深低温冷冻骨的形态学及免疫组织化学检测

背景：深低温冷冻技术能够降低同种异体骨的免疫原性,对修复骨缺损具有重要意义。 目的：观察深低温冷冻对骨材料中骨形态发生蛋白2表达的影响。 方法：18只新西兰兔随机分为3组,深低温冷冻3个月组,深低温冷冻6个月组,新鲜骨对照组,取右侧胫骨中上1/3,分别于-80 ℃保存保存3个月,6个月及不保存,进行苏木精-伊红染色和骨形态发生蛋白2免疫组织化学染色。 结果与结论：深低温冷冻后骨材料形态学,与新鲜骨差异不大;深低温冷冻3个月及6个月骨组织中骨形态发生蛋白2均有表达,但较新鲜骨降低。     

大段异体骨支撑复合自体髂骨移植修复肱骨良性骨缺损

大段骨缺损的临床处理仍是一个棘手的问题。随着同种异体骨免疫学研究的进展和现代冷藏技术的提高,深低温冷冻保存大段同种异体骨日益广泛地应用于修复各种原因引起的大段骨缺损。我们自1996年以来,采用深低温冷冻大段同种异体骨支撑复合自体髂骨移植修复肱骨良性大段骨缺损3例,效果满意。     

同种异体冻干小骨结合解剖型钢板治疗跟骨关节内粉碎性骨折

目的观察和分析同种异体冻干小骨结合解剖型钢板治疗跟骨关节内粉碎性骨折的治疗效果。方法对22例25侧跟骨关节内粉碎性骨折,经跟骨外侧入路在C形臂X线机监控下行解剖型钢板内固定、同种异体冻干小骨植骨术。结果本组22例平均随访22.3个月,按Maryland足部评分系统评估术后功能,优7侧,良11侧,可4侧,优良率81.9%。结论同种异体冻干小骨结合解剖型钢板治疗跟骨关节内粉碎性骨折可以取得满意的疗效。     

头状骨近端关节面的功能解剖

目的：研究近侧列腕骨切除术后头状骨近端关节面的解剖及其和桡尺骨远端关节面的匹配。方法：12例福尔马林固定的尸体手标本，行近侧列腕骨切除术后观测头状骨近端关节面的解剖学特点和活动；纵、横行剖开远侧列腕骨和桡尺远端，描模正常桡骨一月骨关节面和桡尺骨一头状骨关节面的匹配曲线。结果：头状骨近端关节面有桡、尺、背、掌4个子关节面组成，和桡尺骨远端在矢状面上匹配差，冠状面上好，尺侧部分匹配好，桡侧部分匹配差。结论：头状骨近端和桡尺骨远端形成的关节面其解剖学上明显不同千诈常腕美节．     

环氧乙烷处理同种异体皮质骨板移植组织学研究

目的研究环氧乙烷处理同种异体皮质骨板移植组织学表现.方法分别移植经过零下70℃深低温冷冻4周、零下70℃深低温冷冻4周+48℃环氧乙烷灭菌处理的山羊同种异体皮质骨板,对移植骨板以及周围软组织进行组织学检查.结果术后3～6周深低温冷冻组移植骨板周围软组织少量炎性细胞浸润,环氧乙烷+深低温冷冻组移植骨板周围软组织较多炎性细胞浸润,但炎细胞浸润评分差别无统计学意义(P＞0.05);术后6～24周两组移植骨板周围软组织炎性细胞浸润进一步减轻.深低温冷冻组和环氧乙烷+深低温冷冻组术后3周部分移植骨板原哈佛式管扩大,其内有少量间充质组织和新生血管;6周间充质细胞分化为破骨细胞和成骨细胞,破骨细胞使移植骨板哈佛式管进一步扩大以及出现骨吸收隙窝,在哈佛式管和骨吸收隙窝边缘出现成骨细胞贴附和新骨形成;12周移植骨板骨吸收隙窝缩小,成骨细胞和新骨形成增多,部分新骨改建形成板层骨;24周移植骨板基本完成吸收替代和改建塑形.两组移植骨板内无炎细胞浸润,组织学表现相似.结论环氧乙烷灭菌同种异体皮质骨板移植周围软组织出现炎细胞浸润,不影响移植骨板爬行替代和内部改建塑形.     

