Histomorphometric analysis of poly-L/D-lactide 96/4 sutures in the gastrocnemius tendon of rabbits

BACKGROUND: Common Achilles tendon ruptures are not usually fixed by bioabsorbable sutures due to limitations in their strength retention properties. Modern technology has made it possible to develop bioabsorbable sutures with prolonged strength retention. Aims: To evaluate histologically tissue reactions of poly-L/D-lactide (PLDLA) sutures implanted in Achilles tendon of rabbits. MATERIAL AND METHODS: Fifteen rabbits were evaluated at 2, 6 and 12 weeks postoperatively, with five rabbits in each follow-up group. PLDLA monofilament sutures were implanted into the medial gastrocnemius tendon. Polyglyconate monofilament sutures with similar diameter (Maxon 4-0, Cyanamid of Great Britain Ltd., Gosport, UK) were implanted in the contralateral gastrocnemius tendon. The histology was studied in hard-resin embedded samples. The thickness of the formed fibrous tissue capsule was determined histomorphometrically. RESULTS: PLDLA led to formation of significantly thinner fibrous tissue capsule than Maxon sutures of the same diameter. Median thickness (PLDLA vs. Maxon) at two weeks was 5.26 vs.13.22 microm, at six weeks 11.66 vs. 80.97 microm, and at 12 weeks 10.63 vs. 17.59 microm (p<0.01). CONCLUSIONS: During the 12 week follow-up period, PLDLA sutures implanted intratendineously formed thinner fibrous capsule than Maxon sutures of the same diameter. The suture materials were not totally absorbed by 12 weeks.     

Strength retention properties of self-reinforced poly L-lactide (SR-PLLA) sutures compared with polyglyconate (Maxon) and polydioxanone (PDS) sutures. An in vitro study

Recent developments in manufacturing techniques have led to the development of strong bioabsorbable materials such as self-reinforced poly L-lactide (SR-PLLA) sutures. The aim of the study was to investigate the mechanical properties of SR-PLLA sutures in comparison with polyglyconate (Maxon) and polydioxanone (PDS) sutures in vitro. Sutures made of SR-PLLA (0.3, 0.5 and 0.7 mm diameter), Maxon (0.3 and 0.5 mm diameter) and PDS (0.3 and 0.5 mm diameter) were studied by immersion in phosphate-buffered distilled water (pH 7.4) at 37 degrees C for 40 weeks. The breaking force of straight sutures and suture knots was measured. Tensile strength and percentage elongation were calculated. Means, standard deviations, differences between means, and confidence intervals for differences between means were evaluated. SR-PLLA, PDS and Maxon sutures of 0.3 and 0.5 mm diameter were of comparable initial tensile strength. Initial knot tensile strength values were lower than those of their counterpart straight sutures. Maxon sutures had lost their tensile strength by 12 weeks; PDS sutures by 20 weeks. SR-PLLA sutures of 0.3 mm diameter had a strength of 161.6 MPa and those of 0.5 mm diameter had a strength of 134 MPa at 40 weeks. The highest percentage elongation of straight sutures (62.8% and 62%) was exhibited by PDS; the lowest by SR-PLLA (35.6% and 35%). In loop tests, PDS showed the highest percentage elongation (43.7% and 58.1%) and SR-PLLA had the lowest values (19.7% and 33%). SR-PLLA sutures had the most prolonged strength retention in vitro, but the lowest elongation (elasticity). Compared with straight sutures, knots had lower tensile strength and elongation values. SR-PLLA sutures can be applied to the closure of wounds that need prolonged support, such as bone.     

