关节软骨缺损修复的实验与临床

关节软骨的修复一直是骨科领域尚未完全解决的一大难题。现就关节软骨损伤后促进自身修复、组织或细胞移植修复、组织工程修复等方面对关节软骨修复方法作一综述。     

永生化软骨细胞为基础的工程化软骨修复软骨缺损的实验研究

目的探讨用永生化软骨细胞作为种子细胞修复羊关节软骨缺损的可行性.方法把人端粒酶催化亚基(hTERT)转染羊关节软骨细胞,筛选阳性克隆并通过旋转生物反应器-微载体技术进行大量扩增;将扩增的永生化软骨细胞与β-磷酸三钙复合并在体外培育后,植入羊前肢肱骨头关节面软骨缺损处;3和6个月取材,进行形态学和免疫组织化学评价.结果实验组,术后6个月,缺损区被新生的软骨组织所充填,Ⅱ型胶原染色呈强阳性,与对照组差别有显著性差异(P＜0.01).在材料组,缺损区边缘可见部分新生软骨组织;空白对照组,缺损区新生软骨组织形成.结论永生化软骨细胞在软骨组织工程中具有潜在的应用前景.     

Effects of low-intensity pulsed ultrasound in repairing injured articular cartilage

Articularcartilageisakindofavascularmusculoskeletaltissueswitharelativelylowcapacityforintrinsicrepairorregeneration.Intheabsenceofavailablestemcells,thereislittlehopeforcartilagerepairafterinjuryandthetissuesprogress alongadegenerativepathwaythatdestroysthe biochemicalandbiomechanicalproperties.Sorepairof damagedarticularcartilageisoneofthemostdifficult problemsinorthopaedicsandnosatisfactorytreatment methodhasbeenfoundinthisfield.Ultrasoundhas beenwidelyusedinclinicaldiagnosisandtreatment.It…     

The capacity of pig articular cartilage in organ culture to regenerate after breakdown induced by complement-sufficient antiserum to pig erythrocytes

Explants of pig articular cartilage including invading marrow and subchondral bone (together=‘invasion zone’) were cultivated for 10 or 14 days in complement-sufficient rabbit antiserum to pig erythrocytes (AS+C′), and then transferred to heat-inactivated normal rabbit serum (NRS) for a period of recovery. In AS+C′ the cartilage matrix lost first proteoglycan and then collagen, but the cells remained viable. The degradation of collagen was accompanied by a fibroblastic transformation of the chondrocytes, first seen in the region immediately above the invasion zone. Immunohistochemical studies showed that after cultivation in AS+C′, IgG antibodies entered areas in which the matrix was depleted of proteoglycan and reacted strongly with the majority of chondrocytes; those that had undergone fibroblastic transformation exhibited little or no reaction. The degree of recovery in NRS depended on the extent to which the matrix had broken down in AS+C′. If degradation of collagen was confined to the region immediately above the invasion zone, and elsewhere only proteoglycan had been lost, new metachromatic material was regenerated in the non-calcified cartilage, and the fibroblast-like chondrocytes resumed their normal appearance and regained their reactivity with the IgG antibodies of AS; new cartilage and chondroid tissue appeared in the cavities of the invasion zone. If degradation of collagen and fibroblastic transformation of chondrocytes had spread throughout the cartilage, breakdown continued in NRS and cartilage disappeared completely above the invasion zone; new cartilage was sometimes formed in the cavities of the invasion zone.     

An experimental study on transplantation of articular cartilage

Summary Behaviour and fate of the transplanted articular cartilage were studied in 180 adult rabbits, using the scanning electron microscope, histological staining and autoradiographic examination.The junction between the host cartilage and the transplanted cartilage was covered with thin connective tissue layers which extended from the edges 4 weeks after transplantation.After the 24th week of transplantation the cartilage tissue appeared to be degenerating at the periphery of the graft.However, in the middle portion of the graft surviving cartilage cells were observed using ³⁵S autoradiography.

