Rat Achilles Tendon Healing Research Articles

Effect of platelet-rich plasma on rat Achilles tendon healing is related to microbiota

Background and purpose — In 3 papers in Acta Orthopaedica 10 years ago, we described that platelet-rich plasma (PRP) improves tendon healing in a rat Achilles transection model. Later, we found that microtrauma has similar effects, probably acting via inflammation. This raised the suspicion that the effect ascribed to growth factors within PRP could instead be due to unspecific influences on inflammation. While testing this hypothesis, we noted that the effect seemed to be related to the microbiota.Material and methods — We tried to reproduce our old findings with local injection of PRP 6 h after tendon transection, followed by mechanical testing after 11 days. This failed. After fruitless variations in PRP production protocols, leukocyte concentration, and physical activity, we finally tried rats carrying potentially pathogenic bacteria. In all, 242 rats were used.Results — In 4 consecutive experiments on pathogen-free rats, no effect of PRP on healing was found. In contrast, apparently healthy rats carrying Staphylococcus aureus showed increased strength of the healing tendon after PRP treatment. These rats had higher levels of cytotoxic T-cells in their spleens.Interpretation — The failure to reproduce older experiments in clean rats was striking, and the difference in response between these and Staphylococcus-carrying rats suggests that the PRP effect is dependent on the immune status. PRP functions may be more complex than just the release of growth factors. Extrapolation from our previous findings with PRP to the situation in humans therefore becomes even more uncertain.

Mesenchymal Stem Cell Therapy on Tendon/Ligament Healing

A normal healing response after ligament and tendon rupture results in scar formation and an inferior tissue that fails to emulate its original structure, composition, and function. More regenerative healing (closer to the original) can be obtained through early suppression of inflammatory cells and associated cytokines. Examination of the immune mediated response of mesenchymal stem/stromal cells (MSCs) during healing indicates that MSCs reprogram macrophages from a pro-inflammatory M1 phenotype to an anti-inflammatory M2 phenotype. Based on these studies our objective was to treat ligament and tendon injuries with MSCs in order to modulate their inflammatory response. Our initial studies using allogeneic cells demonstrated an in vivo dose dependency of MSCs on ligament healing. Medial collateral ligaments (MCLs) treated with 1 × 106 (low dose) MSCs exhibited less inflammation and a reduced number of M1 macrophages compared to ligaments treated with 4 × 106 (high dose) MSCs. Strength of ligament was also improved with the low dose treatment. We then examined the in vivo effects of MSCs that had been preconditioned to be more anti-inflammatory. Treatment with these preconditioned MSCs was compared with normally processed (unconditioned) MSCs using the rat Achilles tendon and MCL healing models. Pre-conditioned MSCs significantly reduced inflammation by increasing the M2 macrophages and decreasing the M1 macrophages. Most importantly, treatment with pre-conditioned MSCs improved tissue strength to levels comparable to intact tissue. Overall, pre-conditioned MSC-treatment out-performed unconditioned MSCs to improve ligament and tendon healing by stimulating a more robust, paracrine-mediated immunosuppressive response.

Cox-2 inhibition and the composition of inflammatory cell populations during early and mid-time tendon healing.

During early tendon healing, the cells within the regenerating tissue are, to a large part, inflammatory leukocytes (CD45+). In a rat Achilles tendon healing model, the inflammation resolves between 5 and 10 days. In the same model, Cox inhibitors (NSAIDs) impair healing when given during the first 5 days, but have a positive effect if given later. We tested the hypothesis that a Cox inhibitor would exert these effects by influencing inflammation, and thereby the composition of the inflammatory cell subpopulations. Achilles tendon transection was performed in 44 animals. Animals were randomized to either parecoxib or saline injections. Healing was evaluated by mechanical testing day 7 after surgery and by flow cytometry day 3 and 10. Cross-sectional area, peak force and stiffness were reduced by parecoxib 31, 33, and 25% respectively (p=0.005, p=0.002, and p=0.005). By flow cytometry, there was a strong effect of time (p<0.001) on virtually all inflammatory cell subpopulations (CD45, CD11b, CD68, CCR7, CD163, CD206, CD3, CD4), but no significant effect of parecoxib at any time point. The results suggest that the negative effects of Cox inhibitors on tendon healing might be exerted mainly via mechanisms not directly related to inflammatory cells.

