Achilles Tendon Healing Research Articles

Accelerated Healing of the Rat Achilles Tendon in Response to Autologous Conditioned Serum

Background Despite advances in the treatment of ruptured Achilles tendon, imperfections of endogenous repair often leave patients symptomatic. Local administration of autologous conditioned serum (ACS) in patients with inflammatory, degenerative conditions has shown beneficial effects. Purpose Because ACS also contains growth factors that should accelerate tendon healing, we studied the effect of ACS on the healing of transected rat Achilles tendon. Study Design Controlled laboratory study. Methods In preliminary in vitro experiments, rat tendons were incubated with ACS and the effect on the expression of Col1A1 and Col3A1 was assessed by real-time quantitative polymerase chain reaction. To test its effect in vivo, the Achilles tendons of 80 Sprague Dawley rats were transected and sutured back together. Ten rats from each group (ACS group, n = 40; control group, n = 40) were euthanized at 1, 2, 4, and 8 weeks postoperatively for biomechanical (n = 7) and histologic (n = 3) testing. Lysyl oxidase activity was assayed by a flurometric assay. The organization of repair tissue was assessed histologically with hematoxylin and eosin- and with Sirius red-stained sections, and with immunohistochemistry. Results Tendons exposed to ACS in vitro showed a greatly enhanced expression of the Col1A1 gene. The ACS-treated tendons were thicker, had more type I collagen, and an accelerated recovery of tendon stiffness and histologic maturity of the repair tissue. However, there were no differences in the maximum load to failure between groups up to week 8, perhaps because lysyl oxidase activities were unchanged. Conclusion and Clinical Relevance Overall, our study demonstrates that treatment with ACS has the potential to improve Achilles tendon healing and should be considered as a treatment modality in man. However, as strength was not shown to be increased within the parameters of this study, the clinical importance of the observed changes in humans still needs to be defined.

Rat Achilles tendon healing: mechanical loading and gene expression

Injured tendons require mechanical tension for optimal healing, but it is unclear which genes are upregulated and responsible for this effect. We unloaded one Achilles tendon in rats by Botox injections in the calf muscles. The tendon was then transected and left to heal. We studied mechanical properties of the tendon calluses, as well as mRNA expression, and compared them with loaded controls. Tendon calluses were studied 3, 8, 14, and 21 days after transection. Intact tendons were studied similarly for comparison. Altogether 110 rats were used. The genes were chosen for proteins marking inflammation, growth, extracellular matrix, and tendon specificity. In intact tendons, procollagen III and tenascin-C were more expressed in loaded than unloaded tendons, but none of the other genes was affected. In healing tendons, loading status had small effects on the selected genes. However, TNF-alpha, transforming growth factor-beta1, and procollagens I and III were less expressed in loaded callus tissue at day 3. At day 8 procollagens I and III, lysyl oxidase, and scleraxis had a lower expression in loaded calluses. However, by days 14 and 21, procollagen I, cartilage oligomeric matrix protein, tenascin-C, tenomodulin, and scleraxis were all more expressed in loaded calluses. In healing tendons, the transverse area was larger in loaded samples, but material properties were unaffected, or even impaired. Thus mechanical loading is important for growth of the callus but not its mechanical quality. The main effect of loading during healing might thereby be sought among growth stimulators. In the late phase of healing, tendon-specific genes (scleraxis and tenomodulin) were upregulated with loading, and the healing tissue might to some extent represent a regenerate rather than a scar.

