利用脂肪干细胞构建组织工程软骨修复兔膝关节软骨缺损

目的探讨以脂肪于细胞（ADSCs）复合脱细胞软骨基质支架构建组织工程软骨修复兔膝关节软骨缺损的效果。方法以人关节软骨脱细胞基质为支架，复合经诱导的兔ADSCs，体外分别经静态培养和生物反应器培养，构建组织工程软骨。对膝全厚关节缺损进行修复，并与单支架组、空白对照组比较，其中空白对照组12个关节，脱细胞软骨支架组16个关节，静态培养细胞支架组24个关节，生物反应器培养细胞支架组8个关节。分别于术后3、6个月对修复关节进行大体、组织学及免疫组化观察。结果实际完成观察的关节数为44个，其中空白对照组9个，脱细胞软骨支架组11个，静态培养细胞支架组18个，生物反应器培养细胞支架组6个。空白对照组全为纤维组织或纤维软骨样修复；单支架组5个关节为未成熟透明软骨，无成熟透明软骨形成；静态培养细胞支架组83．3％为透明软骨，其中3个关节为成熟透明软骨，12个关节为未成熟透明软骨；生物反应器培养细胞支架组100％为透明软骨，其中2个为成熟透明软骨，4个为未成熟透明软骨。Wakitani评分各组差异有统计学意义（P〈0．05）。结论ADSCs复合脱细胞软骨基质支架能良好地修复兔膝关节全厚软骨缺损，应用生物反应器技术有助于构建组织工程软骨，促进软骨缺损的修复。     

脱细胞软骨支架材料修复兔关节软骨缺损

目的 观察异种异体脱细胞软骨支架材料(ACM)复合同种异体兔骨髓间充质干细胞(rBMSCs)修复兔股骨内髁关节软骨缺损的效果.方法 (1)密度梯度离心和差速贴壁法获得原代兔BMSCs,选择第3代BMSCs作为种子细胞;(2)利用冷冻干燥、胰酶消化和化学去垢剂等方法制备脱细胞软骨支架材料;(3)3个月龄新西兰兔股骨内髁制备直径4 mm,深3 mm砌关节软骨缺损模型,24只新西兰兔以2个时间段随机分为3组,Ⅰ ACM-BMSCs组:第3代BMSCs 1×106个/ml与ACM于37℃5%CO2饱和湿度复合48 h;Ⅱ ACM组;Ⅲ空白对照组.(4)移植6、12周后大体及组织学观察,免疫组织化学染色观察修复组织Ⅱ型胶原,Wakitani评分评估修复效果.结果 (1)大体观察及组织学观察:6和12周Ⅰ组再生组织与正常关节软骨面平齐,修复部位表面较平整,界限模糊,接近正常软骨.Ⅱ组修复组织表面不平整并有明显下陷,修复组织全层可见成纤维样细胞,深层可见极少数透明软骨样细胞.Ⅲ组未见明显修复,肉芽组织形成伴成纤维样细胞增生;(2)Wakitani组织学评分可见在不同的时间段I组和Ⅱ组均低于Ⅲ组,差异有统计学意义(P<0.05),Ⅰ组和Ⅱ组间组织学评分差异无统计学意义(P>0.05).(3)免疫组织化学:ACM-BMSCs组修复组织的细胞为软骨样细胞,可见柱状排列,周围软骨基质Ⅱ型胶原染色阳性.结论 以ACM为支架材料,同种异体BMSCs为种子细胞制备的组织工程化软骨对兔股骨内髁关节软骨缺损有修复作用,形成的新生软骨为透明软骨样组织.     

