骨髓基质细胞移植修复半月板无血运区损伤的实验研究

目的比较自体与同种异体骨髓基质细胞移植对半月板无血运区损伤修复的影响。方法 40只成年新西兰大白兔随机平均分为 A、 B两组。 A组兔的骨髓基质细胞 (MSC)经体外培养后与纤维蛋白凝胶 (FG)混合,自体移植于其一侧的膝关节半月板缺损区,即 FG＋自体 MSC（自体移植组）;另一侧单纯植入 FG(FG植入组 )。于 B组兔的一侧膝关节半月板缺损区移植 FG＋同种异体 MSC(异体移植组 ),另一侧缺损不予修复 (空白对照组 )。分别于术后第 1、 2、 3个月取材,观察半月板损伤部位的组织形态学变化。结果 (1)自体移植组 :术后 1个月缺损区可见纤维组织,内有大量成纤维细胞;术后 2个月见大量软骨细胞并有胶原纤维形成;术后 3个月损伤区呈纤维软骨愈合。 (2)空白对照组 :术后 1～ 3个月缺损区始终未愈合。 (3)单纯 FG植入组 :术后 1～ 3个月缺损区可见纤维组织,内有少量成纤维细胞,没有软骨细胞生长,呈瘢痕样愈合。 (4)同种异体移植组 :与自体移植组所见大致相同,但有 3侧缺损区可见大量淋巴细胞浸润,胶原纤维少。结论骨髓基质细胞移植可促进半月板无血运区损伤的愈合,同种异体骨髓基质细胞移植修复半月板无血运区损伤发生免疫排斥反应的机率较低。     

半月板无血运区损伤治疗进展

半月板是维持膝关节内环境的基本结构,半月板损伤后膝关节力学稳定性发生重大变化,常导致早期骨关节炎.半月板再生潜力很小,尤其是半月板无血运区损伤,以往认为不能自然愈合.如何促进半月板无血运区损伤的愈合,一直是骨科医生面临的最具有挑战性的难题之一.该文就半月板无血运区损伤后的愈合特点、传统治疗和目前治疗现状作一综述.     

微量直流电刺激半月板无血运区裂伤愈合的实验研究

目的　观察微量直流电刺激犬半月板无血运区裂伤修复的组织学特征。方法　将２０ 条成年杂种犬双后肢膝外侧半月板无血运区造成纵形裂伤,每条犬后肢一侧为刺激组,对侧为对照组,用自制微电流刺激器刺激２ 、４ 、８ 、１２ 周后分别取材,行光镜及电镜观察。结果　电刺激２ 周半月板裂伤出现部分愈合,１２ 周完全愈合,而自身对照组均未愈合。结论　电刺激可诱导裂缘纤维软骨细胞分裂增殖;促进毛细血管趋向性增生、胶原合成及排列。半月板无血运区组织细胞并非惰性细胞,采用微量直流电刺激对半月板无血运区裂伤具有明显的促进修复作用。     

电刺激半月板无血区愈合的组织学特征

半月板无血运区的损伤是不能自然愈合的，如何能使半月板无血区愈合已经成为骨科临床上的一大难题。我们根据微电流刺激骨生长的理论，采用自行设计的微量直流电刺激器，用２０只成年杂种狗作为实验动物，在每只狗双后肢膝外侧半月板无血运区造成纵形裂伤，随机分为实验组和对照组。１２周后实验组均出现愈合，自身对照组均未愈合。本文对电刺激半月板愈合光镜、电镜组织学特征进行了观察及讨论，认为半月板无血运区组织细胞并非惰性细胞，它们具备有潜在的再生能力，而电刺激具有诱导软骨细胞分化、增殖作用，并能促进毛细血管趋向性增生和胶原合成及排列。     

