组织工程化骺板软骨修复兔部分骺板损伤的实验研究

目的 利用组织工程骺板软骨修复兔骺板缺损,防止肢体畸形发生.方法 取2周龄兔骺板软骨细胞,体外复合牛关节软骨细胞外基质后植入4周龄兔右侧股骨远端骺板缺损处,左侧仅造成缺损,无填充物,作为自身空白对照.于每周行X线检查,4、8、12、16周时行组织学和免疫组化检查.结果 X线显示移植侧股骨畸形明显比对侧轻,骺板缺损处未见明显骨桥生成,被结构紊乱的软骨组织填充,具有一定的生长能力.空白对照侧骨桥形成,骺板早闭.组织学显示移植侧骺板较窄,排列紊乱.免疫组化结果显示移植处染色阳性.结论 组织工程方法培养出骺板软骨不仅可以阻止骨桥产生,还具有一定的生长能力,有望成为治疗骺板早闭的一种合适的材料.

Abstract：

Objective The aim of this study is to establish the feasibility of acellular cartilage matrix as a scaffold for epiphyseal chondrocytes to form tissue engineering growth plate for the treatment of partial growth plate injury. Methods We collected and cultured the epiphyseal plate chondrocytes from 2 weeks old rabbit in vitro, seeded in acellular cartilage matrix to form a complex. It was then transferred to the rabbit right distal femur which was subjected to growth plate injury. The left femur was subjected to the same injury without further treatment (control). The femur recovery was monitored with weekly X-ray. The morphology was studied with hematoxylin- eosin stain and immunohistochemistry. Results The right femur recovery was significantly better than that of the left radiologically. Histology and immunohistochemistry studies revealed that the growth defects were filled with cartilaginous tissue, which has a similar structure as natural epiphyseal plate . On the left side (control), the epiphyseal defect was filled with bone bridge and epiphyseal plate growth was arrestted. Conclusions Epiphyseal cartilage tissue engineering prevents bone bridge generation and also has some growth potential. It may be a suitable treatment materials for repairing the epiphyseal plate defect     

骺板组织工程方法治疗骺板损伤的实验研究

目的 探索应用骺板组织工程治疗骺板损伤的新方法.方法 将骺板软骨细胞与可降解多孔丝素材料复合,植入骺板损伤的动物模型中,同时设单纯植入可降解多孔丝素材料组和空白对照组.应用组织学Mason三色染色、细胞学BrdU细胞示踪技术、影像学X线检测等方法,观察骺板修复、短缩畸形和成角畸形情况.结果 通过Masson三色染色我们发现:骺板软骨细胞复合可降解多孔丝素材料组(SF+ cell)存在一定的短缩畸形和成角畸形,骺板损伤处发现类似正常的柱状排列细胞；单纯植入可降解多孔丝素材料组(SF)存在明显的短缩畸形和成角畸形,骺板损伤处发现松散的柱状排列细胞；空白对照组存在显著的短缩畸形和成角畸形,骺板损伤处未见柱状排列细胞.通过X线检测我们发现:三组短缩畸形分别为(3.24±0.58)％、(6.32±0.37)％和(8.47±0.51)％.三组成角畸形分别为(4.89±1.27)°、(10.67±1.58)°和(18.44±2.65)°.其短缩畸形和成角畸形的程度均存在统计学意义(均P=0.0001,P＜0.01).结论 骺板组织工程方法治疗骺板损伤是一种可行的方法,骺板软骨细胞复合可降解多孔丝素材料组的疗效优于单纯植入可降解多孔丝素材料组.     

不同自体移植物防止骺板损伤后骨桥形成的实验研究

目的：比较在骺板损伤后分别将自体脂肪，游离骺板，带肌瓣骺板作为移植物来防止骨桥形成，减小畸形发生的效果。方法：将32只未成熟家兔随机分为4组，以A、B、C、D组表示，将A－D组家兔左股骨近端外侧骺板造成人为缺损，A组缺损旷置，B、C、D组分别以自体脂肪，游离腓骨骺板，带肌瓣腓骨骺板作为移植物填塞缺损。术后12周处死，双下肢拍X线片和CT扫描。并对胫骨缺损处进行组织学观察。结果：只有D组（带肌瓣骺板作为移植物组）家兔发生的胫骨畸形较轻，与A组（缺损旷置组）在统计学上差异有显著性意义。结论：自体脂肪作为移植物不能完全避免骨桥的发生；带肌瓣骺板移植较游离骺板易于存活，有一定临床应用前景；损伤后骺板具有一定再生能力。     

