吻合血管同种异体骨移植后存活状况研究

目的报道吻合血管同种异体骨移植术后不同时段存活状况。方法建立吻合血管同种异体股骨干移植动物模型，在术后不同时段进行活体解剖，观察血管的通畅度．并切取移植骨进行组织学、电镜及SDH染色检测。结果术后4周对照组血管基本完全闭塞，而实验组术后血管保持通畅。组织学检查显示对照组术后移植骨骨陷窝内骨细胞缺失，哈弗斯管内血管亦消失；而实验组术后骨陷窝内始终有骨细胞充填。电镜表现为对照组术后移植骨出现骨细胞核浓缩．核碎裂，直至骨陷窝内骨细胞丢失；实验组术后骨细胞超微结构正常、对照组术后2周．骨组织SDH染色已无蓝染的骨细胞，而实验组术后可见同心圆排列蓝染的骨细胞。结论在免疫调控下，吻合血管的同种异体骨移植术后供体始终保持活力状态。     

Scapulohumeral arthrodesis for post-traumatic proximal humeral loss using vascularized fibular transplantation and allograft bone

Loss of the proximal humerus following trauma or tumor ablation results in tremendous functional impairment because of the inability to position the hand in space. Although scapulohumeral athrodesis is described in the literature, insufficient follow-up has prevented full appreciation of functional outcome and patient satisfaction. This report describes favorable restoration of upper extremity function in two patients who sustained traumatic injuries to the proximal humerus following scapulohumeral arthrodesis In both cases, vascularized fibula transplantation and allograft bone were used, and pedicled latissimus muscle transfers were required for soft-tissue closure.     

Experimental study on vascularized fresh whole-joint allograft transplantation

The vascularization of fresh whole knee and hip joints was done as part of an allograft transplant procedure involving forty rats (12 weeks old). Cyclosporin and prednisolone were injected into thirty-five recipient rats; five rats did not receive any drugs and were used as the control group. All control group rats with necrotic limbs died two to three weeks after the operation. Eleven of the twenty rats with transplanted knee joints and ten of the fifteen rats with transplanted hip joints began walking with full weight-bearing and the transplanted joints exhibited good mobility one month after the operation. Autopsy findings four, eight and sixteen weeks after the operation showed no degenerative or necrotic changes of the articular cartilage or subchondral bone of the joints. We concluded that this technique of vascularized whole-joint transplantation in rats was successful. This may be clinically applicable to reconstructive surgery after resection of diseased joints.     

Composite vascularized skin/bone graft model: a viable source for vascularized bone marrow transplantation

In this study, we introduce a new model for vascularized skin and bone marrow transplantation. Twenty-five Lewis (RT1(1)) rats were studied. Anatomic dissection studies were performed in 5 animals. In the experimental group, 10 isograft transplantations were performed between Lewis rats. Combined groin skin and femoral bone flaps were transplanted based on the femoral artery and vein. Transplants were evaluated on a daily basis. All flaps survived without problems over 100 days posttransplant. The skin component remained pink and pliable, and grew new hair. Histological examination of the femoral bone (except the femoral head) revealed active hematopoiesis with a viable compact and cancellous bone components on day 100 posttransplant. This model can be applied to tolerance induction studies across the major Histocompatibility (MHC) barrier, where bone will serve as donor of stem and progenitor cells, and the skin flap will serve as a monitor of graft rejection.     

吻合血管的同种异体骨移植后移植物再血管化的研究

目的 探讨吻合血管的同种异体骨移植后移植物在宿主体内再血管化过程及其规律.方法 建立吻合血管的同种异体骨移植修复兔股骨大段缺损的模型,实验分成两组,实验组(吻合旋股外血管股骨中上段等位异体骨移植组)和对照组(未吻合血管的股骨中上段等位异体骨移植组).对移植物骨膜、皮质骨及骨髓进行组织学切片,同时对微血管内皮细胞进行免疫组化染色,血管计数后.进行统计学分析,制备墨汁灌注标本,观察各部位微血管再生情况.结果 实验组即可见骨膜、皮质骨和骨髓同时出现再血管化现象,而对照组则由浅至深逐渐出现再血管化现象,在术后2、4、8、16周实验组微血管密度分别为10.0±1.8、15.8±1.5、13.8±1.5、13.8±1.5,对照组分别为2.8±0.8、6.0±0.9、5.5±1.0、6.0±1.1,实验组明显高于对照组;墨染显示实验组墨染范围及程度明显强于对照组.结论 吻合血管的异体骨移植能加快移植物再血管化的过程.     