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Résumé Comportement et évolution du cartilage articulaire transplanté ont été étudiés chez 180 lapins adultes par microscopie électronique, colorations histologiques et autoradiographies.Quatre semaines après la transplantation, une fine couche de tissue conjonctif qui s'étend depuis les bords recouvre la jonction entre le cartilage du receveur et le cartilage transplanté.Vingt-quatre semaines après la transplantation, le tissu cartilagineux commence à dégénérer à la périphérie du greffon. Cependant, à la partie centrale de celuici, des cellules cartilagineuses survivantes peuvent être mises en évidence grâce à l'autoradiographie au S³⁵.

     

Ultrastructural changes of TMJ articular cartilage and synovial membrane following occlusal trauma in rabbit

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自体骨膜游离移植的实验研究--不同龄动物大块关节软骨缺损修复的比较

目的研究比较自体骨膜移植软骨再生修复不同龄动物大块关节软骨缺损。方法用52只不同龄家兔自体骨膜游离移植修复大块关节软骨缺损,比较移植骨膜生发层朝向关节腔与松质骨时再生软骨的差别。结果经不同时期肉眼和组织学检查证实,幼年兔和成年兔的骨膜移植都能生成软骨,修复大块关节软骨缺损。在成年兔骨膜再生的软骨与成年兔本身周围正常软骨的厚度、组织结构一样。移植骨膜生发层朝向关节腔与松质骨二者间再生软骨结果无明显差别。结论骨膜具有再生软骨的能力,可用来移植修复关节软骨的缺损。骨膜移植生发层不同朝向对软骨再生无明显影响。成年后骨膜移植修复关节软骨缺损能够生成与自身相适应的软骨。     

Induction of cartilage growth in a rabbit ear model: a pilot study

Identifying the control of cartilage regeneration is important both clinically and in tissue engineering research. A rabbit ear model was used to simulate surgery and trauma to explore the effect of perichondrial stripping on underlying cartilage in vivo. Ten rabbits (20 ears) formed four groups: two controls and two experimental groups. Group 1 served as the unoperated control group and underwent no treatment. Group 2 served as the operated control group and underwent elevation of auricular skin flaps without stripping the perichondrium. Groups 3 and 4 underwent increasing degrees of surgical insult. Group 3 underwent elevation of a skin flap with stripping of the perichondrium on both sides of the cartilage. Group 4 underwent both perichondrial stripping and the insertion of a thin silicone sheet as a barrier between the denuded cartilage and the skin flaps. At 3 months, punch biopsies of the cartilage were performed in each zone of insult, creating multiple thin sections. The results were analyzed using a computerized morphometry system. Histopathological examination of the groups revealed a regenerative layer of neocartilage which showed distinct hypercellular features of regeneration. The thickness of the new layer was proportional to the degree of the insult (p<0.01). A controlled insult to the perichondrium created a regenerative layer of cartilage; it seems that this layer of neocartilage is proportional to the insult. Further studies are in progress to clarify these findings.     

Experimental evaluation of the usefulness of osteochondral allograft for articular cartilage defect

This study was conducted to evaluate the usefulness of osteochondral allografts for articular cartilage defects. Cartilaginous defects measuring 4.5mm in diameter were experimentally prepared in both the weight-bearing and non-weight-bearing regions of the femur in six male miniature pigs (9 months old). Osteochondral grafting was performed using fresh autografts (group AU), fresh allografts (group AL), or frozen allografts (group FA). Untreated cartilaginous defects were used as the control (group D). All the pigs were killed 4 weeks later, and the respective osteochondral grafts were macro- and microscopically evaluated. Hematoxylin and eosin staining, safranin O staining, and immunostaining [matrix metalloprotease-1 (MMP-1) and tissue inhibitor of metalloprotease-2 (TIMP-2)] were used for the histological evaluations of transplanted cartilage. Macroscopic and microscopic findings were assessed according to the criteria proposed by Wakitani et al. Although groups AU and AL showed similar median scores (ranges) for the evaluation of cartilaginous defect restoration, groups FA and D showed unfavorable scores: 3.9 (0–9) in group AU; 4.5 (0–12) in group AL; 10.2 (4–12) in group FA; and 7.0 (5–11) in group D. Immunostaining revealed almost identical results in groups AU and AL. As there were no histologically significant differences in the status of the osteochondral grafts between fresh autografts and fresh allografts, fresh allografts might be useful donor osteochondral grafts.     