Mechano-growth factor E peptide promotes healing of rat injured tendon

To determine the possible beneficial effects of a synthetic mechano-growth factor E peptide (MGF-C25E) on the healing of rat Achilles tendon. A significantly increased Achilles functional index was found in the MGF-C25E-treated tendons relative to the saline-treated tendons. Macroscopic observation showed that the repair of MGF-C25E-treated (1000ng/ml) tendons was superior to that of the saline-treated tendons. The mean value of the stiffness of the repaired tendons was significantly increased in the MGF-C25E-treated (1000ng/ml) tendons relative to the saline-treated tendons. Moreover, histological analysis provided evidence of tissue remodeling and cell alignment. MGF-C25E plays an important role in promoting healing of injured rat tendon and could be used as a potential therapeutic factor for tendon repair.

Controlled release of curcumin from curcumin-loaded nanomicelles to prevent peritendinous adhesion during Achilles tendon healing in rats.

We introduced curcumin-loaded nanomicelles into a tendon-healing model to evaluate their effects on tendon healing and adhesion. Three groups consisting of 36 rats underwent rupture and repair of the Achilles tendon. The treatment group received an injection of curcumin-loaded nanomicelles (gold nanorods [GNRs]-1/curcumin in polymeric nanomicelles [curc@PMs] at a dosage of 0.44 mg curcumin/kg in 0.1 mL saline) into the surgical site and exposed to laser postoperatively at weeks 1, 2, and 3, for three times 10 seconds each, on the surgical site in the rats that underwent tendon rupture and repair, while the other two groups received 0.44 mg curcumin/kg in 0.1 mL saline and 0.1 mL of saline, respectively. The specimens were harvested at 4 weeks and subjected to biomechanical and histological evaluation. The scoring results of tendon adhesion indicated that GNRs-1/curc@PMs group was in the lowest grade of peritendinous adhesions compared to the other groups. Histological assessment further confirmed the preventive effect of GNRs-1/curc@PMs on tendon adhesion. These findings indicated greater tendon strength with less adhesion in the group treated with GNRs-1/curc@PMs combined with laser exposure, and that nanoparticle-based therapy may be applied to prevent adhesion in clinical patients.

Nucleostemin- and Oct 3/4-positive stem/progenitor cells exhibit disparate anatomical and temporal expression during rat Achilles tendon healing.

BackgroundThe recent discovery of residing tendon stem/progenitor cells has triggered a growing interest in stem cells as a useful tool in tendon repair. Our knowledge of their involvement in naturally healing tendons is, however, sparse. The aim of this study was to identify and determine stem/progenitor cells in relation to different healing phases and regions in a rat model of Achilles tendon rupture.MethodsSurgery was performed to create a mid-tendon rupture on the right Achilles tendon of 24 rats, whereas the left tendon was used as a control. Tendons were harvested at one, two, eight and 17 weeks post-rupture and stained with antibodies specific to stem/progenitor cells (Octamer-binding transcription factor 3/4 (Oct 3/4) and nucleostemin), migrating cells (Dynamin 2 (Dyn 2)) and leukocytes (CD45). A histological examination was performed on sections stained with Alcian blue.ResultsAt one and two weeks post-rupture, a large number of stem/progenitor cells were discovered throughout the tendon. Most of these cells were nucleostemin positive, whereas only a few Oct 3/4-positive cells were found, mainly situated inside the injury region (I region). At eight and 17 weeks, the increment in stem/progenitor cells had diminished to equal that in the control tendons. At all time points, Oct 3/4-positive cells were also found in the connective tissue surrounding the tendon and at the muscle-tendon junction in both ruptured and control tendons and were often seen at the same location as the migration marker, Dyn 2.ConclusionsThe whole length of the Achilles tendon is infiltrated by stem/progenitor cells at early time points after a mid-tendon rupture. However, different stem/progenitor cell populations exhibit varying anatomical and temporal expressions during Achilles tendon healing, suggesting distinct reparative implications. Oct 3/4 may thus act as a more local, migrating stem/progenitor cell involved in injury-site-specific regenerative effects, as compared to the more general proliferative role of nucleostemin-positive stem/progenitor cells.

Effect of taurine on rat Achilles tendon healing

Taurine has anti-inflammatory and antioxidant characteristics. We have introduced taurine into a tendon-healing model to evaluate its effects on tendon healing and adhesion formation. Two groups of 16 rats underwent diversion and repair of the Achilles tendon. One group received a taurine injection (200 mg/ml) at the repair site, while the other group received 1 ml of saline. Specimens were harvested at 6 weeks and underwent biomechanical and histological evaluation. No tendon ruptured. Average maximum load was significantly greater in the taurine-applied group compared with the control group (p < 0.05). Similarly, average energy uptake was significantly higher in the taurine-applied group compared with the control group (p < 0.05). We observed no significant differences in stiffness in both groups (p > 0.05). After histological assessment, we found that fibroblast proliferation, edema, and inflammation statistically decreased in the treatment group (p < 0.05). These findings could indicate greater tendon strength with less adhesion formation, and taurine may have an effect on adhesion formation.

