Attenuation of the process of muscle regeneration in the toad gastrocnemius muscle by low energy laser irradiation

The effect of number, frequency, and timing of HeNe (6.0 mW; 31.2J/cm²) and Ga-As-diode (average power at 2.82 Hz-0.005 mW) laser irradiations on the process of muscle regeneration at 14 days following cold injury to the toad gastrocnemius muscle was investigated using histomorphometric methods. The volume fraction (percent of total injured area) of mononucleated cells, myotubes and degenerated fibers was 10 ± 1%, 0%, and 4 ± 1%, respectively, in the HeNe laser irradiated muscles (5 irradiations every alternate day, beginning on the 4th day after injury), whereas in the control nonirradiated muscles, these values were significantly higher comprising 57 ± 2% (P < 0.01), 11 ± 1% and 10 ± 2% (P < 0.05), respectively. The volume fraction of young myofibers in injured areas that were subjected to the same laser irradiation regime was 8.6-fold significantly higher (P < 0.01) than their volume fraction in control muscles. The histomorphometric results were the same for injured zones of muscles that were laser irradiated only once, on the 9th day postinjury, and for those that received five consecutive irradiations every alternate day. Muscle regeneration was equally promoted by single Ga-As-diode laser or HeNe irradiation. Multiple irradiations of Ga-As-diode laser caused some pathological changes in the newly formed muscular structures. It is concluded that the process of skeletal muscle regeneration is markedly promoted by low energy laser irradiations, but that the effect depends on the number, timing, and frequency of irradiations and the type of laser used. © 1994 Wiley-Liss, inc.     

Myosin heavy chain isoform profiles remain altered at 7 months if the lacerated medial gastrocnemius is poorly reinnervated: A study in rabbits

Lacerated skeletal muscles often do not recover full function after repair. Denervated muscles with altered myosin heavy chain isoform (MHC) profiles are known to result in functional impairment. We studied the functional recovery of lacerated muscles, assessing MHC profile changes in association to the involvement of the intramuscular nerve (IM). We tested three lacerated models using the rabbit's medial gastrocnemius where the IM was either cut (NNR), repaired (NR), or preserved intact (NP). Muscles were assessed 7 months after repair for muscle atrophy, isometric contraction (by electrical stimulation), and fibrosis formation at the lesion site. Changes in myofibrillar actomyosin adenosine triphosphatase activity, MHC profile, regenerating myofibers and reinnervation were assessed by Western blot, histology, or immunohistology. Lacerated muscles with a repaired (NR) or an intact (NP) IM showed good recovery, with no significant changes in the MHC profile. Muscles where the IM was not repaired (NNR) resulted in significant scar area at the lesion site (p < 0.05), muscle atrophy (67%, p < 0.05) and loss in contractile properties (63% of the uninjured side, p < 0.05). At 7 months, all muscles were reinnervated. However, the NNR had an inappropriate (polyneural) and poorly distributed reinnervation, the presence of regenerating myofibers, and demonstrated a fast‐to‐slow MHC transition (71%:29% to 44%:56%, ANOVA, p = 0.018). This was associated to the cut IM when the NNR muscle was lacerated. Poor reinnervation in lacerated skeletal muscles alters the myosin heavy chain profile permanently. This study provides a rationale to also consider biological solutions to improve nerve regeneration and reinnervation in the surgical repair of lacerated muscles. © 2009 Orthopaedic Research Society. Published by Wiley Periodicals, Inc. J Orthop Res 28:732–738, 2010     

Correlation between biomechanical and structural changes during the regeneration of skeletal muscle after laceration injury