Effects of transforming growth factor-β1 and vascular endothelial growth factor 165 gene transfer on Achilles tendon healing

Repaired Achilles tendons typically take weeks before they are strong enough to handle physiological loads. Gene therapy is a promising treatment for Achilles tendon defects. The aim of the present study was to evaluate the histological/biomechanical effects of Transforming growth factor-β1 (TGF-β1) and vascular endothelial growth factor 165 ( VEGF 165) gene transfer on Achilles tendon healing in rabbits. Bone Marrow-Derived Mesenchymal Stem Cells (BMSCs) were transduced with adenovirus carrying human TGF-β1 cDNA (Ad-TGF-β1), human VEGF 165 cDNA (Ad-VEGF 165), or both (PIRES-TGF-β1/VEGF 165) Viruses, no cDNA (Ad-GFP), and the BMSCs without gene transfer and the intact tendon were used as control. BMSCs were surgically implanted into the experimentally injured Achilles tendons. TGF-β1 distribution, cellularity, nuclear aspect ratio, nuclear orientation angle, vascular number, collagen synthesis, and biomechanical features were measured at 1, 2, 4, and 8 weeks after surgery. The TGF-β1 and TGFβ1/VEGF 165 co-expression groups exhibited improved parameters compared with other groups, while the VEGF 165 expression group had a negative impact. In the co-expression group, the angiogenesis effects of VEGF 165 were diminished by TGF-β1, while the collagen synthesis effects of TGF-β1 were unaltered by VEGF 165. Thus treatment with TGF-β1 cDNA-transduced BMSCs grafts is a promising therapy for acceleration and improvement of tendon healing, leading to quicker recovery and improved biomechanical properties of Achilles tendons.

The roles of TGF-β1 gene transfer on collagen formation during Achilles tendon healing

Collagen content and cross-linking are believed to be major determinants of tendon structural integrity and function. The current study aimed to investigate the effects of transforming growth factor (TGF)-β1 on the collagen content and cross-linking of Achilles tendons, and on the histological and biomechanical changes occurring during Achilles tendon healing in rabbits. Bone marrow-derived mesenchymal stem cells (BMSCs) transfected with the TGF-β1 gene were surgically implanted into experimentally injured Achilles tendons. Collagen proteins were identified by immunohistochemical staining and fiber bundle accumulation was revealed by Sirius red staining. Achilles tendons treated with TGF-β1-transfected BMSCs showed higher concentrations of collagen I protein, more rapid matrix remodeling, and larger fiber bundles. Thus TGF-β1 can promote mechanical strength in healing Achilles tendons by regulating collagen synthesis, cross-link formation, and matrix remodeling.

Early Achilles tendon healing in sheep

The biomechanics of early tendon healing is important for designing post-injury training, but this has not been described in an animal model, similar to humans in size. We measured elastic and viscoelastic properties of a tendon regenerate in sheep, in a study designed to see the effects of exogenously applying the growth and differentiation factor CDMP-2. This is the first description of early tendon healing in sheep Achilles tendons. Twenty female sheep underwent Achilles tendon transection without suturing or immobilization. Two hours after the operation, 100 mug of CDMP-2 or placebo was injected into the hematoma. The sheep were slaughtered after 3 weeks, and tendon regenerates tested for viscoelastic properties by cyclical loading, before a destructive tensile test. Thereafter, all specimens were examined by high resolution computerized tomography (CT), and histology. The tendon regenerate formed a sleeve, around the tendon stumps. Failure occurred between the regenerate sleeve and the tendon stumps. There was an unexpectedly large variation in force at failure. In the CDMP-2 group, force correlated with regenerate transverse area, but not in the controls. Thus, the variation in maximum stress was smaller in the CDMP-2 group (P = 0.009). Although the force at failure was only a tenth of normal, the capacity to store elastic energy was already near normal (hysteresis 16%). The mean transverse area, force at failure and stiffness were all about 30% larger in the CDMP-2 group, but this was not significant. There were no signs of bone or cartilage formation on CT or histology. Results are compatible with a positive effect of CDMP-2, but the power was too low to demonstrate any such effect. Considering that spontaneous ruptures in humans are likely to have a more variable geometry than in this model, humans can also be expected to vary a lot in early mechanical characteristics. This emphasizes the importance of individualized rehabilitation programs. The low hysteresis suggests that the energy storing capacity is rather easy for the tissues to develop; possibly it is harder to create appropriate energy dissipation, in order to avoid re-rupture.

