Regeneration Of Muscle Injury Research Articles

Pulsed low-level infrared laser alters mRNA levels from muscle repair genes dependent on power output in Wistar rats

Satellite cells are present in skeletal muscle functioning in the repair and regeneration of muscle injury. Activation of these cells depends on the expression of myogenic factor 5 (Myf5), myogenic determination factor 1(MyoD), myogenic regulatory factor 4 (MRF4), myogenin (MyoG), paired box transcription factors 3 (Pax3), and 7 (Pax7). Low-level laser irradiation accelerates the repair of muscle injuries. However, data from the expression of myogenic factors have been controversial. Furthermore, the effects of different laser beam powers on the repair of muscle injuries have been not evaluated. The aim of this study was to evaluate the effects of low-level infrared laser at different powers and in pulsed emission mode on the expression of myogenic regulatory factors and on Pax3 and Pax7 in injured skeletal muscle from Wistar rats. Animals that underwent cryoinjury were divided into three groups: injury, injury laser 25 Mw, and injury laser 75 mW. Low-level infrared laser irradiation (904 nm, 3 J cm−2, 5 kHz) was carried out at 25 and 75 mW. After euthanasia, skeletal muscle samples were withdrawn and the total RNA was extracted for the evaluation of mRNA levels from the MyoD, MyoG, MRF4, Myf5, Pax3, and Pax7 gene. Pax 7 mRNA levels did not alter, but Pax3 mRNA levels increased in the injured and laser-irradiated group at 25 mW. MyoD, MyoG, and MYf5 mRNA levels increased in the injured and laser-irradiated animals at both powers, and MRF4 mRNA levels decreased in the injured and laser-irradiated group at 75 mW. In conclusion, exposure to pulsed low-level infrared laser, by power-dependent effect, could accelerate the muscle repair process altering mRNA levels from paired box transcription factors and myogenic regulatory factors.

Histological analysis of the association of low level laser therapy and platelet-rich plasma in regeneration of muscle injury in rats

ObjectiveMuscle injuries are common, and their treatment requires costs and time off. Platelet rich plasma and low level laser therapy have been shown to be affordable and easy to use. The aim of this study was to evaluate the associated effects of low level laser therapy and platelet rich plasma on the treatment of the soleus muscle injured by strain in rats. MethodsThirty-five rats were randomly allocated into five groups: Control (C), Injury Control (IC), injury PRP (IP), injury LLLT (ILT) and injury LLLT and PRP (ILTP). The strain injury was induced in the soleus muscle and the IP group received application of platelet rich plasma immediately after the lesion, while the ILT group received low level laser therapy. After seven days, all animals were euthanized and the soleus muscle removed for further histological analysis. ResultsThe association of both treatments (ILTP) resulted in better histological aspects than the low level laser therapy and platelet rich plasma alone, when compared with the injury group (IC). The collagen analysis exhibited a significant increase in the ILT group (2.99 SD=1.13) compared to the C (1.88 SD=0.41, p=0.012) and IP (2.04 SD=0.44, p=0.018). ConclusionThe association of low level laser therapy with platelet rich plasma produced better results on muscle injury compared to the isolated use of these therapies. Furthermore, none of the treatments could modulate the collagen deposition in relation to injury group.

MYOD mediates skeletal myogenic differentiation of human amniotic fluid stem cells and regeneration of muscle injury

IntroductionHuman amniotic fluid stem (hAFS) cells have been shown to differentiate into multiple lineages, including myoblasts. However, molecular mechanisms underlying the myogenic differentiation of hAFS cells and their regenerative potential for muscle injury remain to be elucidated.MethodsIn order to induce myogenic differentiation of hAFS cells, lentiviruses for MYOD were constructed and transduced into hAFS cells. Formation of myotube-like cells was analyzed by immunocytochemistry, and expression of molecular markers for myoblasts was analyzed by reverse transcription polymerase chain reaction and Western blotting. For in vivo muscle regeneration, MYOD transduced hAFS cells were injected into left tibialis anterior (TA) muscles injured with cardiotoxin, and muscle regeneration was analyzed using hematoxylin and eosin, immunocytochemistry and formation of neuro-muscular junction.ResultsMYOD expression in hAFS cells successfully induced differentiation into multinucleated myotube-like cells. Consistently, significant expression of myogenic marker genes, such as MYOG, DES, DMD and MYH, was induced by MYOD. Analysis of pre-myogenic factors showed that expression of PAX3, MEOX1 and EYA2 was significantly increased by MYOD. MYOD was phosphorylated and localized in the nucleus. These results suggest that in hAFS cells, MYOD is phosphorylated and localized in the nucleus, thus inducing expression of myogenic factors, resulting in myogenic differentiation of hAFS cells. To test regenerative potential of MYOD-transduced hAFS cells, we transplanted them into injured muscles of immunodeficient BALB/cSlc-nu mice. The results showed a substantial increase in the volume of TA muscle injected with MYOD-hAFS cells. In addition, TA muscle tissue injected with MYOD-hAFS cells has more numbers of neuro-muscular junctions compared to controls, indicating functional restoration of muscle injury by MYOD-hAFS cells.ConclusionsCollectively, our data suggest that transduction of hAFS cells with MYOD lentiviruses induces skeletal myogenic differentiation in vitro and morphological and functional regeneration of injured muscle in vivo.

Electromyography and morphology during regeneration of muscle injury in rats

Healing of the partially ruptured rat gastrocnemius muscle was studied correlating electromyographical findings with morphological changes. Fibrillation potentials and positive sharp waves were seen both proximal and distal to the injury 7 days after the injury and disappeared in the proximal part by day 14 and in the distal part by day 21. Late components of motor action potential were observed from day 14 onwards. Denervation was mainly myogenic, i.e. caused either by rupture of myofibres, whence the abjunctional stump lost its contact with the neuromuscular junction on the adjunctional stump, or by necrosis of the segment of the ruptured myofibre lying underneath the neuromuscular junction. Lesser extent of denervation was neurogenic, i.e. caused by damage to intramuscular nerve fibres. The reinnervation occurs either by regeneration of the necrotized myofibres, by regeneration of the severed nerves, or by collateral innervation of new neuromuscular junctions in the abjunctional stumps. The present study indicates that electromyography may be useful in the diagnosis and follow-up of skeletal muscle injuries.