异种骨复合自体骨髓修复骨缺损的实验研究

目的　观察异种骨复合自体红骨髓对骨缺损的修复能力。方法　选健康纯系 ６ 月龄大耳白兔 ２０ 只,双侧桡骨中段制成 １５～１８ ｃｍ 的骨缺损模型,骨蜡封闭两断端骨髓腔,均植入等长度的异种骨,左侧注射红骨髓作为实验组,右侧不注射作为实验对照组。术后 ２、４、６、８ ｗ 拍 Ｘ 线片,行组织学检查。结果　实验组 ８ ｗ 时, Ｘ 线示骨缺损修复良好、髓腔基本相通,组织学检查见皮质骨形成、骨髓腔相通。而对照组 ８ ｗ 时 Ｘ 线下仍可见未完全吸收的移植骨,光镜示缺损区有残存的坏死骨,纤维组织充填缺损区。结论　异种骨复合自体红骨髓后可明显促进骨融合,提高修复骨缺损的能力     

不同材质异体骨移植治疗创伤性骨缺损临床分析

采用同种异体骨移植治疗创伤性骨缺损38例.所有患者随访10个月以上,37例骨性愈合,其中大段异体骨复合自体骨移植15例骨折愈合,感染性骨缺损感染复发1例,骨折不愈合;局限性骨缺损松质骨颗粒复合自体骨髓移植22例骨折全部愈合.认为同种异体骨为一种良好骨植入材料,长段异体骨复合自体骨后可用于创伤造成的大段骨缺损,异体松质骨颗粒复合自体红骨髓移植可用于修复陈旧性骨折和局限性骨缺损.     

自体红骨髓复合骨膜碎片移植修复骨缺损的实验观察

目的观察自体红骨髓复合骨膜碎片移植修复骨缺损的形态学变化。方法40只健康中国家兔取双侧桡骨中段制作15 mm长的骨缺损模型。随机选择20只行自体红骨髓复合骨膜碎片移植术（移植组）,分别于术后第1、2、4、6、8周行X线摄片后,取出桡骨标本,大体观察、显微镜观察骨缺损区的形态学变化。另20只为对照组,桡骨骨缺损区内不移植任何组织。结果移植组：第1周移植组织增生;第2周移植组织弹性固定桡骨远近两断端;第4周在原始骨小梁间可见大量成纤维细胞;第6周骨小梁密集,成骨细胞排列在骨小梁周围,骨细胞位于骨陷窝内;第8周可见典型内、外环骨板及哈佛系统,骨髓腔形成,腔内可见多种细胞。对照组：第1周两断端边缘有少量新生组织;第2周新生组织覆盖两断端边缘;第4周可见成骨细胞;第6周有骨小梁形成;第8周形成骨密质并封闭缺损区远近两断端。结论自体红骨髓复合骨膜碎片移植可修复桡骨骨缺损,其过程可分为细胞分化增殖期、幼稚骨形成期及骨改造塑形期3个阶段。     

外源性VEGF复合煅烧骨修复骨缺损

目的 探讨血管内皮生长因子（VEGF）复合煅烧骨（TBC）的修复骨缺损作用及临床应用前景。方法 新西兰大白兔36只,建立桡骨干中段10mm骨缺损模型。实验侧植入VEGF—TBC复合物,对照侧单纯植入TBC。术后第1、2、4、6、8和12周行X线、植入材料内部血运重建和新骨生成的定量观察,并测定TBC的降解率。结果 VEGF—TBC复合物修复骨缺损的效果优于单纯的TBC,VEGF可以加速TBC的降解。结论 VEGF—TBC复合物能够满意修复骨缺损,是一种较为理想的骨组织工程材料,具有良好的临床应用前景。     