A standardized and reliable model for muscle laceration injuries was developed. The biomechanical and morphological changes during the process of muscle repair after injury were analysed, and the reproducibility of the methods was evaluated. The soleus muscles of Sprague-Dawley rats were completely transected and were allowed to heal for 5, 7, 10, 14, 21, 28, or 56 days, when the muscles either were pulled to failure to measure different parameters of tensile strength or were removed for morphological analysis. During the repair process, the regenerating myofibers penetrated into the connective-tissue scar and formed new myotendinous junctions, thus restoring the functional continuity across the muscle stumps. The muscle atrophied significantly during the recovery period. Mechanical failure occurred in the scar until day 10, and thereafter it occurred within myofibers. Until day 10, the failure load, strain, and specific energy increased to as much as 46.59. and 36% of the control level, respectively: thereafter, there were only minor changes. Stress (tensile strength per cross-sectional area) reached 86% of the control level by day 21 and further increased to as much as 96% of the control level until day 56. These results indicate that the scar becomes stronger than muscle within 14 days: thereafter, the weakest point is the atrophic muscle. The fact that the stress value was most rapidly normalized suggests that, qualitatively, the regenerated muscle had virtually regained its pretrauma strength by day 56 and that the low values of failure load, strain, and specific energy were mainly due to atrophy of the muscle. Thus, further increase in the tensile strength of the regenerated muscle-tendon unit may require active exercise to reverse muscle atrophy.     

Leukemia inhibitory factor promotes the regeneration of rat uterine horns with full‐thickness injury

Severe uterine injuries may lead to infertility or pregnancy complications. There is a lack of effective methods to restore the structure and function of seriously injured uteri. Leukemia inhibitory factor (LIF), which plays a crucial role in blastocyst implantation, promotes the process of regeneration after injury in several different tissues. In this study, we explored the effect of LIF on the regeneration of rat uterine horns following full‐thickness injury. One hundred and twenty four female Sprague–Dawley rats were assigned to three groups, including a sham‐operated group (n = 34 uterine horns), a PBS/collagen group (n = 90 uterine horns), and a LIF/collagen group (n = 124 uterine horns). The regenerated uterine horns were collected at 1, 2, 4, 8, or 12 weeks after the surgery. The results showed that LIF/collagen scaffolds increased the number of endometrial cells and neovascularization 2 weeks after uterine full‐thickness defect in excision sites (p < 0.001 vs PBS/collagen). Eight weeks after the surgery, the number of endometrial glands was dramatically higher in the LIF/collagen scaffolds group (35.2 ± 4.1/field) than in the PBS/collagen scaffolds (15.1 ± 1.4/field). The percentage of a‐smooth muscle actin (a‐SMA)‐positive areas in the LIF/collagen scaffolds (88.8% ± 9.8%) was also significantly higher than that in the PBS/collagen group (52.9% ± 3.7%). Moreover, LIF improved the pregnancy rate and fetus number. We also found that LIF inhibited the infiltration of inflammatory cells and down‐regulated the pro‐inflammatory cytokine IL‐12 expression while up‐regulating the anti‐inflammatory cytokine IL‐10 expression in the injured part of the uterine horns. Our results indicate that LIF promotes regeneration of the uterus after injury, and this is at least partially due to its immunomodulatory properties. In addition, it is worth to explore further the possibility for LIF/collagen to be an alternative therapeutic approach for uterine damage in the clinic in near future.     

Skeletal muscle fiber type conversion during the repair of mouse soleus: potential implications for muscle healing after injury.

We used a mouse model of cardiotoxin injury to examine fiber type conversion during muscle repair. We evaluated the soleus muscles of 37 wild-type mice at 2, 4, 8, and 12 weeks after injury. We also used antibodies (fMHC and sMHC) against fast and slow myosin heavy chain to classify the myofibers into three categories: fast-, slow-, and mixed (hybrid)-type myofibers (myofibers expressing both fMHC and sMHC). Our results revealed an increase in the percentage of slow-type myofibers and a decrease in the percentage of fast-type myofibers during the repair process. The percentage of hybrid-type myofibers increased 2 weeks after injury, then gradually decreased over the following 6 weeks. Similarly, our analysis of centronucleated myofibers showed an increase in the percentage of slow-type myofibers and decreases in the percentages of fast- and hybrid-type myofibers. We also investigated the relationship between myofiber type conversion and peroxisome proliferator-activated receptor-gamma coactivator-1alpha (PGC-1alpha). The expression of both PGC-1alpha protein, which is expressed in both the nucleus and the cytoplasm of regenerating myofibers, and sMHC protein increased with time after cardiotoxin injection, but we observed no significant differential expression of fMHC protein in regenerating muscle fibers during muscle repair. PGC-1alpha-positive myofibers underwent fast to slow myofiber type conversion during the repair process. These results suggest that PGC-1alpha contributes to myofiber type conversion after muscle injury and that this phenomenon could influence the recovery of the injured muscle.     