胫骨平台骨折对半月板愈合影响的实验研究

[目的]胫骨平台骨折常可合并包括半月板在内的膝关节软组织损伤,而骨折的发生对半月板愈合会产生何种影响尚未见报道。本文通过建立胫骨平台骨折合并半月板损伤的动物模型,试图观察骨折引起的膝关节改变对半月板修复产生的影响。[方法]建立12只新西兰兔外侧胫骨平台骨折,以及半月板非全层撕裂伤和打孔缺损模型,设立无骨折对照,12周后通过大体与光镜观察两组半月板损伤模型修复的差异。[结果]在无血运区,无论有无合并平台骨折,半月板损伤均未见任何修复迹象;两组有血运区的半月板撕裂伤均已修复裂隙,与正常组织难以区分;而在半月板外1/3区的打孔缺损模型,仅胫骨平台骨折组可见部分蛋白凝块的填充。[结论]胫骨平台骨折同时合并半月板损伤时,未见骨折阻碍半月板愈合的证据,相反,骨折能促使缺损区域的组织填充,但是否有益于半月板愈合尚有待进一步研究。     

自体骨髓基质细胞移植治疗兔膝关节半月板无血运区损伤

目的 观察自体骨髓基质细胞在体内分化为软骨细胞的能力及对半月板无血运区损伤修复的影响。方法 将体外培养的自体骨髓基质细胞收集后与纤维蛋白凝胶混合,植入４０只成年兔一侧膝半月板无血运区损伤的裂口内,另侧设空白对照或单纯纤维蛋白凝胶植放。分别于术后１（１２只）、２（１２只）、３（１５只）个月取村,行肉眼、光镜组织学观察。结果 术后２个月１２个膝半月板损伤处在量的软骨细胞并有胶原纤维形成,３个月１３个膝     

骨髓基质干细胞复合藻酸钙凝胶修复全层半月板无血运区缺损

目的 观察骨髓基质干细胞复合藻酸钙凝胶注射式修复全层半月板无血运区缺损的效果.方法 2008年6月至2009年2月制造成年山羊半月板前角无血运区全层缺损模型.以自体骨髓基质干细胞复合可注射藻酸钙凝胶修复半月板缺损(Ⅰ组),同时设立单纯载体组(Ⅱ组)和空白对照组(Ⅲ组).术后4、8、16周处死动物,行大体观察,组织学观察、电镜观察和MRI检查并比较修复效果.结果 Ⅰ组缺损完全被修复组织填充,结合紧密,与正常半月板组织相似,在4～16周效果逐渐改善,大体观察优于其他各组.光镜见细胞随凝胶纤维排列分布,载体纤维间隙大多为细胞分泌基质所充填,细胞排列密集,基质分布均匀.透射电镜见Ⅰ组细胞呈软骨细胞样形态,细胞突起较多,细胞器丰富,细胞为不同走向排列的纤维包绕.MRI检查发现Ⅰ组修复效果较好.结论 骨髓基质干细胞复合藻酸钙凝胶注射可有效地修复全层半月板无血运区缺损.     

关节软骨和半月板组织碎块移植研究进展

关节软骨和半月板无血运区损伤后不能自然愈合,软骨组织碎块移植为之修复提供了新思路。体外和体内实验研究均证实,关节软骨及半月板无血运区切成碎块后细胞能有效地从碎块组织释放,与周围支架材料复合或直接导致组织修复。自体软骨植入系统（CAIS）及同种畀体幼年关节软骨碎块移植物（DeNovoNT）,已应用于,临床。该文就关节软骨及半月板组织碎块移植的基础研究及临床应用研究及其进展作一综述。     

经关节镜半月板部分切除治疗半月板无血管区损伤

目的 分析关节镜下半月板部分切除治疗半月板无血管区损伤的手术疗效，提出较适宜的手术时机。方法 随访276例(侧)半月板无血管区损伤关节镜下部分切除的患者，从手术时间、镜下部分切除术后疗效进行分析、比较。结果 半月板损伤后15d～2个月组行半月板部分切除术后优良率达79％；2～4年组治疗术后优良率为57．2％。结论 半月板损伤应早期诊断、早期治疗；早期关节镜下部分切除无血管区损伤半月板的疗效明显优于晚期手术治疗。     