Addition of Nitric Oxide Through Nitric Oxide-paracetamol Enhances Healing Rat Achilles Tendon

Nitric oxide is an important messenger molecule in many physiological processes. The addition of NO via NO-flurbiprofen enhances the material properties of healing tendon, however, flurbiprofen has a detrimental effect on healing. We asked if NO delivered by a cyclooxygenase 3 inhibitor (paracetamol/acetaminophen) would enhance healing in a rat Achilles tendon healing model. Rats were injected subcutaneously daily with NO-paracetamol, paracetamol or vehicle from two days before surgery to the day of tissue harvesting. Paracetamol had no effect on tendon healing compared with vehicle alone. NO-paracetamol did not change the failure load, but did decrease the water content, enhance the collagen content, reduce the cross-sectional area and improve the ultimate stress of healing tendon compared with paracetamol and vehicle. The collagen organization of the healing tendon in the NO-paracetamol group, as determined by polarized light microscopy, was enhanced. Our data suggests NO-paracetamol increases the total collagen content and enhances organization while decreasing the cross-sectional area of healing rat Achilles tendon and is consistent with human clinical trials where NO has improved the symptoms and signs of tendinopathy.

Modulation of early functional recovery of Achilles tendon to bone unit after transection by BPC 157 and methylprednisolone

In the presented study we compared the effect of stable peptide BPC 157 and methylprednisolone on early functional recovery after Achilles tendon to bone transection in a rat model before collagen healing started. Surgical transection of the right Achilles tendon to bone area was performed in seventy two Wistar Albino male rats. Healing Achilles tendon edges were harvested at days 1-4 following the transection. Using Achilles functional index (AFI), myeloperoxidase activity, histological inflammatory cell influx and vascular index early functional recovery was evaluated. Agents (stable peptide BPC 157 10 microg methylprednisolone 5 mg, normal saline 5 ml) were given alone (/kg b.w., intraperitoneally, once daily, first 30 min after surgery, last 24 h before analysis). Control group received normal saline 5 ml/kg. BPC 157 improved functional recovery (AFI values increased at all time points, p <0.05) by anti-inflammatory (decreased myeloperoxidase (MPO) activity and histological inflammatory cell influx, p <0.05) and increased new blood vessel formation (increased vascular index, p <0.05). Methyprednisolone decreased MPO activity and histological inflammatory cell influx, (p <0.05) but also decreased new blood vessel formation and did not affect early functional recovery. Stable peptide BPC 157 with combined anti-inflammatory action and induction of early new blood vessel formation facilitates early functional recovery in Achilles tendon to bone healing.

Ex vivo adenoviral transfer of bone morphogenetic protein 12 (BMP-12) cDNA improves Achilles tendon healing in a rat model

The aim of our study was to evaluate the histological and biomechanical effects of BMP-12 gene transfer on the healing of rat Achilles tendons using a new approach employing a genetically modified muscle flap. Biopsies of autologous skeletal muscle were transduced with a type-five, first-generation adenovirus carrying the human BMP-12 cDNA (Ad.BMP-12) and surgically implanted around experimentally transected Achilles tendons in a rat model. The effect of gene transfer on healing was evaluated by mechanical and histological testing after 1, 2, 4 and 8 weeks. One week after surgery, the maximum failure load of the healing tendons was significantly increased in the BMP-12 group, compared with the controls, and the tendon stiffness was significantly higher at 1, 2 and 4 weeks. Moreover, the size of the rupture callus was increased in the presence of BMP-12 and there was evidence of accelerated remodeling of the lesion in response to BMP-12. Histological examination showed a much more organized and homogeneous pattern of collagen fibers at all time points in lesions treated with the BMP-12 cDNA muscle graft. Both single fibrils and the collagen fibers had a greater diameter, with a higher degree of collagen crimp than the collagen of the control groups. This was confirmed by sirius red staining in conjunction with polarized light microscopy, which showed a higher shift of small yellow-green fibers to strong yellow-orange fibers after 2, 4 and 8 weeks in the presence of BMP-12 cDNA. There was also an earlier shift from fibroblasts to fibrocytes within the healing tendon, with less fat cells present in the tendons of the BMP-12 group compared with the controls. Treatment with BMP-12 cDNA-transduced muscle grafts thus produced a promising acceleration and improvement of tendon healing, particularly influencing early tissue regeneration, leading to quicker recovery and improved biomechanical properties of the Achilles tendon. Further development of this approach could have clinical applications.

