自体外周血干细胞与脱钙骨复合移植修复骨缺损的临床应用

目的：观察自体外周血干细胞(APBSC)与脱钙骨(DBM)复合移植治疗良性骨肿瘤术后骨缺损的疗效。方法：对12例骨肿瘤患者，手术切除骨肿瘤脱钙骨填充骨缺损，术后第7天行骨髓动员，每天皮下注射特尔立3．0μg／kg，连续3d，第4天静脉滴注10mg地塞米松后采集APBSC。在X线电视透视下将一枚骨穿针准确穿入骨缺损部位，然后抽取自体外周血干细胞即可注入骨缺损部位。通过术后连续X线片，了解其骨缺损修复能力。结果：12例病人术后得到随访，时间为4～16个月，在2～4个月以内均开始有不同程度的骨化，成骨效应满意。结论：自体外周血干细胞(APBSC)／脱钙骨(DBM)复合移植是一种治疗骨缺损有效的新方法。     

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。     

自体外周血及自体红骨髓干细胞与脱钙骨复合移植治疗骨缺损的比较研究

目的 对比观察自体外周血干细胞（APBSC）／脱钙骨（DB）复合移植与自体外红骨髓（ARBM）／DB复合移植治疗骨缺损的疗效。方法 36只家兔双侧桡骨造成1cm骨缺损,随机分为DB组、ARBM／DM级和APBSC／DB组,每组12只,分别进行X线片、生物力学和组织学检查,然后作对比分析。结果 术后第2、4、8、14周,APBSC／DB组和ARBM／DB组X线片、改进的GaryX线评分和光镜观察结果,以及术后第14周整骨破坏载荷和骨缺损修复形态学评分均明显优于DB组;但APBSC／DB组与ARBM／DB组间差异无显著性。结论 APBSC与ARBM都能在DB的骨形态蛋白诱导下促进成骨细胞的形成,复合移植疗效明显优于单纯DB移植。     

自体外周血造血干细胞移植治疗系统性红斑狼疮的临床观察

目的观察自体外周血造血干细胞移植治疗系统性红斑狼疮（SLE）的临床疗效。方法5例对其他治疗效果反应较差的复发SLE患者接受自体外周血造血干细胞移植术，造血干细胞动员方案采用环磷酰胺＋粒细胞集落刺激因子，应用CS-3000 Plus血细胞分离机采集外周血干细胞，预处理方案采用环磷酰胺＋抗胸腺球蛋白抗体方案，常规措施预防移植相关并发症。移植后3个月，复查各项免疫指标，评估疗效。结果5例患者造血干细胞动员均获成功，造血重建顺利回输后外周血中性粒细胞计数≥0．5×10^9／L的天数为8。16d；白细胞总数恢复正常的天数为10-20d：血小板计数〉20×10^9／L的天数为12～20d。无一例出现严重的移植后并发症。临床症状明显改善或消失。3个月后复查各项指标均较移植前有明显改善。结论自体外周血造血干细胞移植治疗SLE的临床疗效好，不良反应少，是治疗难治性SLE的一种较好的方法。     

自体骨髓基质干细胞在齿槽裂骨缺损修复中的应用

目的探讨人自体骨髓基质干细胞（human bone marrow stromal cells，hBMSCs）在治疗齿槽裂骨缺损中的可行性。方法2002至2005年，选择齿槽裂骨缺损患者7例（单侧6例，双侧1例），以患者自体骨髓基质干细胞为种子细胞，部分脱钙骨（partly demineralized bone matrix，pDBM）为支架材料构建组织工程骨，治疗齿槽裂骨缺损。从患者髂前上棘穿刺取骨髓，密度梯度离心法分离hBMSCs，经体外成骨诱导和扩增至第3代。将诱导的hBMSCs，复合部分脱钙骨体外培养1周后，手术回植骨缺损区。分别于术后1、3、6、12、24、36个月进行临床外形和三维CT检查随访。结果6例患者头部三维CT检查，结果示术后3个月能形成组织工程化骨，并修复骨组织缺损。术后1～3年的随访表明组织工程骨稳定存在，无明显骨吸收现象，临床治疗效果稳定。1例患者（双侧齿槽裂）植入物外露感染。结论以自体hBMSCs为种子细胞，部分脱钙骨为支架材料，利用组织工程技术可在人体内形成稳定的组织工程化骨组织，并临床修复齿槽裂骨缺损。     