不同直径骺板贯穿伤对骺板发育影响的实验研究

目的　探讨不同直径克氏针造成骺板贯穿伤对骺板生长的影响。方法　选用封闭群系日本大耳兔 4 8只 ,随机分为两个实验组 ,每组 2 4只 ,采用直径分别为 1.5mm和 3 .5mm的克氏针纵形贯穿兔股骨远端骺板中心 ,于术后 1、3、6及 12周处死实验兔。测量骺板受损面积百分率 ,并从大体、组织学方面观察骺板发育情况。结果　 1.5mm和 3 .5mm组骺板损伤面积百分率分别为( 1.69± 0 .0 9) %和 ( 9.13± 0 .2 0 ) %。术后 12周 3 .5mm组实验侧股骨较对照侧短缩( 2 .0 8± 0 .4 6)mm(P <0 .0 1) ,而 1.5mm组未出现骨生长抑制。组织学观察发现术后 3周骨桥初步形成 ,术后 12周骨桥成熟 ,3 .5mm组骨桥周边区域骺板增殖细胞层明显变薄。 1.5mm组未发现骺板生长阻滞。结论　克氏针造成骺板贯穿伤与骺板受损面积有关 ,超过骺板表面积约 9.0 %就可能导致骨生长抑制     

骨桥切除治疗骺板部分闭合的初步报告

介绍骨桥切除的手术方法及３例术后随访结果。２例骺板骨桥位于股骨远端，另１例位于胫骨远端，均为中心型骨桥，并以肢体短缩为主要畸形。手术采取Ｐｅｔｅｒｓｏｎ所描述的方法，在电视Ｘ线透视下，通过干骺端应用微型电动骨钻作一骨隧道，逐渐磨除骺板骨桥，然后用自体脂肪填塞，以防止新骨形成。术后平均随访１８．６个月，Ｘ线片显示骺板仍保持开放，成角畸形也获得明显矫正，受累侧骺板继续生长。其中１例左股骨远端骺板骨桥，术后２０个月又增长１．８ｃｍ。结果表明，骨桥切除是治疗骺板部分闭合的有效方法，适用于骨桥面积小于受累骺板的５０％，其年龄距生理性能板闭合尚有２～３年的儿童。     

可吸收聚消旋乳酸棒垂直骺板植入对兔骺板生长的影响

目的 探讨可吸收聚消旋乳酸棒(poly-D,L-lactide acid,PDLLA)垂直骺板植入对幼兔股骨远端骺板生长的影响.方法 采用封闭群系5周龄日本大白兔24只,随机分为两组:1.0mm、2.0 mm组,每组12只.在幼兔右侧殴骨远端跨骺板中心分别垂直植入直径为1.0 mm、2.0 mm的PDLLA棒;左侧作为对照,只作膝关节切开,不钻孔,不植入.电脑计算骺板损伤面积,术后6、12、24周处死动物.通过测量双侧股骨长度、双侧膝关节外翻角、组织病理学、骨骺磨片和透射电镜等方法观察PDLLA棒对骺板生长的影响.结果 1.0 mm、2.0 mm的PDLLA棒垂直植入所造成的骺板损伤分别占骺板面积的(1.39±0.09)%、(3.33±0.92)%;术后6、12、24周各组内实验侧和对照侧相比,双侧股骨长度、外翻畸形无明显差异(P＞0.05).结论 可吸收聚消旋乳酸棒垂直骺板植入不会对兔骺板生长造成不良的影响.     

骺板骨桥切除术治疗儿童骺板部分早闭

骺板是儿童骨骺与干骺端之间的生长活跃软骨区,具有纵向和横向生长的功能,骨折、感染、肿瘤、辐射均可导致骺板损伤。骺板损伤后可能在骨骺与干骺端之间的软骨区形成骨性连接即骨桥,使骺板全部或部分提前闭合,导致肢体短缩和（或）成角畸形。传统的治疗方法是针对临床表现的短缩和（或）成角畸形,采取诸如截骨矫形、骨骼延长或健侧骺板阻滞术等方法进行治疗。     