Muscle stem cell activation in a mouse model of rotator cuff injury

Rotator cuff (RC) tears are frequently complicated by muscle atrophy. Muscle stem cells (MuSCs) repair damaged myofibers following injury, but their role in the prevention or pathogenesis of atrophy following RC tears remains undefined. We hypothesized that the RC MuSC population would be affected by supraspinatus (SS) and infraspinatus (IS) tendon transection (TT) compared to uninjured muscle in a mouse model of RC tear. C57BL6/J mice underwent unilateral SS and IS TT and contralateral sham surgery. At 3, 8, or 14 weeks after injury, mice were euthanized, and SS and IS were harvested for FACS sorting of CD31‐/CD45‐/Sca1‐/ITGa7+/VCAM+ MuSCs or histological analysis. Ki‐67+ MuSCs from injured muscle increased 3.4‐fold at 3 weeks (p = 0.03) and 8.1‐fold at 8 weeks (p = 0.04) following TT injury, but returned to baseline by 14 weeks (p = 0.91). Myod1 remained upregulated 3.3‐fold at 3 weeks (p = 0.03) and 2.0‐fold at 14 weeks (p = 0.0003), respectively. Myofiber cross‐sectional area was decreased at both 3 and 14 weeks after injury, but the number of MuSCs per fiber remained relatively constant at 3 (p = 0.3) and 14 (p = 0.6) weeks after TT. In this study, we characterized the longitudinal effect of RC tendon injury on the MuSC population in supraspinatus and infraspinatus muscles. MuSCs are transiently activated, and are not depleted, in spite of persistent muscle atrophy. © 2017 Orthopaedic Research Society. Published by Wiley Periodicals, Inc. J Orthop Res 36:1370–1376, 2018.

     

Microcurrent Electrical Neuromuscular Stimulation Facilitates Regeneration of Injured Skeletal Muscle in Mice

Conservative therapies, mainly resting care for the damaged muscle, are generally used as a treatment for skeletal muscle injuries (such as muscle fragmentation). Several past studies reported that microcurrent electrical neuromuscular stimulation (MENS) facilitates a repair of injured soft tissues and shortens the recovery period. However, the effects of MENS on the regeneration in injured skeletal muscle are still unclear. The purpose of this study was to investigate the effect of MENS on the regenerative process of injured skeletal muscle and to elucidate whether satellite cells in injured skeletal muscle are activated by MENS by using animal models. Male C57BL/6J mice, aged 7 weeks old, were used (n = 30). Mice were randomly divided into two groups: (1) cardiotoxin (CTX)-injected (CX, n = 15) and (2) CTX-injected with MENS treatment (MX, n=15) groups. CTX was injected into tibialis anterior muscle (TA) of mice in CX and MX groups to initiate the necrosis-regeneration cycle of the muscle. TA was dissected 1, 2, and 3 weeks after the injection. Muscle weight, muscle protein content, the mean cross-sectional areas of muscle fibers, the relative percentage of fibers having central nuclei, and the number of muscle satellite cells were evaluated. MENS facilitated the recovery of the muscle dry weight and protein content relative to body weight, and the mean cross-sectional areas of muscle fibers in CTX-induced injured TA muscle. The number of Pax7-positive muscle satellite cells was increased by MENS during the regenerating period. Decrease in the percentages of fibers with central nuclei after CTX-injection was facilitated by MENS. MENS may facilitate the regeneration of injured skeletal muscles by activating the regenerative potential of skeletal muscles.