放射性核素骨显像动态检测组织工程化骨血管化的价值

[目的] 放射性核素骨显像动态检测评价组织工程化骨血管化与骨代谢的价值.[方法] 取大白兔24只,抽取骨髓,行骨髓间充质干细胞分离、培养与骨向诱导.双侧股骨髁制作0.6 cm×1.2 cm的骨缺损,将诱导后的骨髓间充质干细胞与珊瑚羟基磷灰石复合植入左侧骨缺损处为实验组,右侧单纯植入珊瑚羟基磷灰石为对照组.术后4、8、12周分别行静态放射性核素骨显像检测,用ROI计数计算不同时间点的ROI平均计数.[结果] 结果术后4、8、12周实验组ROI计数(单位像素)均较对照组有显著性增高.实验组ROI计数随时间的延长,呈明显的上升趋势,术后8周开始增长放缓;对照组ROI计数术后8周增长缓慢,8周后开始增长加快,两组均在12周达到峰值.[结论] 骨髓间充质干细胞诱导后与珊瑚羟基磷灰石复合可有效的修复股骨髁松质骨缺损.放射性核素骨显像可动态检测组织工程化骨血管化及骨代谢变化.     

自体外周血干细胞与脱钙骨复合移植治疗兔桡骨缺损的实验研究

目的：观察自体外周血干细胞／脱钙骨（APBSC／DB）复合移植治疗骨损损的疗效,方法：24只家兔双侧桡骨做成长度1cm的骨缺损,随机分为APBSC／DB组和DB组,每组12只,分别进行X线,生物力学以及组织学的对比研究,结果：术后第2,4,8,14周,X线观察,改进的Gay评分及光镜观察结果显示,APBSC／DB组优于DB组,术后第14周,APBSC／DB组整骨破坏载荷和骨缺损修复形态学评分均明显优于DB组。结论：APBSC在DB的骨形态蛋白诱导下具有明显的成骨作用,因此APBSC／DB复合移植治疗骨缺损的疗效优于单纯DB移植。     

造血干细胞动员中外周血CD34+细胞的变化趋势及其对采集的影响

目的 探讨外周血造血干细胞动员过程中外周血CD34+细胞的变化趋势及其对采集时机和采集结果的影响.方法 2010年4月至2011年12月外周血造血干细胞动员及采集27例,其中自体13例,采用化疗联合粒细胞集落刺激因子(G-CSF) 10μg· kg-1·d-1动员方案;健康供者14例,单独应用G-SCF 7.5μg· kg-1·d-1动员方案.监测外周血CD34+细胞数,并与采集产物中获得的单个核细胞(MNC)及CD34+细胞量进行相关性分析.结果 健康供者中获得MNC(5.84±1.48)×108/kg,CD34+细胞(3.93±2.16)×106/kg.自体动员共获得MNC(6.58±3.72)×108/kg,CD34+细胞(3.98±3.06)×106/kg,仅1例自体动员、采集失败.自体动员外周血CD34+细胞在第4天达峰值,健康供者第5天仍处于上升阶段.外周血CD34+细胞百分比和绝对值与采集产物CD34+细胞呈明显正相关.采集当天CD34+细胞绝对值≥20/μ1时,单次采集获得CD34+细胞≥2×106/kg者的比例达76.2％(16/21).结论 外周血CD34+细胞计数是造血干细胞采集过程中的重要监测指标,对于把握采集时机和预测采集效果具有重要意义,CD34+细胞绝对值≥20/μ1可作为采集阈值.     