组织工程化人工骨种植体修复兔骨缺损效果研究

目的探讨珊瑚羟基磷灰石（CHA）复合重组人胰岛素样生长因子-Ⅰ（IGF-Ⅰ）及自体红骨髓（ABM）修复骨缺的效果。方法选取36只家兔,随机分A、B、C、D、E、F共6组,各组种植体分别为CHA/IGF-Ⅰ/ABM、CHA/IGF-Ⅰ、CHA/ABM、CHA、ABM、空白。术后于4、8、12周分别对动物的一般情况及伤口进行观察,每次观察后随机处死家兔2只并取材,以组织学方法观察成骨情况。成骨情况按Lane-Sandh组织学评分标准进行评分。结果术后4、8、12周大体观察及病理组织切片结果显示A组与其他各组比较有统计学意义。结论CHA/IGF-Ⅰ/ABM构建的组织工程化人工骨种植体成骨能力强,是修复骨缺损的良好材料。     

柞蚕丝素蛋白-羟基磷灰石-BMSCs组织工程化骨修复老龄动物骨缺损

目的 探讨柞蚕丝素蛋白-羟基磷灰石-骨髓间充质干细胞(TSF-HA-BMSCs)构成的组织工程骨对兔桡骨缺损的修复作用.方法 将TSF-HA与成骨诱导后兔骨髓间充质干细胞(BMSCs)进行复合,构建新的组织工程骨.取36只12月龄老龄日本大耳白兔于左侧桡骨中段造15mm长的骨缺损区.实验分为3组(A、B、C组各12只).A组:TSF-HA+ BMSCs,B组:HA+ BMSCs,C组:无支架组.分别在术后8和12w摄取X线片,观察骨缺损区的修复及骨塑性情况,骨缺损区组织标本采取HE染色进行组织学观察,比较8和12 w各组的骨修复情况.结果 成功培养出了BMSCs,且细胞在材料表面生长状态良好.骨缺损修复术后8、12wX线检测显示,A组骨缺损的修复效果最好,B组次之,C组几乎没有修复作用,各组间差异有统计学意义(P＜0.05);HE染色表明,8w时与B组或C组相比,A组新骨生成最多,骨修复最快,差异有统计学意义(P＜0.05);12 w后取桡骨观察证实:实验组支架大部分被吸收,骨折处密度增高,骨质发白,实验对照组可见部分支架未被吸收,但骨折愈合良好;空白组骨缺损处被结缔组织填充覆盖.结论 TSF-HA组织程化骨具有很好的骨缺损修复能力,有望成为新骨缺损修复的替代材料.     

Bio-Oss结合Bio-Gide修复牙槽临界骨缺损的效果

目的 观察Bio-Oss骨粉复合Bio-Gide骨膜在兔牙槽骨缺损修复过程中引导成骨的效果.方法 建立兔下颌牙槽骨临界骨缺损模型,采取自身对照,随机选择一侧下颌骨以Bio-Oss/Bio-Gide修复骨缺损,作为实验侧;另外一侧作为空白对照侧.分别于术后1、4、7、10 w通过组织学切片观察以及新生骨占缺损面积定性及定量分析骨缺损区的成骨效果.结果 术后1 ～10w组织学切片观察显示,实验侧和对照侧均有不同程度的新骨形成以及植骨材料的降解吸收;随时间延长,新生骨量增加,并与种植体表面形成骨性结合.在术后各时间点,实验侧新生骨占面积百分比均大于对照侧(P＜0.05).结论 可吸收性胶原膜Bio-Gide结合Bio-Oss应用于牙种植体周围骨缺损中,可以引导骨组织再生,重建缺损的骨组织,新生骨与种植体形成骨性结合.     

