This study evaluated the anxiolytic effects of cannabidiol (CBD) in male rodents using the Elevated Zero Maze test. Rodents were orally administered 20 mg/kg of 99% pure CBD or a sesame oil vehicle daily for 10 days. On day 10, the CBD group traveled less distance and spent less time in the open arms, suggesting sedation rather than anxiolysis. Future studies should test lower CBD doses and consider sex differences to better assess anxiolytic potential while avoiding sedation.

1. Oral Cannabidiol (CBD) Administration: Assessing Anxiolytic Properties of CBD in a
Rodent Model
INTRODUCTION RESULTS
Usmaan Zunnu Rain*, Nejra Kulaglic*, Jordan Skully, Chela Wallin, Dr. Susanne Brummelte, Dr. Scott E. Bowen
Department of Psychology, Wayne State University. Detroit, Michigan
Cannabidiol (CBD), a constituent in cannabis that is non-
psychoactive, is claimed to be an effective anxiolytic by
activation of the 5HT1a receptor (Crippa et al., 2011, Blessing et
al., 2015). CBD has been popularized due to its therapeutic
properties, yet there is still a need for additional scientific
literature. This study analyzed the anxiolytic properties of a 99%
pure CBD compound, supplied by Ellipse Analytics. Anxiety
responses (distance traveled and time spent within open arm),
after CBD administration, were tested and measured on male
rodents via Elevated Zero Maze.
This study was conducted to evaluate the effectiveness of
CBD’s anxiolytic properties on male rodents in the
Elevated Zero Maze.
Drug and Administration: Pure cannabidiol (CBD), derived
from hemp product, was diluted in sesame oil (vehicle) to create
a 20 mg/kg dosage group. Administration of CBD to rodents via
oral gavage (P.O.) given once daily for 10 days.
*Funding for this project was provided by Ellipse Analytic (Denver, Colorado).
References:
Blessing, E. M., Steenkamp, M. M., Manzanares, J., & Marmar, C. R. (2015). Cannabidiol as a potential treatment for anxiety disorders. Neurotherapeutics, 12(4), 825-836.
Crippa, J. A. S., Derenusson, G. N., Ferrari, T. B., Wichert-Ana, L., Duran, F. L., Martin-Santos, R., ... & Hallak, J. E. C. (2011). Neural basis of anxiolytic effects of cannabidiol (CBD) in
generalized social anxiety disorder: a preliminary report. Journal of psychopharmacology, 25(1), 121-130.
METHODS
Figure 1: Elevated Zero Maze: After
CBD administration, Noldus Ethovision
was used to measure distance traveled and
time spent within the arms of the maze.
Rats were placed on an initial start point
within the open arm of the maze. After 5
minutes, they were taken off.
Administer lower dosage of CBD prior to EZM testing
Address potential sex differences in response to CBD
Assess a potential entourage effect by comparing CBD isolate against other cannabinoids, and CBD in
combination with other cannabinoids
FUTURE DIRECTIONS
ACKNOWLEDGEMENTS & REFERENCES
A special thanks to Alixandria Trombley, Ava Palopoli, Lauren
Richardson, Kristy Tilson, Yusuf Yasarlar, Saadia Taj, and Brandon
Brown for their support in data collection and imputation.
Figure 2. Timelines of CBD administration and
corresponding cohort assessments. Anxiety and stress testing
were assessed together (anxiety tested on the 1st and 10th day
and stress tested on the 5th day). CBD dosing for behavioral
testing included vehicle (n= 16), and 20 mg/kg (n= 16).
0 and 20 mg/kg Distance Traveled by Day
OBJECTIVE
DISCUSSION
There was a significant difference between dose groups on Day 10, where the 20mg/kg dose group traveled less
and spent less time within the open arms of the EZM. This signifies a potential sedation effect due to the lack of
movement within the dosage group. Anxiolytic potential of CBD is difficult to assess due to possible sedation.
Figure 3. Means displayed represent distance traveled (cm) in the entirety
of the Elevated Zero Maze by dose group by day. Dose groups evaluated
are control (sesame oil, n=16) and 20mg/kg (n=16). There was no
significant effect of dose groups related to the total distance traveled on
Day 1. On Day 10, there was a significant difference between the dose
groups [P=.017]. Error bars shown are mean ± SEM.
Figure 4. Means displayed represent the cumulative time (s) spent in the
open arm of the EZM by dose by day. Dose groups evaluated are control
(sesame oil, (n=16) and 20mg/kg (n=16). There was no significant effect
of dose group on Day 1. There was a significant difference between dose
groups on Day 10 [P=.043]. Error bars shown are mean ± SEM.
0 and 20 mg/kg Open Arm Duration (s) by Day

