Injection of high dose botulinum-toxin A leads to impaired skeletal muscle function and damage of the fibrilar and non-fibrilar structures

Botulinum-toxin A (BoNT/A) is used for a wide range of conditions. Intramuscular administration of BoNT/A inhibits the release of acetylcholine at the neuromuscular junction from presynaptic motor neurons causing muscle-paralysis. The aim of the present study was to investigate the effect of high dose intramuscular BoNT/A injections (6 UI = 60 pg) on muscle tissue. The gait pattern of the rats was significantly affected 3 weeks after BoNT/A injection. The ankle joint rotated externally, the rats became flat footed, and the stride length decreased after BoNT/A injection. Additionally, there was clear evidence of microstructural changes on the tissue level by as evidenced by 3D imaging of the muscles by Synchrotron Radiation X-ray Tomographic Microscopy (SRXTM). Both the fibrillar and the non-fibrillar tissues were affected. The volume fraction of fibrillary tissue was reduced significantly and the non-fibrillar tissue increased. This was accompanied by a loss of the linear structure of the muscle tissue. Furthermore, gene expression analysis showed a significant upregulation of COL1A1, MMP-2, TGF-b1, IL-6, MHCIIA and MHCIIx in the BoNT/A injected leg, while MHVIIB was significantly downregulated. In conclusion: The present study reveals that high dose intramuscular BoNT/A injections cause microstructural damage of the muscle tissue, which contributes to impaired gait.

[1]  F. Heinen,et al.  Botulinum Toxin A , 1997 .

[2]  B. Ertl-Wagner,et al.  How doctors think – and treat with botulinum toxin , 2010, Developmental medicine and child neurology.

[3]  M. Gough.  Does botulinum toxin prevent or promote deformity in children with cerebral palsy? , 2009, Developmental medicine and child neurology.

[4]  A. Judge,et al.  Botulinum neurotoxin type A causes shifts in myosin heavy chain composition in muscle. , 2005, Toxicon : official journal of the International Society on Toxinology.

[5]  Olga Veksler,et al.  Fast approximate energy minimization via graph cuts , 2001, Proceedings of the Seventh IEEE International Conference on Computer Vision.

[6]  M. Wiberg,et al.  Questionnaire about the Adverse Events and Side Effects Following Botulinum Toxin A Treatment in Patients with Cerebral Palsy , 2015, Toxins.

[7]  Christoph Lepper,et al.  An absolute requirement for Pax7-positive satellite cells in acute injury-induced skeletal muscle regeneration , 2011, Development.

[8]  C. David,et al.  Tomographic Hard X-ray Phase Contrast Micro- and Nano-imaging at TOMCAT , 2010 .

[9]  Anders Brahme,et al.  Development of phase-contrast X-ray imaging techniques and potential medical applications. , 2008, Physica medica : PM : an international journal devoted to the applications of physics to medicine and biology : official journal of the Italian Association of Biomedical Physics.

[10]  Katrin Deinhardt,et al.  The journey of tetanus and botulinum neurotoxins in neurons. , 2003, Trends in microbiology.

[11]  S. Wilkins,et al.  Simultaneous phase and amplitude extraction from a single defocused image of a homogeneous object , 2002, Journal of microscopy.

[12]  S. Schiaffino,et al.  Muscle type and fiber type specificity in muscle wasting. , 2013, The international journal of biochemistry & cell biology.

[13]  T. Ryan,et al.  The three-dimensional structure of trabecular bone in the femoral head of strepsirrhine primates. , 2002, Journal of human evolution.

[14]  A. Shortland,et al.  Could muscle deformity in children with spastic cerebral palsy be related to an impairment of muscle growth and altered adaptation? , 2012, Developmental medicine and child neurology.

[15]  M. El-Domyati,et al.  The use of Botulinum toxin‐A injection for facial wrinkles: a histological and immunohistochemical evaluation , 2015, Journal of Cosmetic Dermatology.

[16]  Hyun-Ho Jung,et al.  Skeletal Muscle Atrophy Induced by Intramuscular Repeated Dose of Botulinum Toxin Type A in Rats , 2007, Drug and chemical toxicology.

[17]  J. Carruthers,et al.  Botulinum toxin in facial rejuvenation: an update. , 2009, Dermatologic clinics.

[18]  Philip K. Russell,et al.  Botulinum toxin as a biological weapon: medical and public health management. , 2001, JAMA.

[19]  Paola Coan,et al.  X-ray phase-contrast imaging: from pre-clinical applications towards clinics , 2013, Physics in medicine and biology.

[20]  Richard Baker,et al.  The effects of botulinum toxin injection frequency on calf muscle growth in young children with spastic cerebral palsy: a 12-month prospective study , 2013, Journal of children's orthopaedics.

