不同来源间充质干细胞在膝关节软骨修复中的临床研究进展

近年来由于器官修复干细胞治疗技术的提出,使得不能自身修复的关节软骨成为研究热点之一,间充质干细胞对膝关节软骨修复已表现出明显的治疗优势。学者们临床上初步研究了不同来源的间充质干细胞膝关节软骨修复作用,并通过结合移植、软骨组织工程等技术提高了骨髓、脂肪、滑膜、脐带血等来源干细胞修复人类软骨效果,均取得了较好的临床疗效。不同干细胞来源不同各自会有一定的优缺点。目前临床研究仍处于试验阶段,尚无确切定论何种干细胞及何种技术处理最适合于人类软骨的修复。大规模和或结合新处理技术的临床试验以及远期疗效的验证,需要进一步临床深入研究。     

MicroRNAs在骨髓间充质干细胞成软骨分化中的作用

软骨组织由细胞外基质与分散其间的软骨细胞共同构成，由于缺乏血管?神经和淋巴系统，损伤后自身修复能力差。目前各种促进软骨损伤修复的方法效果都不理想，诱导骨髓间充质干细胞向软骨细胞分化修复软骨损伤已成为当下研究的热点。多种MicroRNA参与并调控骨髓间充质干细胞成软骨分化过程，本文就MicroRNA调控骨髓间充质干细胞成软骨分化及其机制的研究进展做一综述。     

脂肪间充质干细胞治疗膝关节骨关节炎应用进展

膝关节骨关节炎(KOA)的特征是关节透明软骨局灶性丧失,滑膜组织炎症伴骨赘形成.目前KOA的治疗方法虽可在一定程度上减轻患者疼痛并改善关节活动度,但不能促进受损关节的软骨再生.脂肪间充质干细胞作为种子细胞,可用于KOA治疗中修复受损的关节软骨,为KOA治疗提供了一种新思路.该文对脂肪间充质干细胞治疗KOA的研究进展进行...     

间充质干细胞成软骨分化相关研究进展

软骨细胞移植疗法已成功应用于软骨损伤修复和软骨组织工程研究,但软骨细胞取材有限,因此探索软骨细胞替代物成为研究热点。间充质干细胞具有分化为骨、软骨等组织的潜能,在软骨损伤、骨关节炎修复及软骨组织工程等方面起着重要作用。该文就间充质干细胞成软骨分化相关研究进展作一综述。     

关节液来源间充质干细胞在组织工程中应用研究进展

间充质干细胞是治疗软骨损伤的新方法,间充质干细胞的来源选择是目前研究重点。关节液来源间充质干细胞(SF-MSC)是从关节液中分离而来,具有间充质干细胞的生物学特性。SF-MSC易于获取,与脂肪和骨髓来源的间充质干细胞相比,其具有更强的软骨分化能力,被认为是治疗软骨损伤的理想种子细胞。体内研究证实,SF-MSC复合支架材料或关节内注射SF-MSC对修复软骨损伤有很好的效果。目前研究表明,SF-MSC有望成为治疗软骨损伤的候选间充质干细胞。该文就SF-MSC在组织工程中应用的研究进展作一综述。     

