致交感型颈椎病发病的重要因素;颈椎高位硬膜外封闭可有短期疗效因此具有重要的诊断价值;颈椎前路植骨融合术是治疗交感型颈椎病的有效方法。     

Postoperative instability of cervical OPLL and cervical radiculomyelopathy

The presence of cervical spine instability with respect to preoperative and postoperative changes in angular, horizontal, and rotational displacement of the vertebral body were studied. With the anterior approach, the instability in the remaining unfused segments, and their relation to the kyphotic or lordotic fused segment were studied. With the posterior approach, postoperative ROM (range of motion) could be better maintained, and horizontal displacement was improved in more cases by laminoplasty compared with laminectomy. With the anterior approach, the compensatory function for the loss of motion of the segments resulting from fusion was most remarkable at the levels of C2-3 and C6-7. In the alignment of the anterior fused segments, it appears important that the physiologic lordotic position be maintained.     

Anterior cervical foramenotomy for cervical radiculopathy

Anterior cervical discectomy (ACD) is standard practice for cervical radiculopathy. Irrespective of the precise method used, it involves more or less complete disc removal with resultant anatomical and biomechanical derangements, and frequently the insertion of a bone or prosthetic graft. Anterior cervical foramenotomy is an alternative procedure that allows effective anterior decompression of the nerve root and lateral spinal cord, whilst conserving the native disc, preserving normal anatomy and movement, and protecting against later degeneration at adjacent spaces as far as possible. The aim of the study was to determine the safety and efficacy of anterior cervical foramenotomy in the treatment of cervical radiculopathy and took the form of a prospective study of 21 cases under the care of a single surgeon. All patients had a single level or two level anterior cervical foramenotomy. All had pre- and postoperative visual analogue scores for arm and neck pain, arm strength, sensation and overall use. A comparison between patients' perceptions and surgeon's observations was also made. Patients were followed up for between 10 and 36 months. Sixty-eight per cent completed full pre- and postoperative assessments. Twenty-eight per cent of the responders had complete arm pain resolution. There were statistically significant reductions in arm and neck pain, and overall disability. The surgeon's impression of improvement paralleled that of the patients. There was one complication with discitis. Anterior cervical foramenotomy is a safe and effective treatment for cervical radiculopathy caused by posterolateral cervical disc prolapse or uncovertebral osteophyte, and might also reduce adjacent segment degeneration.     

Percutaneous cervical nucleoplasty and percutaneous cervical discectomy treatments of the contained cervical disc herniation

Background  

There were no studies in literature to compare the clinical outcomes of percutaneous nucleoplasty (PCN) and percutaneous cervical discectomy (PCD) in contained cervical disc herniation.     

颈横动脉颈段皮支皮瓣修复颈部瘢痕挛缩

目的 观察颈横动脉颈段皮支皮瓣修复颈部瘢痕挛缩的临床效果.方法 笔者单位1988-2011年收治颈前区烧伤后瘢痕挛缩患者66例.采用颈横动脉颈段皮支皮瓣修复患者颈部瘢痕,包括岛状皮瓣55例(其中9例行预扩张)、非岛状皮瓣11例(其中1例行预扩张).术中先切除、松解患者颈部瘢痕,在锁骨上、下及前胸区设计颈横动脉颈段皮支皮瓣,其轴心血管为颈横动脉在胸锁乳突肌、肩胛舌骨肌交界处穿出的皮动脉.皮瓣后界达斜方肌前缘,外侧界达三角肌中段,内侧界达胸骨中线,下界达乳头下3.0 ～4.0 cm处.术中先切开皮瓣外、下、内缘,锐性分离达锁骨平面后改为钝性剥离,分离到蒂部后,分离深度以皮瓣旋转后可无张力覆盖创面为度.其中预扩张的皮瓣供区直接拉拢缝合,非预扩张皮瓣供区植皮封闭.结果 本组患者中64例术后皮瓣成活良好;2例术后皮瓣下血肿致尖端部分坏死,经补充植皮后治愈;供区均愈合.所有皮瓣色泽、质地与周围组织匹配良好;皮瓣感觉功能术后初期恢复为胸部感觉,6个月后完全恢复为颈部感觉.结论 颈横动脉颈段皮支皮瓣血供恒定,解剖操作相对简便,皮瓣色泽、质地与颈部相近,是修复颈部严重瘢痕挛缩的良好选择.     

经皮髓核成形术与椎间盘切除术治疗退变性颈椎间盘突出症

目的比较经皮髓核成形术与经皮椎间盘切除术治疗退变性颈椎间盘突出症的临床疗效及对颈椎稳定性的影响。方法2002年7月至2004年12月共收治退变性颈椎间盘突出症患者80例,行经皮髓核成形术42例(PCN组),经皮椎间盘切除术38例(PCD组)。回顾性分析两组的临床资料,比较两组在手术时间、临床效果及颈椎稳定性等的差异。结果所有病例随访6~26个月,PCN组平均(12±5)个月;PCD组平均(12±4)个月。两组手术均获成功。两组手术时间有显著差异(t=-21·70,P=0·000);两组手术临床效果(JOA评分)经自身配对t检验显示均有显著性差异(PCN:t=14·05,P=0·000;PCD:t=-14·79,P=0·000),即两组均有效;两组手术临床效果(Williams评分)经Kruskal-Wallis检验无显著差异(z=-0·377,P=0·706,>0·05),即两组临床效果相似。两组手术后均无颈椎不稳病例发生,颈椎稳定性手术前后均无显著差异(P>0·05)。结论经皮髓核成形术与经皮椎间盘切除术治疗颈椎间盘突出症的临床疗效优良,对颈椎稳定性影响小,不会造成颈椎失稳的发生。     