骺板损伤早闭治疗的研究进展

骺板是儿童骨骼所特有的结构,它是骨骺与干骺端间生长活跃的软骨区,呈波浪状薄板样,具有纵向和横向生长的功能。其损伤会导致儿童长管状骨骨骺与干骺端之间形成骨性连接即骨桥,使骺板全部或部分提前闭合,造成肢体短缩和（或）成角畸形。近年来,由于对其病理、诊断和实验研究的不断深入及关节镜、显微外科等新技术的不断发展,在临床治疗方面已相继取得了新的进展。现将其作一综述。骨折、感染、肿瘤、辐射等因素均会导致骺板的损伤。而骺板损伤所致早闭往往在受累肢体出现成角畸形或短缩时才被发现。根据Ｘ线平片上骨桥位于骺板的部位…     

骺部分早闭再开放自体骺软骨充填术

介绍骺开放自体骺软骨移植填充治疗儿童骺早闭的手术技术并评价其疗效，两例分别为胫骨远端和胫骨近端骺早闭患儿，其闭合范围〈５０％，行骺开放及自体髂骨骺软骨填充治疗。分别随访１０个月和１４个月，结果纵向生长能力恢复，成角畸形部分矫正，表明生长期儿童行骺开放和自体骺软骨移植不但可防止骨桥形成，而且可恢复纵向生长能力，并矫正成角畸形。     

儿童骺板损伤组织工程学治疗研究进展

儿童骺板损伤非常常见,骨折、感染、恶性肿瘤或医源性损伤等原因均可导致,损伤后易引起生长阻滞、成角或旋转畸形,严重影响儿童的身心健康。目前临床骨桥切除术结合相应材料填充的治疗方法成功率低。采用软骨组织工程技术构建具有生物活性的骺板软骨成为治疗儿童骺板损伤新的研究方向。因此,现主要从组织工程三要素：种子细胞、生长因子、组织...     

压力和张力对骨骺板软骨细胞的影响

新西兰幼兔１９只，在其右股骨下端安装带有拉力和压力传感器的外固定装置，向骺板近端分别施压力和张力，左侧股骨作为对照。４周后两侧股骨作长度测量和组织学检查。结果表明压力使骺板厚度全层变薄，肥大细胞数减少，骨生长表现为抑制状态。张力使骺板增殖层和肥大层厚度增加，肥大细胞数增多，骨生长活跃。机械力使骺板软骨细胞在组织学上的变化具有统计学意义。     

Achondrogenesis type II, abnormalities of extracellular matrix

Immune and lectin histochemical and microchemical methods were employed to study growth cartilage from seven cases of achondrogenesis type II (Langer-Saldino). The normal architecture of the epiphyseal and growth plate cartilage was replaced by a morphologically heterogeneous tissue. Some areas were comprised of vascular canals surrounded by extensive fibrous tissue and enlarged cells that had the appearance and histochemical characteristics of hypertrophic chondrocytes. Other areas contained a mixture of cells ranging from small to the enlarged chondrocytes. The extracellular matrix in the latter areas was more abundant and had characteristics of both precartilage mesenchymal matrix and typical cartilage matrix; it contained types I and II collagen, cartilage proteoglycan, fibronectin, and peanut agglutinin binding glycoconjugate(s). Peptide mapping of cyanogen bromide cartilage collagen peptides revealed the presence of types I and II collagen. These observations could be explained by a defect in the biosynthesis of type II collagen or in chondrocyte differentiation.     

纤维蛋白胶对骨骺损伤后骨桥形成的预防作用

目的 利用未成熟大鼠制作胫骨近端骨骺损伤模型,比较不同方法 修复生长板损伤的治疗效果,评价纤维蛋白胶作为一种新型生物材料应用于微创治疗方法 的可行性.方法 实验动物分为4组,A组为未损伤组,作为正常对照;B组为单纯损伤组,按照模型损伤后未予特殊处理;C组为脂肪组织填充治疗组,按照模型损伤后取自体腹股沟区脂肪组织填充损伤区,骨蜡封闭骨洞;D组为纤维蛋白胶填充治疗组,按照模型损伤后以预先配置好的纤维蛋白胶(广州倍绣生物技术有限公司)填充损伤区,骨蜡封闭骨洞.通过组织化学染色观察损伤模型的建立、损伤愈合过程以及骨桥形成情况.取材后先进行显微CT检查,再行组织学检测.比较纤维蛋白胶、脂肪作为填充材料预防治疗大鼠胫骨近端骨骺损伤后骨桥形成的情况,以及大鼠胫骨长度和近端角度的变化.结果 组织学检查显示,纤维蛋白胶和脂肪填充组损伤区域逐渐被纤维组织替代,单纯损伤组损伤区域逐渐被骨桥组织替代.纤维蛋白胶填充治疗组在避免损伤区出现不可逆的骨桥方面,与脂肪填充治疗组效果接近(P>0.05).纤维蛋白胶填充组与单纯损伤组之间存在明显差异(P<0.05),脂肪填充组与单纯损伤组之间存在显著性差异(P<0.01).结论 显微CT可以用于骨骺损伤后骨桥形成的观察研究中.纤维蛋白胶制作来源广泛,术中应用简便,效果与传统的脂肪填充治疗接近,因而推测可能作为一种新的治疗方法 引入临床,预防骨骺损伤后的骨桥形成.     