环氧乙烷处理同种异体皮质骨板移植组织学研究

目的 研究环氧乙烷处理同种异体皮质骨板移植组织学表现。方法 分别移植经过零下70℃深低温冷冻4周、零下70℃深低温冷冻4周+48℃环氧乙烷灭菌处理的山羊同种异体皮质骨板,对移植骨板以及周围软组织进行组织学检查。结果 术后3～6周深低温冷冻组移植骨板周围软组织少量炎性细胞浸润,环氧乙烷+深低温冷冻组移植骨板周围软组织较多炎性细胞浸润,但炎细胞浸润评分差别无统计学意义(P>0.05);术后6～24周两组移植骨板周围软组织炎性细胞浸润进一步减轻。深低温冷冻组和环氧乙烷+深低温冷冻组术后3周部分移植骨板原哈佛式管扩大,其内有少量间充质组织和新生血管;6周间充质细胞分化为破骨细胞和成骨细胞,破骨细胞使移植骨板哈佛式管进一步扩大以及出现骨吸收隙窝,在哈佛式管和骨吸收隙窝边缘出现成骨细胞贴附和新骨形成;12周移植骨板骨吸收隙窝缩小,成骨细胞和新骨形成增多,部分新骨改建形成板层骨:24周移植骨板基本完成吸收替代和改建塑形。两组移植骨板内无炎细胞浸润,组织学表现相似。结论 环氧乙烷灭菌同种异体皮质骨板移植周围软组织出现炎细胞浸润,不影响移植骨板爬行替代和内部改建塑形。     

同种异体肌腱解剖重建修复慢性踝关节不稳

背景：同种异体肌腱解剖重建应用于踝关节修复重建的报道目前较少。 目的：分析运用深低温冷冻保存同种异体肌腱解剖重建修复慢性踝关节不稳的临床疗效。 方法：运用深低温冷冻保存同种异体肌腱解剖重建修复慢性踝关节不稳26例,其中跟腓韧带和距腓前韧带同时损伤或松弛18例,距腓前韧带单独损伤或松弛8例。采用美国足踝外科协定(AOFAS)评分及Good评级评估踝关节功能,并进行患侧与健侧踝关节背伸、跖屈活动度、后足活动度比较。 结果与结论：所有患者治疗后均获得随访,随访时间9-24个月,平均15个月。所有患者均未出现复发性踝关节外侧不稳,美国足踝外科协定(AOFAS)评分：同时修复跟腓韧带和距腓前韧带组,治疗前(48.4±3.7)分,治疗后(88.2±3.8)分,治疗后较治疗前平均提高39.8分;单独修复距腓前韧带组治疗前(50.0±6.4)分,治疗后(89.5±3.4)分,治疗后较治疗前平均提高39.5分。Good评级优 19例,良 6例,可 1例,优良率 96%。患者均无严重并发症。结果提示应用深低温冷冻保存同种异体肌腱解剖重建踝关节外侧韧带治疗踝关节慢性外侧不稳,增大了腱骨接触面积,增加了骨腱愈合的概率,增强了踝关节的稳定性,其远期疗效仍待进一步评估。 中国组织工程研究杂志出版内容重点：肾移植;肝移植;移植;心脏移植;组织移植;皮肤移植;皮瓣移植;血管移植;器官移植;组织工程全文链接：     