种植床自体骨复合珊瑚羟基磷灰石在牙种植手术中的临床应用研究

目的 评价种植床自体骨复合珊瑚羟基磷灰石在牙种植手术中的临床应用效果.方法 32例(37颗)种植牙病例分成A、B两组.A组24例(27颗种植体),植入德国Ankylos螺纹种植体后,在部分暴露的植体处植入珊瑚羟基磷灰石.B组8例(10颗种植体),采用经牙槽嵴顶入路,使用德国费亚丹公司的FRIOSR空心取骨钻进行定位定向和定深度后制备圆柱状骨,游离取出后即刻碾磨成碎骨颗粒,与珊瑚羟基磷灰石混合后植入到种植体周骨缺损区.植入术后4~6个月暴露种植体,最终完成烤瓷冠修复,二期手术时根据成骨情况评估成骨效果.结果 两组病例均获得了良好的骨修复效果,单纯珊瑚羟基磷灰石植骨后成骨效果满意及基本满意率共为85.19%.自体骨与珊瑚羟基磷灰石混合植骨后的成骨效果满意及基本满意率共为100.00%.结论 种植床自体骨可作为有效的植骨材料,手术创伤小,无需开辟第2手术区,值得推广.     

BMSCs-BMP2-煅烧牛骨复合物的制作及其在兔尺骨骨缺损修复中的应用

目的制作兔骨髓间充质干细胞(BMSCs)-骨形态蛋白2(BMP2)-煅烧牛骨复合物,并观察其在兔尺骨骨缺损修复中的应用。方法将兔BMSCs分离、培养及成骨诱导后,接种于BMP2-煅烧牛骨复合物,扫描电镜下观察复合情况。将54只新西兰大白兔随机分为观察组、对照组、空白组各18只,于无菌条件下造成右侧尺骨中段10mm长缺损。观察组植入BMSCs-BMP2-煅烧牛骨复合物,对照组植入BMP2-煅烧牛骨复合物,空白组不植入任何材料,术后均不做任何内外固定。各组术后4、8、12周每组随机处死6只,分别行骨缺损处的放射学和组织学检查。结果成骨诱导后的BMSCs均匀分布在煅烧骨表面及孔洞内,与煅烧骨表面贴附紧密,细胞呈圆形,表面光滑,胞体较大,直径15μm。随着术后时间的延长,观察组和对照组Lane-Sandhu放射学评分、组织学评分均逐渐升高,P均<0.05;空白组无变化,P均>0.05。术后4、8、12周,观察组和对照组Lane-Sandhu放射学评分、组织学评分均高于空白组,观察组升高更明显,P均<0.05。结论 BMSCs能够较好地与BMP2-煅烧牛骨复合物进行体外复合培养,BMSCs-BMP2-煅烧牛骨复合物植入有助于兔尺骨骨缺损的修复。     

天然型无机骨复合骨形成蛋白修复颌骨缺损的临床研究

报告２４例颌骨缺损患者，采用植入相应形态天然型无机骨复合骨形成蛋白（ＮＮＢ／ＢＭＰ）修复１１例，采用单纯天然型无机骨（ＮＮＢ）修复１３例，两组均有术后局部组织水肿、均无全身反应，２１例Ｉ期愈合，ＮＮＢ组中３例Ⅰ期愈合，ＮＮＢ／ＢＭＰ组骨缺损修复早，启动快，成骨多，Ｘ线示１２周颌骨缺损完全修复。ＮＮＢ组术后０．５￣１年形成骨性愈合，密度趋于一致。术后颌骨外形和功能良好，避免了自体骨移植缺点，并发症少     

Effect of beta-tricalcium phosphate/poly-l-lactide composites on radial bone defects of rabbit

ObjectiveTo explore the effect ofβ-TCP/PLLA scaffold in repairing rabbit radial bone defects.MethodsThirty New Zealand rabbits were divided into β-TCP /PLLA group (group A), pure PLLA group (group B) and contrast group (group C) randomly. The rabbits were sacrificed respectively after 4, 8, 12, 24 weeks and the X-ray film was performed at the same time to evaluate the repair effect in different groups.ResultsX-ray film showed there was uneven low density bone callus development in defect region after 4 weeks in group A. The defect region was filled with neonate osseous tissue completely during 12-24 weeks. X-ray score revealed that repair of bone defect results significantly better than group B and group C.ConclusionsThe β-TCP /PLLA composite is capable of repairing radial bone bone defects. β-TCP/PLLA scaffold is significant because of rapid degradation ability, good histocompatibility and osteogenic action.     