兔骨髓间充质干细胞修复膝关节软骨缺损的实验研究

目的研究由同种异体兔骨髓间充质干细胞(MSCs)诱导分化的软骨细胞和聚乳酸-聚乙醇酸共聚物(PLGA)双层支架构建复合体对兔软骨缺损的修复作用。方法用密度梯度离心法和贴壁培养法获得5月龄兔骨髓来源间充质干细胞,在体外培养并进行分化诱导后作为种子细胞复合双层PLGA构建成复合体。36只兔在股骨髁间制造骨软骨缺损模型,分成三组,A组植入复合体,B组植入单纯双层PLGA支架,C组植入自体骨软骨。第24周取材进行大体观察、组织学检查和Wakitani评分。结果 A组及C组植入物愈合良好,组织学检查见Ⅰ、Ⅱ型胶原纤维形成。A组可见透明软骨修复。Wakitani评分A组2.75,B组7.00,C组1.98,A、C组与B组间统计学分析单项指标得分差异有统计学意义(P0.05),A组与C组间差异无统计学意义(P0.05)。结论诱导兔MSCs+PLGA双层支架能较好地修复兔膝关节骨软骨损伤。     

Ⅰ型胶原负载骨髓基质细胞修复兔膝关节软骨缺损

目的探讨同种异体骨髓基质细胞(MSCs)-Ⅰ型胶原复合移植修复关节软骨缺损的疗效。方法构建Ⅰ型胶原支架,将体外培养的兔MSCs吸附于该支架上培养,检测支架对MSCs的吸附及增殖的影响,再移植于兔膝关节全层软骨缺损处,分批取材,进行大体、组织学观察。结果MSCs在Ⅰ型胶原支架内贴附良好,分泌胞外基质,MSCs-胶原复合移植组软骨缺损处在术后12周为透明软骨样修复,空白对照组为纤维瘢痕样修复。结论Ⅰ型胶原支架与MSCs相容性良好,MSCs-胶原复合移植可达到透明软骨样修复软骨缺损。     

关节软骨源性多孔支架复合自体软骨细胞修复兔关节软骨缺损的长期疗效观察

[目的]对关节软骨源性多孔支架复合自体软骨细胞复合体修复兔膝关节软骨缺损的长期效果进行观察和评价。[方法]实验动物新西兰大白兔共10只,分两组:(1)关节软骨源性支架对照组:缺损内置入关节软骨源性支架;(2)关节软骨源性支架复合细胞组:缺损内置入关节软骨源性支架-自体软骨细胞复合体。手术后15个月取材做大体摄像,甲醛固定、10%EDTA脱钙,石蜡切片,采用四种组织化学染色法(HE、奥新兰(AB)、甲苯胺兰(TO)、藩红花"O"染色)和Ⅱ型胶原免疫组化染色,观察修复后的兔关节软骨组织细胞形态特征。[结果]关节软骨源性支架对照组,大体观察关节表面有凹陷。石蜡切片HE染色见未修复缺损处为梭形细胞,纤维软骨,缺损处AB、甲苯胺兰、藩红花"O"染色为阴性,Ⅱ型胶原染色为阳性;关节软骨源性支架复合细胞组,大体见关节软骨表面修复平整、光滑。常规HE染色镜下见软骨全层修复良好,为透明软骨细胞,可见软骨陷窝潮线排列结构。细胞外基质特染:AB、甲苯胺兰、藩红花"O",Ⅱ型胶原特种染色均为阳性。[结论]组织学观察结果发现,关节软骨源性支架组为纤维软骨组织结构,未能完全修复膝关节软骨缺损;而关节软骨源性支架复合细胞组关节软骨缺损修复良好,未见退变,具有软骨组织的特征。     

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。     

纳米左旋聚乳酸/聚己内酯复合骨髓基质源性软骨细胞修复犬膝关节软骨缺损

目的探讨纳米左旋聚乳酸/聚己内酯(纳米PLLA-b-PCL)复合骨髓基质源性软骨细胞修复犬膝关节软骨缺损的可行性。方法将纳米PLLA-b-PCL支架/骨髓基质源性软骨细胞复合物植入犬膝关节软骨缺损,其为实验组;另一组膝关节造成缺损后旷置,作为空白对照。术后12周,24周取材,行大体及组织学观察,组织学评分。结果术后12周、24周大体形态学和组织学观察均表明实验组得到较好修复,而对照组缺损未修复。结论骨髓基质细胞源性软骨细胞是修复关节软骨缺损较理想的种子细胞;纳米PLLA-b-PCL在新生软骨形成的同时,逐渐降解吸收,是组织工程修复关节软骨缺损的适宜支架材料,其具有良好的应用前景。     