慢病毒介导的Sox9基因在兔骨髓间充质干细胞的过表达促进软骨损伤修复

目的:明确在体内Sox9基因过表达的兔骨髓间充质干细胞对于关节软骨损伤修复的作用。方法:以慢病毒介导的Sox9基因转染兔骨髓间充质干细胞(BMSCs),体外检测软骨特异性分子,将新西兰大白兔24只48个膝关节随机分为3组,动物麻醉后,双侧股骨滑车处的关节面上用直径4 mm的钻头钻孔,深度3 mm,穿透软骨下骨,造成全层关节软骨损伤,将转染后的细胞植入体内用以修复全层关节软骨损伤,实验组植入BMSCs-(Lenti-Sox9-EGFP)-藻酸钙复合物,实验对照组植入BMSCs-藻酸钙复合物,空白对照组只钻孔。术后6、12周分别进行光镜、电镜观察,以及HE、免疫组织化学染色检测软骨的修复程度。结果:经Sox9基因转染后的细胞在3 d时,Sox9基因表达最高,随后下降。转染后3 d,Ⅱ型胶原开始表达,到14 d时达到最高。表明Sox9过表达启动了兔骨髓间充质干细胞的软骨分化。组织学观察显示,实验组术后6周缺损处有透明软骨样组织填充,术后12周缺损处软骨和软骨下骨修复良好。两对照组,缺损处由纤维组织填充。免疫组织化学显示,修复组织内Ⅱ型胶原,免疫组化染色结果阳性强于两对照组。组织学评分结果显示实验组软骨损伤修复各时间点效果明显优于两对照组,差异有统计学意义。结论:Sox9基因过表达的兔骨髓间充质干细胞(BMSCs)促进软骨损伤的修复。     

膝关节骨关节炎的保膝治疗进展

目的基于遵循阶梯性治疗理念,对膝关节骨关节炎(knee osteoarthritis,KOA)保膝治疗进展作一综述。方法查阅近年国内外相关文献,对KOA各阶段治疗方法进行总结分析。结果 KOA的治疗旨在减轻症状、延缓关节结构性改变、维持关节功能和提高生活质量,因此保守治疗仍是早期KOA的首选治疗方法。关节镜手术可协助诊断和分级,并同期行关节内刺激物清理和软骨的有限修复。截骨术适用于下肢负重力线、关节线异常的KOA,能有效平衡膝关节负重应力,改善症状,缓解疾病进展。关节牵伸术治疗KOA能改善膝关节力学环境,一定程度修复缺损软骨,缓解症状。随着技术进步,近年来单髁关节置换(unicompartmental knee arthroplasty,UKA)重新兴起,UKA属于真正的关节表面置换,是前内侧KOA患者保膝治疗的重要手段。结论目前KOA保膝治疗方法较多,主要针对患者病程以及病情严重程度进行选择。截骨术和UKA是临床应用最多且保膝成功率高的重要外科技术,但应注意恰当选择适应证,术者需积累足够临床经验,以取得良好效果。     

膝关节骨关节炎的保膝治疗进展北大核心CSCD

目的基于遵循阶梯性治疗理念,对膝关节骨关节炎(knee osteoarthritis,KOA)保膝治疗进展作一综述。方法查阅近年国内外相关文献,对KOA各阶段治疗方法进行总结分析。结果 KOA的治疗旨在减轻症状、延缓关节结构性改变、维持关节功能和提高生活质量,因此保守治疗仍是早期KOA的首选治疗方法。关节镜手术可协助诊断和分级,并同期行关节内刺激物清理和软骨的有限修复。截骨术适用于下肢负重力线、关节线异常的KOA,能有效平衡膝关节负重应力,改善症状,缓解疾病进展。关节牵伸术治疗KOA能改善膝关节力学环境,一定程度修复缺损软骨,缓解症状。随着技术进步,近年来单髁关节置换(unicompartmental knee arthroplasty,UKA)重新兴起,UKA属于真正的关节表面置换,是前内侧KOA患者保膝治疗的重要手段。结论目前KOA保膝治疗方法较多,主要针对患者病程以及病情严重程度进行选择。截骨术和UKA是临床应用最多且保膝成功率高的重要外科技术,但应注意恰当选择适应证,术者需积累足够临床经验,以取得良好效果。     