Key points

• Microcurrent electrical neuromuscular stimulation (MENS) facilitated the recovery of the relative muscle dry weight, the relative muscle protein content, and the mean cross-sectional areas of muscle fibers of injured TA muscle in mice.
• The number of satellite cells was increased by MENS during the regenerating phase of injured skeletal muscle.
• Decrease in the percentages of fibers with central nuclei was facilitated by MENS.
• MENS may facilitate the regeneration of injured skeletal muscles.

Key words: Muscle injury, regenerative potential, muscle satellite cell, central nuclei, physiotherapy, sports injury     

Rat skin wound healing induced by alternagin-C, a disintegrin-like, Cys-rich protein from Bothrops alternatus venom

Alternagin‐C (ALT‐C) is a disintegrin‐like, Cys‐rich protein isolated from Bothrops alternatus snake venom, which has been shown to induce in vivo angiogenesis. Therefore, this protein could be interesting as a new approach for tissue regeneration studies. Here the effects of ALT‐C on fibroblasts and inflammatory cells, collagen type III and type I and TGF‐α expression in a rat wounded skin model were studied. Thirty‐five male Wistar rats (weight 270 ± 20 g) were divided into seven groups with five animals in each of the following groups: a control group which wounded animals received treatment with natrozol^® gel only; ALT‐C10, ALT‐C60 and ALT‐C100 groups of wounded animals that were treated with the same amount of gel containing 10, 60 and 100 ng of ALT‐C, respectively. Animals were treated once a day with 20 µl of gel associated or not with ALT‐C for 1, 3, 5 or 7 days. ALT‐C treatment increased the fibroblast density, collagen deposition and accelerated the inflammatory process, mostly in the ALT‐C60 group. These results indicate that ALT‐C improves wound repair process in rat skin. Thus, ALT‐C could be a candidate to the development of a novel therapeutic strategy for wounded skin repair.     

GaAs 904-nm laser irradiation improves myofiber mass recovery during regeneration of skeletal muscle previously damaged by crotoxin

This work investigated the effect of gallium arsenide (GaAs) irradiation (power: 5?mW; intensity: 77.14?mW/cm(2), spot: 0.07?cm(2)) on regenerating skeletal muscles damaged by crotoxin (CTX). Male C57Bl6 mice were divided into six groups (n?=?5 each): control, treated only with laser at doses of 1.5?J or 3?J, CTX-injured and, CTX-injured and treated with laser at doses of 1.5?J or 3?J. The injured groups received a CTX injection into the tibialis anterior (TA) muscle. After 3?days, TA muscles were submitted to GaAs irradiation at doses of 1.5 or 3?J (once a day, during 5?days) and were killed on the eighth day. Muscle histological sections were stained with hematoxylin and eosin (H&E) in order to determine the myofiber cross-sectional area (CSA), the previously injured muscle area (PIMA) and the area density of connective tissue. The gene expression of MyoD and myogenin was detected by real-time PCR. GaAs laser at a dose of 3?J, but not 1.5?J, significantly increased the CSA of regenerating myofibers and reduced the PIMA and the area density of intramuscular connective tissue of CTX-injured muscles. MyoD gene expression increased in the injured group treated with GaAs laser at a dose of 1.5?J. The CTX-injured, 3-J GaAs laser-treated, and the CTX-injured and treated with 3-J laser groups showed an increase in myogenin gene expression when compared to the control group. Our results suggest that GaAs laser treatment at a dose of 3?J improves skeletal muscle regeneration by accelerating the recovery of myofiber mass.     

