Systematic Review of Photobiomodulation Therapy (PBMT) on the Experimental Calcaneal Tendon Injury in Rats.

This systematic review analyzed the light parameters and the effects of photobiomodulation therapy (PBMT) through low-level laser therapy (LLLT) and/or LED (light-emitting diode) on tendon repair of rats submitted to calcaneal injury. This study was conducted in accordance with the guidelines of the Preferred Reporting Items for Meta-Analysis, and PubMed and MEDLINE databases were accessed to search eligible studies published in English. The search terms were as follows: "Achilles tendon" or "Calcaneal tendon" or "tendon injuries" or "soft tissue injuries" and "tendinopathy" or "tendinitis" and "low-level light therapy" or "low-level laser therapy" or "low intensity power therapy" or "light-emitting diode" or photobiomodulation." The SYRCLE (SYstematic Review Center for Laboratory animal Experimentation) risks of bias was used to assess the risk of bias for selected studies. A total of 225 studies were found based on the descriptors used, and only 33 studies were eligible. Light parameters identified per point of irradiation were approximately 60 mW (continuous mode at infrared spectra), 2 W cm-2 , 2 J and 45 J cm-2 . Light parameters at red spectra, continuous versus pulsed mode, and PBMT combined or compared with other therapies such as ultrasound, and studies using unhealthy rats (ovariectomized and/or diabetic models) were also identified and grouped according to these similarities. The main effects found were decreased inflammatory markers and signs of inflammatory process. PBMT (laser/LED) has positive effects in reducing the inflammatory and time for tissue repair in animal models of tendon injury and/or tendinitis using parameters identified.

Effects of Therapy with Light Emitting Diode (LED) in the Calcaneal Tendon Lesions of Rats: A Literature Review.

The objective of this review was to analyze original articles about the effects of therapy with LED in experimental models of calcaneal tendon lesions of rats. The search was performed in the period from February to May 2018, in the following electronic databases: MEDLINE, SciELO, and LILACS, besides the Google Scholar, using the descriptors "Achilles tendon", "Rats", "LED", "Tendinopathy", and "Low-level Light Therapy", as well as their matching parts in the Portuguese and Spanish languages, related to and in association with the relevant terms to the content sought. From the descriptors used 215 works were found. After application of eligibility criteria 8 works were selected, in which positive results were found after the application of the LED. Regarding the main results found with phototherapy, we observed a significant reduction in inflammation. Only one article mentioned little reduction of inflammation. In relation to the number of sessions, there was wide variation, with an average of approximately 5 sessions every 24 hours. Studies in this review pointed out, therefore, positive results in the repair of the calcaneal tendon after therapy with irradiation LED; however, carrying out more experimental studies that help the standardization of parameters to be used in this therapy for further clinical studies becomes necessary.

Low-Level Laser Therapy (780 nm) on VEGF Modulation at Partially Injured Achilles Tendon.

OBJECTIVE: The aim of this study was to analyze the modulatory effects of near infrared (780 nm) low-level laser therapy (LLLT) on the presence of the vascular endothelial growth factor (VEGF) in the partially injured Achilles tendons of rats. BACKGROUND: LLLT stimulates the healing process for Achilles tendon injuries, although the extent of the modulatory effect of LLLT on the VEGF levels found in the injured tendons remains unclear. METHODS: Sixty-five male Wistar rats were distributed in the following seven groups: LASER 1, 3, and 7 (10 partially injured Achilles tendons in each group, which were treated with LLLT for 1, 3, and 7 days, respectively); Sham 1, 3, and 7 (same injury, with simulated LLLT); Control group containing the five remaining animals and in which no procedures were performed. LLLT was applied once a day for 10 sec, with a mean power of 70 mW and fluency of 17.5 J/cm(2). After euthanasia, all of the Achilles tendons were surgically removed and the VEGF levels were analyzed using immunohistochemistry. RESULTS: The VEGF levels remained close to normal (p > 0.05) when comparing the experimental groups (LASER and Sham: 1, 3, and 7) with the Control group. CONCLUSION: LLLT did not stimulate the expression of VEGF in the treated Achilles tendons.

Low-level laser therapy modulates pro-inflammatory cytokines after partial tenotomy.