结缔组织生长因子促进半月板无血管区损伤愈合

目的：探讨结缔组织生长因子（CTGF）对纤维软骨细胞外基质分泌、VEGF表达及促进半月板无血管区的损伤修复中的作用。方法：将新西兰大白兔半月板白区,经胶原酶消化、离心分离后,提取半月板纤维软骨细胞,培养至第2代。用流式细胞仪鉴定该细胞表面CD31,CD44,CD45和CD105标志物,并用Ⅱ型胶原抗体做免疫细胞化学鉴定,以证明所培养、传代的细胞是纤维软骨细胞。分别将细胞培养在浓度100 ng/ml的CTGF培养基中3、14 d后,通过实时定量聚合酶链反应,检测Ⅰ型胶原、Ⅱ型胶原和VEGF基因的表达变化情况。造模,在兔半月板中央区,制作长3 mm的纵行撕裂。将45只大白兔随机分为3组,处理方式为：半月板单纯缝合术,缝合术加填充PBS-纤维蛋白胶,缝合术加填充1．5滋g CTGF-纤维蛋白胶。术后第1、4、10周用荧光免疫组织化学分析法显示Ⅰ型、Ⅱ型胶原和VEGF在损伤处的表达与分布情况,直观撕裂处的愈合情况。结果：体外实验第14天,定量RT-PCR结果显示,100 ng/ml CTGF组中的Ⅰ型、Ⅱ型胶原和VEGF mRNA表达比PBS对照组,明显增加。体内实验的荧光免疫组织化学结果显示,在术后第10周,CTGF治疗组中的Ⅰ型、Ⅱ型胶原和VEGF已完全填充撕裂损伤处。 PBS-蛋白胶组中,损伤处仍有明显裂隙。结论：CTGF可促进半月板无血管区重要的细胞外基质（Ⅰ型、Ⅱ型胶原）的合成,同时损伤处VEGF的表达活性明显增强,有利于促进无血管区半月板撕裂损伤的愈合。     

Results of rasping of meniscal tears with and without anterior cruciate ligament injury as evaluated by second-look arthroscopy

Purpose: Meniscal rasping without suturing has been experimentally shown to stimulate vascular induction in tears in the avascular zone of menisci, resulting in meniscal healing. The goals of this study were to arthroscopically assess the results of meniscal rasping and analyze the factors affecting meniscal healing. Type of Study: Retrospective cohort study. Methods: Forty-eight torn menisci in 47 patients (age range, 14-47 years; average, 24 years) treated arthroscopically with the meniscal rasping technique were evaluated by second-look arthroscopy. The interval between the injury and the time of surgery ranged from 3 weeks to 13 years. There were 35 lateral and 13 medial meniscal tears associated with 44 anterior cruciate ligament injuries; 28 of the menisci had a full-thickness longitudinal tear and the other 20 had a partial-thickness tear. The length of the tears ranged from 10 to 33 mm (mean, 14.4 mm). The distance from the capsule to the tear ranged from 1 to 9 mm (mean, 5.0 mm). Results: Thirty-four menisci (71%) healed completely (without a marked visible unhealed area), 10 (21%) healed incompletely, and 4 (8%) showed no evidence of healing. There were no relationships between outcome and age, gender, injured side, or time from injury and rasping. Both the distance from the capsule to the tear and the length of the tear were longer in the unhealed menisci. Stable tears had a high healing rate after meniscal rasping. Conclusions: Meniscal rasping without suturing is an easy procedure to perform and seems to be a reliable way to repair longitudinal tears in the avascular region of the meniscus, although the healing potential of the procedure is affected by the distance from the capsule to the tear site and the length and the stability of the tear.Arthroscopy: The Journal of Arthroscopic and Related Surgery, Vol 19, No 5 (May-June), 2003: pp 463–469     

Repair of avascular meniscal injuries using juvenile meniscal fragments: An in vitro organ culture study

We investigated whether the implantation of juvenile allograft and minced meniscal fragments could improve the healing of avascular meniscal injuries, which cannot heal spontaneously. Concentric cylindrical explants were excised from the inner two‐thirds of swine medial menisci. The inner cylinder consisted of a “sandwich” structure, with minced juvenile meniscal fragments, juvenile meniscal columns, minced mature meniscal fragments, or mature meniscal columns implanted in the middle. The explants were cultured in vitro for 2, 4, or 6 weeks. Interfacial meniscal repair was assessed by histology, immunohistochemistry, biomechanical testing, and confocal laser scanning microscopy. Histology and confocal microscopy results revealed that tissue repair and cell accumulation at the interface were best at all time points in the juvenile meniscal fragments group, followed by the juvenile columns, minced mature fragments, and mature columns groups, respectively. At 6 weeks, the implantation of juvenile allograft and minced meniscal fragments increased the shear strength, peak force, and energy to failure in the peripheral interface. Picosirius red/polarized light microscopy and immunohistochemistry results showed concurrent expression of type I and II collagen in the interfacial repair tissue. In conclusion, implantation of juvenile allograft and minced meniscal fragments could increase the healing of avascular meniscal injury in vitro. © 2013 Orthopaedic Research Society Published by Wiley Periodicals, Inc. J Orthop Res 31:1514–1519, 2013     

