带血管蒂肌腿移植修复跟腱缺损的实验研究

目的：为带血供肌腱移植修复跟腿,缺损提供生物力学和组织学依据。方法：选用新西兰大白兔15只,其中12只分两组：一侧行带因管蒂趾长屈肌腱转位修复跟腿缺损,对侧为游离肌腿移植对照组,术后12周取材,分别行组织学检查和生物力学测试。结果：带血管蒂肌腿组移植跟腿组织学形态近似正常跟腿,肌腿最大拉伸力为正常跟腿的67．7％,而游离肌腿组移植跟腿的腱纤维为瘢痕包裹,最大拉伸力为跟腿的35．3％,两者的差异性非常显著（P＜0．01）,结论：带血管蒂肌腿移植修复跟腿缺损优于游离肌腿移植。     

带血管蒂腓骨长肌腱转位修复跟腱缺损的临床应用

目的：报道带血管蒂腓骨长肌腱转位修复跟腱缺损的应用解剖、生物学力学和手术方法。方法：在４０侧动脉灌注红色乳胶的成人下肢标本上，对腓骨长肌腱进行显微解剖学观测及生物力学研究。在解剖学研究的基础上，设计并应用带血管腓骨长肌腱及皮瓣转位修复跟腱。结果：采用该术式修复跟腱及皮肤缺损９例。经１０个月～１．５年随访，移植的肌腱及皮瓣全部成活８例，愈合良好，跟腱功能恢复。１例皮瓣边缘性坏死，经短期换药愈合。结论：带血管蒂腓骨长肌腱和皮瓣转位是跟腱及皮肤缺损较理想的修复方法。     

带血管蒂排骨长肌腱转位修复跟腱缺损的临床应用

目的：报道带血管蒂腓骨长肌腱转位修复跟腱缺损的应用解剖、生物学力学和手术方法。：在４０例动脉灌注红色乳胶的成人下肢标本上，对腓骨长肌腱进行显微解剖学观测及生物力学研究。在解剖学研究的基础上，设计并应用带血管腓骨长肌腱及皮瓣转位修复跟腱。结果：采用该术式修复跟腱及皮肤缺损９例。经１０个月￣１．５年随访，移植的肌腱及皮瓣全部成活８例，愈合良好，跟腱功能恢复。１例皮瓣边缘性坏死，经短期换药愈合。结论：带     

带血管蒂游离腓骨复合组织瓣移植一期修复小腿下段骨与组织缺损的临床应用

目的 探讨带血管蒂游离腓骨复合组织瓣移植一期修复小腿下段骨与组织缺损的临床疗效.方法 选择2015年2月至2019年6月宝鸡市中医医院采用带血管蒂游离腓骨复合组织瓣移植一期修复小腿下段骨与组织缺损患者9例.所有患者均为外伤后小腿下段皮肤软组织缺损合并胫骨大段缺损(其中1例合并有胫前肌腱缺损).皮肤软组织缺损面积4 cm...     

异种肌腱修复肌腱缺损微观实验:验证可作为临床肌腱修复的生长支架

背景:自体肌腱修复肌腱缺损因可用肌腱有限且形成供区功能障碍,同种异体肌腱同样来源有限,并且价格昂贵,在临床上很难满足其需要。目的:观察不同时期异种肌腱修复肌腱缺损的微观变化,为异种肌腱作为临床组织工程化肌腱生长支架提供理论依据。方法:取6月龄的Leghorn鸡屈趾肌腱经化学去细胞处理后作为异种肌腱移植供体,健康成熟日本大耳白兔36只,建立双后肢跟腱中间束2 cm缺损模型,随机分为异种肌腱移植组和自体肌腱移植组,每组18只。肌腱移植缝合用4-0无创伤肌腱缝合线行双"8"字缝合,移植后伸直位管型石膏固定2周,对供体肌腱行去细胞前后大体观察、生物力学测定、组织学光镜及电镜观察,术后2,4,9周每组取6只兔对标本行组织学光镜及电镜检测。结果与结论:1肌腱经过化学去细胞处理后色泽变白,质地较前柔软,去细胞前可见细胞与胶原纤维交替紧密排列,去细胞后胶原排列相对松散,且无细胞及细胞碎片,去细胞后肌腱的力学强度较术前减弱。2由电镜图片直观看到:随移植时间的延长,移植的粗大鸡肌腱胶原纤维逐渐被再生的兔肌腱胶原纤维最终替代,而新生成的纤细胶原纤维经改造塑形变为粗细相等的较粗大纤维,排列方向逐渐趋于平行,在结构和功能上达到正常肌腱水平。结果表明,去细胞后的最大抗拉力是去细胞前的83.44%,能够满足肌腱移植生物力学的要求。最终肌腱修复是再生胶原纤维形成的结果,异种肌腱经过理化方法处理后可作为临床肌腱修复的生长支架使用。     