兔骨髓间充质干细胞复合纳米羟基磷灰石/左旋聚乳酸修复兔软骨缺损的实验研究

目的评价兔骨髓间充质干细胞复合纳米羟基磷灰石/左旋聚乳酸(Nano-HA/PLLA)修复兔膝关节全层软骨缺损的效果。方法体外分离培养兔骨髓间充质干细胞(rBMSCs),取第3代将其与Nano-HA/PLLA在体外构建组织工程软骨(tissue engineering cartilage,TEC)。将18只新西兰大白兔股骨内髁关节面造一直径4.5 mm、深度5 mm的软骨缺损模型,随机分成3组,即实验组、支架组和空白对照组,每组6只,分别予复合细胞的Nano-HA/PLLA、Nano-HA/PLLA及空白填充。术后12、24周取材行大体标本、组织学、免疫组织化学观察及Wakitani组织学评分。结果实验组显示缺损有骨软骨组织形成,软骨下骨基本达到生理整合,组织学及免疫组化显示有大量细胞外基质形成;支架组及对照组显示出有限的再生重建。以上三组12周Wakitani组织学评分分别为5.5±1.401,9.3±1.304和10.2±1.052,24周时为2.2±0.837,7.4±1.144和8.2±1.225,两个时间点实验组均优于支架组及空白组,差异有统计学意义(P<0.05);且实验组24周时优于12周(P<0.05)。结论复合骨髓间充质干细胞的多孔Nano-HA/PLLA能提高成年兔膝关节承重部的骨软骨缺损的修复。     

应用自体脂肪干细胞修复猪关节软骨缺损的初步实验研究

目的 探索自体脂肪干细胞修复猪关节软骨缺损的可行性.方法 从猪背部脂肪组织中获取脂肪干细胞,经过体外培养扩增,以50×106/ml的浓度将脂肪干细胞接种于PLGA(polylacticacid/polyglycolicacid,PLA/PGA,PLGA)中,细胞材料复合物在体外成软骨诱导2周.于猪膝关节软骨非负重区形成2个直径8mm的环形、全层软骨缺损,实验组回植经诱导后的细胞材料复合物,对照组放置单纯支架材料.术后12周取材,缺损修复区行大体观察、组织学HE及藏红花染色、免疫组化检测.结果 术后12周实验组缺损区大部分被修复,缺损被软骨组织填充,修复区表面光滑.组织学染色显示有典型的透明软骨样结构.藏红花染色发现修复组织表达丰富的聚合蛋白多糖.对照组则未能修复关节软骨缺损,缺损区面积增大,表面覆盖薄层纤维组织.结论 猪自体脂肪干细胞可以作为组织工程种子细胞,修复猪关节软骨缺损.     

含自体富集骨髓干细胞松质骨移植与自体松质骨镶嵌移植修复关节软骨缺损的对比研究

目的：评价含富集骨髓干细胞松质骨镶嵌移植与自体松质骨镶嵌移植两种方法修复兔膝关节骨软骨缺损的修复效果,为临床应用提供实验依据。方法：选健康3月龄新西兰兔16只,随机分成A、B两组,每组8只16膝。A组采用含富集自体骨髓干细胞的自体松质骨移植,B组采用自体松质骨移植。术后第12周处死动物取材,分别进行大体观察、光镜观察,并对观察指标采用Wakitani组织学评分法进行评分和秩和检验。结果：大体观察及光镜观察显示含富集骨髓干细胞松质骨镶嵌移植组大体观察关节面大多呈乳白色,表面比较平整,与周围软骨融合较好,未发现裂隙;光镜检查见关节软骨的厚度为正常软骨的2/3左右,表面较光滑,与周围组织接合较好,大多为透明软骨细胞;组织学评分为（4.44±1.41）分。自体松质骨镶嵌移植组大体观察见关节面呈灰白色,表面稍有凹陷,欠光整;光镜检查见关节软骨的厚度较薄,为正常软骨的1/3～1/2,移植物中央区域厚度更薄,见成纤维细胞和极少量透明软骨细胞;组织学评分为（8.93±1.18）分。两组组织学评分比较差异有统计学意义（P〈0.01）。结论：含富集骨髓干细胞松质骨镶嵌移植能以类透明软骨组织修复全层关节软骨缺损,可用于较大面积软骨缺损的修复。自体松质骨移植修复软骨缺损的能力较差。     