Mechanical Load and BMP Signaling During Tendon Repair: A Role for Follistatin?

Healing of the rat Achilles tendon is sensitive to mechanical loading, and the callus strength is reduced by 3/4 after 14 days, if loading is prevented. Exogenous GDFs stimulate tendon healing. This response is influenced by loading: without loading, cartilage and bone formation is initiated. This implies BMP signaling is crucial during tendon healing and influenced by mechanical loading. We therefore asked if mechanical loading influences the gene expression of the BMP signaling system in intact and healing tendons, and how the BMP signaling system changes during healing. The genes were four BMPs (OP-1/BMP-7, GDF-5/CDMP-1/BMP-14, GDF-6/CDMP-2/BMP-13, and GDF-7/CDMP-3/BMP-12), two receptors (BMPR1b and BMPR2), and the antagonists follistatin and noggin. The Achilles tendon was transected in rats and left to heal. Half of the rats had one Achilles tendon unloaded by injection of Botox in the calf muscles. Ten tendons were analyzed before transection and for each of four time points. All genes except noggin were expressed at all time points, but followed different patterns during healing. Loading strongly decreased the expression of follistatin, which could lead to increased signaling. The BMP system appears involved in tendon maintenance and healing, and may respond to mechanical loading.

High-dose vitamin C supplementation accelerates the Achilles tendon healing in healthy rats

This experimental study was performed to assess, whether or not, vitamin C, required during the collagen synthesis, would influence the Achilles tendon healing in a healthy rat model. The right Achilles tendons of 42 healthy female Wistar Albino rats were completely ruptured. The rats were randomly divided into the vitamin C and control groups and both groups included third, tenth and twenty-first day subgroups. One hundred and fifty milligrams (1.5 cc) of vitamin C and 1.5 cc % 0.9 NaCl were injected once for every 2 days for the vitamin C and control groups, respectively. Qualitative and quantitative microscopic comparisons of the repair tissues of both groups were made on the mentioned days. Angiogenesis was more evident on the third day in the vitamin C group. There was a significant difference between the control and vitamin C groups regarding the type I collagen production on the tenth day. The structure of the repair tissue was almost in the form of regular dense connective tissue at the end of twenty-first day in the vitamin C group. Mean collagen fiber diameter was considerably higher, and the number of active fibroblasts in the repair tissue was slightly elevated in the vitamin C group during the entire healing process. High-dose vitamin C supplementation once for every 2 days has stimulating effects on the Achilles tendon healing because of early angiogenesis and increased collagen synthesis in a healthy rat model. Further studies are needed to make clear the mentioned encouraging effects of the vitamin C on the Achilles tendon healing.

Mechanical properties during healing of Achilles tendon ruptures to predict final outcome: A pilot Roentgen stereophotogrammetric analysis in 10 patients