骨形态发生蛋白12诱导骨髓间充质干细胞构建组织工程肌腱

背景：以往肌腱损伤的修复方法有端端吻合、自体肌腱移植、同种异体肌腱或人工肌腱移植等,但均有其各自缺点。 目的：探讨以家兔骨髓间充质干细胞为种子细胞,骨形态发生蛋白12诱导,聚羟基乙酸复合材料作为支架材料,预构组织工程化肌腱重建家兔跟腱缺损的可能性。 方法：家兔抽取股骨近端骨髓,分离所得细胞传代至第2代,加入10 μg/L骨形态发生蛋白12诱导分化,并与Ⅰ型胶原溶液按一定比例移植于预张的聚羟基乙酸缝线上预制组织工程化肌腱备用。将家兔制备成跟腱缺损模型,分别采用不同方法修复跟腱缺损：组织工程化肌腱修复,Ⅰ型胶原-聚羟基乙酸缝线修复,丝线行端端修复。修复12周对各组肌腱行形态学、力学及组织病理学观察。 结果与结论：修复12周家兔肌腱组织病理切片：组织工程化肌腱组可见大量梭形纤维母细胞顺应力学方向排列均匀分布于胶原中,纤维细胞明显增多,胶原致密;Ⅰ型胶原-聚羟基乙酸缝线组可见部分纤维组织增生伴少许肉芽组织形成,胶原纤维呈松散网丝状,细胞排列紊乱分布不均;丝线组可见纤维组织旁见大量肉芽组织形成。修复12周家兔肌腱生物力学强度：骨形态发生蛋白12+聚羟基乙酸重建肌腱的力学强度明显优于Ⅰ型胶原-聚羟基乙酸组,与丝线缝合组差异无显著性意义;但骨形态发生蛋白12+聚羟基乙酸重建肌腱的力学强度低于正常肌腱。提示以自体骨髓间充质干细胞作为种子细胞,骨形态发生蛋白12诱导,以聚羟基乙酸为支架,可望构建组织工程化肌腱。构建的组织工程肌腱具有一定的生物力学特性,能用于修复跟腱缺损。     

氯仿/甲醇处理同种异体肌腱移植的生物力学变化

目的 :探讨氯仿 /甲醇 (CM)处理的同种异体肌腱移植后的生物力学性质。方法 :2 4只新西兰兔随机选择 1侧跟腱行CM腱移植 ,另一侧行自体移植。按术后 1、2、6、12周行生物力学测试。结果 :自体腱移植组和CM肌腱移植组的拉伸强度和刚度在术后 1、2周 ,6周与术前、术后 12周分别有显著性差异(P <0 .0 1)。自体腱移植组与CM腱移植组术后 12周的拉伸强度和刚度无显著性差异 (P >0 .0 5 )。结论 :CM腱移植组与自体腱移植组在各个时间点的力学变化相近 ,无统计学变异 ,可以用来替代自体腱。     

复合组织移植一期修复复杂指背腱膜损伤

目的:报道应用复合组织移植一期修复复杂指背腱膜损伤的临床疗效。方法:采用掌背动脉为蒂带肌腱手背逆行岛状皮瓣、足背动脉为蒂带趾长伸肌腱复合组织皮瓣及肌腱移植锁骨下带蒂皮瓣3种方法修复,为36例合并皮肤缺损的复杂指背腱膜损伤进行急诊修复。结果:36例全部成活,随访1￣5年,按TA M功能评定法:优11例、良20例、差5例,优良率86.1%。结论:(1)对合并皮肤缺损的复杂指背腱膜损伤根据伤情,较大面积的手背手指皮肤肌腱缺损,以足背动脉为蒂带趾长伸肌腱复合组织皮瓣修复;(2)手指近节皮肤肌腱缺损,以掌背动脉为蒂带肌腱手背逆行岛状皮瓣修复;(3)手指中远节皮肤肌腱缺损,以肌腱移植锁骨下带蒂皮瓣修复,可获得良好疗效。     