Enhancement of bone defect healing in old rats by TGF-beta and IGF-1

Bone defects are often created in order to repair bone pathologies. In the aging population, the healing of such defects is very limited. Bone healing in aging depends on the availability of various hormone and growth factors. The ability of growth factors to enhance bone formation in femoral defects in old rats was tested. Bone defects were induced in femurs of old rats. A single dose of transforming growth factor-beta (TGF-beta), IGF-1, TGF-beta+IGF-1 or saline was inserted in the defect and bones were tested after 2 and 4 weeks. Radiology revealed that mineralization appeared in the 2 weeks group in defects treated with TGF-beta and in defects treated with TGF-beta, TGF-beta+IGF-1 in the 4 weeks groups. Computerized tomography (CT) coronal and axial images revealed that 4 weeks after treatment with TGF-beta+IGF-1, a complete bone bridge was observed. Morphology revealed that these defects were filled with trabecular bone. A less pronounced bone healing was observed after TGF-beta or IGF-1, while control specimens revealed partial healing of the bone defect. Biomechanical tests indicated that treatment with TGF-beta, IGF-1 or TGF-beta+IGF-1 resulted in a significant increase of bone bending rigidity compared to control in the 4 weeks group and that TGF-beta+IGF-1 was the most inductive in this respect. The ability to induce bone healing in aging by TGF-beta+IGF-1 is of a great clinical importance for restoration of bone strength and biomechanical properties of bone defects in aging.     

Effects of zinc compound on body weight and recovery of bone marrow in mice treated with total body irradiation

This study aimed to investigate if zinc compound would have effects on body weight loss and bone marrow suppression induced by total body irradiation (TBI). ICR mice were divided randomly into two groups and treated with test or control compounds. The test compound contained zinc (amino acid chelated with bovine prostate extract), and the control was reverse osmosis pure water (RO water). One week after receiving the treatment, mice were unirradiated, or irradiated with 6 or 3 Gy by 6 MV photon beams to the total body. Body weight changes were examined at regular intervals. Three and 5 weeks after the radiation, animals were sacrificed to examine the histologic changes in the bone marrow. Lower body weight in the period of 1-5 weeks after radiation and poor survival rate were found after the 6 Gy TBI, as compared with the 3 Gy groups. The median survival time after 6 Gy and 3 Gy TBI for mice given the test compound were 26 and 76 days, respectively, and the corresponding figures were 14 and 70 days, respectively, for mice given the control compound (p < 0.00001). With zinc supplement, the mean body weight in mice which received the same dose of radiation was 7-8 g heavier than in the water-supplement groups during the second and third weeks (p < 0.05). Hence, there was no statistically significant difference in survival rate between zinc and water supplement in mice given the same dose of irradiation. Histopathologically, there was less recovery of bone marrow cells in the 6 Gy groups compared with the 3 Gy groups. In the 3 Gy water-supplement group, the nucleated cells and megakaryocytes were recovered in the fifth week when recovery was still not seen in the 6 Gy group. With zinc supplement, these cells were recovered in the third week. In this study, we found that zinc is beneficial to body weight in mice treated with TBI. Histologic examination of bone marrow showed better recovery of bone marrow cells in groups of mice fed with zinc. This study suggests that zinc can be used as supplements in cancer patients receiving radiotherapy to reduce radiation-induced complications.     

Pure red cell aplasia–evidence for an inhibitory action of the bone marrow adherent cell layer

Summary. Evidence is presented for an inhibitory action of the bone marrow adherent cell layer on erythropoiesis in a case of pure red cell aplasia. Cultures of the patient's bone marrow with autologous and normal plasma showed normal numbers of CFU-C, but total absence of BFU-E. There was no inhibition of BFU-E when normal bone marrow was cultured with the patient's plasma. T-cell depletion of the patient's bone marrow did not affect these results, but after removal of the adherent cells, normal numbers of BFU-E were cultured. These observations suggest that the adherent cells, which are predominantly macrophages, were responsible for the inhibitory action on erythropoiesis.     