颈椎不稳致交感型颈椎病的诊断和治疗

目的 探讨交感型颈型病的发病机制及有效的治疗方法。方法 回顾了1989-1998年应用颈前路间盘切除加植骨融合术治疗的交感型颈椎病患者18例,分析了患者产及术后颈椎伸、屈侧位X光片。结果 18例患者术前均有颈椎不稳,不稳定节段为1个者6例,2个者9例,3个者3例;颈椎不稳主要发生于C3-C4和C4-C5,偶见于C5-C6和C6-C7。14例患者术前行颈椎高位硬膜外封闭,11例有效;于不稳定节段行颈前路间盘切除加植骨融合术,18例均获随访,平均随访时间为1年9个月,术后有效率为88.9%,结论 颈椎不稳定是交感型颈椎病发病的重要因素。颈椎高位硬膜外封闭具有重要的诊断价值。颈前路间盘切除加植骨融合术是治疗交感型颈椎病的有效方法。     

颈椎自锁PEEK椎间融合器在颈椎病治疗中的应用

目的评价应用颈椎自锁PEEK椎间融合器前路减压治疗颈椎病的疗效。方法应用颈椎自锁PEEK椎间融合器治疗颈椎病患者18例,观察椎间融合器的稳定性和融合情况,采用Borden法测量椎间隙高度、颈椎曲度,对术前和术后JOA评分、椎间隙高度、颈椎曲度等指标进行统计学分析。结果 18例患者均获随访,时间6～42个月,未见严重并发症。术后6个月提示椎间骨性融合。椎间高度:术前为(3.26±0.68)mm,术后1周为(6.03±0.89)mm(P<0.01);术后6个月为(5.89±0.78)mm,与术后1周比较变化不大(P>0.05)。颈椎生理弧度:术前为(2.55±0.48)mm,术后1周为(3.24±0.67)mm(P>0.05);术后6个月为(8.14±1.17)mm,与术后1周比较明显改善(P<0.01)。末次随访根据JOA评分法进行疗效评价:优4例,良9例,可3例,差2例。结论颈椎自锁PEEK椎间融合器可以有效恢复颈椎生理曲度及椎间隙高度,可以获得满意的融合率,改善颈脊髓功能。     

Comparison of cervical kinematics between patients with cervical artificial disc replacement and anterior cervical discectomy and fusion for cervical disc herniation

Background contextAlthough anterior cervical discectomy and fusion (ACDF) is an effective treatment option for patients with cervical disc herniation, it limits cervical range of motion, which sometimes causes discomfort and leads to biomechanical stress at neighboring segments. In contrast, cervical artificial disc replacement (ADR) is supposed to preserve normal cervical range of motion than ACDF. A biomechanical measurement is necessary to identify the advantages and clinical implications of ADR. However, literature is scarce about this topic and in those available studies, authors used the static radiological method, which cannot identify three-dimensional motion and coupled movement during motion of one axis.PurposeThe purpose of this study was to compare the clinical parameters and cervical motion by three-dimensional motion analysis between ACDF and ADR and to investigate the ability of ADR to maintain cervical kinematics.Study designThis was a prospective case control study.Patient samplePatients who underwent ADR or ACDF for the treatment of single-level cervical disc herniation.Outcome measuresVisual analog scale (VAS), Korean version of Neck Disability Index (NDI, %), and three-dimensional motion analysis were used.MethodsThe patients were evaluated by VAS and the Korean version of the NDI (%) to assess pain degree and functional status. Cervical motions were assessed by three-dimensional motion analysis in terms of sagittal, coronal, and horizontal planes. Markers of 2.5 cm in diameter were attached at frontal polar (Fpz), center (Cz), and occipital (Oz) of 10–20 system of electroencephalography, C7 spinous process, and both acromions. These evaluations were performed preoperatively and 1 month and 6 months after surgery.ResultsThe ACDF and ADR groups revealed no significant difference in VAS, NDI (%), and cervical range of motion preoperatively. After surgery, both groups showed no significant difference in VAS and NDI (%). In motion analysis, significantly more range of motion was retained in flexion and extension in the ADR group than the ACDF group at 1 month and 6 months. There was no significant difference in lateral tilt and rotation angle. In terms of coupled motion, ADR group exhibited significantly more preserved sagittal plane motion during right and left rotation and also showed significantly more preserved right lateral bending angle during right rotation than ACDF group at 1 month and 6 months. There was no significant difference in other coupled motions.ConclusionThree-dimensional motion analysis could provide useful information in an objective and quantitative way about cervical motion after surgery. In addition, it allowed us to measure not only main motion but also coupled motion in three planes. ADR demonstrated better retained cervical motion mainly in sagittal plane (flexion and extension) and better preserved coupled sagittal and coronal motion during transverse plane motion than ACDF. ADR had the advantage in that it had the ability to preserve more cervical motions after surgery than ACDF.     

Posterior cervical laminoplasty formultilevel cervical spine stenosis

Cervical laminoplasty for treating multilevel spinal stenosis appears to be a good surgical alternative to the more traditional laminectomy or anterior decompression and fusion. This procedure avoids the morbidity associated with extensive anterior procedures and also appears not to be associated with late kyphosis, which can be seen in patients after a laminectomy. This review outlines the rationale, indications, contraindications, and early clinical results for patients undergoing a posterior laminoplasty.