小腿有关肌腱转位复跟腱的生物力学评价

为临床选用自体肌腱转位修复跟腱提供生物力学依据。选用防腐固定成年和新鲜青年下肢各７例，制备拉伸试件在ＳＷＤ－１０型材料试验机上，进行单向拉伸破坏实验。结论：腓骨长肌腱是跟腱缺损修复术较理想的自体材料。     

碳化二亚胺交联改性脱细胞异种猪肌腱修复跟腱缺损

背景：异体肌腱移植是目前修复肌腱缺损的理想方法,但移植后的排斥反应使其使用受到限制。 目的：观察碳化二亚胺交联改性脱细胞处理的版纳近交系微型猪肌腱移植修复兔跟腱缺损的效果。 方法：横向切除40只日本大白兔双侧跟腱,随机分组：实验组以碳化二亚胺交联改性脱细胞版纳近交系微型猪肌腱移植修复,对照组以自体肌腱移植修复。 结果与结论：①组织学观察：两组新生组织内细胞主要都是单核的成纤维细胞和纤维细胞,术后1-4周主要是呈椭圆形或圆形的成纤维细胞,细胞聚集区的细胞周围有新生胶原形成,这些胶原的走向较紊乱;局灶性条索状成熟胶原呈岛状分布,形成所谓“胶原岛”;术后12周时细胞越来越拉长,成为梭形和长条形的纤维细胞。②实验室检测：两组白细胞、C-反应蛋白水平、羟脯氨酸含量及抗拉强度差异均无显著性意义。说明碳化二亚胺交联改性的脱细胞版纳近交系微型猪肌腱能成功修复兔跟腱缺损,且具有组织相容性好、移植排斥反应轻、生物力学性能强的优点。     

小腿有关肌腱转位修复跟腱的生物力学评价

目的：为临床选用自体肌腱转位修复跟腱提供生物力学依据。方法：选用防腐固定成年和新鲜青年下肢各７例。制备拉伸试件，在ＳＷＤ－１０型材料试验机上，进行单向拉伸破坏实验。结果：跟腱、腓骨长肌腱、腓骨短肌腱、胫骨后肌腱及展肌腱固定组分别为２２９２．６Ｎ、１０２０．５Ｎ、７５２．０Ｎ、９３８．９Ｎ和７２１．３Ｎ，新鲜组分别为１９２７．１Ｎ、８１９．５Ｎ、３４６．７Ｎ、６９９．７Ｎ、３０３．８Ｎ。结论：腓骨长肌腱是跟腱缺损修复术较理想的自体材料。     

Repair of Achilles tendon defect with autologous ASCs engineered tendon in a rabbit model

Adipose derived stem cells (ASCs) are an important cell source for tissue regeneration and have been demonstrated the potential of tenogenic differentiation in vitro. This study explored the feasibility of using ASCs for engineered tendon repair in vivo in a rabbit Achilles tendon model. Total 30 rabbits were involved in this study. A composite tendon scaffold composed of an inner part of polyglycolic acid (PGA) unwoven fibers and an outer part of a net knitted with PGA/PLA (polylactic acid) fibers was used to provide mechanical strength. Autologous ASCs were harvested from nuchal subcutaneous adipose tissues and in vitro expanded. The expanded ASCs were harvested and resuspended in culture medium and evenly seeded onto the scaffold in the experimental group, whereas cell-free scaffolds served as the control group. The constructs of both groups were cultured inside a bioreactor under dynamic stretch for 5 weeks. In each of 30 rabbits, a 2 cm defect was created on right side of Achilles tendon followed by the transplantation of a 3 cm cell-seeded scaffold in the experimental group of 15 rabbits, or by the transplantation of a 3 cm cell-free scaffold in the control group of 15 rabbits. Animals were sacrificed at 12, 21 and 45 weeks post-surgery for gross view, histology, and mechanical analysis. The results showed that short term in vitro culture enabled ASCs to produce matrix on the PGA fibers and the constructs showed tensile strength around 50 MPa in both groups (p > 0.05). With the increase of implantation time, cell-seeded constructs gradually form neo-tendon and became more mature at 45 weeks with histological structure similar to that of native tendon and with the presence of bipolar pattern and D-periodic structure of formed collagen fibrils. Additionally, both collagen fibril diameters and tensile strength increased continuously with significant difference among different time points (p < 0.05). In contrast, cell-free constructs failed to form good quality tendon tissue with fibril structure observable only at 45 weeks. There were significant differences in both collagen fibril diameter and tensile strength between two groups at all examined time points (p < 0.05). The results of this study support that ASCs are likely to be a potential cell source for in vivo tendon engineering and regeneration.     