Heat stress facilitates the regeneration of injured skeletal muscle in rats

Background Skeletal muscle stem cells, so-called muscle satellite cells, are responsible for the repair and the regeneration of adult skeletal muscle tissues. Heat stress can facilitate the proliferation and the differentiation of myoblasts in vitro and can enhance their proliferative potential, which may stimulate the regrowth of atrophied skeletal muscle. The purpose of this study was to investigate the effect of heat stress on the regeneration of skeletal muscle injury induced by cardiotoxin. Methods Male Wistar rats, aged 7 weeks, were randomly divided into six groups: a nonheated control group that received a physiological saline injection, a group heat stressed before physiological saline injection, a group heat stressed after physiological saline injection, a group injected with cardiotoxin without heat stress, a group heat stressed before cardiotoxin injection, and a group heat stressed after cardiotoxin injection (25 in each group). To initiate muscle injury and regeneration, 0.5 ml of 10 μM cardiotoxin was injected into the left tibialis anterior muscle. Conscious rats in some groups were exposed to environmental heat stress (41°C for 60 min) in a heat chamber 24 h before or immediately after cardiotoxin or physiological saline injection. The heating protocol in the present study causes an increase in the colonic temperature to 41°C. The left tibialis anterior muscles were dissected 1, 3, 7, 14, and 28 days after injection of cardiotoxin or physiological saline. Results The wet weight and water content of muscles increased 1 day after cardiotoxin injection regardless of the application of heat stress, but normalized after 7–14 days. The muscle protein content in control rats had increased 7 days after heat stress. Although the muscle protein content decreased on cardiotoxin injection, heat stress caused a significant recovery in protein level. Expression of heat shock protein 72 (HSP72) and the number of Pax7-positive nuclei decreased after cardiotoxin injection but increased on the application of heat stress in both normal control and cardiotoxin-injected groups. Conclusions Heat stress stimulated not only the proliferation of satellite cells but also protein synthesis during the regeneration of injured skeletal muscle. It is thus strongly suggested that the heating of injured skeletal muscle may facilitate recovery. There was no direct relationship between the level of HSP72 expression and muscle protein content, suggesting that HSP72 expression may not be the key signal for protein synthesis in the necrosis–regeneration process.     

Improvement of peripheral nerve regeneration in acellular nerve grafts with local release of nerve growth factor

Previous studies have demonstrated the potential of growth factors in peripheral nerve regeneration. A method was developed for sustained delivery of nerve growth factor (NGF) for nerve repair with acellular nerve grafts to augment peripheral nerve regeneration. NGF‐containing polymeric microspheres were fixed with fibrin glue around chemically extracted acellular nerve grafts for prolonged, site‐specific delivery of NGF. A total of 52 Wister rats were randomly divided into four groups for treatment: autografting, NGF‐treated acellular grafting, acellular grafting alone, and acellular grafting with fibrin glue. The model of a 10‐mm sciatic nerve with a 10‐mm gap was used to assess nerve regeneration. At the 2nd week after nerve repair, the length of axonal regeneration was longer with NGF‐treated acellular grafting than acellular grafting alone and acellular grafting with fibrin glue, but shorter than autografting (P < 0.05). Sixteen weeks after nerve repair, nerve regeneration was assessed functionally and histomorphometrically. The percentage tension of the triceps surae muscles in the autograft group was 85.33 ± 5.59%, significantly higher than that of NGF‐treated group, acellular graft group and fibrin‐glue group, at 69.79 ± 5.31%, 64.46 ± 8.48%, and 63.35 ± 6.40%, respectively (P < 0.05). The ratio of conserved muscle‐mass was greater in the NGF‐treated group (53.73 ± 4.56%) than in the acellular graft (46.37 ± 5.68%) and fibrin glue groups (45.78 ± 7.14%) but lower than in the autograft group (62.54 ± 8.25%) (P < 0.05). Image analysis on histological observation revealed axonal diameter, axon number, and myelin thickness better with NGF‐treated acellular grafting than with acellular grafting alone and acellular grafting with fibrin glue (P < 0.05). There were no significant differences between NGF‐treated acellular grafting and autografting. This method of sustained site‐specific delivery of NGF can enhance peripheral nerve regeneration across short nerve gaps repaired with acellular nerve grafts. © 2009 Wiley‐Liss, Inc. Microsurgery, 2009.     