In vivo and in vitro degradation of monofilament absorbable sutures, PDS and Maxon

Two new absorbable monofilament suture materials polydioxanone and Maxon are being employed increasingly in abdominal surgery because of increased strength retention and decreased tissue reactivity compared with previously available materials. As part of our investigation of the behaviour of suture materials, 3-0 sutures of polydioxanone and Maxon were enclosed in nylon pouches, a technique developed for in vivo experiments to prevent cellular interaction with implanted devices. The pouched sutures were gas sterilized, then implanted in either the extrafascial space or peritoneal cavity for periods of 1-5 wk. Sterilized sutures were also incubated in Ringer's lactate at 37 degrees C. Tensile strength of the exposed sutures was measured. For a given suture material and duration of incubation, there was no significant difference in tensile strength degradation among the three test environments. Although the strength of unexposed Maxon is greater than that of polydioxanone, the residual strength of Maxon decreases more rapidly in use, so that, after 2 wk, the strength of polydioxanone is greater. Scanning electron microscope examination of the suture surfaces reveals that polydioxanone develops surface crazing with time, whereas the surface morphology of Maxon remains relatively unaltered.     

带血管蒂肌腱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腱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只分两组一侧行带血管蒂趾长屈肌腱转位修复跟腱缺损,对侧为游离肌腱移植对照组,术后12周取材,分别行组织学检查和生物力学测试.结果带血管蒂肌腱组移植跟腱组织学形态近似正常跟腱,肌腱最大拉伸力为正常跟腱的67.7%,而游离肌腱组移植跟腱的腱纤维为瘢痕包裹,最大拉伸力为跟腱的35.3%,两者的差异性非常显著(P＜0.01).结论带血管蒂肌腱移植修复跟腱缺损优于游离肌腱移植.     

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。     

A Growth and Differentiation Factor-5 (GDF-5)-coated Suture Stimulates Tendon Healing in an Achilles Tendon Model in Rats

Growth and differentiation factor-5 (GDF-5) is essential for normal skeletal development and induces tendon-and ligament-like structures at ectopic sites. Therefore, we investigated the influence of a GDF-5-coated suture on the healing Achilles tendon in rats. The right Achilles tendon in 80 rats was transected and sutured with an absorbable polyglactin suture. Animals were randomized to an uncoated-suture control and a GDF-5-coated suture group. At 1, 2, 4 and 8 weeks after surgery the repair tissue was evaluated biomechanically and histologically. Biomechanical testing revealed significantly thicker tendons, which were stiffer at 1, 2, and 4 weeks, in the experimental group than in the control group. The maximum tensile strength was significantly increased at 2 weeks after surgery. Histologically we found cartilage-like cell nests 4 weeks after tendon repair, which were positive for type II collagen. In conclusion, local growth factor delivery by a coated suture material showed a promising beneficial effect on tendon repair. The appearance of cartilage-like structures may demonstrate the chondroinductive capacity of GDF-5, which in these circumstances, however, might be overcome by modifications of the GDF-5 dose and/or the suture material.     

Age-related changes in biomechanical properties of the Achilles tendon in rabbits

We investigated age-related changes in the mechanical properties of rabbit Achilles tendon. The animals used were immature (age 3 weeks, body mass 380 g), young adult (age 8–10 months, body mass 4.1 kg) and old (age 4–5 years, body mass 5.1 kg) rabbits. The cross-sectional area of the tendon increased with growth and the tensile strength of the young adult [67.3 (SEM 4.2) MPa] and old [66.7 (SEM 3.8) MPa] tendon was significantly higher than that of the immature tendon [23.9 (SEM 3.8) MPa]. However, there was no statistically significant difference in tensile strength between mature and old tendons. These differences may be attributable to the change in body mass. The gradient of the stress-strain curves, that is, the tangent modulus of the mature tendon [618.0 (SEM 87.0) MPa], was higher than that of the immature [281.0 (SEM 104.6) MPa] and old [530.5 (SEM 91.0) MPa] tendon, although the difference was not significant. The elongation at failure was approximately 16% for all age groups. These results would suggest that rabbit Achilles tendon is highly compliant during growth.     