Weft-knitted silk-poly(lactide-co-glycolide) mesh scaffold combined with collagen matrix and seeded with mesenchymal stem cells for rabbit Achilles tendon repair

Abstract

Natural silk fibroin fiber scaffolds have excellent mechanical properties, but degrade slowly. In this study, we used poly(lactide-co-glycolide) (PLGA, 10:90) fibers to adjust the overall degradation rate of the scaffolds and filled them with collagen to reserve space for cell growth. Silk fibroin-PLGA (36:64) mesh scaffolds were prepared using weft-knitting, filled with type I collagen, and incubated with rabbit autologous bone marrow-derived mesenchymal stem cells (MSCs). These scaffold–cells composites were implanted into rabbit Achilles tendon defects. At 16 weeks after implantation, morphological and histological observations showed formation of tendon-like tissues that expressed type I collagen mRNA and a uniformly dense distribution of collagen fibers. The maximum load of the regenerated Achilles tendon was 58.32% of normal Achilles tendon, which was significantly higher than control group without MSCs. These findings suggest that it is feasible to construct tissue engineered tendon using weft-knitted silk fibroin-PLGA fiber mesh/collagen matrix seeded with MSCs for rabbit Achilles tendon defect repair.     

跟腱复合组织缺损的显微外科修复

目的 :探讨跟腱复合组织缺损的显微外科治疗。方法 :应用膝降血管大收肌腱、骨、皮复合组织瓣移植一期修复跟腱伴皮肤和跟骨缺损。结果 :施术 3侧 ,取得满意效果。结论 :该术式可修复不同类型的跟腱复合组织缺损 ,并具有操作简便、易行和供区损伤小等特点。     

Knitted poly-lactide-co-glycolide scaffold loaded with bone marrow stromal cells in repair and regeneration of rabbit Achilles tendon

The objectives of this study were to evaluate the morphology and biomechanical function of Achilles tendons regenerated using knitted poly-lactide-co-glycolide (PLGA) loaded with bone marrow stromal cells (bMSCs). The animal model used was that of an adult female New Zealand White rabbit with a 10-mm gap defect of the Achilles tendon. In group I, 19 hind legs with the created defects were treated with allogeneic bMSCs seeded on knitted PLGA scaffold. In group II, the Achilles tendon defects in 19 hind legs were repaired using the knitted PLGA scaffold alone, and in group III, 6 hind legs were used as normal control. The tendon-implant constructs of groups I and II were evaluated postoperatively at 2, 4, 8, and 12 weeks using macroscopic, histological, and immunohistochemical techniques. In addition, specimens from group I (n = 7), group II (n = 7), and group III (n = 6) were harvested for biomechanical test 12 weeks after surgery. Postoperatively, at 2 and 4 weeks, the histology of group I specimens exhibited a higher rate of tissue formation and remodeling as compared with group II, whereas at 8 and 12 weeks postoperation, the histology of both group I and group II was similar to that of native tendon tissue. The wound sites of group I healed well and there was no apparent lymphocyte infiltration. Immunohistochemical analysis showed that the regenerated tendons were composed of collagen types I and type III fibers. The tensile stiffness and modulus of group I were 87 and 62.6% of normal tendon, respectively, whereas those of group II were about 56.4 and 52.9% of normal tendon, respectively. These results suggest that the knitted PLGA biodegradable scaffold loaded with allogeneic bone marrow stromal cells has the potential to regenerate and repair gap defect of Achilles tendon and to effectively restore structure and function.     

Repair of tendon defect with dermal fibroblast engineered tendon in a porcine model