The protective effect of hepatic dysfunction on vascularized allograft survival

Recent studies have demonstrated that hepatic dysfunction, induced by experimental biliary ligation (EBL), impairs lymphocytic responsiveness to PHA stimulation in vitro and to cellular antigens in vivo. This suppression appears to be selective for T-cell mechanisms while B-cell-mediated functions remain intact. The purpose of this study was to determine whether coexisting hepatic insufficiency could exert a protective effect on vascularized or nonvascularized allograft survival in the transplanted recipient. Female Wistar-Furth (Rtlw) 225 g rats were assigned randomly to three groups: EBL, sham operation (Sham) and normal control (NC). Fourteen days following operation animals received heterotopic cardiac or skin allografts from Buffalo (Rtlb) donors. Cardiac and skin graft survival was determined daily, rejection was confirmed histologically, and technical failures were omitted from analysis. Allograft survival was expressed as median survival time +/- SEM. Serum total bilirubin (mean +/- SEM) was significantly elevated at Day 14 in EBL animals compared to Sham and NC groups (15.1 +/- 1.0 vs 0.1 +/- 0 and 0.2 +/- 0.1 mg/dl, respectively, P less than 0.01). Median cardiac allograft survival time by Probit was 10.6 +/- 2.6 vs 5.6 +/- 0.7 and 6.0 +/- 0.9 days, respectively (P less than 0.03). Skin graft survival (mean and range) was similar in all groups. These results demonstrate that EBL in the rat suppresses T-cell function and significantly prolongs vascularized allograft survival, but not skin allograft survival across the Rtl histocompatibility barrier. The mechanism whereby coexisting hepatic dysfunction exerts a protective effect on vascularized allograft survival warrants further investigation.     

一种大鼠带血管的骨髓移植模型

目的 构建一种简单可靠的带血管的骨髓移植模型.方法 将供体带血管蒂的股骨通过显微血管吻合的方式移植到受体腹股沟区,20只Lewis近交系大鼠随机分3组:同基因移植组Lewis→Lewis;排斥组Lewis→BN;免疫抑制组Lewis→BN,术后给予环孢素A.通过大体和病理学检查观察各组移植物存活情况,流式细胞计数监测外周血嵌合水平.结果 术后30 d,同基因移植组股骨存活良好,骨髓苏木素-伊红(HE)染色与正常骨髓无异,而排斥组第7天即发生显著的排斥反应,骨髓细胞明显减少、坏死,外周血嵌合水平几乎为0;和同系移植组一样,免疫抑制组术后30 d移植物存活良好,术后1、2、3、4周外周血中可检测到供体特异性嵌合,嵌合水平分别为(4.7±2.0)％、(2.2±1.2)％、(1.8±0.9)％、(1.5±0.3)％.结论 带血管蒂的股骨移植是一种简便可靠的骨髓移植模型,可作为一种新的骨髓移植方法诱导免疫耐受.     

吻合血管同种异体骨移植的解剖学研究

目的 为选用吻合血管同种异体骨移植的供区提供解剖学依据。方法 选用经动脉灌注红色乳胶的成人上、下肢标本各４０侧,着重对肱骨、桡骨、股骨和胫骨的滋养血管、进行观测;另选上述供骨区成人干燥骨各５０根,作滋养孔（或称管）观察。结果 肱骨上端有４～１２个滋养也,旋肱前、后动脉是其滋养血管主要来源;肱骨干滋养孔多位于肱骨中段前内侧面,滋养动脉多发自肱动脉;桡骨下端滋养血管主要由骨间前动脉及桡动脉供应;股骨干滋养１～３个,滋养动脉来自股深动脉的穿支;胫骨干滋养孔恒定位于胫骨中、上１／３段后面,滋养血管来自胫后动脉分支。结论 根据长骨大段骨缺损,选用在外形上与受区相匹配的供骨,上述血管长度、口径均适合显微外科的吻接要求。     

Bilateral vascularized femoral bone transplant: a new model of vascularized bone marrow transplantation in rats, part I

We present a new model of vascularized bone marrow transplantation-bilateral vascularized femoral bone (BVFB) isograft transplant based on abdominal aorta and inferior vena cava. A total of 7 BVFB isograft transplants were performed between Lewis (RT1) rats. In the donor, both femoral bones were harvested based on the abdominal aorta and inferior vena cava. In the recipient, the harvested isograft transplants were transferred into the inguinal region (in 3 animals) and into the abdominal cavity (in 4 animals). The mean operation time was 3 hours and 35 minutes. The mean warm ischemic time was 35 minutes. The vascular pedicles of the transplants that were transferred into the inguinal region were thrombosed at day 7 posttransplantation. The vascular pedicles of transplants into the abdominal cavity were patent and the bones were viable during the follow-up period of 63 days posttransplant. We have confirmed the feasibility of BVFB transplantation based on abdominal aorta and inferior vena cava.     