Can transcutaneous electrical nerve stimulation improve achilles tendon healing in rats?

BACKGROUND: Tendon injury is one of the most frequent injuries in sports activities. TENS is a physical agent used in the treatment of pain but its influence on the tendon's healing process is unclear. OBJECTIVE: To evaluate the influence of TENS on the healing of partial rupture of the Achilles tendon in rats. METHOD: Sixty Wistar rats were submitted to a partial rupture of the Achilles tendon by direct trauma and randomized into six groups (TENS or Sham stimulation) and the time of evaluation (7, 14, and 21 days post-injury). Burst TENS was applied for 30 minutes, 6 days, 100 Hz frequency, 2 Hz burst frequency, 200 µs pulse duration, and 300 ms pulse train duration. Microscopic analyses were performed to quantify the blood vessels and mast cells, birefringence to quantify collagen fiber alignment, and immunohistochemistry to quantify types I and III collagen fibers. RESULTS: A significant interaction was observed for collagen type I (p=0.020) where the TENS group presented lower percentage in 14 days after the lesion (p=0.33). The main group effect showed that the TENS group presented worse collagen fiber alignment (p=0.001) and lower percentage of collagen III (p=0.001) and the main time effect (p=0.001) showed decreased percentage of collagen III at 7 days (p=0.001) and 14 days (p=0.001) after lesion when compared to 21 days. CONCLUSIONS: Burst TENS inhibited collagen I and III production and impaired its alignment during healing of partial rupture of the Achilles tendon in rats.

The effect of organic silicon injection on Achilles tendon healing in rats.

The aim of this study was to assess the efficacy of a soluble absorbable silicon compound on healing of the Achilles tendon. The Achilles tendons of 21 Wistar albino rats were cut and repaired. A 0.01 ml organic silicon solution (silanol) was injected peri/intratendinously into the left leg of all rats and the same dose of saline into the right leg postoperatively. Rats were randomly divided into 3 groups for biomechanical testing on Day 10 (7 rats) and Day 20 (7 rats) and histological and immunohistochemical assessment on Day 20 (7 rats). Fibroblast cell count and diameter, tissue vascularity and blood vessel diameter were evaluated by histomorphometry. Basic fibroblast growth factor (bFGF) immunoreactivity was analyzed with immunohistochemistry on Day 20. Failure load and stiffness of the repaired tendons were measured on Days 10 and 20. The number of fibroblasts per area, average fibroblast diameter, number of vessels parallel to collagen bundles per area and average vessel diameter were significantly higher in the organic silicon group than in the control group (p<0.05). Strong immunoreactivity of bFGF in the silicon group was detected. Failure load was significantly higher in the silicon group than in the control group on Day 10 (p=0.041). On Day 20, while a difference still existed, this difference was not significant. There was no effect of the silicon injection on stiffness of healing tendons. Organic silicon appears to have a positive effect on tendon healing and is suitable for further studies on host healing response modification.

Microtrauma stimulates rat Achilles tendon healing via an early gene expression pattern similar to mechanical loading

Mechanical loading increases the strength of healing tendons, but also induces small localized bleedings. Therefore, it is unclear if increased strength after loading is a response to mechanotransduction or microtrauma. We have previously found only five genes to be up-regulated 15 min after a single loading episode, of them four were transcription factors. These genes are followed by hundreds of genes after 3 h, many of them involved in inflammation. We now compared healing in mechanically unloaded tendons with or without added microtrauma induced by needling of the healing tissue. Nineteen rats received Botox into the calf muscle to reduce loading, and the Achilles tendon was transected. Ten rats were randomized to needling days 2-5. Mechanical testing on day 8 showed increased strength by 45% in the needling group. Next, another 24 rats were similarly unloaded, and 16 randomized to needling on day 5 after transection. Nineteen characteristic genes, known to be regulated by loading in this model, were analyzed by qRT-PCR. Four of these genes were regulated 15 min after needling. Three of them (Egr1, c-Fos, Rgs1) were among the five regulated genes after loading in a previous study. Sixteen of the 19 genes were regulated after 3 h, in the same way as after loading. In conclusion, needling increased strength, and there was a striking similarity between the gene expression response to needling and mechanical loading. This suggests that the response to loading in early tendon healing can, at least in part, be a response to microtrauma.

