Indicate the nature of the ethical review permissions, relevant licences (e.g., Animal (Scientific Procedures) Act 1986), and national or institutional guidelines for the care and use of animals, that cover the research
+
+
+
+
+
+
Study design
6
For each experiment, give brief details of the study design including the following
6a
The number of experimental and control groups
+
+
+
+
+
+
6b
Any steps taken to minimise the effects of subjective bias when allocating animals to treatment (e.g., randomisation procedure) and when assessing results (e.g., if performed, describe who was blinded and when)
+
Randomization was not described for the allocation of sham group
Not described
No blinding procedure was described
Not described
Not described
6c
The experimental unit (e.g., a single animal, group, or cage of animals)
+
+
+
+
+
+
Experimental procedures
7
For each experiment and each experimental group, including controls, provide precise details of all procedures carried out. For example:
7a
How (e.g., drug formulation and dose, site and route of administration, anaesthesia and analgesia used (including monitoring), surgical procedure, method of euthanasia). Provide details of any specialist equipment used, including supplier(s)
The full formulation of -tocopherol was not described
Method of euthanasia was not described The supplier of the fracture device was not reported
+
Method of euthanasia was not described
+
Methods of analgesia and euthanasia were not described
7b
When (e.g., time of day)
Not described
Not described
Not described
Not described
Not described
Not described
7c
Where (e.g., home cage, laboratory, water maze)
+
+
Not described
+
+
Not described
7d
Why (e.g., rationale for choice of specific anaesthetic, route of administration, drug dose used)
Not described
Not described
+
Not described
Not described
Not described
Experimental animals
8
8a
Provide details of the animals used, including species, strain, sex, developmental stage (e.g., mean or median age plus age range), and weight (e.g. mean or median weight plus weight range)
Sex and age were not described
Age was not described
+
Age was not described
Age was not described
Sex was not described
8b
Provide further relevant information such as the source of animals, international strain nomenclature, genetic modification status (e.g., knock-out or transgenic), genotype, health/immune status, drug or test naïve, previous procedures, and so forth
Not described
Not described
+
Not described
Not described
Not described
Housing and husbandry
9
Provide details of
9a
Housing (type of facility e.g., specific pathogen free (SPF); type of cage or housing; bedding material; number of cage companions; tank shape and material, etc., for fish).
Cage type was not fully described
Cage type was not fully described
Cage type was not fully described
Cage type was not fully described
Cage type was not fully described
Cage type was not fully described
9b
Husbandry conditions (e.g., breeding programme, light/dark cycle, temperature, quality of water, etc., for fish, type of food, access to food, and water, environmental enrichment)
Specific type of food given was not described
+
Light/dark cycle and temperature were not described
Specific type of food given was not described
Not described (only access to food was described)
Not described (only access to food and water was described)
9c
Welfare-related assessments and interventions that were carried out prior to, during, or after the experiment
Not described
+
+
Not described
Analgesia was not given after the fracture procedure (just depended on cast immobilization)
Not described
Sample size
10
10a
Specify the total number of animals used in each experiment, and the number of animals in each experimental group
+
+
+
+
+
+
10b
Explain how the number of animals was arrived at. Provide details of any sample size calculation used
Not described
Not described
Not described
Not described
Not described
Not described
10c
Indicate the number of independent replications of each experiment, if relevant
Not described
Not described
Not described
Not described
Not described
Not described
11
Allocating animals to experimental groups
11a
Give full details of how animals were allocated to experimental groups, including randomisation or matching if performed
Randomization procedure was not fully explained
Randomization procedure was not fully explained for the allocation of treatment groups
Not explained
Randomization procedure was not fully explained
Not explained
Not explained
11b
Describe the order in which the animals in the different experimental groups were treated and assessed
Not described
Not described
Not described
Not described
Not described
Not described
Experimental outcomes
12
Clearly define the primary and secondary experimental outcomes assessed (e.g., cell death, molecular markers, behavioural changes)
+
+
+
+
+
+
Statistical methods
13
13a
Provide details of the statistical methods used for each analysis
+
+
+
+
+
+
13b
Specify the unit of analysis for each dataset (e.g., single animal, group of animals, single neuron)
+
+
+
+
+
+
13c
Describe any methods used to assess whether the data met the assumptions of the statistical approach
+ Not relevant as nonparametric test was used
+ Not relevant as nonparametric test was used
+ Not relevant as nonparametric test was used
+ Not relevant as nonparametric test was used
+ Not relevant as nonparametric test was used
+ Not relevant as nonparametric test was used
Results
Baseline data
14
For each experimental group, report relevant characteristics and health status of animals (e.g., weight, microbiological status and drug or test naive) prior to treatment or testing (this information may often be tabulated)
Not fully described (i.e., only baseline weight was reported)
Not fully described (i.e., only baseline weight was reported)
Not fully described (i.e., only baseline weight was reported)
Not fully described (i.e., only baseline weight was reported)
Not fully described (i.e., only baseline weight was reported)
Not fully described (i.e., only baseline weight was reported)
Numbers analysed
15
15a
Report the number of animals in each group included in each analysis. Report absolute numbers (e.g., 10/20, not 50%)
+
+
Not described
+
+
Not described
15b
If any animals or data were not included in the analysis, explain why
? (no exclusion was reported)
? (no exclusion was reported)
? (no exclusion was reported)
? (no exclusion was reported)
? (no exclusion was reported)
? (no exclusion was reported)
Outcomes and estimation
16
Report the results for each analysis carried out, with a measure of precision (e.g., standard error or confidence interval)
+ (see full commentary in reanalysis table)
Some statistical analyses were not reported fully (see full commentary in reanalysis table)
Not described. (see full commentary in reanalysis table)
+ (see full commentary in reanalysis table)
+ (see full commentary in reanalysis table)
The measure of precision was not reported in the result (see full commentary in reanalysis table)
17
Adverse events
17a
Give details of all important adverse events in each experimental group
+ (no complication reported)
Not described
Not described
Not described
Not described
Not described
17b
Describe any modifications to the experimental protocols made to reduce adverse events