Wound repair during arm regeneration in the red starfish Echinaster sepositus

Starfish can regenerate entire arms following their loss by both autotomic and traumatic amputation. Although the overall regenerative process has been studied several times in different asteroid species, there is still a considerable gap of knowledge as far as the detailed aspects of the repair phase at tissue and cellular level are concerned, particularly in post‐traumatic regeneration. The present work is focused on the arm regeneration model in the Mediterranean red starfish Echinaster sepositus; to describe the early cellular mechanisms of arm regeneration following traumatic amputation, different microscopy techniques were employed. In E. sepositus, the repair phase was characterized by prompt wound healing by a syncytial network of phagocytes and re‐epithelialisation followed by a localized subepidermal oedematous area formation. Scattered and apparently undifferentiated cells, intermixed with numerous phagocytes, were frequently found in the wound area during these first stages of regeneration and extensive dedifferentiation phenomena were seen at the level of the stump, particularly in the muscle bundles. A true localized blastema did not form. Our results confirm that regeneration in asteroids mainly relies on morphallactic processes, consisting in extensive rearrangement of the existing tissues which contribute to the new tissues through cell dedifferentiation, redifferentiation, and/or migration.     

Stromal‐derived factor‐1 delivered via hydrogel drug‐delivery vehicle accelerates wound healing in vivo

Topical treatment of superficial wounds has many advantages including decreased cost and increased ease of application compared with systemic treatments. Many of the advantages, however, are lost when it is necessary for repeated doses of topical medications to be given over an extended period of time. Therefore, a drug‐delivery vehicle that delivers biologically appropriate doses in a sustained fashion would prove valuable. In this study, an alginate hydrogel scaffold impregnated with the angiogenic chemokine stromal‐derived factor‐1 was used to provide targeted, though short‐term, delivery directly into the wound bed. Wounds were created on the dorsum of mice, and either a stromal‐derived factor‐1‐impregnated or a saline‐impregnated scaffold was applied. Wounds were explanted after 1, 3, 7 days, wound area was measured, and histology and immunohistochemistry for endothelial markers were performed. The remaining wound area in stromal‐derived factor‐1‐treated wounds vs. controls was not significant 1 day after wounding (96.7±0.1 vs. 97.5±1.1%, p=0.317), but was significant after 3 days postwounding (46.7±0.1 vs. 82.3±2.4%, p=0.046) and 7 days postwounding (2.3±1.3 vs. 32.0±4.0%, p=0.049). Immunohistochemistry revealed a greater degree of endothelial cell invasion into the wound bed infiltration compared with controls. The results of this study suggest significant clinical promise for our hydrogel‐delivery vehicle in the treatment of wounds.     

Muscle precursor cell autografting in a murine model of urethral sphincter injury

OBJECTIVE: To determine whether muscle precursor cells (MPCs) harvested from limb skeletal muscle can enhance the regeneration process of the striated urethral sphincter after injury. MATERIAL AND METHODS: Striated urethral sphincters of male mice were injured by an injection of a myotoxic substance (notexin). In the experimental group, 2 days after injury, MPCs were enzymatically harvested from striated muscles of the lower limbs and labelled with PKH 26, then immediately re-injected into the injured urethral sphincter of the same animal. In the control group, saline buffer was injected instead of MPCs. Animals were killed 7 days or 1 month after injury and the sphincters removed for histological study (the presence of PKH 26-labelled myofibres, measurement of myofibre diameter and total number of myofibres). RESULTS: MPC autografting accelerated sphincter muscle repair, as shown by a higher myofibre diameter (P = 0.03) and number (P = 0.01) in the experimental group than in the controls at 7 days. One month after their injection MPCs were still detectable in the regenerating sphincters and participated in the formation of new myofibres. CONCLUSION: This study provides the experimental basis for a new therapeutic approach to urethral sphincter insufficiency after surgical or obstetrical injury, based on MPC autografting.     