Injury and repair of articular cartilage: Related scientific issues

Articular cartilage (AC) is the soft tissue lining the ends of bones in diarthrodial joints. It is responsible for providinglubrication and compressive stiffness to the joint during articulation while responding viscoelastically to mechanical loading. Injury of the tissue caused by trauma or disease can be devastating to joint function as these mechanisms fail. Because of this, it is essential to review the basic science underlying the mechanical roles AC plays while healthy, the biomechanical and biological perspectives of the injury and repair processes, and the current repair techniques available for injured AC. This is done in an effort to further our understanding of the healing capacity of AC and facilitate new efforts into AC repair.     

耳软骨整体再造鼻侧软骨的可行性分析

目的：以耳软骨为供区,探索整体再造鼻侧软骨的方法。方法：20具（40侧）尸体标本,取下耳软骨40枚,鼻侧软骨40枚。CT扫描后重建三维图像,测量鼻侧软骨各解剖分区的相关数据。设计耳软骨3个供区可整体移植再造同侧鼻侧软骨,测量相关的形态数据。结果：耳软骨3个供区的相关形态数据大于同侧鼻侧软骨的相应数据。结论：耳甲腔、耳屏及两者之间连接的峡部（CVIT区域）,三角窝、耳轮及两者的连接部（TFH区域）,耳甲艇、耳轮及两者之间的连接部分（CBH区域）可整体取下,修整后整体再造同侧鼻侧软骨。     

The Fate of Diced Cartilage Grafts of Traumatized Versus Nontraumatized Origin

Background Diced cartilage grafts have long been used in rhinoplasty. Along with their various reported advantages, they also have some disadvantages. The irregular resorption rate of the engrafted mass is one of the major issues noted with diced cartilage grafts. An explanation for the unpredictable resorption rate has not yet been elucidated. This study aimed to determine the role of traumatized versus nontraumatized cartilage as the source of diced cartilage grafts. Methods This study included the noses of 32 patients (19 traumatized noses and 13 nontraumatized noses) who underwent surgery using Surgicel-wrapped diced cartilage grafts. Results The most remarkable result noted in this study was that a Surgicel-wrapped diced cartilage graft, if prepared from traumatized cartilage (in 19 patients), failed to maintain a stable long-term volume. Partial volume loss was noted in 100% of these patients. However, the rate of this partial resorption was different for every individual and could not be predicted. The highest retention of cartilage graft volume was seen primarily over the nasal osseocartilaginous junction. In nontraumatized patients, however, the engrafted mass maintained long-term volume stability. These patients evidenced no visible external irregularities, and only a few very slight imperfections were noted with finger palpation. Conclusion Diced cartilage of traumatic origin is not recommended for any purpose. In such cases, the authors’ engrafting algorithm consists of autobone or ear conchal cartilage grafts for dorsal augmentation, columellar struts from the nontraumatized part of the septum, and spreader grafts from the nontraumatized part of the septum or ear concha.     

Surgical technique for articular cartilage transplantation to full-thickness cartilage defects in the knee joint

Focal arthritic defects in the knee lead to pain, swelling, and dysfunction. Treatment of the defects has includeddrilling, abrasion, and grafting. This report describes our surgical technique of autogenous articular cartilage grafting of arthritic and traumatic articular cartilage lesions. Articular cartilage grafting can be performed as a single arthroscopic outpatient procedure. The mixture of articular cartilage and cancellous bone appears to provide a supportive matrix for cartilage formation. Pain relief is excellent if careful surgical technique and a defined rehabilitation program is followed. Further collagen typing data and additional biopsies will reveal more about the durability of the newly formed cartilage.     