Harvesting autologous tenocytes for tendon engineering may cause secondary tendon defect at the donor site. Dermal fibroblasts are an easily accessible cell source and do not cause major donor site defect. This study aims to explore the possibility of tendon engineering using dermal fibroblasts. A total of 45 hybrid pigs were randomly divided into three groups: experimental group (n = 15)--repair of tendon defect with a dermal fibroblast engineered tendon; control group 1 (n = 15)--repair of defect with a tenocyte engineered tendon; and control group 2 (n = 15)-repair of defect with a scaffold alone. Both autologous dermal fibroblasts and tenocytes were seeded on polyglycolic acid (PGA) unwoven fibers to form a cell-scaffold construct and cultured in vitro for 7 days before in vivo implantation to repair a defect of flexor digital superficial tendon. Specimens were harvested at weeks 6, 14, and 26 for gross, histological, and mechanical analyses. Microscopy revealed good attachment of both dermal fibroblasts and tenocytes on PGA fibers and matrix production. In vivo results showed that fibroblast and tenocyte engineered tendons were similar to each other in their gross view, histology, and tensile strength. At 6 weeks, parallel collagen alignment was observed at both ends, but not in the middle in histology, with more cellular components than natural tendons. At weeks 14 and 26, both engineered tendons exhibited histology similar to that of natural tendon. Collagens became parallel throughout the tendon structure, and PGA fibers were completely degraded. Interestingly, dermal fibroblast and tenocyte engineered tendons did not express type III collagen at 26 weeks, which remained observable in normal pig skin and control group 2 tissue using polarized microscopy, suggesting a possible phenotype change of implanted dermal fibroblasts. Furthermore, both fibroblast and tenocyte engineered tendons shared similar tensile strength, about 75% of natural tendon strength. At 6 weeks in control group 2, neo-tissue was formed only at the peripheral area by host cells. A cord-like tissue was formed at weeks 14 and 26. However, the formed tissue was histologically disorganized and mechanically weaker than both cell-engineered tendons (p < 0.05). These results suggest that dermal fibroblasts may have the potential as seed cells for tendon engineering.     

肿瘤坏死因子α拮抗剂对应力屏蔽导致的跟腱挛缩形态学影响

背景：跟腱应力屏蔽后肿瘤坏死因子α明显升高,应用肿瘤坏死因子α拮抗剂干预肿瘤坏死因子α的作用是否会干预跟腱挛缩尚不清楚。 目的：通过观察重组人Ⅱ型肿瘤坏死因子受体-抗体融合蛋白(益赛普)干预应力屏蔽后大鼠跟腱形态学变化,了解肿瘤坏死因子α在肌腱挛缩中的影响及肿瘤坏死因子α拮抗剂对肌腱挛缩的干预作用。 方法：20只健康雄性SD大鼠左后肢行跟腱应力屏蔽后随机数字表法均分为实验组和模型组,两组中随机各抽取5只大鼠以右后肢为正常对照组。建模后即刻实验组大鼠使用0.6 mg/kg重组人Ⅱ型肿瘤坏死因子受体-抗体融合蛋白、模型组使用1 mL磷酸缓冲盐溶液于大鼠皮下注射,后根据重组人Ⅱ型肿瘤坏死因子受体-抗体融合蛋白半衰期两组分别再注射3次,干预2周时大体下及透射电镜下观察跟腱形态学改变。 结果与结论：大体观察实验组跟腱明显比模型组细小、光滑,但较正常对照组粗。电镜下模型组胶原纤维束较实验组组疏松、紊乱;实验组横切面和纵切面均与正常对照组差异无显著性意义。说明,在2周时,肿瘤坏死因子α拮抗剂能明显预防应力屏蔽所致的肌腱挛缩。 中国组织工程研究杂志出版内容重点：组织构建;骨细胞;软骨细胞;细胞培养;成纤维细胞;血管内皮细胞;骨质疏松;组织工程全文链接：     

改良膝降动脉为蒂大收肌腱骨皮瓣修复跟腱复合组织缺损

目的　探讨改良膝降动脉为蒂的大收肌腱骨皮瓣修复跟区跟腱复合组织缺损方法及效果。 方法　对3例外伤致足跟区跟腱止点伴跟骨及皮肤软组织缺损的患者,采用以膝降动脉为蒂的隐动脉穿支皮瓣修复跟区皮肤缺损,以膝降动脉关节支为蒂的大收肌腱骨瓣移植修复跟区复合组织缺损。其中跟区皮肤缺损面积为7 cm×9 cm～5 cm×7 cm ,跟腱缺损长度为6～8 cm,切取皮瓣面积为8 cm×10 cm～6 cm×8 cm ,大收肌腱切取长度为7～9 cm。 结果　3例复合组织瓣全部顺利成活,术后随访6～12个月,皮瓣感觉恢复S3～4,双足提踵试验及单足提踵试验均阴性,步态恢复均正常。 结论　应用隐动脉穿支设计皮瓣,皮瓣设计于大腿内侧, 改良了隐动脉皮瓣设计于小腿内侧,造成切口过大,分离血管蒂较长等缺点,疗效可靠,是目前较理想修复方法。