Achilles Tendon Healing in Rats is Improved by Intermittent Mechanical Loading during the Inflammatory Phase

Achilles Tendon Healing in Rats is Improved by Intermittent Mechanical Loading during the Inflammatory Phase

Paper # 216: The Effect of Organic Silicon Injection on Achilles Tendon Healing in Rats

Organic silicon compounds can affect the activity of fibroblasts and growth factor production but the exact mechanisms are unknown. The purpose of this study was to investigate the effect of organic silicon (monomethyltrisilanol) on healing in a rat Achilles tendon healing model. Organic silicon compounds can affect the activity of fibroblasts and growth factor production but the exact mechanisms are unknown. The purpose of this study was to investigate the effect of organic silicon (monomethyltrisilanol) on healing in a rat Achilles tendon healing model. We hypothese that organic silicon would enhance the healing process in achilles tendon repairs. Twenty-one female Wistar-Albino rats weighting between 350 and 450 grams were used for this study. All left legs prepared as silicon group (21 tendons) and all right legs prepared as control group (21 tendons). Rats were randomly divided into three groups for histological assessment at day 20 (7 rats), biomechanical testing at day 10 (7 rats) and 20 (7 rats). After general anestesia full thickness transverse tenotomy with a number 15 blade was made at a point 0.5 cm proximal to distal insertion of the tendons. Achilles tendon was repaired with modified Kessler technique by using 5/0 monofilament polypropylene sutures. All left legs prepared as silicon group (n:21) and all right legs prepared as control group (n:21) thus, comparison of silicon injected tendons and saline injected tendons were achieved on the same rat. In order to optimise silicon effect on the collagen synthesizing fibroblasts, injection was started at day 3, the end of the inflammatory phase. An 0.01 ml sterile monomethyltrisilanol solution (Conjonctyl®, Sedifa Laboratoire, Monaco) was injected peri/intratendinously on the postoperative day 3,5,7,9,11,13,16 and 19 at the each left leg as silicon group. Same dose of sterile saline solution was injected on same days and same manner at the each right leg as control group.Sacrification process was planned at postoperative day 10 (7 rats/14 tendons for biomechanical testing) and 20 (7 rats/14 tendons for biomechanical testing and 7 rats/14 tendons for histologic assessment). For histologic assesment, fibroblast cell count, fibroblast cell diameter, tissue vascularity and diameter of vessels were evaluated with the morphometric analysis at day 20. For biomechanical assesment, tensile strength of the repaired tendons was also measured biomechanically at day 10 and 20. Biomechanical tests were completed in a computer assisted loading device. (LF plus, Lloyds, UK) The Student t test and Mann-Whitney U test were used to evaluate the significance of differences. A difference was considered to be statistically significant when p <0.05 and 0.01. An increased infiltration of active fibroblasts per area was observed with morphometric analysis on tendon repair site of Silicon group at day 20 (Silicon:76,5±25,4; Control:45,5±16,1 p:0,018*) . The fibroblast cell diameter was also statistically high in Silicon group (Silicon:25,3±1,6 A0 ; Control:17,49±1,8 A0 p:0,001*). Vascularization per area (Silicon:39±8,7; Control:27,2±4,9 p:0,011*) and diameter of vessels (Silicon:27,3±3,7 A0; Control:19,4±1,8 A0 p:0,001*) in Silicon group were increased significantly than in control. The collagen fibrils around the active fibroblasts and extracellular matrix in Silicon group exhibited better organisation and stainability. The cross-sectional area of healing tendon in Silicon group was statistically high at day 10 (Silicon: 17±5,8 mm2 ; Control:10,5±4,5 mm2 p:0,039*) and 20 (Silicon: 15,3±2,6 mm2 ; Control:10,4±2,3 mm2 p:0,003*). The healing tendon constructs in the Silicon group had better failure loads than control group at day 10 (Silicon:45,4±5,7 N; Control:34,7±10,8 N p:0,041*). The difference between two groups was not significant at day 20 (Silicon: 44±22,9 N ; Control:35,4±14,7 N p:0,42). The failure site in all samples was in the tendinous portion of the healing tendon. Our results are in agreement with our initial hypothesis, organic silicon enhances the tendon healing in early phase and suitable for further studies on early mobilization after tendon surgery.