2. Oral Cannabidiol (CBD) Administration: Assessing Anxiolytic Properties of CBD in a Rodent Model
Usmann Zunnu Rain*, Nejra Kulaglic*, Jordan Skully, Chela Wallin, Dr. Susanne Brummelte, Dr. Scott E. Bowen
Department of Psychology, Wayne State University. Detroit, Michigan
• Cannabidiol (CBD) is a non-psychoactive constituent in cannabis
• Anxiolytic and panicolytic properties
• Lack of extensive scientific literature
• 99% pure compound of CBD provided by Ellipse Analytics (Denver, CO)
Evaluate the effectiveness of CBD’s anxiolytic properties on male rodents in
the Elevated Zero Maze.
CBD could provide a safer alternative to a variety of current therapeutic
drugs
INTRODUCTION
OBJECTIVE
WHY?

3. Oral Cannabidiol (CBD) Administration: Assessing Anxiolytic Properties of CBD in a Rodent Model
Usmann Zunnu Rain*, Nejra Kulaglic*, Jordan Skully, Chela Wallin, Dr. Susanne Brummelte, Dr. Scott E. Bowen
Department of Psychology, Wayne State University. Detroit, Michigan
METHODS
Figure 2: Timelines of CBD administration and corresponding cohort
assessments.
Figure 1: EZM Training Apparatus

4. Oral Cannabidiol (CBD) Administration: Assessing Anxiolytic Properties of CBD in a Rodent Model
Usmann Zunnu Rain*, Nejra Kulaglic*, Jordan Skully, Chela Wallin, Dr. Susanne Brummelte, Dr. Scott E. Bowen
Department of Psychology, Wayne State University. Detroit, Michigan
RESULTS
Figure 3. Means displayed represent the distance traveled
(cm) in the entirety of the Elevated Zero Maze by dose group
by day. Beta weights show that the 20mg/kg CBD dosage
group traveled 283.261 cm less than the vehicle on Day 10.
Figure 4. Means displayed represent the cumulative time (s)
spent in the open arm of the EZM by dose by day. Based on
beta weights, the 20mg/kg CBD dosage group spent 16.4
seconds less in the open arms of EZM compared to the
vehicle on Day 10.

5. There was a significant difference between dose groups on Day 10, where the 20mg/kg
dose group traveled less and spent less time within the open arms of the EZM. This
signifies a potential sedation effect due to the lack of movement within the dosage group.
Anxiolytic potential of CBD is difficult to assess due to possible sedation.
Administer lower dosages of CBD before EZM testing
Address potential sex differences in response to CBD
Assess a potential entourage effect by comparing CBD isolate against other cannabinoids, and
CBD in combination with other cannabinoids
DISCUSSION
Oral Cannabidiol (CBD) Administration: Assessing Anxiolytic Properties of CBD in a Rodent Model
Usmaan Zunnu Rain*, Nejra Kulaglic*, Jordan Skully, Chela Wallin, Dr. Susanne Brummelte, Dr. Scott E. Bowen
Department of Psychology, Wayne State University. Detroit, Michigan
FUTURE DIRECTIONS