Tendon injuries give rise to substantial morbidity, and current understanding of the mechanisms involved in tendon injury and repair is limited. This lesion remains a clinical issue because the injury site becomes a region with a high incidence of recurrent rupture and has drawn the attention of researchers. We already demonstrated that low-level laser therapy (LLLT) stimulates the synthesis and organization of collagen I, MMP-9, and MMP-2 and improved the gait recovery of the treated animals. The aim of this study was to evaluate the effects of LLLT in the nitric oxide and cytokines profile during the inflammatory and remodeling phases. Adult male rats were divided into the following groups: G1--intact, G2-- injured, G3--injured + LLLT (4 J/cm(2) continuous), G4--injured + LLLT (4 J/cm(2)-20 Hz--pulsed laser). According to the analysis, the animals were euthanized on different dates (1, 4, 8, or 15 days after injury). ELISA assay of TNF-α, IL-1ß, IL-10, and TGF-ß was performed. Western blotting of isoform of nitric oxide synthase (i-NOS) and nitric oxide dosage experiments was conducted. Our results showed that the pulsed LLLT seems to exert an anti-inflammatory effect over injured tendons, with reduction of the release of proinflammatory cytokines, such as TNF-α and the decrease in the i-NOS activity. Thanks to the pain reduction and the facilitation of movement, there was a stimulation in the TGF-ß and IL-1ß release. In conclusion, we believe that pulsed LLLT worked effectively as a therapy to reestablish the tendon integrity after rupture.

Extracorporeal Shock Wave Treatment (ESWT) enhances the in vitro-induced differentiation of human tendon-derived stem/progenitor cells (hTSPCs).

Extracorporeal shock wave therapy (ESWT) is a non-invasive and innovative technology for the management of specific tendinopathies. In order to elucidate the ESWT-mediated clinical benefits, human Tendon-derived Stem/Progenitor cells (hTSPCs) explanted from 5 healthy semitendinosus (ST) and 5 ruptured Achilles (AT) tendons were established. While hTSPCs from the two groups showed similar proliferation rates and stem cell surface marker profiles, we found that the clonogenic potential was maintained only in cells derived from healthy donors. Interestingly, ESWT significantly accelerated hTSPCs differentiation, suggesting that the clinical benefits of ESWT may be ascribed to increased efficiency of tendon repair after injury.

Evaluation of low-level laser therapy, platelet-rich plasma, and their combination on the healing of Achilles tendon in rabbits.

Tendon repair is still one of the challenges for rehabilitation. Various treatments for tendon injuries have been used in recent decade. This study was established to investigate the effects of low-level laser therapy (LLLT), platelet-rich plasma (PRP) treatment alone, and using combined method on the healing of Achilles tendon in rabbits. Seventy-two healthy mature male white New Zealand rabbits were divided randomly into four groups of 18 animals each: control: partial tenotomy with no treatment, only 1 mL normal saline was injected on days 1, 8, and 15 at the site of splitting; PRP: partial tenotomy with PRP treatment on days 1, 8, and 15 at the site of splitting; LLLT: partial tenotomy with LLLT (K30 hand-held probe, AZOR, Technica, Russia, 650 nm, 30 mW, surface area = 1 cm(2), 60 S/cm(2), energy density = 1.8 J/cm(2)) for 15 consecutive days; LLLT + PRP: partial tenotomy with LLLT + PRP. At the end of trial, the rabbits were euthanatized and tendon specimens were harvested and were submitted for histopathological evaluation, hydroxyproline levels, and biomechanical measurement. The Tukey post hoc test was performed. The results for these parameters showed that PRP or LLLT alone has significant advantages over untreated animals (P < 0.05). Furthermore, it was found that the combined treatment with PRP and LLLT is even more efficient. There was no significant difference (P > 0.05) between the two groups of LLLT and PRP. However, the treatments combining PRP and LLLT showed significant results in comparison of PRP or LLLT alone (P < 0.05). Our results demonstrate that the healing time of injured tendon decreases by using the two therapies combined.

Analysis of the effect of phototherapy in model with traumatic Achilles tendon injury in rats.