Microsurgical reconstruction with vascularized fibula and massive bone allograft for bone tumors

European Journal of Orthopaedic Surgery & Traumatology - Combining massive bone allograft and vascularized fibula in intercalary reconstruction following resection of bone tumors represents a...     

A novel rat full‐thickness hemi‐abdominal wall/hindlimb osteomyocutaneous combined flap: influence of allograft mass and vascularized bone marrow content on vascularized composite allograft survival

Vascularized bone marrow transplantation (VBMT) appears to promote tolerance for vascularized composite allotransplantation (VCA). However, it is unclear whether VBMT is critical for tolerance induction and, if so, whether there is a finite amount of VCA that VBMT can support. We investigated this with a novel VCA combined flap model incorporating full‐thickness hemiabdominal wall and hindlimb osteomyocutaneous (HAW/HLOMC) flaps. Effects of allograft mass (AM) and VBMT on VCA outcome were studied by comparing HAW/HLOMC VCAs with fully MHC‐mismatched BN donors and Lewis recipients. Control groups did not receive treatments following transplantation. Treatment groups received a short course of cyclosporine A (CsA), antilymphocyte serum, and three doses of adipocyte‐derived stem cells (POD 1, 8, and 15). The results showed that all flaps in control allogeneic groups rejected soon after VCAs. Treatment significantly prolonged allograft survival. Three of eight recipients in HLOMC treatment group had allografts survive long‐term and developed donor‐specific tolerance. Significantly higher peripheral chimerism was observed in HLOMC than other groups. It is concluded that the relative amount of AM to VBMT is a critical factor influencing long‐term allograft survival. Accordingly, VBMT content compared with VCA mass may be an important consideration for VCA in humans.     

Promotion of pancreatic islet allograft survival by intrathymic transplantation of bone marrow.

An important goal in the treatment of insulin-dependent diabetes by pancreatic islet transplantation is the development of strategies that allow permanent survival of islet allografts without continuous host immunosuppression. In this study, we demonstrate that inoculation of allogeneic bone marrow into the thymus of adult rats treated with a single dose of anti-lymphocyte serum induces an unresponsive state that permits survival of subsequent pancreatic islet allografts transplanted to an extrathymic site. This effect is donor specific, cannot be reproduced by systemic administration of bone marrow, and is associated with persistence of chimeric cells in the thymus of the recipient. In addition, lymph node cells from long-term recipients of intrathymic bone marrow display markedly reduced proliferative responses to donor alloantigens in mixed lymphocyte culture. Interaction of maturing thymocytes with foreign alloantigens may produce the unresponsiveness. This model offers a potential approach for establishing donor-specific allograft acceptance in adult recipients.     

Half-joint allograft transplantation in human bone tumours

Summary Seven patients with malignant or aggressive bone tumours involving the end of a long bone have been treated by wide resection and an allograft providing a half-joint transplantation. The observation period is from 2 to 9 years (average 5 years). The allografts were preserved at –70 °C for a period ranging from 1 week to more than 2 years. Computed tomography was very useful in the preoperative determination of the grade of infiltration tumour growth in bone. The host's own ligaments were used in the reconstruction. Immunosuppression was not used.The fate and metabolism of the allografts were followed by clinical examination, conventional radiographs, angiography, bone scintigraphy, cell-mediated immunity tests, biopsies for histology and by determining the excretion of the urinary products of bone metabolism (duHYPro, dUCa, dUPi). Positive isotope labelling was demonstrated as early as 2 weeks after transplantation and this increased and stabilised at about 6 months. Simultaneously, the collagen/bone matrix turnover (HYPro excretion) showed a slightly raised level for up to 3–6 months, stabilising thereafter indicating a moderate, but prolonged, regenerative activity of the graft. On the basis of these and the histological studies, weight-bearing was gradually started at 6 months, which is earlier than described in previous reports. The patients had knee flexion up to 90° and walked well. Studies on whole-blood cell-mediated immunity showed only slight non-significant changes. The results indicate that the grafts incorporated well with early, and subsequently prolonged, evidence of metabolic activity.