骨髓干细胞转分化为肾祖细胞并参与修复急性肾脏损伤的研究

目的 观察骨髓干细胞是否可以向肾祖细胞转分化,成为肾脏祖细胞库的肾外来源;验证粒细胞集落刺激因子(G-CSF)是否可以促进骨髓干细胞向肾脏祖细胞的转分化,提高肾脏修复的效能.方法 6周龄全身表达绿色荧光蛋白(GFP)的C57BL/6J转基因小鼠提供骨髓,6~8周龄同种无荧光标记的C57BL/6J小鼠40只作为骨髓受体.骨髓移植前,受体小鼠接受致死剂量的γ放射线137Cs照射,骨髓重建情况经流式细胞仪检测确认.骨髓重建完毕后所有小鼠均接受单侧肾脏缺血再灌注损伤.干细胞动员效果及向肾脏归巢情况经流式细胞仪检测鉴定.损伤4、8周后取肾脏标本行免疫荧光组织化学染色,观察骨髓来源的肾脏祖细胞数以及骨髓细胞在微血管形成中的作用.损伤4周后通过组织切片免疫荧光组织化学方法观察并计数微血管细胞数.结果 G-CSF动员1 d后,分别为CD29、CD34、Sca-1、c-Kit、Flk-1阳性的干细胞占外周血非红系细胞的比例均高于对照组(P<0.05).损伤4周后,G-CSF动员组的肾脏中,骨髓来源并且分别表达Sca-1/GFP、CD29/GFP的干细胞的比例均高于对照组(P<0.05);在损伤4周及8周后,肾脏切片免疫荧光组织化学显示G-CSF动员肾脏中骨髓来源的肾祖细胞即Sca-1/GFP双阳性的细胞数量高于对照组.损伤4周后,动员组肾脏中表达CD31的微血管密度高于对照组(P<0.05).损伤4周后肾脏组织中存在CD105/GFP及α-SMA/GFP双阳性的细胞.结论 ①骨髓干细胞可以转分化为器官特异性干细胞-肾脏祖细胞;②G-CSF可以加速这一转分化的过程,并使损伤肾脏得到更好的修复.     

自体外周血干细胞／脱钙骨复合移植治疗兔桡骨缺损的实验研究

目的：观察自体外周血干细胞（Autologous Pexipheral Blood Stem Cell-APBSC)／脱钙骨（Decalcified Bone Matrix-DBM)复合移植治疗骨缺损的疗效。方法：36只家兔桡骨做成1cm骨缺损，随机分为DBM组自体红骨髓（Autologous red bone marrow-ARBM)/DBM组APBSC／DBM组，每组12只。分别进行X线观察、生物力学检查和组织学观察的对比研究。结果：术后第2、4、8、14周APBSE／DBM组X线观察改进的Gary评分和光镜观察均优于DBM组，与ARBM／DBM组相似，术后第14周APBSC／DBM组整骨破坏载荷和骨缺损修复，形态学评分也优于DBM组，与ARBM／DBM组比较，前没有显性差异，后有显性差异。结论：APBSC／DBM复合移植治疗骨缺损的疗效优于单纯DBM移植，与ARBM／DBM组相似。结合临床我们认为具有良好的应用前景。     

Osteogenesis after Bone and Bone Marrow Transplantation: II. The Initial Cellular Events Following Transplantation of Decalcified Allografis of Cancellous Bone

An experimental study was done in rabbits to investigate the fate of allogeneic iliac cancellous bone, both non-decalcified and decalcified with hydrochloric acid, transplanted to a muscular site for up to 14 days. Some of the treated allografts were impregnated with autologous bone marrow cells, obtained from the femoral medulla by aspiration, and each was compared with allografts alone. Combined myelo-osseous grafts produced bone after 7 to 8 days implantation, as did marrow autografts alone. In addition non-decalcified implants stimulated the production of multinucleated giant cells. Three different types of wash solution were used but these did not influence the cell population seen, nor the new bone formation. It is concluded that the critical events in bone formation after transplantation occur less than 8 days after the transplantation and that marrow cells have osteogenic capacity. This has relevance to the clinical aspects of bone grafting.     

Bone cement with reduced proportion of monomer in total hip arthroplasty: Preclinical evaluation and randomized study of 47 cases with 5 years' follow-up

Bone cement with reduced amount of monomer and low curing temperature may improve implant fixation due to reduced toxicity. We analyzed the mechanical, chemical and thermal properties of such a cement (Cemex Rx) using Palacos R as control. The in vivo performance of the 2 cements was also evaluated in a prospective randomized study of 47 hips, where either of the cement types was used to fixate Lubinus SP2 prostheses with the stem made of titanium alloy. Cemex Rx had a reduced tensile strength, probably because this cement was manually mixed, as recommended by the manufacturer. A standardized laboratory test showed lower curing temperature for Cemex, but measurements at 37° and with prechilled Palacos R and Cemex Rx, as in clinical work, showed no difference. In the clinical study radiostereometric measurements of cup and stem migration showed similar values in the 2 groups up to 5 years after the operation. The cement mantle was stable in both groups, but the stems migrated similarly inside the cement mantle regardless of the type of cement used. Proximal wear was low (0.04-0.05 mm/year) and tended to be lower in the Cemex group (p = 0.02). Aluminum and vanadium levels in serum increased 5 years after the operation, but no difference was noted between the 2 groups. Collagen markers (PICP, ICTP) showed similar increases in bone turnover 6 weeks and 6 months after operation in both groups.     