血管冷冻保护剂的临床应用

背景：冷冻保存的同种异体血管在血管移植修复中蕴藏着广泛的临床应用前景。深低温冻存是实现同种异体血管长期保存的常用方法。在低温保存过程中,有效的冷冻保护剂对于防止血管组织低温损伤和保持细胞活力至关重要,但对于最佳保护剂的选择并未达成共识。目的：总结近年来各大血管组织库中使用的冷冻保护剂,比较其临床应用效果,以期为将来深低温冻存同种异体血管的应用提供参考。方法：由第一作者检索中国知网、万方、PubMed和Google Scholar数据库2015-2022年发表的相关文献,中文检索词为“同种异体血管,动脉,静脉,深低温冻存,冷冻保护剂,冷冻损伤,血管移植”,英文检索关键词为“cryopreservation,blood vessels,vascular allografts,arteries,veins,cryoinjury,cryoprotectants,cryoprotective agents”,检索国内外有关应用于临床的血管冷冻保护剂及其近远期结果的相关文献,最终选取54篇文献进行综述。结果与结论：(1)在血管的深低温冻存研究中,常使用包含渗透性和非渗透性的冷冻保护剂组合,通过程序...     

骨钉和脱钙骨在骨创伤和骨缺损中的应用

目的：探讨同种异体冻干骨钉固定关节内骨折和脱钙骨移植治疗骨缺损的实用价值。方法：应用骨钉治疗关节内骨折７例，其中股骨头骨折４例，距骨骨折１例，内踝骨折２例；应用脱钙骨皮质骨条、松质骨粒移植治疗四肢骨折骨不连６例、骨良性病变切除术后骨缺损６例。结果：术后随访３～１２个月，骨钉固定者８个月后骨钉与宿主骨融合，脱钙骨皮质骨条和松质骨粒植骨者２～６个月愈合，无一例发生免疫排斥反应，近期效果满意。结论：认为骨钉为临床治疗关节内骨折提供了一种新的内固定材料，避免再次手术取内固定物，脱钙骨皮质骨条或骨粒适宜于骨不连和骨缺损的填充植骨治疗。     

同种异体骨移植后T淋巴细胞的分布与变化

背景：研究表明体液免疫在同种异体骨移植免疫排斥反应中不起主要作用,目前一般认为细胞免疫在排斥反应中起主导作用。 目的：概述同种异体骨移植后T淋巴细胞的分布与变化。 方法：采用电子检索的方式,在万方数据库(http://www.wanfangdata.com.cn/)、Pubmed数据库(http:// www.ncbi.nlm.nih.gov.pubmed/)中检索1990-01/2011-12关于同种异体骨移植后T淋巴细胞的分布与变化的文章,关键词为“骨移植,同种异体,T淋巴细胞”。排除重复研究、普通综述或Meta分析类文章,筛选纳入31篇文献进行评价。 结果与结论：同种异体骨移植后,体内抑制T淋巴细胞被激活,对免疫排斥反应起负反馈调节作用,T淋巴细胞发生显著变化;但深低温冷冻、冷冻干燥、γ射线辐照、复合免疫抑制剂等方法可以有效降低其免疫原性及疾病传播的风险。大量研究认为,同种异体骨具有骨组织的完整性结构、机械稳定性、生物学活性、骨诱导能力和极低的免疫性以及与宿主骨有较强的愈合能力等优点,植入体内后通过全方位再血管化、新骨形成、异体骨与宿主骨连接而实现生物学骨掺入过程。提示同种异体骨是一种可行的移植材料,可作为目前修复骨缺损的重要手段之一。      

Incorporation of perforated and demineralized cortical bone allografts. Part II: A mechanical and histologic evaluation