Feasibility of a self‐assembling peptide hydrogel scaffold for meniscal defect: An in vivo study in a rabbit model

The inner avascular zone of the meniscus has limited healing capacity as the area is poorly vascularized. Although peptide hydrogels have been reported to regenerate bone and cartilage, their effect on meniscus regeneration remains unknown. We tested whether the self‐assembling peptide hydrogel scaffold KI24RGDS stays in the meniscal lesion and facilitates meniscal repair and regeneration in an induced rabbit meniscal defect model. Full‐thickness (2.0 mm diameter) cylindrical defects were introduced into the inner avascular zones of the anterior portions of the medial menisci of rabbit knees (n = 40). Right knee defects were left empty (control group) while the left knee defects were transplanted with peptide hydrogel (KI24RGDS group). Macroscopic meniscus scores were significantly higher in the KI24RGDS group than in the control group at 2, 4, and 8 weeks after surgery. Histological examinations including quantitative and qualitative scores indicated that compared with the control group, the reparative tissue in the meniscus was significantly enhanced in the KI24RGDS group at 2, 4, 8, and 12 weeks after surgery. Immunohistochemical staining showed that the reparative tissue induced by KI24RGDS at 12 weeks postimplantation was positive for Type I and II collagen. KI24RGDS is highly biocompatible and biodegradable, with strong stiffness, and a three dimensional structure mimicking native extracellular matrix and RGDS sequences that enhance cell adhesion and proliferation. This in vivo study demonstrated that KI24RGDS remained in the meniscal lesion and facilitated the repair and regeneration in a rabbit meniscal defect model.     

Effects of abrasion therapy on tears in the avascular region of sheep menisci

This study was designed to test the hypothesis that abrasion of the parameniscal synovium aids healing of a stable tear in the avascular region of the meniscus in a sheep model. In six sheep, a 5-7-mm longitudinal full-thickness tear was made in the avascular inner half of the anterior part of the lateral meniscus. The parameniscal synovium was abraded superiorly and inferiorly from the meniscus periphery to the lesion. Three animals in a control group received identical meniscal tears but no abrasion treatment. A harness prevented weight bearing and maintained the knee fully flexed after surgery. Twelve weeks after the operation, no healing was seen in any tears. Histologic examination revealed in both groups increased numbers of dividing chondrocytes on either side of the tear. In the test group, several layers of fibroblasts, which appeared in two menisci to derive from the upper meniscal surface, were seen covering both cut surfaces of the tear. It is concluded that the distance from the periphery to the defect is too far for abrasion therapy to stimulate sufficient cellular ingrowth to facilitate repair of tears in the avascular region of the meniscus.     

Expression of cytokines after meniscal rasping to promote meniscal healing

Purpose: We have developed a new surgical procedure of rasping a meniscal surface to repair a tear in the avascular zone. This procedure stimulates vascular induction to the tear, resulting in meniscal healing. The purpose of this study was to elucidate the mechanism of vascular induction and meniscal healing. Type of Study: Randomized trial. Methods: A full-thickness longitudinal tear of 5 mm in length was created in the avascular zone of the anterior segment of both medial menisci of rabbits. Meniscal rasping of about 0.5 mm in depth was then done on the femoral surface of the left meniscus from the parameniscal synovium to the inner portion including the longitudinal tear, and the right meniscus was left untreated as a control. After surgery, at 1, 7, 14, 28, 56, and 112 days, 4 rabbits were killed, both medial menisci were resected, and immunohistochemical staining with monoclonal antibodies was used to quantify expression of interleukin-1α (IL-1α), transforming growth factor-β1 (TGF-β1), platelet-derived growth factor (PDGF), and proliferating-cell nuclear antigen (PCNA) on the femoral surface of the menisci. A positive ratio of immunostaining was encountered. Results: The positive ratio of IL-1α, TGF-β1, PDGF, and PCNA on the rasped surface area reached its peak at 1, 7, 14, and 7 days, respectively, after surgery, and thereafter gradually declined. Although the time course of the positive ratio was different among these cytokines, the positive ratio on the rasped surface was significantly higher than that on the control surface at the early stage of the observation period. Conclusions: The cytokine network on the rasped meniscal surface appears to be the key to explaining the mechanism of vascular induction and meniscal healing by meniscal rasping.Arthroscopy: The Journal of Arthroscopic and Related Surgery, Vol 17, No 7 (September), 2001: pp 724–731     