Ultrastructural Determinants of Murine Achilles Tendon Strength During Healing

The mechanisms by which tendon strength is established during growth and development and restored following injury are not completely understood and are likely to be complex, multifactorial processes. Several studies examining the relationship between mechanical behavior and ultrastructural characteristics of tendons and ligaments during growth and maturation suggest that collagen fibril diameter is strongly correlated with tendon strength. Because of the similarities between development and repair processes of musculoskeletal tissues, increases in tendon strength during healing may be related to increases in fibril ultrastructural parameters such as fibril size, numerical density, and area fraction. In this study, we compared murine Achilles tendons at various time points after tenotomy with sham-operated controls in tensile tests to failure and examined tendons using electron microscopy to assess collagen fibril ultrastructure. We found that in the 6-week period following Achilles tenotomy, fibril mean diameter remained significantly smaller than sham-side diameter by a factor of 2–3. Despite the persistently small fibril size, increasing numerical density resulted in a gradual increase in fibril area fraction. Biomechanical strength did not reach that of intact tendons until some time between 5 and 7 weeks, approximately the same time period when fibril area fraction began to approach sham values. These data suggest that parameters other than collagen fibril size are most responsible for increased tendon strength during healing.     

Ultrastructural determinants of murine achilles tendon strength during healing

The mechanisms by which tendon strength is established during growth and development and restored following injury are not completely understood and are likely to be complex, multifactorial processes. Several studies examining the relationship between mechanical behavior and ultrastructural characteristics of tendons and ligaments during growth and maturation suggest that collagen fibril diameter is strongly correlated with tendon strength. Because of the similarities between development and repair processes of musculoskeletal tissues, increases in tendon strength during healing may be related to increases in fibril ultrastructural parameters such as fibril size, numerical density, and area fraction. In this study, we compared murine Achilles tendons at various time points after tenotomy with sham-operated controls in tensile tests to failure and examined tendons using electron microscopy to assess collagen fibril ultrastructure. We found that in the 6-week period following Achilles tenotomy, fibril mean diameter remained significantly smaller than sham-side diameter by a factor of 2-3. Despite the persistently small fibril size, increasing numerical density resulted in a gradual increase in fibril area fraction. Biomechanical strength did not reach that of intact tendons until some time between 5 and 7 weeks, approximately the same time period when fibril area fraction began to approach sham values. These data suggest that parameters other than collagen fibril size are most responsible for increased tendon strength during healing.     

The role of graft materials in suture augmentation for tendon repairs and reattachment

Various biomaterials have been used to augment sutures for the repair and reattachment of tendons. This study examined four different graft materials in a simple and reproducible model using chicken Achilles tendons to determine the strength and mechanism of suture reinforcement of tendon repairs. The graft materials tested were Gore-Tex(R) Soft Tissue Patch, Graftjacket, bovine pericardium, and an experimental graft material from Xylos Corporation. Testing was performed in shear to simulate forces on a torn tendon repair and pull-off to simulate those on a tendon reattachment to bone. Compared to unaugmented suture, grafts increased suture fixation strength from 10% to 60% in shear and from 0% to 36% in pull-off with the bovine pericardium graft, providing significant improvement in both tests. In no cases (even unaugmented) did the suture pull directly through the tendon, but instead sliced along it, demonstrating that the interface between the suture and the tendon determines fixation strength. Grafts function by increasing the area, friction, and nature of this interface, not by acting as a barrier for suture pull-through.     

Tensile properties of fresh human calcaneal (Achilles) tendons

The purpose of this study was to measure the tensile properties of fresh human calcaneal (Achilles) tendons. Twenty fresh cadaveric (age range = 57-93 years) bone-Achilles tendon complexes were harvested within 24 hr postmortem. The calcaneus together with 15 cm of the Achilles tendon extending proximally from the insertion on the calcaneus was clamped and biomechanically tested. Each tendon was firmly fixed in clamps in an MTS Systems Corporation MTS testing machine and tension was applied at a displacement rate of 8 cm per minute until the tendon failed. The tensile force and tensile strain (as measured using an extensometer) were recorded and plotted using onboard software. The narrow age range of our donors prevented any meaningful correlation between age and tensile properties; however, the results showed that: 1) the average ultimate tensile strength (UTS) of the human Achilles tendon was 1189 N (range = 360-1,965), 2) there was a correlation between left and right legs for UTS, 3) there was a correlation between left and right legs in regard to cross sectional area, and 4) there was no correlation between UTS and cross-sectional area.     

