3D-Microscopic Muscle Architecture in Cerebral Palsy
ConditionSpastic Cerebral Palsy
Trial statusRecruiting
Trial phaseNot listed
Trial typeObservational
Biological sexAll
Age2-9
SponsorUniversitaire Ziekenhuizen KU Leuven
About this trial
The focus of this study is to understand and define the mechanisms of the altered muscle development and growth on a microscopic level within a long-term perspective in children with cerebral palsy and to relate these findings to muscle macroscopic properties defined by muscle imaging, to neuromuscular symptoms and to treatment.
This study aims to (1) evaluate intrinsic microscopic muscle properties of young growing children with CP, and (2) to evaluate these muscle properties in relation to macroscopic properties, neuromuscular symptoms and to treatment.
Improved understanding of changes in microscopic muscle properties, and how they relate to macroscopic properties and to the neuromuscular symptoms as well as how they are influenced by treatment, has the potential to delineate CP phenotypes prone to intervention and to optimize treatment protocols or develop new treatments, leading to new avenues for improving function in CP.
The method to study microscopic muscle properties involves analysis of muscle biopsies (histological / immunohistochemistry analysis, SC and IC culture, gene expression). Biopsies will be collected using the minimally invasive percutaneous needle microbiopsy technique, suitable for collecting repeated samples over time in the same individual while still leading to sufficient tissue of good quality for subsequent analysis1,2. For the children with CP, the local hospital's tradition of applying general anesthesia for delivering BTX injections will be exploited to collect the muscle samples prior to the BTX session and the one-year follow-up, or general anesthesia planned for orthopedic surgery or diagnostic imaging such as MRI, etc. For the collection of the samples 3 months before the BTX session, as well as 3 months and 6 months after BTX injections, the common approach for microbiospy collection will be applied, with local sedation on the skin (Rapydan©) and fascia (Xylocaine©) and local anesthesia by using Kalinox© (nitrous oxide in oxygen) under supervision of the University Hospital PROSA team. Biopsies of TD muscles will be collected in children with no history of neurological disorder, nor musculoskeletal problems at the level of the gastrocnemius or semitendinosus, at the time of upper limb orthopedic or trauma surgery and thus always under general anesthesia. Ultrasound guided percutaneous muscle biopsy has been performed in children (2 months-18 years)3,4 and has proven to be safe and well-tolerated. A pilot study (S61110) was conducted to confirm that the microbiopsy technique is suitable for the analysis of microscopic muscle properties and is well-tolerated in children with CP.
Two specific research goals are planned, with hypotheses emerging from literature.
Eligibility criteria
This trial accepts healthy volunteersQualifiers
Children (boys/girls) diagnosed with predominantly spastic type of CP
Uni- or bilateral involvement
Gross Motor Function Classification Scale (GMFCS) Level I-III50
2 to 9 years of age
Disqualifiers
Presence of dystonia or ataxia
Previous surgery less than 6 months at the investigated muscles
Severe co-morbidities (that are likely to prevent proper assessment, such as severe cognitive problems)
Children (boys/girls)
Trial population
Children with spastic cerebral palsy, who have routine follow-up care at the CP reference center of the University Hospitals Leuven.
Trial design
Design model
Cohort
Time perspective
Prospective
Treatments tested in this trial
Not listed
Trial groups
130 Participants
are grouped into 4 trial groupsGroup A: Children with spastic cerebral palsy
Group B: Typical developing children
Group C: Adolescents and adults with HSP
Group D: Typically developing adolescents and adults
Trial outcomes
Primary outcomes
1
Muscle fiber size
Cross sectional area in µm of the fiber. Muscle fiber size will be assessed on cryosection stained with an antibody cocktail specific to laminin, myosin heavy chain (MHC)-I, MHC-IIA, MHC-IIB, MHC-embryonal and immunofluorescent detection will be done using appropriate secondary antibodies.
Time frame
Through a study participation of 1,2 years depending on the trajectory: 1 evaluation moment at baseline, 1 follow-up 1,2 years after, or 5 evaluations: 3 months pre, baseline, 3 months after, 6 months after and 1,2 years after baseline2
Muscle fibre type proportion
For each muscle, muscle fiber proportion will be assessed on cryosection stained with an antibody cocktail specific to laminin, myosin heavy chain (MHC)-I, MHC-IIA, MHC-IIB, MHC-embryonal and immunofluorescent detection will be done using appropriate secondary antibodies.
Time frame
Through a study participation of 1,2 years depending on the trajectory: 1 evaluation moment at baseline, 1 follow-up 1,2 years after, or 5 evaluations: 3 months pre, baseline, 3 months after, 6 months after and 1,2 years after baseline3
Capillary density
Capillaries /mm2 fiber and capillary to fiber ratio. Capillaries will be detected in-situ. Capillaries will be assessed using CD31 staining.
Time frame
Through a study participation of 1,2 years depending on the trajectory: 1 evaluation moment at baseline, 1 follow-up 1,2 years after, or 5 evaluations: 3 months pre, baseline, 3 months after, 6 months after and 1,2 years after baseline4
Satellite cell density
Number of satellite cells/mm2. Satellite cells will be detected in-situ. Co-localization of satellite cells will be assessed using Pax7 staining in conjunction with MHC-I/laminin, DAPI and CD31. Abundance of satellite cell content (Pax7+/DAPI+) will be counted and their association with the different fiber types will be identified.
Time frame
Through a study participation of 1,2 years depending on the trajectory: 1 evaluation moment at baseline, 1 follow-up 1,2 years after, or 5 evaluations: 3 months pre, baseline, 3 months after, 6 months after and 1,2 years after baselineSecondary outcomes
1
Overall changes in collagen content
Estimation of the collagen content (µg/mg) collected from a micro biopsy from the medial gastrocnemius muscle and the semitendinosus muscle.
Time frame
Through a study participation of 1,2 years depending on the trajectory: 1 evaluation moment at baseline, 1 follow-up 1,2 years after, or 5 evaluations: 3 months pre, baseline, 3 months after, 6 months after and 1,2 years after baseline2
Overall change in muscle composition of the medial gastrocnemius muscle and the semitendinosus muscle by means of echo-intensity
Muscle composition can be indirectly estimated based on echo-intensity. Echo-intensity represents the gray scale of the ultrasonography image and can be related to the muscle content of contractile and non-contractile elements. The echo-intensity is extracted and expressed on an 8bit gray scale in arbitrary units (the tones of a grayscale image with a bit depth of 8 ranges from 0 (black) to 255 (white) and all the 254 shades of gray in between).
Time frame
Through a study participation of 1,2 years depending on the trajectory: 1 evaluation moment at baseline, 1 follow-up 1,2 years after, or 5 evaluations: 3 months pre, baseline, 3 months after, 6 months after and 1,2 years after baseline3
Overall change in muscle tendon length of the medial gastrocnemius muscle and the semitendinosus muscle
Through a study participation of 1,2 years, depending on the trajectory: one evaluation moment at baseline (BTX injection), or 1 evaluation moment extra 1,2 years after baseline (combined with a second BTX injection), or 5 evaluations 3 months pre baseline, baseline, 3 months after baseline, 6 months after baseline and 1,2 years after baseline.
Time frame
Through a study participation of 1,2 years depending on the trajectory: 1 evaluation moment at baseline, 1 follow-up 1,2 years after, or 5 evaluations: 3 months pre, baseline, 3 months after, 6 months after and 1,2 years after baselineSponsors and contacts
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Source ClinicalTrials.gov