Achilles tendon healing in rats is improved by intermittent mechanical loading during the inflammatory phase

Tendons adapt to changes in mechanical loading, and numerous animal studies show that immobilization of a healing tendon is detrimental to the healing process. The present study addresses whether the effects of a few episodes of mechanical loading are different during different phases of healing. Fifty female rats underwent Achilles tendon transection, and their hind limbs were unloaded by tail suspension on the day after surgery. One group of 10 rats was taken down from suspension to run on a treadmill for 30 min/day, on days 2-5 after transection. They were euthanized on day 8. Another group underwent similar treadmill running on days 8-11 and was euthanized on day 14. Continuously unloaded groups were euthanized on days 8 and 14. Tendon specimens were then evaluated mechanically. The results showed that just four loading episodes increased the strength of the healing tendon. This was evident irrespective of the time point when loading was applied (early or late). The positive effect on early healing was unexpected, considering that the mechanical stimulation was applied during the inflammatory phase, when the calluses were small and fragile. A histological study of additional groups with early loading also showed some increased bleeding in the loaded calluses. Our results indicate that a short episodes of early loading may improve the outcome of tendon healing. This could be of interest to clinical practice.

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.

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.

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.

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.

Effect of vascular endothelial growth factor on rat Achilles tendon healing.

This study evaluated the effect of exogenous vascular endothelial growth factor (VEGF) on tendon healing and regulation of other growth factors in a rat Achilles tendon model. Fifty Sprague-Dawley rats were used. In the experimental group, the left Achilles tendon was transected and repaired with the modified Kessler suture technique, and the right Achilles tendon was transected and repaired with resection of plantaris tendon. VEGF, 100 mul (50 mug/ml), was injected into each tendon at the repair site. The same surgical procedures were performed in the control group, with the same volume of saline injected into the repair sites. At intervals of 1, 2, and 4 weeks, the animals were killed and the tendons were harvested and evaluated for tensile strength (1, 2, and 4 weeks) and gene expression (postoperative day 4). At 1 week postoperatively, when plantaris tendon was preserved, the tensile strength of the repaired tendons with VEGF treatment (3.63 +/- 0.62 MPa) was significantly higher than the tensile strength of the repaired tendons with saline treatment (2.20 +/- 0.36 MPa). There was no difference in tensile strength between the two groups without the plantaris tendon support. At 2 weeks postoperatively, the tensile strength was 11.34 +/- 3.89 MPa in the group with VEGF treatment and plantaris tendon preservation, which was significantly higher than the tensile strength in the other groups. There was no significant difference in tensile strength among the groups at 4 weeks postoperatively. The gene expression showed that transforming growth factor-beta in the VEGF-treated tendon was up-regulated in the early stage of tendon healing, whereas expression of platelet-derived growth factor, basic fibroblast growth factor, and insulin-like growth factor-1 was not significantly different among the groups. In conclusion, administration of exogenous VEGF can significantly improve tensile strength early in the course of the rat Achilles tendon healing and was associated with increased expression of transforming growth factor-beta.

Gastric pentadecapeptide BPC 157 accelerates healing of transected rat Achilles tendon and in vitro stimulates tendocytes growth

In studies intended to improve healing of transected Achilles tendon, effective was a stable gastric pentadecapeptide BPC 157 (GEPPPGKPADDAGLV, M.W. 1419). Currently in clinical trials for inflammatory bowel disease (PLD-116, PL 14736, Pliva), it ameliorates internal and external wound healing. In rats, the right Achilles tendon transected (5 mm proximal to its calcaneal insertion) presents with a large tendon defect between cut ends. Agents (/kg b.w., i.p., once time daily) (BPC 157 (dissolved in saline, with no carrier addition) (10 μg, 10 ng or 10 pg) or saline (5.0 ml)), were firstly applied at 30 min after surgery, the last application at 24 h before autopsy. Achilles functional index (AFI) was assessed once time daily. Biomechanical, microscopical and macroscopical assessment was on day 1, 4, 7, 10 and 14. Controls generally have severely compromised healing. In comparison, pentadecapeptide BPC 157 fully improves recovery: (i) biomechanically, increased load of failure, load of failure per area and Young’s modulus of elasticity; (ii) functionally, significantly higher AFI-values; (iii) microscopically, more mononuclears and less granulocytes, superior formation of fibroblasts, reticulin and collagen; (iv) macroscopically, smaller size and depth of tendon defect, and subsequently the reestablishment of full tendon integrity. Likewise, unlike TGF-β, pentadecapeptide BPC 157, presenting with no effect on the growth of cultured cell of its own, consistently opposed 4-hydroxynonenal (HNE), a negative modulator of the growth. HNE-effect is opposed in both combinations: BPC 157 + HNE (HNE growth inhibiting effect reversed into growth stimulation of cultured tendocytes) and HNE + BPC 157(abolished inhibiting activity of the aldehyde), both in the presence of serum and serum deprived conditions. In conclusion, these findings, particularly, Achilles tendon transection fully recovered in rats, peptide stability suitable delivery, usefully favor gastric pentadecapeptide BPC 157 in future Achilles tendon therapy.