BackgroundThere are presently few methods described for in vivo monitoring of the mechanics of healing human tendon ruptures, and no methods for prediction of clinical outcome. We tested if Roentgen stereophotogrammetric analysis (RSA) can be used to follow the restoration of mechanical properties during healing of ruptured Achilles tendons, and if early measurements can predict clinical results.MethodsAchilles tendon repair was studied with RSA in 10 patients with a total rupture. Tantalum beads were implanted in conjunction with surgical repair. The patients were evaluated at 6, 12 and 18 weeks, and after 1 year. RSA was performed with two different mechanical loadings, and the strain induced by increasing load was measured. The transverse area was determined by ultrasound. CT scan at 12 weeks confirmed that the tantalum beads were located within the tendons. Functional testing was done after 1 year. A heel raise index was chosen as primary clinical outcome variable.ResultsThe strain was median 0.90, 0.32 and 0.14 percent per 100 N tendon force at 6 weeks, 18 weeks and one year respectively. The error of measurement was 0.04 percent units at 18 weeks. There was a large variation between patients, which appears to reflect biological variation. From 6 to 18 weeks, there was a negative correlation between increase in transverse area and increase in material properties, suggesting that healing is regulated at the organ level, to maximize stiffness. Modulus of elasticity during this time correlated with a heel raise index at one year (Rho = 0.76; p = 0.02).ConclusionWe conclude that the RSA method might have potential for comparing different treatments of Achilles tendon ruptures.

The Effect of Therapeutic Ultrasound Intensity on the Ultrastructural Morphology of Tendon Repair

This study evaluated the effects of ultrasound intensity on the ultrastructural morphology of Achilles tendon healing. Twenty Sprague-Dawley rats with surgically hemi-transected Achilles tendons were randomly assigned into four groups of 0, 0.5, 1.2 and 2 W/cm 2 for ultrasound treatment, with five rats in each group. The treatments were administered with 1 MHz continuous ultrasound daily starting from day 5 after injury. On day 30, ultrathin slides of the Achilles tendons were prepared and examined with transmission electron microscopy. Results showed that the mean collagen fibril size of all treatment groups was higher than the control ( p < 0.05). There was no significant difference in the collagen fibril size among the treatment groups. These findings suggest that therapeutic ultrasound can enhance the maturation of collagen fibrils of repairing tendons, and this was not dependent on the intensity of ultrasound applied. (E-mail: rsgng@polyu.edu.hk)

Oxygen free radicals and tendon healing

Nitric oxide is a small free radical generated by a family of enzymes, the nitric oxide synthases (NOSs). In a series of experiments performed over the last 15 years, we showed that nitric oxide is induced by all 3 isoforms of NOS during tendon healing and that it plays a crucial beneficial role in restoring tendon function. In normal tendons, very little NOS activity was found, whereas in injured rat and human tendons, NOS activity was expressed in healing fibroblasts in a temporal fashion. In healing rat Achilles tendon fibroblasts, the first isoform to be expressed was endothelial NOS, followed by inducible NOS and then brain or neuronal NOS. Systemic inhibition of NOS activity decreased the cross-sectional area and mechanical properties of the healing rodent Achilles tendons. The addition of nitric oxide via nitric oxide-flurbiprofen enhanced rat Achilles tendon healing. The addition of nitric oxide to cultured human tendon cells via chemical means and via adenoviral transfection enhanced collagen synthesis, suggesting that one mechanism for the beneficial effect of nitric oxide on tendon healing might be via matrix synthesis. Most recently, 3 randomized, double-blind clinical trials evaluated the efficacy of nitric oxide donation via a patch in the management of the tendinopathy. In all 3 clinical trials, there was a significant positive beneficial effect of nitric oxide donation to the clinical symptoms and function of patients with Achilles tendinopathy, tennis elbow, and supraspinatus tendinitis.