细胞移植及软骨组织工程技术在关节软骨损伤临床应用进展

细胞移植和软骨组织工程技术是治疗关节软骨损伤的新方法,也是当今国内外关节骨科的研究热点。本文综述了国际上各种细胞移植和组织工程技术在关节软骨损伤临床的应用,包括软骨细胞移植技术、联合组织工程支架的软骨细胞移植技术和干细胞移植技术,着重介绍了各种技术的特点和临床应用现状及进展,以期为国内软骨损伤治疗技术选择提供思路。     

关节软骨修复与细胞因子

关节软骨损伤后的修复非常有限，其对创伤、炎症的反应是由软骨细胞、滑膜组织分泌或关节液中含有的细胞因子所介导的。随着现代分子生物学的发展，人们已发现多种细胞因子参与并调节软骨的生长过程，对软骨损伤的修复起积极的促进作用。下面就近年来较受国内外学者关注的几种细胞因子与软骨损伤修复的关系作一综述。     

修复关节软骨大面积缺损的实验研究

目的 比较和评价筋膜软骨细胞和骨膜、关节软骨移植修复关节软骨大面积缺损的能力和生物特性。方法 用冻存和新鲜的异体筋膜上培养的软骨细胞、骨膜和关节软骨移植修复大面积关节软骨缺损,通过大体标本、光学显微镜、扫描和透射电镜、放射身显影、微量元素和柱层析氨基酸定量测定、一氧化氮含量测定等多种观察方法进行评价。结果 新鲜和冻存的筋膜软骨细胞移植在结构、形成新的软骨细胞能力、代谢活性方面均优于游离软骨细胞移植     

Effects of varying intensities of laser energy on articular cartilage: a preliminary study

The effects of laser energy on articular cartilage were studied utilizing the neodymium YAG laser. Partial-thickness cartilage defects were surgically attempted in the femoral condyles of knee joints in guinea pigs. The defects were exposed to laser energy of varying intensities [group I, 25 J (5 W X 5 sec); group II, 75 J (15 W X 5 sec); group III, 125 J (25 W X 5 sec)]. A fourth group was studied, in which the defect was not lased. Animals were killed at weekly intervals from 1 to 6 weeks and the knee joints were subjected to histological analysis. At 5 weeks, the knees exposed to 25 and 75 J demonstrated a reparative process with chondral proliferation. The knees exposed to 125 J demonstrated fibrotic tissue and tissue necrosis that resulted in fibrosis. In the knees not exposed to laser energy, numerous foci of granulation tissue were present at all stages with the end point of healing being one of fibrosis with disorganized patchy cartilage islands.     

The effect of low-level laser therapy during orthodontic movement: a preliminary study