[21]  Mitchell F. Brin,et al.  Botulinum neurotoxin type A-cleaved SNAP25 is confined to primary motor neurons and localized on the plasma membrane following intramuscular toxin injection , 2017, Neuroscience.

[22]  B. Jabbari,et al.  Treatment of Chronic Migraine with Focus on Botulinum Neurotoxins , 2015, Toxins.

[23]  F. Tok,et al.  Botulinum Toxin A Treatment in Children with Cerebral Palsy: Its Effects on Walking and Energy Expenditure , 2012, American journal of physical medicine & rehabilitation.

[24]  Chantal Couture,et al.  Mechanisms of action , 1994, Schizophrenia Research.

[25]  Jack R Engsberg,et al.  Relationships between spasticity, strength, gait, and the GMFM-66 in persons with spastic diplegia cerebral palsy. , 2007, Archives of physical medicine and rehabilitation.

[26]  R. Lieber,et al.  Passive mechanical properties and related proteins change with botulinum neurotoxin A injection of normal skeletal muscle , 2012, Journal of orthopaedic research : official publication of the Orthopaedic Research Society.

[27]  T. Takala,et al.  Regulation of synthesis of fibrillar collagens in rat skeletal muscle during immobilization in shortened and lengthened positions. , 2001, Acta physiologica Scandinavica.

[28]  E. Vuorio,et al.  Localization of type I and III collagen and fibronectin production in injured gastrocnemius muscle. , 1991, Laboratory investigation; a journal of technical methods and pathology.

[29]  K. Aoki,et al.  The rat Digit Abduction Score (DAS) assay: a physiological model for assessing botulinum neurotoxin-induced skeletal muscle paralysis. , 2013, Toxicon : official journal of the International Society on Toxinology.

[30]  D M Simpson,et al.  Assessment: Botulinum neurotoxin for the treatment of movement disorders (an evidence-based review) , 2008, Neurology.

[31]  Botulinum Toxin and Muscle Atrophy: A Wanted or Unwanted Effect. , 2016, Aesthetic surgery journal.

[32]  R. Stein,et al.  Skeletal muscle fibre type transformation following spinal cord injury , 1997, Spinal Cord.

[33]  L. V. van Loon,et al.  Reduced satellite cell numbers with spinal cord injury and aging in humans. , 2012, Medicine and science in sports and exercise.

[34]  A P Cosgrove,et al.  BOTULINUM TOXIN IN THE MANAGEMENT OF THE LOWER LIMB IN CEREBRAL PALSY , 1994, Developmental medicine and child neurology.

[35]  S. Christiansen,et al.  Botulinum toxin treatment of extraocular muscles in rabbits results in increased myofiber remodeling. , 2005, Investigative ophthalmology & visual science.

[36]  Jin Han,et al.  Proteomic Changes in Rat Gastrocnemius Muscle After Botulinum Toxin A Injection , 2013, Annals of rehabilitation medicine.

[37]  R. Lieber,et al.  Systems analysis of transcriptional data provides insights into muscle's biological response to botulinum toxin , 2014, Muscle & nerve.

[38]  A. Martinuzzi,et al.  Safety profile of incobotulinum toxin A [Xeomin(®)] in gastrocnemious muscles injections in children with cerebral palsy: Randomized double-blind clinical trial. , 2016, European journal of paediatric neurology : EJPN : official journal of the European Paediatric Neurology Society.

[39]  Thomas C. Südhof,et al.  Botulinum neurotoxin A selectively cleaves the synaptic protein SNAP-25 , 1993, Nature.

[40]  Jennifer A. Lawson,et al.  Satellite cells , connective tissue fibroblasts and their interactions are crucial for muscle regeneration , 2022 .

[41]  C. Das,et al.  Botulinum neurotoxin for the treatment of movement disorders. , 2018, Australian journal of general practice.

[42]  A. Papavasiliou,et al.  Safety of Botulinum Toxin A in Children and Adolescents with Cerebral Palsy in a Pragmatic Setting , 2013, Toxins.

[43]  T. Ryan,et al.  Quantification and visualization of anisotropy in trabecular bone , 2004, Journal of microscopy.

[44]  R. Lieber,et al.  Dramatic changes in muscle contractile and structural properties after 2 botulinum toxin injections , 2015, Muscle & nerve.

[45]  A. B. Scott,et al.  Botulinum toxin injection of eye muscles to correct strabismus. , 1981, Transactions of the American Ophthalmological Society.

[46]  Stan Z. Li,et al.  Markov Random Field Modeling in Image Analysis , 2001, Computer Science Workbench.

[47]  D. Dressler,et al.  Botulinum Toxin: Mechanisms of Action , 2005, European Neurology.

[48]  K. Ohlendieck,et al.  Drastic increase of myosin light chain MLC-2 in senescent skeletal muscle indicates fast-to-slow fibre transition in sarcopenia of old age. , 2009, European journal of cell biology.