基于膝关节MRI的人工智能重建软骨模型对膝关节软骨损伤的评价

目的探讨基于膝关节薄层MRI数据的人工智能(artificial intelligence, AI)重建模型对膝关节软骨损伤评价的可行性。方法选取2021年5月至2022年4月在北京清华长庚医院以膝关节重度骨关节炎住院且拟行全膝关节置换术的33例患者(共41膝), 男15例, 年龄(71±5)岁;女26例, 年龄(71±9)岁。左膝19例, 右膝22例。术前对患侧膝关节进行薄层MR检查, 并对膝关节薄层MRI数据进行AI建模, 选取模型中软骨部分利用主成分分析(principal component analysis, PCA)进行模型摆正, 将术中截取的膝关节胫骨平台软骨依据国际软骨修复协会(International Cartilage Repair Society, ICRS)软骨损伤分级进行分级, 并与膝关节AI重建软骨模型及膝关节MRI人工识别的ICRS分级结果进行比较。结果 AI重建软骨模型的软骨损伤分级与术中截取实物标本的软骨损伤分级相比较, AI重建软骨模型对于ICRS分级4级软骨损伤诊断的敏感性、特异性、阳性预测值和阴性预测值分别为93.1%、91.4%、92.2%和...     

骨髓间充质干细胞成软骨分化的影响因素研究进展

软骨组织工程的研究发展为关节软骨损伤的修复带来了新的希望.骨髓间充质干细胞(BMSCs)具有分化为骨、软骨、肌腱、脂肪等组织的多分化潜能[1],也是目前软骨组织工程的研究热点[2].BMSCs在修复软骨损伤时,无论在体内体外,都须经历向软骨细胞分化的过程,在此过程中有着诸多影响因素,现就目前研究影响BMSCs向软骨细胞分化的文献进行综述分析如下.     

关节镜治疗膝关节骨性关节炎78例临床分析

目的:总结关节镜下行关节清理术治疗膝关节骨性关节炎的临床效果与应用价值。方法:回顾分析2010年4月至2013年2月局麻下为78例膝关节骨性关节炎患者经关节镜行关节清理术的临床资料,其中男29例,女49例;平均(61±10)岁;关节镜下关节清理术包括刨削清理突入髌股关节、胫股关节及髁间窝内增生肥厚的炎性滑膜组织,修整老化退变、破损的半月板,解除交锁因素,清除剥脱的软骨碎屑,修平缺损的软骨边缘,松解关节腔内的炎性索带及粘连带,切断滑膜皱襞,清理髌下脂肪,去除关节腔内的游离体。术后随访患侧膝关节的功能状态,患者日常活动能力及对手术的满意度。结果:手术时间平均(50±10)min,术中出血量极少。术后平均随访(15±9)个月,78例患者中优48例,良23例,差7例,优良率达91%。结论:局麻下经关节镜行关节清理术治疗膝关节骨性关节炎可有效解除临床症状,改善功能,延缓病变发展,严格把握手术适应证可取得满意疗效。     

Human mesenchymal stem cells in synovial fluid increase in the knee with degenerated cartilage and osteoarthritis

We investigated whether mesenchymal stem cells (MSCs) in synovial fluid (SF) increased in the knee with degenerated cartilage and osteoarthritis. SF was obtained from the knee joints of 22 patients with anterior cruciate ligament (ACL) injury during ACL reconstruction, and cartilage degeneration was evaluated arthroscopically. SF was also obtained from the knee joints of 6 healthy volunteers, 20 patients with mild osteoarthritis, and 26 patients with severe osteoarthritis, in which the grading was evaluated radiographically. The cell component in the SF was cultured for analyses. Synovium (SYN) and bone marrow (BM) were also harvested during total knee arthroplasties. The MSC number in SF was correlated with the cartilage degeneration score evaluated by arthroscopy. The MSC number in the SF was hardly noticed in normal volunteers, but it increased in accordance with the grading of osteoarthritis. Though no significant differences were observed regarding surface epitopes, or differentiation potentials, the morphology and gene profiles in SF MSCs were more similar to those in SYN MSCs than in BM MSCs. We listed 20 genes which were expressed higher in both SYN MSCs and SF MSCs than in BM MSCs, and 3 genes were confirmed by quantitative RT-PCR. MSCs in SF increased along with degenerated cartilage and osteoarthritis.     