Laser perforated and partially demineralized cortical bone allografts were orthotopically transplanted into sheep tibiae. This paper reports results of the mechanical testing of the transplanted bones, which was done at nine months postoperatively. Animals were divided into three groups of eight according to the type of cortical allograft used: group 1, no treatment; group 2, laser hole grid; and group 3, laser hole grid and partial demineralization. Thus, changes in flexural rigidity of 24 transplanted whole tibiae were investigated. Starting in the anterior direction at the tibial tuberosity, the flexural rigidity was determined using a nondestructive 4-point bending test. The elliptical distribution of the flexural rigidity was compared to the untreated contralateral control bone of each animal. Mechanical parameters were defined as percentage rates for comparative analysis between groups. Flexural rigidity measurement showed that bones transplanted with untreated allografts were stiffer than contralateral control bones. Partial demineralization of allografts reduced the flexural rigidity of transplanted bones below the level of contralateral control bones. Flexural rigidities of test bones transplanted with laser perforated and partially demineralized allografts were higher than those seen in bones transplanted with partially demineralized allografts. These results were corroborated by the histologic analysis which showed that untreated allografts, although surrounded by a periosteal bone cuff that effectively increased their outer diameter. In contrast, excessive bone resorption was observed in partially demineralized allografts. Laser-perforated and partially demineralized allografts showed histologic evidence of complete incorporation into the host bone. Based on this mechanical evaluation, it was concluded that processing of cortical bone allografts by the combination of perforation and partial demineralization resulted in improved mechanical strength of the transplanted bones as compared to processing by partial demineralization alone.     

Incorporation of perforated and demineralized cortical bone allografts. Part I: radiographic and histologic evaluation

Massive cortical bone allografts have been found to incorporate slowly into host bone. They are subject to complications such as nonunion, fatigue fracture and infection. In an attempt to improve osteoinduction in cortical bone allografts, laser perforated and partially demineralized cortical bone allografts were orthotopically transplanted into the sheep tibia. In this model, mid-shaft tibial bone allografts from out-bred sheep donor animals were prepared by partial demineralization and drilling of 0.33-mm diameter holes with a pulsed, 2.94-microm wavelength Erbium:Yttrium-Aluminum-Garnet laser. Recipient animals of the same out-bred strain were divided into three groups of eight according to the type of cortical allograft used: group 1, fresh-frozen, no treatment; group 2, laser hole grid; and group 3, laser hole grid and partial demineralization. Plain films were taken in two standard views at monthly intervals. Incorporation was evaluated at nine months postoperatively. Longitudinal radiographic data was correlated to a histologic and morphometric evaluation of each bone graft. Computer tomography was used for the latter analysis. Results showed that untreated allografts, although surrounded by a periosteal bone cuff, were poorly incorporated. Partial demineralization lead to excessive resorption of allografts, but little new bone formation. Laser perforation and partial demineralization induced complete incorporation of allografts into the host bone. Based on the results of the radiographic, histologic and morphometric evaluation, the development of laser-perforated and partially demineralized bone allografts was proposed for clinical use.     

腓骨骨膜瓣移植在儿童四肢骨肿瘤瘤段骨射线灭活回植术中的应用观察

目的 探讨腓骨骨膜瓣移植覆盖灭活瘤段骨断端在儿童四肢骨肿瘤射线灭活再植术中应用的可行性和临床效果。方法 回顾性分析2016年6月—2017年6月郑州大学附属肿瘤医院骨与软组织科8例原发四肢恶性或侵袭性骨肿瘤患儿临床资料,男4例、女4例,年龄9~17(13.6±2.5)岁;骨肉瘤5例,尤文肉瘤、软骨肉瘤、反复复发的侵袭性朗格汉斯组织细胞增生症各1例。均采用瘤段骨扩大切除+术中灭活自体瘤段骨回植术治疗,回植的瘤段骨断端均应用带蒂或游离腓骨骨膜瓣覆盖。肿瘤位于胫骨中上段5例,应用同侧带腓血管蒂腓骨膜瓣逆行转位覆盖瘤段骨远侧断端;肿瘤位于股骨中下段2例、肱骨中段1例,应用游离腓血管蒂腓骨膜瓣移植覆盖近侧断端。瘤段骨为单纯骨干5例,包含关节面3例,长度13~26(18.1±3.9)cm。灭活方法均为术中离体射线灭活。术后定期随访,拍摄X线、CT观察记录瘤段骨体内愈合情况,并观察患肢长度变化。结果 本组8例患儿术后随访12~24(18.8±3.5)个月。8例患儿共有13处断端均获得骨愈合,腓骨膜覆盖的断端平均愈合时间6~10(7.3±1.3)个月,且骨膜覆盖区域可观察到成骨现象。末次随访时,5例骨干灭活和1例骨关节灭活的患儿未发生双侧肢体不等长,2例骨关节灭活患儿患侧下肢长度较对侧短缩1.5 cm和2.0 cm。结论 在儿童四肢骨肿瘤手术治疗中,应用腓骨骨膜瓣移植覆盖灭活瘤段骨断端的方法可行,能够有效促进瘤段骨的愈合和防止骨不愈合、骨不连发生,且操作简便,并发症较少,推荐临床开展应用。     