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Résumé 7 malades atteints de tumeurs osseuses malignes ou aggressives situées au niveau de l'extrémité d'un os long ont été traités par résection large et allogreffe, réalisant la transplantation d'une hémi-articulation. La durée d'observation est de 2 à 9 ans (en moyenne 5 ans). Les allogreffes ont été conservées à –70° pendant une période allant d'une semaine à plus de 2 ans. La tomographie computerisée a été très utile pour la détermination pré-opératoire du degré d'infiltration de la croissance tumorale osseuse. Les ligaments du sujet receveur ont été utilisés pour la reconstruction. On n'a pas eu recours à l'immuno-suppression.L'évolution et le métabolisme des allogreffes ont été suivis grâce à l'examen clinique, aux radiographies standard, à l'angiographie, à la scintigraphie osseuse, aux tests d'immunité cellulaire, aux biopsies pour examen histologique et en déterminant l'excrétion urinaire des produits du métabolisme osseux (hydroxyproline) calcium et phosphates). Des réactions isotopiques positives se sont manifestées dès la deuxième semaine après la transplantation, puis ont augmenté et se sont stabilisées vers le 6ème mois. Simultanément, le »turnover« collagène/matrice osseuse (excrétion de l'hydroxyproline) a montré une légère augmentation pendant 3 à 6 mois, indiquant une activité de régénération de la greffe, modérée mais prolongée. Du fait de ces constatations et des résultats des examens histologiques, la reprise de l'appui a été progressivement autorisée à partir du 6ème mois, c'est à dire plus tôt qu'il n'est dit dans les publications plus anciennes. Les malades ont récupéré une flexion du genou atteignant 90° et marchent correctement. L'étude de l'immunité cellulaire sanguine n'a montré que de minimes changements, non significatifs. Ces résultats prouvent que les greffes s'incorporent bien, avec des signes d'activité métabolique précoce et longtemps prolongée.

     

Chimerism after vascularized limb versus bone marrow transplantation

This study used quantitative PCR in the murine model to compare the ability of a limb allograft vs. a comparable dose of marrow suspension to induce chimerism. Female C57Bl/6 mice received a vascularized hindlimb allograft, a comparable dose of 5 x 10 (6) donor bone marrow cells, or a standard dose (20 x 10 (6)) of marrow suspension from male Balb/c donors. All recipients were treated with a regimen based on CD40 costimulation blockade and T cell depletion. Y chromosome-specific quantitative PCR was used to measure chimerism. Most recipients of limb allografts demonstrated low levels of chimerism after 1 week (3/4) and 1 month (3/4). Most recipients of 5 x 10 (6) marrow cells had low levels of chimerism at 1 week (4/6) and only 1/5 after 1 month. All recipients of 20 x 10 (6) cells except one demonstrated either low or high levels of chimerism after 1 week (5/5) and 1 month (5/6). The marrow component of a limb allograft is thus more effective at inducing microchimerism compared to a comparable dose of bone marrow suspension.     

山羊血管化骨髓移植的解剖学基础研究

目的通过对山羊股骨、髂骨、胸骨、肋骨4个较为常见的血管化骨髓移植供区部位相关结构的测量观察,从移植手术的角度,分析其解剖特点,为下一步建立山羊血管化骨髓移植模型提供一定的解剖依据。方法取6只（共12侧）体质量相当的山羊做活体解剖研究。观测4个供区血液供应情况,并对比4种移植物体质量、体积、骨髓干细胞密度等因素。结果髂骨可见主要滋养血管为髂内动脉及其分支,另外紧贴骨面的臀部肌肉也会有细小动脉进入骨膜,髂内血管为较理想的血管蒂;股骨的主要滋养动脉有3条：股深动脉、旋股外侧动脉及膝降动脉,取髂外血管为血管蒂;双侧胸廓内动脉及伴行静脉为胸骨的主要滋养血管,双侧胸廓内动静脉的近端可作为游离血管蒂;肋骨以第7肋为例,其肋间后动脉及其伴行静脉为主要滋养血管,可以取到一定长度的游离血管蒂。结论山羊的髂骨及股骨是较为理想的血管化骨髓移植术供区,其游离血管蒂长度及其动静脉外径均可满足异体移植术的需求。     

A new method of bone marrow transplantation leads to extension of skin allograft survival

Tolerance induction through allogeneic bone marrow transplantation is an alternative method to chronic immunosuppression in maintaining long-term allograft survival. In this article, we introduce a new method of bone marrow allotransplantation, which preserves its natural microenvironment and does not require marrow processing or recipient conditioning. A total of 43 skin graft transplantations were performed in nine experimental groups between isogeneic [Lewis to Lewis (LEW, RT1(1))] and allogeneic [Lewis x Brown Norway (LBN --> F1, RT1(1+n)) to Lewis] rats under 35-day protocol of alphabeta T-cell receptor (TCR) monoclonal antibody (mAb) and cyclosporine (CsA) protocol. Monotherapies combined with "crude" bone marrow transplantation resulted in extended survival up to 21 days under CsA and up to 10 days under alphabeta-TCR mAb protocol. The use of combined protocol of alphabeta-TCRmAb/CsA with crude bone marrow transplantation resulted in the extension of skin allograft survival up to 65 days (P < .05). This new simple method of "crude" bone marrow allotransplantation without recipient conditioning is a promising, minimally invasive technique with a potential for direct clinical application.