Injectable bone

Tissue engineering applies the principles of biology and engineering to the development of functional substitutes,for example 1) cells, 2) scaffolds, 3) some cytokines, for lost tissues. The meaning of this bone regeneration is that it decreases the needed tissues and burden of patients. In this time, we applied to mesenchymal stem cells (MSCs) from the own bone marrow as cell sources. MSCs are thought to be multipotent cells that can replicate. And we also used a beta-tricalcium phosphate (beta-TCP) as a scaffold and fibrin glue as materials to regenerate a injectable bone and we injected into the subcutaneous space on the dorsum of the rat. After 8 weeks of implantation, it could be confirmed newly formed bone and fibrin glue functioned as a injectable materials without loosing the cell activity and the proliferation of MSCs. Next we applied Platelet Rich Plasma (PRP) to improve the ability of osteogenesis. PRP contains some cytokines and are expected to promote the increase of osteogenesis. The merit is not immunity rejection from autologous blood collected in the immediate preoperative period. The admixtures of PRP or fibrin glue have fluidity and gel-like consistency as the thrombin mixing. They can be injected with a syringe in tissues. We named this "Injectable Bone". According to the histological observations, the MSCs with PRP were well formed mature bone and neovascularization compared with control (defect only) after 8 weeks implantation. These results demonstrated that the mixture of MSCs and PRP were useful as injectable bone substitute and its ability of bone regeneration is almost equal to autogenous particulate cancellous bone and marrow (PCBM).     

Lymphocyte function after autologous bone marrow transplantation (BMT): a comparison with patients treated with allogeneic BMT and with chemotherapy only

T- and B-cell function after autologous bone marrow transplantation (ABMT) was assessed and compared with that found after allogeneic BMT and after chemotherapy only. In all 16 patients with acute leukaemia in remission treated with high-dose chemotherapy and single or double ABMT, T-helper numbers and function in an assay measuring PWM-induced Ig synthesis were grossly defective and closely resembled the defects observed after allo-BMT involving chemoradiotherapy (six patients). T-helper activity was more variable after chemotherapy only (eight patients), but in individual patients the defect was as great as that observed after BMT. In contrast, suppressor activity was comparably increased in all patient groups and increased numbers of Leu 15+ Dr+ Tac- suppressor T cells were consistently observed, suggesting chronic activation of suppressor T cells, the cause of which remains unknown. B-cell function was also uniformly impaired in all patients tested. It is therefore concluded that defective immune function after BMT is not due to alloimmune or radiotherapy-mediated effects. Furthermore, since many patients were studied a prolonged period after BMT and had no T cells with a thymic phenotype and no evidence of infection, it is unlikely that the defects are secondary to cellular immaturity following marrow regeneration or to superadded infection. The gross immune defects observed after various forms of BMT are likely, therefore, to be directly attributable to the chemotherapy involved.     

Bone Augmentation in Rabbit Tibia Using Microfixed Cobalt-Chromium Membranes with Whole Blood and Platelet-Rich Plasma

Background: Bone augmentation is a subject of intensive investigation in regenerative bone medicine and constitutes a clinical situation in which autogenous bone grafts or synthetic materials are used to aid new bone formation. Method: Based on a non-critical defect, Co-Cr barrier membranes were placed on six adult Fauve de Bourgogne rabbits, divided into two groups: whole blood and PRP. Three densitometric controls were performed during the experiment. The animals were euthanized at 30, 45, 60, and 110 days. The presence of newly formed bone was observed. Samples for histological studies were taken from the augmentation center. Results: External and internal bone tissue augmentation was observed in almost all cases. Significant differences between PRP- and whole blood–stimulated bone augmentation were not observed. At 60 days, bones with PRP presented higher angiogenesis, which may indicate more proliferation and cellular activity. Conclusion: PRP activates the bone regeneration process under optimized conditions by stimulation of osteoblast proliferation after six weeks, when a significant difference in cellular activity was observed. Membranes could stimulate bone augmentation at the site of placement and in the surrounding areas.     