颈椎椎间隙测量及同种异体骨颈椎融合骨垫的研制

目的　根据MRI测量正常国人颈椎椎间隙数据,研制符合国人解剖学特征的同种异体骨颈椎椎间融合骨垫。 方法　通过MRI测量100例C2,3至C6,7每个椎间隙的前缘、中点、后缘高度值及横径宽度值、矢状径长度值,其中男性有50人(20~55岁,平均年龄38.7岁),女性有50人(20~55岁,平均年龄36.1岁)。并根据该数据设计同种异体骨颈椎椎间融合骨垫。 结果　同节段颈椎椎间隙正中矢状面后缘高度<前缘高度<中间高度（P<0.05）,椎间隙横径大于矢状径（P<0.05）,同节段同部位的测量值,男性组大于女性组（P<0.05）;同种异体骨颈椎椎间融合骨垫设计为前缘高度为6.0、7.0、8.0、9.0 mm 4个型号的楔状,其表面具有倒齿结构,中央有脱钙骨基质（DBM）置入孔,用于置入DBM粉末。 结论　依据MRI测量的颈椎椎间隙数据设计的同种异体骨颈椎融合骨垫符合国人解剖学要求。     

保留BMP的微脱矿骨条在脊柱融合术中的应用研究

目的探讨保留BMP的微脱矿骨条在脊柱融合术中的临床疗效。方法自2005年6月~2010年12月应用保留BMP的微脱矿骨条行脊柱融合术26例﹙实验组﹚,其中颈椎8例,胸腰椎18例,术后对所有患者行切口愈合情况、异物反应及脊柱融合情况进行观察,对照组为同期应用普通同种异体骨条行脊柱后路融合术26例。结果实验组与对照组伤口均一期愈合,未发生感染。颈椎实验组、对照组均现脊柱融合;胸腰椎实验组坚强融合16例,可能融合2例,对照组坚强融合13例,可能融合5例。随访6~24月,平均15.2月,内固定系统无松动及断钉断棒。结论保留BMP的微脱矿骨条应用在脊柱融合术中,脊柱融合可,无排异、感染等并发症,临床疗效满意。     

初次全膝置换术中非包容性骨缺损的处理

目的：通过对初次全膝关节置换术中非包容型骨缺损的重建,介绍自体或异体骨移植重建的方法和体会。方法：对16例（20膝）非包容性骨缺损的膝关节行全膝关节置换及自体或同种异体移植术。其中13例（17膝）为自体股骨髁或胫骨平台修整后移植,2例为股骨远端冻干异体半关节骨移植,1例为胫骨近端冻干异体半关节骨移植;螺钉固定4膝,钢板固定16膝。结果：术后随访1—8a,平均3．7a。所有患者术后均弃拐行走,术后疼痛完全消失,下肢畸形及关节功能明显改善,HSS膝关节评分由术前17．2（11～29分）提高到术后85．2（76～94）分。1例股骨远端异体半关节移植患者术后1a出现异体骨骨折。其他患者经X线复查时未见假体松动和感染征象,无明显的植骨块移位、骨折、骨吸收、不愈合。结论：在非包容型骨缺损的初次全膝关节置换术中,采用自体或异体骨移植重建可保持假体的长期稳定性,同时为较年轻的患者将来翻修而保存骨量。     