The aim of this study was to investigate the effect of low-intensity laser (LILT) infrared (830 nm) therapy in tendon inflammation, tendinitis induced by mechanical trauma in rat Achilles tendon. For this, we used 65 young male Wistar rats, weighing ± 300 g divided into different groups: C = control (n = 5) and experimental (n = 10/group), with two different times of sacrifice such as treated with L = laser, D = treated with diclofenac, and T = untreated injured. The tendon inflammation was induced by controlled contusion in the medial region of the Achilles tendon of the animals. The treated groups received some kind of intervention every 24 h, all groups were sacrificed on the 7th or 14th day after the trauma. The tendons were dissected, extracted, and sent for analysis. Histological analysis of the L group showed a decrease in the number of inflammatory cells in relation to other groups in both periods studied. The comparative results between the number of inflammatory cells in the control and treated groups at 7 and 14 days showed statistically significant differences. Qualitative analysis findings obtained by the picrosirius red technique under polarized light showed that in 7 days, the T group presented collagen types I and III in the same proportion; group D presented a predominance of type III fibers, while in group L, type I collagen predominated. The 14-day group D showed collagen types I and III in the same proportion, while in group L, there was a predominance of type I fibers. Biomechanical analysis showed that 7-day groups L and C showed similar stiffness and increased breaking strength. The 14-day groups L and C showed greater rupturing strength as well as increased stiffness angle. Group D showed a decrease of maximum traction strength and degree of rigidity. It was concluded that treatment with LIL in the parameters used and the times studied reduces migration of inflammatory cells and improves the quality of repair while reducing the functional limitations.

Pulsed LLLT improves tendon healing in rats: a biochemical, organizational, and functional evaluation.

In the last decades, the tendon injuries have increased substantially. Previous results suggested that low-level laser treatment (LLLT) promotes synthesis of extracellular matrix and improves the functional properties of the tendon. The aim of this study was to evaluate the effects of different protocols of LLLT on partially tenotomized tendons. Adult male rats were divided into the following: G1-intact, G2-injured, G3-injured + LLLT (4 J/cm(2) continuous), G4-injured + LLLT (4 J/cm(2) at 20 Hz). G2, G3, and G4 were euthanized 8 days after injury. G5-injured, G6-injured + LLLT (4 J/cm(2) continuous), and G7-injured + LLL (4 J/cm(2) at 20 Hz until the seventh day and 2 kHz from 8 to 14 days). G5, G6, and G7 were euthanized on the 15th day. Glycosaminoglycan (GAG) level was quantified by dimethylmethylene blue method and analyzed on agarose gel. Toluidine blue (TB) stain was used to observe metachromasy. CatWalk system was used to evaluate gait recovery. Collagen organization was analyzed by polarization microscopy. The GAG level increased in all transected groups, except G5. In G6 and G7, there was a significant increase in GAG in relation to G5. In G3 and G4, the presence of dermatan sulfate band was more prominent than G2. TB stains showed intense metachromasy in the treated groups. Birefringence analysis showed improvement in collagen organization in G7. The gait was significantly improved in G7. In conclusion, pulsed LLLT leads to increased organization of collagen bundles and improved gait recovery.

Low-level laser therapy combined with platelet-rich plasma on the healing calcaneal tendon: a histological study in a rat model.

The objective of this study was to investigate the effects of low-level laser therapy (LLLT) treatment alone (λ = 660 nm and λ = 830 nm) or associated with platelet-rich plasma (PRP). We used 54 male rats divided into six groups, with nine animals each: group 1, partial tenotomy; group 2 (GII), PRP; group 3 (GIII): λ660 nm; group 4 (GIV), λ830 nm; group 5 (GV), PRP + λ660 nm; and group 6 (GVI), PRP + λ830 nm. The protocol used was power density 0.35 W/cm(2), energy 0.2 J, energy density 7.0 J/cm(2), time 20 s per irradiated point, and number of points 3. Animals in groups GII, GV, and GVI received treatment with PRP, consisting of a single dose of 0.2 mL directly into the surgical site, on top of the tenotomy. Animals were killed on the 13th day post-tenotomy and their tendons were surgically removed for a quantitative analysis using polarization microscopy. The percentages of collagen fibers of types I and III were expressed as mean ± SD. Higher values of collagen fibers type I were obtained for groups GV and GVI when compared with all other groups (p < 0.05), whereas groups GIII and GIV showed no significant difference between them (p > 0.05). For collagen type III, a significant difference was observed between GII and all other groups (p < 0.5), but no significant difference was found between GIII and GIV and between GV and GVI. Results showed that the deposition of collagen type I was higher when treatment with PRP and LLLT was combined, suggesting a faster regeneration of the tendon.