骨关节炎患者滑液中COMP水平与病情严重程度的相关性研究

背景：软骨寡聚基质蛋白（cartilage oligomeric matrix protein,COMP）是软骨中非胶原蛋白的主要成分,软骨的损伤、修复和代谢变化均可能影响COMP的表达水平。目的：研究骨关节炎（osteoarthritis,OA）患者关节滑液中COMP水平与病变严重程度的相关性,探讨注射玻璃酸钠对关节滑液中COMP的影响。方法：52例膝关节0A患者接受关节内注射玻璃酸钠治疗,治疗前、治疗后6个月行X线片检查,按Kellgren放射学诊断标准评级,记录治疗前和治疗开始后5周、6个月时患者膝关节的WOMAC评分。采用ELISA方法测定在接受透明质酸钠治疗前、治疗开始后5周关节液中的COMP水平。19例因半月板或韧带损伤接受关节镜手术的患者作为正常对照组。结果：OA组患者滑液中COMP水平明显高于对照组,有统计学差异（P=0．036）。OA组患者滑液中COMP水平与OA严重程度（WOMAC评分）呈正相关（P=0．001）,与影像学Kellgren分级标准无相关性（P=0．12）。结论：滑液中COMP水平与OA病变严重程度呈正相关,测定OA患者血中COMP水平对进一步研究OA发病机理、早期发现并采取有效防治策略及监测防治效果具有极其重要的意义。     

手术范围对膝关节术后纤维化形成的影响

膝关节纤维化是术后常见的并发症,为影响手术疗效的关键因素之一,常引起关节活动度降低、慢性关节痛、软骨损伤以及关节废用性病变,严重降低了患者的生活质量。因此,如何降低术后膝关节纤维化的发生是提高手术预期疗效的关键,也是临床医学上需要亟待解决的重要问题。笔者团队结合自身前期及最新的研究发现,膝关节纤维化的程度与手术范围大小有重要的关联,利用微创和关节镜方法治疗膝关节三室内的疾病可有效减少滑膜、软骨和脂肪垫组织的损伤,同时结合术后早期康复训练是未来预防术后膝关节纤维化的有效手段。     

Transcriptomic analysis of synovial extracellular RNA following knee trauma: A pilot study

Traumatic knee injuries often result in damage to articular cartilage and other joint structures. Such trauma is a strong risk factor for the future development and progression of osteoarthritis (OA). The molecular mechanisms and signaling pathways modulating response to knee joint trauma remain unclear. Moreover, investigations of biomarkers influencing responses have been targeted rather than broad, unbiased discovery studies. Herein, we characterize the complete complement of extracellular RNA (exRNA) in the synovial fluid of 14 subjects following knee injury. Fluid was collected during surgery from the injured knees, and from the contralateral knee in a subset, undergoing surgical repair of the ACL and/or meniscal repair/debridement. Arthroscopic grading of chondral damage in four knee compartments was performed using the Outerbridge classification. exRNA was extracted and subjected to massively parallel total RNA sequencing. Differential abundance of RNA was calculated between the subject cohorts of injured and non‐injured knee, average Outerbridge score ≥0.5 and less, and chronic and acute injury duration defined as ≤4 months till surgery or longer. Overall, expression of several thousand genes was identified in the synovial fluid. Furthermore, differential expression analysis suggests a role of exRNA fragments of matrix metalloproteinases and skeletal muscle fiber genes in the response to traumatic injury. Together, these data suggest that high‐throughput approaches can indicate exRNA molecular signatures following knee trauma. Future studies are required to more fully characterize the biological roles of these exRNA and the cadence of their respective release that may lead to translational treatment options for post‐traumatic OA. © 2017 Orthopaedic Research Society. Published by Wiley Periodicals, Inc. J Orthop Res 36:1659–1665, 2018.