Meniscal repair enhanced by an interpositional free synovial autograft: An experimental study in rabbits

This report describes the methods and effectiveness of interpositional free synovial autografts (IPFSAs) in promoting the healing of lesions in the avascular portion of the knee joint meniscus in rabbits. Thirty-four specimens were divided into two groups: 17 in the study group and 17 in the control group. The medial meniscus of the left knee joint was excised in each rabbit, and an artificial longitudinal tear was created in the avascular zone of the meniscus. In the study group, an IPFSA was sewn into the tear with a single horizontal suture. In the control group, the tear was repaired without synovium in the same manner. The menisci were then implanted into the right knee joint in the respective animals. At intervals the animals were killed, and the menisci were examined grossly and microscopically. Three menisci were eliminated because they had become attached to the synovial wall. In the study group, the lesion completely healed by 4 weeks, except for one. The menisci in the control group never completely healed, particularly at the deeper levels of the lesion. At 8–16 weeks, autolysis of the specimens made microscopic examination difficult and unreliable in both groups. Based on these results, it has been concluded that an IPFSA can promote the healing process in the avascular zone of a torn meniscus in rabbits and that systemic vascularity to the synovium or the meniscus is not essential for healing to occur.     

前十字韧带断裂继发半月板损害的临床研究

目的研究前十字韧带(anteriorcruciateligament,ACL)断裂对半月板的影响。方法回顾分析1984年12月～1999年12月间收治的419例ACL断裂患者半月板的损伤情况及其与软骨损伤的关系。结果外侧半月板的损伤率随病程增加无显著变化,而内侧半月板的损伤率随病程增加显著增加,由急性期的31.1%增至亚慢性期的48.2%(P<0.01),又增至慢性期的78.8%(P<0.001)。内侧半月板后角损伤率较前角高,差异有显著性意义(P<0.05)。损伤形态以纵裂最常见,随着病程的增加,半月板损伤也越发复杂。内侧半月板损伤患者的内髁软骨损伤的发生率要高于内侧半月板正常者的内髁软骨损伤发生率,但差异无显著性意义(P>0.05);而外侧半月板损伤患者的外髁软骨损伤的发生率却显著高于外侧半月板正常者,差异有非常显著性意义(P<0.01)。结论ACL断裂可伴发和继发半月板的损害,ACL断裂时伴发的多为外侧半月板的损伤,而继发的半月板损害却以内侧为重。内髁软骨损害主要由股胫关节前后向不稳、异常活动增加造成,而与内侧半月板的损伤关系不大。     

Vascular response of the meniscus to injury: effects of immobilization.

Failed meniscal healing may lead to degenerative osteoarthritis of the knee. Healing is thought to be dependent upon an adequate blood supply, yet "normal" vascular changes during healing are not well understood. In this study we have quantified vasoactive and angiogenic responses to medial meniscal injury in a rabbit model under clinically relevant conditions, and related these to histological criteria of healing. Twenty-six adult rabbits were given a standardized meniscal injury; 12 of these had the hind limb immobilized by pinning. Eight normal controls and 12 sham-operated animals were also studied. After 4 weeks, animals underwent either vascular volume (vascular index) determination, or blood flow measurement using coloured microspheres. Histological analysis was also performed to assess meniscal healing. In injured animals, blood flow to the menisci was increased fivefold 4 weeks post-operative; this increase was prevented by immobilization. The vascular index of the menisci was also increased threefold by injury, but not significantly reduced by immobilization. Histological examination of injured menisci showed examples of healing and non-healing tears in both mobile and immobile groups. Meniscal injuries are associated with characteristic changes in vascularity and perfusion, and these changes likely play a significant role in the healing process. Characterization of the vascular responses to meniscal injury may lead to techniques that can promote reliable healing of meniscal tears and thereby improve clinical outcomes.