Integrins in growth plate cartilage

The growth plate is a highly specialized layer of cartilage where chondrocytes proliferate and differentiate in order to bring forth longitudinal bone growth. Interaction of cells with the extracellular matrix significantly influences differentiation and growth. These processes involve integrin as well as nonintegrin cell surface receptors, like annexin V and CD44. In chondrocytes, which are surrounded by abundant matrix, tissue integrity and cell survival depend on well-tuned interactions of cells and ECM. In the field of growth research, besides classical growth stimulating factors acting via endocrine pathways, investigations on local determinants of cell differentiation and survival play an important role. This review describes the role and function of integrins, their occurrence in growth plate cartilage and their possible involvement in differentiation during growth.     

骨骺发育障碍与生长

正常的骨骼对维系人体正常体型及活动非常重要,任何骨骼疾病都会危及健康。骨骼生长发育是一个复杂的过程,受诸多通路的调控及因素影响。骨骺软骨承载的生长板是骨生长的重要部位,其发育障碍直接影响骨骼生长。该文就骨骺的正常生长发育及其障碍所引起的代表性骨病--脊柱骨骺发育不良及软骨发育不全综合征进行阐述。     

A review of the actual knowledge of the processes governing growth and development of long bones

Autoptic samples of human bones (from 8 weeks of gestation to 12 years of age) and a second group of serial, skeletal x-rays (required for pathologies not related to bone dysplasia in children from 4 months to 17 years of age) provided the material for the analysis of the physes normal growth mechanism presented in this review. Before the appearance of the ossification centers epiphyseal growth rests exclusively on chondrocytes proliferation (interstitial growth), without any detectable differentiated cellular organization. When endochondral ossification starts a defined spatial disposition of chondrocytes and a corresponding organization of the intercellular matrix is set up, so that it is possible to identify a growth vector corresponding to the columns of piled chondrocytes with direction from hypertrophic toward the proliferative cell layers. The complexity of the tubular bones growth process is well represented by the spatial arrangement of the growth vectors. In the late epiphyseal growth another mechanism is active in addition to endochondral ossification, namely, articular cartilage interstitial growth and subchondral remodelling. The knowledge of the normal mode of organization of the physis and its temporal sequence can help to better understand of the deviaton from the normal development of metaphyseal and epiphyseal dysplasias.     

Kniest dysplasia. A histochemical study of the growth plate

Chondro-osseous tissue from four patients with the Kniest dysplasia was studied histochemically using a new plastic embedding technique. Extensive vacuolar changes were observed p--1 throughout the endochondral growth plate and adjacent resting cartilage. These changes occurred within the cartilage matrix and also in the lacunae of degenerating chrondrocytes. The septa of the lesions contained chondroitin sulfate, but little keratan sulfate or collagen. Resting cartilage not adjacent to the growth plate stained irregularly and showed few of the vacuolar lesions, and chondrocytes were enlarged and contained cytoplasic inclusions, but no vacuolar material. Thus, there appears to be a sequence of events initiated by cellular accumulation of a substance and progressing to cellular and matrix degeneration.     

儿童正常生长规律及其调控机制

生长是一个连续非线性的过程,不同时期的调控机制及生长特点不尽相同。骨骼的生长受多种因素的影响,而软骨生长板是骨骼生长和发育的关键。包括营养、内分泌激素、炎症细胞因子、旁分泌信号、细胞外基质等在内的多种调控机制通过调节生长板软骨细胞的成熟与分化在骨骼的生长过程中发挥作用。掌握儿童正常生长规律和调控机制对矮身材儿童的诊断、鉴别诊断及此后的规范化治疗具有极其重要的作用。