     

New perspectives for articular cartilage repair treatment through tissue engineering: A contemporary review

In this paper review we describe benefits and disadvantages of the established methods of cartilage regeneration that seem to have a better long-term effectiveness. We illustrated the anatomical aspect of the knee joint cartilage, the current state of cartilage tissue engineering, through mesenchymal stem cells and biomaterials, and in conclusion we provide a short overview on the rehabilitation after articular cartilage repair procedures. Adult articular cartilage has low capacity to repair itself, and thus even minor injuries may lead to progressive damage and osteoarthritic joint degeneration, resulting in significant pain and disability. Numerous efforts have been made to develop tissue-engineered grafts or patches to repair focal chondral and osteochondral defects, and to date several researchers aim to implement clinical application of cell-based therapies for cartilage repair. A literature review was conducted on PubMed, Scopus and Google Scholar using appropriate keywords, examining the current literature on the well-known tissue engineering methods for the treatment of knee osteoarthritis.     

Effective repair of a fresh osteochondral defect in the rabbit knee joint by articulated joint distraction following subchondral drilling.

PURPOSE: Joint distraction has been used to treat osteoarthritis and was found to delay the need for arthrodesis or joint replacement. However, there has been little basic research on articulated joint distraction for the repair of osteochondral defects. We investigated the effects of joint distraction with motion after drilling on a fresh osteochondral defect in the weight bearing area of the rabbit knee joint. METHODS: A full thickness osteochondral defect was created in the weight bearing area of both medial femoral condyles of an adult Japanese white rabbit. After drilling of the defect, the experimental knee joint was distracted for 1.5 mm using a pair of external fixators to decrease compression force. The contralateral knee joint was used as a control with no apparatus. Gross findings and histological evaluation were assessed to study morphology of the repaired cartilage. RESULTS: A partial repair with cartilage-like tissue was observed in the joints of the experimental group at 4 weeks. While cartilage-like tissue stained with Safranin O was found in the experimental group at 8 and 12 weeks, destructive changes were observed in the control joints. Morphological changes were evaluated using the histological grading scale [Wakitani S, Goto T, Pineda SJ, et al. Mesenchymal cell-based repair of large, full-thickness defect of articular cartilage. J Bone Joint Surg Am 1994;76(4):579-92]. There was no significant difference between experimental and control groups at 4 weeks (mean 11.2 and 13.8 points, respectively). However, the mean scores of the experimental groups at 8 and 12 weeks (mean 6.8 and 7.5, respectively) were significantly better than those of the control groups at the same time points (mean 14 points each). Between the experimental groups, the scores at 8 and 12 weeks were both significantly better than those at 4 weeks. CONCLUSION: A combination of subchondral drilling, joint motion and distraction by an articulated external fixator promoted repair of a fresh osteochondral defect in the weight bearing area. Although distraction for 4 weeks was not a long enough period to repair the defect, distraction for 8 and 12 weeks resulted in a good outcome.     