自研全光纤光学相干计算机断层摄影术灵敏检测早期牙釉质龋

目的探索新型生物组织高分辨率成像技术——光学相干计算机断层摄影术（OCT）用于龋病早期诊断。方法选用化学酸蚀法在离体人恒牙釉质光滑面制备0、12、24、48、72、96、120h不同时间的早期脱矿人工龋模型,采用自行研发的口腔全光纤OCT（AF-OCT）进行离体人恒牙早期牙釉质人工龋的体外检测．并与同一牙齿样本的显微数码摄影、扫描电子显微镜和偏振光显微镜的检测结果进行比较．检验AF—OCT体外检测早期龋变的效能。结果AF—OCT可早期准确检测到脱矿12h以内的牙釉质表面组织学水平的改变．开窗区釉质表面脱矿深度与脱矿时间成正比。与3种常用体内外检测手段相比．AF—OCT可早于显微数码摄影观察到牙釉质表面的形貌改变．与扫描电子显微镜和偏振光显微镜检测结果高度相关。结论自研AF—OCT可早期、精确、无创地检测出脱矿12h以内的早期牙釉质表面人工龋变,并可实现量化测定和分析．获得的龋变深度量化测量数据与偏振光显微镜下组织学测量结果高度相关。     

A study of the humoral immune response to massive osteoarticular allografts in sheep

The production of antibodies against the major transplantation antigens has been investigated in sheep receiving osteoarticular allografts consisting of the distal portion of the femur. Grafts were used fresh, frozen or freeze-dried. It was shown that grafts of fresh or frozen tissues elicited a rapid humoral immune response with broad spectrum of anti-leucocyte antibodies in all recipients. In contrast, 36% of animals receiving allografts of freeze-dried bone never developed antibodies. In those recipients that did develop antibodies the time course of the response was similar to that in the recipients of fresh or frozen grafts; the spectrum of antibodies achieved was, however, significantly narrower in the freeze-dried graft recipients. These patterns are compared with those previously obtained in recipients of smaller osteoarticular grafts. The possible significance of these findings is discussed.     

The use of unicondylar osteoarticular allografts in reconstructions around the knee

Unicondylar osteoarticular allografts (UOA) of the knee are mainly used after bone tumour resections for benign aggressive tumours or small malignant tumours with clearly defined margins. They are also used less often in large posttraumatic condylar defects. Between 1989 and 2004, 12 deep-frozen UOA reconstructions (in 11 patients) were performed at our Institute. The diagnosis was chondrosarcoma in four cases, giant cell tumour in three, osteosarcoma in three, posttraumatic defect in one, and one failed UOA. The involved site was the medial femoral condyle in six cases, the lateral femoral condyle in three, the medial side of the tibial plateau in two, and the lateral in one case. One allograft was removed after 29 months because of an intra-articular displaced fracture, and substituted with a new UOA. One patient died of metastatic disease at 24 months. We report the functional and radiographical outcome of the remaining 10 UOAs with a minimum follow-up of 4 years (average 11 years). Two of the 10 patients had excellent results, five were good and three were fair. Radiographically, five patients had "mild" and five had "severe" degenerative changes. One patient with severe degenerative changes had pain and stiffness, therefore the UOA was converted into a prosthesis allograft composite, using a conventional total knee prosthesis. In selected cases of distal femoral and proximal tibial tumours, UOA reconstructions give good functional outcomes with relatively few major complications.     

支撑钢板在股骨远端骨折中的应用

目的 探讨外侧支撑钢板治疗股骨远端骨折的方法及疗效.方法 2004年2月～2006年12月共收治23例股骨远端骨折患者.全部采用切开复位外侧支撑钢板内固定加一期自身骼骨植骨治疗.结果 23例病人切口一期愈合,骨折复位良好,无切口感染、无内固定松动及断裂、无畸形愈合及延迟愈合或者不愈合;全部患者经1～2年(平均1.5年)的随访,骨折均骨性愈合,膝关节功能恢复良好.结论 对于股骨远端AO分型中的A2～C3型骨折,切开复位支撑钢板内固定、一期植骨是一种较好的治疗方法,能减少、预防各种并发症,提高患者生存质量.