伞式经跟骨缝合组合修复跟腱断裂的实验研究

目的 评价伞式经跟骨组合缝合法修复跟腱断裂的生物力学特性。方法 采用伞式经跟骨缝合法，组合应用0-薇乔线(Poliglactin-910)、腱周采用交叉缝合法修复8只家兔单侧跟腱断裂模型，术后固定3天，功能锻炼25天后行离体生物力学实验。结果术后28天组达正常组跟腱生物力学特性的90％。结论 采用伞式经跟骨组合缝合法修复跟腱允许早期康复活动，早期功能锻炼可以迅速提高跟腱的生物力学特性。     

Healing parameters in a rabbit partial tendon defect following tenocyte/biomaterial implantation

Although rabbits are commonly used as tendon repair model, interpretative tools are divergent and comprehensive scoring systems are lacking. Hence, the aim was to develop a multifaceted scoring system to characterize healing in a partial Achilles tendon defect model. A 3 mm diameter defect was created in the midsubstance of the medial M. gastrocnemius tendon, which remained untreated or was filled with a polyglycolic-acid (PGA) scaffold + fibrin and either left cell-free or seeded with Achilles tenocytes. After 6 and 12 weeks, tendon repair was assessed macroscopically and histologically using self-constructed scores. Macroscopical scoring revealed superior results in the tenocyte seeded PGA + fibrin group compared with the controls at both time points. Histology of all operated tendons after 6 weeks proved extracellular matrix (ECM) disorganization, hypercellularity and occurrence of irregular running elastic fibres with no significance between the groups. Some inflammation was associated with PGA implantation and increased sulphated proteoglycan deposition predominantly with the empty defects. After 12 weeks defect areas became hard to recognize and differences between groups, except for the increased sulphated proteoglycans content in the empty defects, were almost nullified. We describe a partial Achilles tendon defect model and versatile scoring tools applicable for characterizing biomaterial-supported tendon healing.     

Adenovirus-mediated gene transfer of growth and differentiation factor-5 into tenocytes and the healing rat Achilles tendon

Growth and differentiation factor-5 (GDF-5) is known to induce tendon tissue and stimulate tendon healing. The hypothesis was that adenoviral GDF-5 transfer leads to transitory transgene expression and improves Achilles tendon healing. In vitro experiments were first performed with rat tenocytes. Transgene expression was evaluated by RT-PCR, Western blotting and GDF-5-ELISA. In vivo virus dosage and transgene expression were examined by a marker gene transfer (LacZ and luciferase). In the main experiment in 131 rats, adenovirus particles (3 x 10(10)) were injected into transected Achilles tendons. The time course of GDF-5 mRNA expression was assessed by real-time RT-PCR. Histology and biomechanical testing were used to evaluate tendon healing and tensile strength. In vitro GDF-5 was secreted with a maximum after 2 weeks (330 ng GDF-5/10(6) cells per 24 hr). In vivo GDF-5 transgene expression showed a maximum at 4 weeks. At 8 weeks, GDF-5 specimens were thicker (p<0.05) with a trend to higher strength (p=0,064). Histology showed greater cartilage formation in type II collagen stains than in controls. Injection of adenovirus particles successfully can deliver the GDF-5 gene in healing tendons and leads to thicker tendon regenerates after 8 weeks. This technique might become a new approach for nonsurgical treatment of tendon injuries.     

脱细胞近交系微型猪肌腱修复兔跟腱缺损

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是其使用受到限制的主要原因。 目的：观察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疗效,以及其作为异种肌腱移植支架材料的可行性。 方法：将40只日本大白兔制作双后肢跟腱缺损实验动物模型后,随机均分为2组,脱细胞猪肌腱组用脱细胞版纳近交系微型猪肌腱修复,自体肌腱组用自体肌腱修复,术后用3-0肌腱线改良HEMI-KESSLER法进行端端原位吻合。 结果与结论：①移植后2周内,脱细胞猪肌腱组与自体肌腱组白细胞计数、C-反应蛋白测量结果差异无显著性意义(P > 0.05)。②两组移植后局部反应小,伤口一期愈合,屈踝功能恢复正常。③组织学检查均未见明显淋巴细胞浸润,肌腱缝合处胶原纤维相互衔接。结果说明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、移植排斥反应轻的优点。     