It has been emphasized that one of the most valuable treatment objectives in dental practice is to afford the patient a pain-free treatment. By the evolution of the laser applications, the dental committee aimed to achieve this goal without analgesic drugs and painful methods. Orthodontic treatment is one of these concerns, that one of the major components of patient to reject this treatment is the pain accompanied during the different treatment phases. Another great concern of the patient is not to get through prolonged periods of treatment. The aim of this study is to evaluate the effect of the low-level (GaAlAs) diode laser (809 nm, 100 mW) on the canine retraction during an orthodontic movement and to assess pain level during this treatment. A group of 15 adult patients with age ranging from 14 to 23 years attended the orthodontic department at Dental School, Damascus University. The treatment plan for these patients included extraction of the upper and lower first premolars because there was not enough space for a complete alignment or presence of biprotrusion. For each patient, this diagnosis was based on a standard orthodontic documentation with photographs, model casts, cephalometric, panorama, and superior premolar periapical radiographies. The orthodontic treatment was initiated 14 days after the premolar extraction with a standard 18 slot edgewise brackets [Rocky Mountain Company (RMO)]. The canine retraction was accomplished by using prefabricated Ricketts springs (RMO), in both upper and lower jaws. The right side of the upper and lower jaw was chosen to be irradiated with the laser, whereas the left side was considered the control without laser irradiation. The laser was applied with 0-, 3-, 7-, and 14-day intervals. The retraction spring was reactivated on day 21 for all sides. The amount of canine retraction was measured at this stage with a digital electronic caliper (Myoto, Japan) and compared each side of the relative jaw (i.e., upper left canine with upper right canine and lower left canine with lower right canine). The pain level was prompted by a patient questionnaire. The velocity of canine movement was significantly greater in the lased group than in the control group. The pain intensity was also at lower level in the lased group than in the control group throughout the retraction period. Our findings suggest that low-level laser therapy can highly accelerate tooth movement during orthodontic treatment and can also effectively reduce pain level.     

滑膜关节与关节软骨的形成

膜关节是人体的重要组成部分,虽然数量很少,但是它们都有独特的生物力学结构和功能。近几十年来滑膜关节一直是骨科领域研究的热点,这充分体现了关节对维持人类机体功能和生活质量的重要性,但是目前对胚胎发育过程中滑膜关节的形成机制依然知之甚少。本文对滑膜关节与关节软骨形成机制的研究进展作一综述。     

An experimental study of the repair of injured articular cartilage using 3H-thymidine autoradiography

Summary The mechanism of repair of injuries to articular cartilage of the distal femur in young, young adult and adult rats was investigated using ³H-Thymidine autoradiography and microscopic observation. It was found that in young rats the defect was repaired by the reactive proliferation of the cartilage cells, that in the young adult rats the wound was repaired by hyaline cartilage which was differentiated from undifferentiated mesenchymal cells (U.M.C.), and that in the adult rats repair with cartilage did not occur. The superficial part of the defect was filled by connective tissue derived from the U.M.C. while the deeper layers were repaired by tissue resembling bone and also derived from the U.M.C. Repair of the injured articular cartilage thus varied with the degree of maturity of each animal, suggesting that age largely influences the mechanism of response to injury.

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Résumé Le mécanisme de la réparation du cartilage articulaire après traumatisme a été étudié chez le rat au niveau de l'extrémité distale du fémur. Les expériences ont porté sur des animaux jeunes, adultes jeunes et adultes et ont été contrôlées par autoradiographies à la thymidine tritiée et observations microscopiques. On a trouvé que chez le jeune rat la perte de substance est réparée par prolifération des cellules cartilagineuses, que chez le jeune adulte la réparation est faite de cartilage hyalin qui se différencie à partir des cellules mésenchymateuses et que chez l'adulte il n'y a pas de réparation par du cartilage. La partie superficielle de la perte de substance est comblée de tissu conjonctif dérivé des cellules mésenchymateuses, tandis que sa partie profonde est réparée par du tissu ressemblant à de l'os, également dérivé des cellules du mésenchyme. La réparation du cartilage articulaire varie donc avec le degré de maturité de chaque animal, ce qui permet de penser que l'âge influence notablement le mécanisme de réponse au traumatisme.

     

骨髓基质干细胞修复兔关节软骨缺损的实验研究

目的研究以多聚乙醇酸(PGA)为支架的骨髓基质干细胞(BMSCs)复合物修复兔膝关节软骨缺损的情况。方法体外培养扩增的自体BMSCs种植于PGA支架并培养72h,然后将支架-细胞复合物植入兔关节软骨缺损模型。术后12周处死动物,标本行大体观察、组织学检查及Ⅱ型胶原免疫组化染色。结果BMSCs-PGA复合物植入后形成丰富的透明软骨样修复组织,新生软骨无明显退变。对照组主要为纤维组织及软骨下骨修复。结论BMSCs-PGA复合物可修复关节软骨缺损。