经肝动脉自体骨髓或脐带血干细胞移植治疗失代偿期肝硬化患者的安全性和近期疗效比较

比较经肝动脉途径注入自体骨髓干细胞或脐带血干细胞治疗失代偿期肝硬化患者的安全性,以及对患者肝功能和凝血酶原活动度的近期改善作用。方法选择失代偿期肝硬化患者65例,随机分为骨髓组33例和脐血组32例;骨髓组患者经股动脉插管至肝固有动脉注入自体骨髓干细胞移植治疗,脐血组经同样途径注入脐带血干细胞治疗;治疗后8周检测血清谷丙转氨酶、总胆红素、凝血酶原活动度、白蛋白和甲胎蛋白水平变化,同时观察临床症状的改善情况及术后的不良反应。结果治疗后第3天两组患者乏力、纳差症状均有改善,两组间差异无显著性。治疗后8w,骨髓组和脐血组白蛋白水平分别由（31.0±4.6） g/L 上升至（34.6±7.1）g/L和由（34.6±7.1） g/L上升至（37.8±8.3） g/L,凝血酶原活动度上升由（48.8±13.4）%上升至（55.5±11.2）%和由（47.5±12.5）上升至（58.9±14.0）%,但两组间改善程度的差异无显著性;在治疗8w末血清谷丙转氨酶、总胆红素和甲胎蛋白在骨髓组分别为（45.6±12.3） IU/L、（28.1±13.5）μmol/L和（11.3±4.1）μg/L,在脐血组分别为（47.2±11.8） IU/L、（30.7±14.8）μmol/L和（9.8±3.5）μg/L,两组间与基础水平相比改善程度的差异也无显著性。结论自体骨髓干细胞或脐带血干细胞经肝动脉途径移植治疗失代偿期肝硬化患者的近期疗效及安全性均良好,但两种细胞治疗的改善水平无显著差异。     

自体骨髓干细胞移植治疗失代偿期肝硬化患者的疗效观察

目的探讨自体骨髓干细胞经肝动脉移植治疗失代偿期肝硬化的安全性和疗效。方法选择失代偿期肝硬化患者425例,在常规治疗的前提下抽取骨髓100～150ml,在体外分离纯化单个核细胞并制成细胞悬液,经肝动脉将其移植入肝脏,术后2、4、8、12周观察患者临床症状、体征及实验室指标。结果患者术中和术后均未出现严重的并发症和不良反应。术后第2周,213例（59%）患者双下肢浮肿明显减轻,第8周时,297例（70%）患者精神、体力、食欲恢复正常,第12周时,ALB由26.6±5.2 g/L升至37.5±4.9g/L（P〈0.05）,ALT由150.7±19.5U/L降至30.7±7.7U/L（P〈0.05）,AST由85.6±32.5U/L降至20.5±15.7U/L（P〈0.05）,TBIL由154.4±20.2μmol/L降至52.3±17.4μmol/L（P〈0.05）,PT由20.1±1.7s降至15.4±1.5s（P〈0.05）,肝脏CT值由35.32±6.68Hu升至42.32±3.26Hu（P〈0.05）。结论经肝动脉行自体骨髓干细胞移植治疗失代偿期肝硬化安全有效。     

Aspergillus pancarditis following bone marrow transplantation for chronic myelogenous leukemia

A 34-year-old man with chronic myelogenous leukemia developed hemoptysis, pain in the left side of the chest, and a systolic heart murmur eight weeks following an allogeneic bone marrow transplant. His clinical status deteriorated, and he died ten weeks after transplantation. Autopsy revealed unsuspected disseminated aspergillosis, including the unusual finding of Aspergillus pancarditis and pericarditis. Cardiac aspergillosis is a uniformly lethal disease in immunocompromised persons and must be aggressively diagnosed following early symptoms.