The Weight-Rearing Capacity of Structural Elements in Femoral Necks

1. Up to the age about 60 the resistance of the femoral neck to compression forces is the same on the left and the right side. After that age the mechanical resistance may decrease considerably on one side. This might be of clinical significance in the explanation of neck fractures.

2. The inferior cortical layer of the femoral neck has a weight-bearing capacity of 40 per cent in adults.

3. The superior cortical layer has a weight-bearing capacity of 20 per cent.

4. The superior and inferior cancellous tissue are of approximately equal value and each takes care of 15 per cent of the weight-bearing function.

5. If both cancellous layers are destroyed at the same time, the weight- bearing capacity is reduced by 30 per cent.

6. If the inferior compact layer and the inferior cancellous layer are eliminated or the superior cortical and trabecular system are removed, the weight-bearing capacity will be 50 per cent of normal.

7. The measurements sem to indicate that the trabecular tissue corresponds to a reinforcing system which helps to increase stability. It might be compared with cables in a bridge construction, the actual foundation of which is represented by the compact tissue.

     

Assessment of leech therapy for knee osteoarthritis: A randomized study

Background and purpose?Symptomatic treatment of osteoarthritis of the knee with leeches is presently undergoing a renaissance. Previous studies have shown methodical weaknesses. In the present study patients were blinded regarding the treatment, and a control group was included to explore possible differences in various subjective clinical scores and intake of pain medication over time between leech therapy and placebo control.

Patients and methods?113 patients with advanced osteoarthritis of the knee were included. The patients were randomized to a single treatment group, group I (single leech application, n = 38), a double treatment group, group II (double application, n = 35), and a control group (n = 40). The second treatment in group II took place after an interval of 4 weeks. The treatment in the control group was simulated with the help of an “artificial leech”. Results were documented with the KOOS and WOMAC scores and also a visual analog scale (VAS) for pain. Changes in the use of pain medication were monitored over 26 weeks.

Results?An improvement in KOOS and WOMAC scores, and also in VAS, was found in all 3 groups following treatment. These improvements were statistically significant for treatment groups I and II during the complete follow-up period. The reduction in individual requirements for pain medication was also statistically significant. The greatest improvement was seen in the group treated twice with the leeches, with a long-term reduction of joint stiffness and improved function in the activities of daily living.

Interpretation?Leech therapy can reduce symptoms caused by osteoarthritis. Repeated use of the leeches appears to improve the long-term results. We have not determined whether the positive outcome of the leech therapy is caused by active substances released during the leeching, the placebo effect, or the high expectations placed on this unusual treatment form.     

Osteogenesis After Bone and Bone Marrow Transplantation: Studies of Cellular Behaviour Using Combined Myelo-osseous Grafts in the Subscorbutic Guinea Pig

In order to study factors influencing osteogenesis after bone and bone marrow transplantation, we have caused guinea pigs to become scorbutic, and looked at the cell morphology at sites of bone formation. We had previously studied normal guinea pigs and found that autologous marrow in intermuscular implants was associated with bone production by the ninth day, regardless of the type of stored allogeneic bone transplanted with it. in subscorbutic guinea pigs, using identical implants, bone did not appear within the first 13 days, and the cell population around the implants was different. These experiments support the dominant role of bone marrow cells in osteogenesis and cast further doubt on the primary role of devitalised bone as an inducer of bone formation. Interference with cell function by deprivation of a single essential molecule, Vitamin C, produces great change in the ability of cells to synthesise bone, or pre-osseous matrix.     