[49]  J. Nielsen,et al.  Botulinum toxin injection causes hyper-reflexia and increased muscle stiffness of the triceps surae muscle in the rat. , 2016, Journal of neurophysiology.

[50]  A. Diaz,et al.  Improving organic tandem solar cells based on water-processed nanoparticles by quantitative 3D nanoimaging. , 2015, Nanoscale.

[51]  S. Skinner,et al.  Botulinum toxin therapy for chronic anal fissures: where are we at currently? , 2017, ANZ journal of surgery.

[52]  L. V. van Loon,et al.  Satellite cells in human skeletal muscle plasticity , 2015, Front. Physiol..

[53]  L. Pearce,et al.  Histologic assessment of dose‐related diffusion and muscle fiber response after therapeutic botulinum a toxin injections , 1994, Movement disorders : official journal of the Movement Disorder Society.

[54]  H. Moriyama,et al.  Interaction between gastrocnemius muscle weakness and moderate exercise deteriorates joint integrity in rat knee , 2015, Scandinavian journal of medicine & science in sports.

[55]  S. Swaminathan,et al.  Structural analysis of the catalytic and binding sites of Clostridium botulinum neurotoxin B , 2000, Nature Structural Biology.

[56]  W. Herzog,et al.  A clinically relevant BTX-A injection protocol leads to persistent weakness, contractile material loss, and an altered mRNA expression phenotype in rabbit quadriceps muscles. , 2015, Journal of biomechanics.

[57]  J. Malmsten.  Time course of segmental reflex changes after chronic spinal cord hemisection in the rat. , 1983, Acta physiologica Scandinavica.

[58]  Po-Fan Hsieh,et al.  Botulinum toxin A for the Treatment of Overactive Bladder , 2016, Toxins.

[59]  S. Tågerud,et al.  Effects of botulinum toxin induced muscle paralysis on endocytosis and lysosomal enzyme activities in mouse skeletal muscle , 1986, Pflügers Archiv.

[60]  P. London.  Injury , 1969, Definitions.

[61]  M. Waugh,et al.  Adverse events and health status following botulinum toxin type A injections in children with cerebral palsy , 2011, Developmental medicine and child neurology.

[62]  Françoise Peyrin,et al.  Synchrotron Radiation X-Ray Phase Micro-computed Tomography as a New Method to Detect Iron Oxide Nanoparticles in the Brain , 2013, Molecular Imaging and Biology.

[63]  Scott Ab.  Botulinum toxin injection of eye muscles to correct strabismus. , 1981 .

[64]  H. Teive,et al.  Pain Relief in Cervical Dystonia with Botulinum Toxin Treatment , 2015, Toxins.

[65]  Kenneth S. Campbell,et al.  Effective fiber hypertrophy in satellite cell-depleted skeletal muscle , 2011, Development.

[66]  Franz Pfeiffer,et al.  Evaluation of phase-contrast CT of breast tissue at conventional X-ray sources - presentation of selected findings. , 2013, Zeitschrift fur medizinische Physik.

[67]  K. Johnson.  An Update. , 1984, Journal of food protection.

[68]  Sîan A Williams,et al.  Muscle volume alterations in spastic muscles immediately following botulinum toxin type‐A treatment in children with cerebral palsy , 2013, Developmental medicine and child neurology.

[69]  N. Stott,et al.  Effects of botulinum toxin A injection and exercise on the growth of juvenile rat gastrocnemius muscle. , 2002, Journal of applied physiology.

[70]  The effect of botulinum toxin A on gastrocnemius length: magnitude and duration of response , 1999, Developmental medicine and child neurology.

[71]  Xinyan Zhao,et al.  Visualization of microvasculature and thrombi by X-ray phase-contrast computed tomography in hepatocellular carcinoma. , 2016, Journal of synchrotron radiation.

[72]  C. Fairhurst,et al.  Botulinum toxin and cerebral palsy: time for reflection? , 2005, Developmental medicine and child neurology.

[73]  B. Pedersen.  IL-6 signalling in exercise and disease. , 2007, Biochemical Society transactions.

[74]  Cosmetic Surgery National Data Bank Statistics. , 2016, Aesthetic surgery journal.

[75]  J. Reichenberg,et al.  Treatment of major depressive disorder using botulinum toxin A: a 24-week randomized, double-blind, placebo-controlled study. , 2014, The Journal of clinical psychiatry.

[76]  James A. Nathan,et al.  Muscle wasting in disease: molecular mechanisms and promising therapies , 2014, Nature Reviews Drug Discovery.

[77]  M. Tagliati,et al.  Systematic Literature Review of AbobotulinumtoxinA in Clinical Trials for Blepharospasm and Hemifacial Spasm , 2015, Tremor and other hyperkinetic movements.