不同跟腱修复材料特性的临床意义

目的　跟腱断裂术后制动弊端多 ,而早期功能锻炼优点明确 ,探讨各种缝合材料在打结后材料力学特性及滑结情况 ,为临床创新设计 ,术后无须制动的缝合组合提供依据。方法　将不同型号薇乔线 (CV)、慕丝线 (Mersilk)、攀状尼龙线 (Nylon)、普迪思 (PDS)各 1 6条剪断后各打 4个结 ,进行材料力学特性测试。结果　缝合材料的最大载荷、刚度、强度、比能分别为 :PDS >CV >Nylon >Mersilk(P <0 .0 5 )。结论　尽管PDS肌腱缝合线易滑结 ,但 1 0 -PDS ,1 0 -CV、1 0Nylon作端对端跟腱修复强度足够 ,相应 5 0PDS、5 0CV缝合材料作腱周修复也可选择 ,且打 4个以上结可靠。尽管Mersilk不易滑结 ,但不适合跟腱修复。不过好的缝合材料应选择最佳的缝合方法才更有临床意义     

Engineering of functional tendon

Surgical tendon repair is limited by the availability of viable tissue for transplantation. Because of its relatively avascular nature, tendon is a prime candidate for engineered tissue replacement. To address this problem, cells isolated from rat Achilles tendon were grown to confluence in culture and allowed to self-assemble into a cylinder between two anchor points. The resulting scaffold-free tissue was composed of aligned, small-diameter collagen fibrils, a large number of cells, and an excess of noncollagenous extracellular matrix; all characteristics of embryonic tendon. The stress-strain response of the constructs also resembles the nonlinear behavior of immature tendons, and the ultimate tensile strength is approximately equal to that of embryonic chick tendon, roughly 2 MPa. These physical and mechanical properties indicate that these constructs are the first viable tendons engineered in vitro, without the aid of artificial scaffolding.     

A novel biodegradable PCL film for tendon reconstruction: Achilles tendon defect model in rats

This study aims to investigate applicability of poly(epsilon-caprolactone) (PCL) biodegradable films for repair of gaps in Achilles tendons in a rat model, also comparing surgical repair versus no repair approaches. PCL was synthesized with tailor-made properties, then, PCL films were prepared by solvent casting. Seventy-five outbred Sprague-Dawley rats were randomly allocated into five groups: (i) sham operated (skin incision only); (ii) no repair (complete division of the Achilles tendon and plantaris tendon without repair); (iii) Achilles repair (with a modified Kessler type suture); and (iv) plasty of Achilles tendon defects with the biodegradable PCL films, and (v) animals subjected to 1 cm mid-substance defect with no repair. Functional performance was determined from the measurements of hindpaw prints utilizing the Achilles functional index. The animals were killed 8 weeks after surgery and histological and biomechanical evaluations were made. All groups subjected to Achilles tendon division had a significant functional impairment that gradually improved so that by day 28 there were no functional impairments in any group whereas animals with a defect remained impaired. The magnitude of the biomechanical and morphological changes at postoperative 8 weeks were similar for no repair group (conservative), Achilles repair group and tendonplasty group (biodegradable PCL film group). The initial rate of functional recovery was significantly different for primary suture, Achilles repair group and PCL film group (p>0.01). But, at the 28th day, functional recovery was quite similar to the other groups. In summary, our results suggest that the PCL film can be an alternative biomaterial for tendon replacement.     

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.     

不同性别的兔髌腱,跟腱及屈趾肌腱生物力学特性测试

本文以３．５％的应变值对雌、雄性成年日本大耳白兔髌腱、跟腱肌腱进行２５次循环加载和应力松驰以及断裂试验，对不同性别三种肌腱的归一化滞后环面积、循环蠕变、归一化应力松驰、应力松驰衰减因子、弹性特征、结构及材料力学参数进行了测定和比较；发现三种肌腱性别之间的粘弹性及断裂特性无明显差异，因此作者认为在选用兔肌腱作生物力学实验时可不考虑性别；而肌腱之间的粘弹性和结构力学、材料力学特性相差甚远，作用认为这是