Bone Marrow Cell Content and Osteoclasts in Crista Biopsies

The bone marrow cell content was investigated by counting the percentage of cells in the areas between the trabeculae in crista biopsies. There was a decrease in the percentage of cells in bone marrow in osteoporotic women compared with our normal material. in patients undergoing haemodialysis there was a positive correlation between bone marrow cell content and osteoclasts, which could indicate a possible connection between osteoclasts and bone marrow cells.     

骨髓基质干细胞与生物衍生骨复合修复山羊胫骨缺损的研究

目的　探讨骨髓基质干细胞 (marrow stromal stem cells,MSCs)与生物衍生骨复合修复山羊胫骨的长段骨 -骨膜缺损的可行性。　方法　将 12月龄山羊 35只双侧胫骨制备成中段 2 0 mm的骨 -骨膜缺损 ,其中 33只 ,将MSCs与生物衍生骨于体外复合培养后 ,将其植入右侧缺损处 ,常规钢板螺钉内固定作为实验组 ,以单纯生物衍生骨植入左侧作为对照组 ,另 2只为空白组不植入任何材料 ,在 2、4、6、8、12、16及 2 4周各时间点分别行大体标本、组织学观察 ,以及生物力学测试 ,比较 3组骨缺损修复的能力。　结果　 4周时实验组支架材料部分吸收 ,植入物表面有纤维骨痂形成 ,对照组支架材料少量吸收 ,植入物表面有少量骨样组织形成 ;8周时 ,大体标本、组织学观察 ,实验组中的支架材料已完全降解 ,骨缺损部分修复 ,对照组中植入物两端少量新骨形成 ,材料中为纤维骨样组织 ,但 8周时 ,实验组与对照组的生物力学强度差异无统计学意义 (P>0 .0 5 ) ;12周时 ,实验组骨缺损完全修复 ,对照组植入物两端有新骨包绕 ,与骨端连接紧密。 12～ 2 4周实验组弯曲应力大于对照组有统计学意义 (P<0 .0 5 ) ;2 4周时空白组骨缺损未修复。　结论　所构建的组织工程骨修复能力较强 ,成骨迅速 ,且量大 ,同期新生骨组织的生物力学强度优于单纯     

阿伦膦酸钠对骨髓生成破骨细胞及骨吸收作用的影响

[目的]研究阿伦膦酸钠对体外培养的小鼠骨髓生成破骨细胞及其骨吸收作用的影响。[方法]收集小鼠骨髓细胞于含有10^-8mol／L的1，25二羟基维生素D3[1，25-（OH）2D3]的α-MEM完全培养基中体外培养，设置不同浓度的阿伦膦酸钠给药，并于培养的第6、9、12d观察记录抗酒石酸酸性磷酸酶（tartrate-resistant acidphosphatase，TRAP）阳性多核巨细胞[破骨样细胞（osteoclast-like cell，OLC）]生成数，反映破骨细胞生成情况。记数培养12d骨磨片上骨吸收陷窝数及吸收面积，反映骨吸收情况。[结果]随着阿伦膦酸钠浓度的增高，TRAP阳性的细胞数减少，骨吸收陷窝数及面积均减少。[结论]阿伦膦酸钠可抑制骨髓细胞体外培养中破骨样细胞的形成，体外可抑制破骨细胞骨吸收作用。     

Evaluation of the ability of bone marrow derived cells to engraft the kidney and promote renal tubular regeneration in mice following exposure to cisplatin

It has been suggested that bone marrow derived stem cells have the ability to engraft the kidney and improve the outcome of severe acute kidney injury (AKI) in mice exposed to high doses of cisplatin, providing hope for cancer patients in whom irreversible renal damage occasionally occurs following the use of this highly effective anti-tumor drug. We tested the therapeutic potential of bone marrow derived cells injected during the acute phase (day 3 after cisplatin administration) of experimentally-induced AKI in C57Bl6/J mice, characterized by massive tubular necrosis, apoptosis, and a low proliferation capacity. We failed to show any benefit of bone marrow derived cells versus a regular homogenate of intact renal cells, or normal saline. Using cell tracers and flow cytometry, we demonstrated that bone marrow derived cells did indeed home to the bone marrow of the recipients but failed to settle in the kidney. Conversely, renal cells homed to injured kidneys. However, neither cell therapy protected the animals against cisplatin-induced death. We therefore question the short-term efficacy of bone marrow derived cells used to repair established injuries of the tubular epithelium.