富血小板血浆联合间充质干细胞治疗膝骨关节炎的疗效

目的观察关节腔内联合注射富血小板血浆与脐带间充质干细胞治疗轻中度膝关节骨关节炎患者的近期疗效。 方法收集聊城市人民医院膝关节骨关节炎患者200例,纳入标准：Kellgren-Lawrence分级1～3级膝关节骨关节炎患者;排除标准：Kellgren-Lawrence分级4级骨关节炎患者及炎症性关节炎患者等。根据电脑产生的伪随机数字,将研究对象随机分为富血小板血浆(PRP)与脐带间充质干细胞(MSC)联合注射组(PRP＋MSC)、单纯PRP注射组(PRP组)、单纯脐带MSC注射组(MSC组)、玻璃酸钠(SH)注射组(SH组),每组各50例。PRP ＋ MSC组关节腔内注射5 ml富血小板血浆与脐带间充质干细胞混合液;PRP组关节腔内注射5 ml富血小板血浆;MSC组关节腔内注射5 ml脐带间充质干细胞;SH组关节腔内注射2.5 ml的SH及曲安奈德注射液。分别在治疗后1、3、6、12个月进行随访,通过视觉疼痛模拟评分(VAS),美国膝关节协会评分( AKS )评价治疗后膝关节功能的改善情况;采用酶联免疫吸附法检测治疗前后关节液中细胞因子的变化情况;利用MRI的T2 Map序列评价治疗前后关节软骨的变化情况。患者年龄、病程、VAS评分、AKS评分、细胞因子含量等计数资料分析采用单因素方法分析,患者性别比例、病情程度等计数资料分析采用卡方检验。 结果200例患者中,175例完成随访,PRP ＋ MSC组45例,PRP组44例,MSC组43例,SH组43例。4组患者治疗前VAS、AKS及关节液中细胞因子的表达无统计学意义(P >0.05)。治疗后1个月,4组患者的VAS评分均较前降低,AKS评分较前升高,组内差异具有统计学意义(t ＝6.45,P <0.01),组间差异无统计学意义(P >0.05)。3个月后,SH组VAS、AKS趋向于治疗前状态,其余3组未见反弹,PRP ＋ MSC联合注射组优于单纯PRP与MSC治疗组,4组间的差异具有统计学意义(F ＝7.84/4.35,P <0.01)。治疗后3周,SH组关节液中细胞因子未见明显改变,PRP ＋ MSC组关节液中细胞因子变化水平高于单纯PRP与MSC组,差异具有统计学意义(F ＝39.21,P <0.01)。治疗后6个月,MRI的T₂Map序列显示PRP＋MSC组、PRP组、MSC组、SH组的平均软骨T2值分别为(54.07±2.64)ms、(56.59±1.40)ms、(56.40±1.57)ms、(57.98±1.95)ms,差异具有统计学意义(F＝30.63,P<0.05)。 结论关节腔内联合注射富血小板血浆与脐带间充质干细胞明显改善膝关节骨关节炎患者的症状,延缓骨关节炎的进展。     

膝骨性关节炎合并骨质疏松症调节机制的研究进展

膝骨性关节炎和骨质疏松症都是与年龄增长相关的骨与关节退行性疾病,两者在发病机制、病理变化、调节机制、临床治疗等方面存在着诸多的共同点,在膝骨关节炎和骨质疏松症的进程中通过复杂的调节机制维持骨代谢平衡、减轻炎症、保护关节软骨、稳定膝关节,进而延缓膝关节的退变;虽然两者间复杂的调节机制已有研究,但是仍然不完善。本研究从炎症、生物力学、激素与骨代谢、遗传、蛋白等方面对膝骨性关节炎并骨质疏松症的调节机制进行了综述,以期为临床防治该病提供新的思路和理论依据。     

膝关节骨关节炎滑液和软骨骨桥蛋白水平与其病变程度的相关性

目的 探讨膝关节骨关节炎(osteoarthritis,OA)患者滑液和关节软骨骨桥蛋白(osteopontin,OPN)水平及其与病变严重程度的关系.方法 随机选取50例膝关节OA患者[男15例,女35例;年龄48～81岁,平均(61.8±7.4)岁]和10名健康对照者[男4例,女6例;年龄59～68岁,平均年龄(63.2±6.0)岁]作为研究对象.采用Mankin评分评价疾病严重程度,Kellgren-Lawrence标准进行放射学分级,酶联免疫吸附法测定关节滑液OPN水平,免疫组化方法测定关节软骨OPN光密度值.结果 OA患者与对照者相比,关节滑液OPN水平[(4519.60±1830.37)pg/ml:(1179.70±303.39)pg/ml)和关节软骨OPN光密度值[(0.60±-0.06):(0.43±0.07)]均明显升高.关节滑液OPN水平与关节软骨OPN表达呈正相关(r=0.411,P=0.003).关节滑液OPN水平与OA病变严重程度(KL分级)呈正相关(r=0.581,P＜0.001).关节软骨OPN表达与OA病变严重程度(Mankin评分)呈正相关(r=0.675,P＜0.001).结论 关节滑液和关节软骨OPN水平与病变严重程度相关.