自体肋软骨移植构建鼻尖软骨复合体在鼻整形中的临床应用

目的：通过应用自体肋软骨重塑鼻尖软骨支架结构,形成稳定的鼻尖软骨复合体,来达到完美、立体的鼻尖外形,同时应用膨体聚四氟乙烯或硅胶假体支架抬高鼻背,从而达到理想的鼻部整形美容效果。方法：以鼻小柱基底部＂几＂字形切口和鼻孔内鼻翼软骨外侧缘切口,彻底分离皮肤达鼻翼基底部,对鼻尖短小朝天者可松解到达梨状孔边缘,显露两侧鼻翼软骨及侧鼻软骨,同时暴露鼻中隔软骨游离端;雕刻自体肋软骨,移植、固定到鼻中隔软骨上,贯穿缝合鼻翼软骨、移植的软骨,形成鼻尖软骨支架结构,构建鼻尖软骨复合体。雕刻膨体聚四氟乙烯或硅胶假体支架放置到鼻背鼻骨骨膜下抬高鼻背。结果：本组96例手术者均采用自体肋软骨移植构建鼻尖软骨复合体行鼻整形,术后7天拆线,切口Ⅰ/甲愈合。随诊6～12个月,95例术后鼻尖表现点明显,鼻形立体、挺拔,自然美观,鼻尖活动度好,效果满意。1例术者感觉鼻小柱下垂,通过修复移植软骨,达到满意效果。结论：应用自体肋软骨重塑鼻尖软骨支架结构,构建鼻尖软骨复合体,同时应用膨体聚四氟乙烯或硅胶假体支架抬高鼻背,可以达到理想的鼻部整形美容效果。     

Tragal Cartilage Grafts in Aesthetic Rhinoplasty

Ear cartilage is an important source of grafts for rhinoplasty. The majority of cartilage grafts is harvested from the concha of the ear. We describe indications in which the tragal cartilage is a more favorable graft source than conchal cartilage. The technique is fast, simple, and does not require special dressings.     

Simplified harvesting of the ear cartilage graft

A simplified technique of harvesting of the ear cartilage graft is described which (1) uses a postauricular incision to minimize the visible scars, (2) removes the whole floor of the conchal fossa to avoid the irregularities, and (3) uses a tie-over dressing to eliminate the need for a head dressing. This technique has been used on 88 patients with one visible and tender medial irregularity, and three minor palpable, but invisible irregularities.     

异体软骨细胞复合Pluronic修复关节软骨缺损

目的 探讨运用同种异体软骨细胞复合Pluronic修复关节软骨缺损的可行性，并应用^3H—TdR放射自显影方法鉴别软骨缺损修复的细胞来源。方法 取同种异体软骨细胞体外培养至第2代，用^3H—TdR标记后复合Pluronic植入兔关节软骨缺损区作为实验组，并采用单纯Pluronic植入作为材料对照组，不作任何处理组为空白对照组，分别于4、8及16周取材，观察其修复效果，并应用放射自显影方法鉴别修复组织的细胞来源。结果 实验组术后8周，缺损表面可见新生软骨形成，术后16周缺损完全修复，表面光滑，与周围界限模糊，放射自显影证实所修复组织的细胞来源于植入细胞。材料对照组及空白对照组缺损均未见明显修复。结论 ①同种异体软骨细胞复合Pluronic修复关节软骨缺损是可行的；②^3H—TdR标记细胞可作为鉴别细胞来源的一种简便可行的方法。     

Augmentation rhinoplasty with soft tissue and cartilage

Augmentation rhinoplasty using soft tissue and cartilage was performed on 120 patients and the results were reexamined. They were found to be satisfactory and without complication. To narrow a round tip, a resection of two-thirds of the lateral crus cephalad portion and a transection of the caudal portion with a strip resection was done. To elevate the tip, septal cartilage was sutured to one-third of the upper part of medial crus to form a columella cartilage strut. To maintain the strut and to prevent pointing, a fibrous muscle tissue stretching from the medial crus to the upper cartilage or a dermis was transplanted into the area surrounding the septal cartilage tip. For a simple elevation of the dorsum, an onlay graft of dermis was applied, but where further elevation was required, further dermis and conchal cartilage was added for suture and attachment to the dermis.     

关节软骨缺损修复研究进展

关节软骨是滑膜关节重要的结构和功能单位,一旦关节软骨发生损伤,由于软骨组织本身缺乏血管,缺乏修复损伤和缺损的未分化细胞,软骨细胞包埋于致密的胶原一蛋白多糖基质中,限制了细胞的增殖和迁移能力,所以自身修复能力非常有限,若治疗不及时或不恰当,难以形成正常的关节软骨,因而无法满足正常关节的功能需求,将会导致严重的关节功能障碍。