Histologic evaluation of skin-derived and collagen-based substrates implanted in palatal wounds

Tissue shortage complicates the surgery of cleft lip and palate anomalies and the healing of defects on the palate impairs growth of the dento-alveolar complex due to scar tissue formation. Implantation of substitutes into the wound area might overcome this adverse effect. The aim of this study was to compare the tissue response to three collagen-based (collagen type I substrate alone, or collagen coated with elastin or chondroitin-6-sulfate) and two skin-derived substrates (unprocessed dermis and AlloDerm) after implantation into 12 dogs. Histology was performed at 3, 10, and 20 days postsurgery. We showed that all substrates were well tolerated. However, it is unclear whether AlloDerm was rapidly degraded or if it was sequestrated. There was no elastin or collagen present in these wounds. All collagen-based substrates showed good epithelial regeneration, although heparan sulfate (JM 403) was absent. Wounds treated with the collagen-based substrates contained fewer myofibroblasts at 20 days postsurgery and the type III collagen fibers in the immature scar tissue were more randomly oriented than in an untreated wound. In conclusion, palatal wounds with a dermal substrate heal with fewer indications of scar tissue formation and evoke only a mild inflammatory reaction, which is preferred over the tissue reaction in an untreated wound.     

High final energy of gallium arsenide laser increases MyoD gene expression during the intermediate phase of muscle regeneration after cryoinjury in rats

The aim of this study was to determine the effects of gallium arsenide (GaAs) laser on IGF-I, MyoD, MAFbx, and TNF-α gene expression during the intermediate phase of muscle regeneration after cryoinjury 21 Wistar rats were divided into three groups (n?=?7 per group): untreated with no injury (control group), cryoinjury without GaAs (injured group), and cryoinjury with GaAs (GaAs-injured group). The cryoinjury was induced in the central region of the tibialis anterior muscle (TA). The region injured was irradiated once a day during 14 days using GaAs laser (904 nm; spot size 0.035 cm², output power 50 mW; energy density 69 J cm^?2; exposure time 4 s per point; final energy 4.8 J). Twenty-four hours after the last application, the right and left TA muscles were collected for histological (collagen content) and molecular (gene expression of IGF-I, MyoD, MAFbx, and TNF-α) analyses, respectively. Data were analyzed using one-way ANOVA at P?<?0.05. There were no significant (P?>?0.05) differences in collagen density and IGF-I gene expression in all experimental groups. There were similar (P?<?0.05) decreases in MAFbx and TNF-α gene expression in the injured and GaAs-injured groups, compared to control group. The MyoD gene expression increased (P?=?0.008) in the GaAs-injured group, but not in the injured group (P?=?0.338), compared to control group. GaAs laser therapy had a positive effect on MyoD gene expression, but not IGF-I, MAFbx, and TNF-α, during intermediary phases (14 days post-injury) of muscle repair.     

Inhibition of ADAM10 in satellite cells accelerates muscle regeneration following muscle injury

Muscle injury is one of the most common orthopedic and sports disorders. For severe cases, surgical repair may be indicated; however, other than immobilization and the administration of anti‐inflammatory drugs there is currently no effective conservative treatment for this condition. Satellite cells (SCs) are muscle‐specific stem cells and are indispensable for muscle regeneration after muscle injury. SCs are activated upon muscle injury to proliferate and differentiate into myoblasts, which subsequently fuse into myofibers and regenerate the damaged muscle. We have previously shown that ADAM10, a membrane‐anchored proteolytic enzyme, is essential for the maintenance of SC quiescence by activating the Notch signaling pathway in SCs. Because suppression of ADAM10 activity in SCs can activate SC differentiation, we asked whether inactivation of ADAM10 in SCs after muscle injury could enhance muscle regeneration. Using Adam10 conditional knockout mice, in which ADAM10 activity can specifically be suppressed in SCs, we found that partial inactivation of ADAM10 accelerates muscle regeneration after muscle injury. Nearly identical results were obtained by the administration of GI254023X, a selective ADAM10 inhibitor. The findings of the present study thus indicate that transient enhancement of SC differentiation after muscle injury expedites muscle regeneration and that ADAM10 can be a potential molecular target in treating muscle injuries. © 2018 Orthopaedic Research Society. Published by Wiley Periodicals, Inc. J Orthop Res 36:2259–2265, 2018.