An in-vivo experimental evaluation of He–Ne laser photostimulation in healing Achilles tendons

There is no method of treatment that has been proven to accelerate the rate of tendon healing or to improve the quality of the regenerating tendon. Low level laser photostimulation has gained a considerable attention for enhancing tissue repair in a wide spectrum of applications. However, there is controversy regarding the effectiveness of laser photostimulation for improvement of the healing process of surgically repaired tendons. Accordingly, the present study was conducted to evaluate the role of helium-neon (He-Ne) laser photostimulation on the process of healing of surgically repaired Achilles tendons. Thirty unilateral Achilles tendons of 30 Raex rabbits were transected and immediately repaired. Operated Achilles tendons were randomly divided into two equal groups. Tendons at group A were subjected to He-Ne laser (632.8 nm) photostimulation, while tendons at group B served as a control group. Two weeks later, the repaired Achilles tendons were histopathologically and biomechanically evaluated. The histopathological findings suggest the favorable qualitative pattern of the newly synthesized collagen of the regenerating tendons after He-Ne laser photostimulation. The biomechanical results support the same favorable findings from the functional point of view as denoted by the better biomechanical properties of the regenerating tendons after He-Ne laser photostimulation with statistical significance (p <or= 0.01) at most of the biomechanical parameters. He-Ne laser photostimulation reported a great value after surgical repair of ruptured and injured tendons for a better functional outcome. It could be applied safely and effectively in humans, especially with respect to the proposed long-term clinical outcome.

The effect of subcutaneously injected nicotine on achilles tendon healing in rabbits

The objective of this study was to evaluate the effect of subcutaneously injected nicotine on transversely transected and sutured achilles tendon healing in an experimental rabbit model. Adult New Zealand rabbits (n=22) weighting 3,000-3,500 g were used in this experimental study. Rabbits were randomly divided into two groups. Achilles tendon was transversely incised and repaired in all animals. In the experiment group subcutaneous injection of Nicotine tartrate 3 mg/kg/day was done. In the control group Serum physiologic injection was done at the same dosage. The injections were made three times a day in equal dosages. Nicotine and SF injections were made until the end of the 8-week, and then all animals were euthanized. Both light microscopic and electron microscopic evaluations were made on 14 animals. In N group light microscopic evaluation showed a visible gap in repair site. The total tendon score represented in N group was less than in SF group. The statistical analysis of the groups was significantly different for total tendon scores (P=0.002). Beside this electron microscopic examination showed inactive and degenerated fibroblasts and irregular collagen fibrils around them as well as collagen synthesis interruption in N group. Biomechanical evaluation was made on eight animals. The average tensile strength values in Group N (139.47+/-44.55 N) were significantly lower than those in Group SF (265.9+/-39.01 N) (z=2.309, P=0.029). Nicotine is the major chemical component common to all cigarettes and previously has been shown to affect wound and fracture healing adversely. The results of this study show that nicotine impairs achilles tendon healing after a surgical repair.

Spontaneous recovery of injured Achilles tendon in inducible nitric oxide synthase gene knockout mice

To determine if inducible nitric oxide synthase (iNOS) gene could affect Achilles tendon healing using iNOS gene knockout mice. 21 iNOS knockout (iNOS(-/-)) mice and 8 of the wild type (iNOS(+/+)) mice were utilized in this study. Group 1: iNOS(+/+) mice (n = 8), group 2: iNOS(-/-) mice (n = 11) and group 3: iNOS(-/-) with a NOS inhibitor, (aminoguanidine, 500 mg/kg/day, via an intraperitoneal mini-osmotic pump for 7 days, n = 10). The right Achilles tendon was transected in all mice and harvested on day 7 for cross-sectional area and biomechanical properties. Serum nitrate concentration of the mice was measured by gas chromatography mass spectrometry (GC/MS). A significant reduction in cross-sectional area of the healing Achilles tendon was observed in group 3 mice compared to group 2 mice (p < 0.01). The serum nitrate concentration in both group 2 and group 3 mice was lower than that in group 1 mice (p < 0.01) iNOS gene deletion and inhibition of NOS did not affect the biomechanical properties of the healing tendons. iNOS gene is not solely responsible for the beneficial effects of nitric oxide (NO) on tendon healing.