LLLT improves tendon healing through increase of MMP activity and collagen synthesis.

The Achilles tendon has a high incidence of rupture, and the healing process leads to a disorganized extracellular matrix (ECM) with a high rate of injury recurrence. To evaluate the effects of different conditions of low-level laser (LLL) application on partially tenotomized tendons, adult male rats were divided into the following groups: G1, intact; G2, injured; G3, injured + LLL therapy (LLLT; 4 J/cm(2) continuous); G4, injured + LLLT (4 J/cm(2), 20 Hz); G5, injured; G6, injured + LLLT (4 J/cm(2) continuous); and G7, injured + LLLT (4 J/cm(2), 20 Hz until the 7th day and 2 kHz from 8 to 14 days). G2, G3, and G4 were euthanized 8 days after injury, and G5, G6, and G7 were euthanized on the 15th day. The quantification of hydroxyproline (HOPro) and non-collagenous protein (NCP), zymography for matrix metalloproteinase (MMP)-2 and MMP-9, and Western blotting (WB) for collagen types I and III were performed. HOPro levels showed a significant decrease in all groups (except G7) when compared with G1. The NCP level increased in all transected groups. WB for collagen type I showed an increase in G4 and G7. For collagen type III, G4 presented a higher value than G2. Zymography for MMP-2 indicated high values in G4 and G7. MMP-9 increased in both treatment groups euthanized at 8 days, especially in G4. Our results indicate that the pulsed LLLT improved the remodeling of the ECM during the healing process in tendons through activation of MMP-2 and stimulation of collagen synthesis.

Effect of low-level laser therapy (660 nm) on acute inflammation induced by tenotomy of Achilles tendon in rats.

In this study, we aimed to analyze the effects of low-level laser therapy (LLLT; 660 nm) on levels of protein expression of inflammatory mediators after cutting Achilles tendon of rats. Thirty Wistar male rats underwent partial incisions of the left Achilles tendon, and were divided into three groups of 10 animals according to the time of euthanasia after injury: 6, 24 and 72 h. Each group was then divided into control group and LLLT group (treated with 100 mW, 3.57 W cm(-2), 0.028 cm(2), 214 J cm(-2), 6 J, 60 s, single point). In LLLT group, animals were treated once time per day until the time of euthanasia established for each group. The group treated with LLLT showed a significant reduction of IL-1ß compared with control groups at three time points (6 h: P=0.0401; 24 h: P=0.0015; 72 h: P=0.0463). The analysis of IL-6 showed significant reduction only in the LLLT group at 72 h compared with control group (P=0.0179), whereas IL-10 showed a significant increase in the treated group compared with control group at three experimental times (6 h: P=0.0007; 24 h: P=0.0256; 72 h: P<0.0001). We conclude that LLLT is an important modulator of inflammatory cytokines release after injury in Achilles tendon.

Different power settings of LLLT on the repair of the calcaneal tendon.

OBJECTIVE: The purpose of this study was to evaluate the effect of an 830-nm GaAlAs diode laser operating at output powers of 40, 60, 80, and 100 mW and energy density of 30 J/cm(2) on the repair of partial calcaneal tendon ruptures in rats. METHODS: A partial tendon rupture was induced in all animals, which were treated with laser irradiation for 5 consecutive days. Six days after injury, the injured tendons were removed and examined by polarized light microscopy. Collagen fiber organization was evaluated by birefringence measurements, and collagen content was determined by Picrosirius Red staining. RESULTS: It was observed that the higher the output power (60-100 mW) the greater the amount of type III collagen (p<0.01). The amount of type I collagen was significantly greater (p=0.05) in the 80 mW group than in the control group (sham stimulation). A non-statistically significant improvement in the realignment of collagen fibers was observed in the irradiated groups. CONCLUSIONS: Low-level laser therapy resulted in significantly greater amounts of type III collagen (output powers of 60 mW or more) and type I collagen (output power of 80 mW), however, no significant differences between groups were found in the realignment of collagen fibers.