     

ACL injury reduces satellite cell abundance and promotes fibrogenic cell expansion within skeletal muscle

Anterior cruciate ligament (ACL) injuries are associated with significant loss of strength in knee extensor muscles that persists despite physical therapy. The underlying mechanisms responsible for this protracted muscle weakness are poorly understood; however, we recently showed significant myofiber atrophy and altered muscle phenotype following ACL injury. We sought to further explore perturbations in skeletal muscle morphology and progenitor cell activity following an ACL injury. Muscle biopsies were obtained from the injured and non‐injured vastus lateralis of young adults (n = 10) following ACL injury, and histochemical/immunohistochemical analyses were undertaken to determine collagen content, abundance of connective tissue fibroblasts, fibrogenic/adipogenic progenitor (FAP) cells, satellite cells, in addition to indices of muscle fiber denervation and myonuclear apoptosis. The injured limb showed elevated collagen content (p < 0.05), in addition to a greater abundance of fibroblasts and FAPs (p < 0.05) in the injured limb. Fibroblast content was correlated with increased accumulation of extracellular matrix in the injured limb as well. A higher frequency of interstitial nuclei were positive for phospho‐SMAD3 in the injured limb (p < 0.05), providing some evidence for activation of a fibrogenic program through transforming growth factor β following an ACL injury. The injured limb also displayed reduced satellite cell abundance, increased fiber denervation and DNA damage associated with apoptosis (p < 0.05), indicating alterations within the muscle itself after the ligament injury. Injury of the ACL induces a myriad of negative outcomes within knee extensor muscles, which likely compromise the restorative capacity and plasticity of skeletal muscle, impeding rehabilitative efforts. © 2017 Orthopaedic Research Society. Published by Wiley Periodicals, Inc. J Orthop Res 35:1876–1885, 2017.

     

Acceleration of wound healing by topical drug delivery via liposomes

Background: Despite intensive research, impaired wound healing remains a considerable complication. Therefore, topically applied liposome-encapsulated buflomedil hydrochloride was investigated for its ability to improve wound repair in normal (n=16) and ischemic (n=16) skin tissue. Methods: Experiments were performed using the wound healing model of the ear of hairless mice. Standardized skin wounds (4.25 mm²) were created by circular excision of the epidermal layer and the subcutaneous tissue. Liposomes were applied daily until complete neovascularization of the wound occurred. Tissue regeneration by complete epithelialization and neovascularization of the wound area, microcirculatory parameters, and leukocyte–endothelium interaction were investigated by means of intravital microscopy. Microvascular perfusion was assessed by laser-Doppler flowmetry. Results: Topical application of buflomedil liposomes led to a significantly (P<0.05) accelerated wound closure in both normal (9.6±0.7 days) and ischemic (13.4±0.1 days) skin tissue compared with animals that were treated with unloaded liposomes (controls; 13.1±0.8 days; 15.3±0.6 days). Complete neovascularization of the wound was also enhanced (P<0.05) in buflomedil-treated animals (normal tissue 18.8±0.4 days; ischemic tissue 19.6±0.7 days) compared with controls (20.6±0.6 days; 22.6±1.2 days). Conclusion: These data suggest that buflomedil-loaded liposomes might be of beneficial use for clinical wound care. Received: 20 January 1999; in revised form: 18 August 1999 Accepted: 20 August 1999