Exogenously Administered Substance P and Neutral Endopeptidase Inhibitors Stimulate Fibroblast Proliferation, Angiogenesis and Collagen Organization during Achilles Tendon Healing

In the last few years much research has been conducted in methods to promote tendon healing. The aim of this study was to determine if the healing process after operative repair of rat Achilles tendons could be stimulated by the paratendinous injection of a sensory peptide, substance P (SP). Ninety-six male Sprague-Dawley rats were randomly allocated to four groups: (I) control buffer injections, (II) injections of SP 10(-6) mol/kg body weight combined with a carrier, (III) injections of SP 10(-8) mol/kg BW with the carrier, and (IV) injections with the carrier only (thiorphan 1 micromol/kg BW and captopril 30 micromol/kg BW, both neutral endopeptidase inhibitors). The influence on tissue repair was determined from the histologic measurement of fibroblast proliferation, angiogenesis, and collagen organization. On days 7, 14, 28 and 42, animals were sacrificed and histologic evaluations were performed on the injured Achilles tendon constructs. The two groups subjected to SP injections showed a significant initial fibroblast proliferation on day 7 (p < 0.05), which rapidly declined by day 14 to the level of cellular proliferation observed with the use of thiorphan and captopril. Capillary proliferation showed a similar evolution, except that in the second week angiogenesis in the treated groups was below the level of the control group. Strikingly, collagen orientation increased faster in the groups injected with SP. This was obvious from the second week already and the difference remained until the completion of the study. This is the first study to demonstrate that paratendinous injections of SP after operative repair of the Achilles tendon in rats appears to provide a boost to the initial stages of healing and significantly accelerate the reparative phase of the healing process.

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× 1010) 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-5/106 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.

The influence of ultrasound on the mechanical properties of healing tendons in rabbits

Background Therapeutic ultrasound is commonly used for treatment of injuries to tendons, ligaments and joint capsules. Opinions still differ, however, as to the beneficial effects of ultrasound treatment of the above-men-tioned conditions.Methods We studied the effect of various ultrasound intensities on the healing of tenotomized, sutured and immobilized Achilles tendons of adult rabbits. Different intensities of pulsating ultrasound (0, 50, 100, 200, 500, 750, 1 000 and 2 000 mW/cm2, frequency 3 MHz) were given over the healing tendons for 5 min daily, using a gel as the coupling agent between the ultrasound probe and the skin. Eleven days after the tenotomy, the healing tendons were analyzed in a materials testing machine.Results The extensibility of the healing tendons was greater after sonication at an intensity of 2 000 mW/cm2than after 50 mW/cm2. We found no significant effect on the load at rupture and normalized load at rupture of the ultrasound-treated healing tendons compared with mock-sonicated healing tendons. A gradual decline was observed, however, in the stiffness and collagen content of the healing tendons with increasing intensity of the ultrasound treatment.Interpretation The pulsating ultrasound treatment applied did not improve the mechanical properties of healing Achilles tendons at day 11 after the operation. On the other hand, a slight decline in stiffness was observed with increasing intensity of ultrasound treatment.

Comparison of therapeutic ultrasound and exercises for augmenting tendon healing in rats

We compared therapeutic ultrasound (US) and exercises on Achilles tendon healing in 49 mature male rats. The medial Achilles tendon of the right leg was transected and the rats were divided into five groups: 1. control ( n = 9), 2. 1 W/cm 2 US ( n = 9), 3. 2 W/cm® US ( n = 10), 4. running ( n = 11) and 5. swimming ( n = 10). All animals were treated daily, starting from postsurgical day 5, according to their group assignment, except for group 1 control. Achilles functional index (AFI) was recorded pre- and postoperatively. On day 30, the Achilles tendons were biomechanically tested. Results revealed that groups 3 and 4 had higher tensile strength than the controls ( p = 0.037 and 0.034), but groups 2 and 5 were not different from the controls. No significant difference in stiffness ( p = 0.21) or AFI ( p = 0.22) was found. These findings suggest that therapeutic US treatment at 2 W/cm 2 and running exercise improved the strength of repairing Achilles tendon. (E-mail: rsgng@polyu.edu.hk)