Effect of In-Ga-Al-P diode laser irradiation on angiogenesis in partial ruptures of Achilles tendon in rats.

OBJECTIVE: This study was conducted to analyze the effect of different irradiances of low-level laser therapy (LLLT) on angiogenesis after partial rupture of Achilles tendon of rats. METHODS: Ninety-six animals were divided into three groups subject to treatment during 3, 5, and 7 days post-lesion. Thirty-two animals were used in each group. The groups were further divided into four subgroups with eight animals in each, receiving In-Ga-Al-P laser (660 nm) treatment at (1) mean output of 10 mW, (2) 40 mW during 10 sec, (3) a sham subgroup, and (4) a non-treatment subgroup. Each animal was subjected to a lesion of the Achilles tendon by dropping a 186-g weight from a 20-cm height over the tendon. Treatment was initiated 6 h post-injury for all the groups. Blood vessels were colored with India ink injection and were examined in a video microscope. RESULTS: Laser exposure promoted an increase in blood vessel count when compared to controls. The 40-mW group showed early neovascularization, with the greatest number of microvessels after three laser applications. The 10-mW subgroup showed angiogenesis activity around the same time as the sham laser group did, but the net number of vessels was significantly higher in the former than in the controls. After seven irradiations, the subgroup receiving 40 mW experienced a drop in microvessel number, but it was still higher than in the control groups. CONCLUSIONS: LLLT of different intensities seems to promote neovascularization in damaged Achilles tendons of rats after partial rupture compared to controls.

Investigation of the supplementary effect of GaAs laser therapy on the rehabilitation of human digital flexor tendons.

OBJECTIVE: To investigate the effect of laser photostimulation in rehabilitation of human digital flexor tendons with a placebo-controlled double-blind prospective study model. BACKGROUND DATA: Low-energy laser therapy has been applied in several rheumatoid and soft tissue disorders with a varying rate of success and it has also been shown to have a positive effect on tendon healing in animal experiments, but no clinical study on laser photostimulation in the treatment of human tendons has been reported to date. MATERIALS AND METHODS: This study was performed in a total of 25 patients with 41 digital flexor tendon injuries in five anatomical zones. In Group I (21 digits in 13 patients), whirlpool and infrared GaAs diode laser with a frequency of 100 Hz. was applied between the 8th and 21st days postoperatively and all patients were given the Washington rehabilitation program until the end of the 12th week. In Group II (20 digits in 12 patients), the same treatment protocol was given but the laser instrument was switched off during applications. RESULTS: The results of the study showed a significant improvement in the laser-treated group only for the parameter of edema reduction (p < 0.01) but the difference between the two groups was non-significant for pain reduction, hand grip strength, and functional evaluation performed according to Strickland and Buck-Gramcko systems using total active motion and fingertip-to distal palmar crease distance parameters (p > 0.05). CONCLUSIONS: Significant improvement obtained in edema reduction both immediately and 12 weeks after supplementary GaAs laser application in our study has been interpreted as an important contribution to the rehabilitation of human flexor tendon injuries because edema is known to have a detrimental effect on functional recovery during both early and late stages of tendon healing. However, our study has failed to show a significant positive effect of supplementary GaAs laser application on the other functional recovery parameters of human flexor tendon injury rehabilitation and we suggest further clinical study in this topic be done using different laser types and dosages in order to delineate the role of this promising treatment modality.

The use of ionising radiation for the treatment of injuries to flexor tendons and supporting ligaments in horses.

A technique was developed using radioactive isotopes as a source of radiation for the treatment of injuries to the superficial and deep flexor tendons and the associated ligaments in the horse. The treatment area was sub-divided so that different dosages could be applied over the limb as necessary. A plaster of Paris impression was taken of the whole area to be treated. In the isotope laboratory a plaster negative was made and loaded with the dose of radioactive isotope. The loaded cast was then strapped to the horse's limb for the calculated time, usually about three days. A total of 42 horses were treated and follow up information was obtained from 28. Twenty-five animals raced again; two relapsed before racing and one was destroyed with navicular disease. Ten of the 42 horses had been treated by firing before irradiation. Five of these returned to racing but the history of four of them was not known.