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 Comedogenicity test (blackheads) 
Report on Emu Oil in the Rabbit Ear
Comedogenicity Testing

University of Texas Medical School at Houston
Dermatology Department. 
Occupational Dermatology Laboratory
Many agents are capable of causing comedo formation when applied to the skin.  Comedones are plugged hair follicle and are comedogenicity in the rabbit ear assay also tend to produce comedones in human follicles.
In this study, we have used both the rabbit ear histological assay and subjective scoring assay to assess the comedogenic properties of Emu oil provided to us by Atlantis Laboratories.
Mat erials and methods:
1.     Test chemicals.  Emu oil (EO): 25% (EO25), 50% (EO50), 75% (EO75), and 100% (EO100) solution.
2.     Animal treatment.  The ears of male albino rabbits weighting 2-4kg were used in this study
Applications were made according to the following table
                Rabbit No.             Left Ear Right Ear
                1-1#                                                        None
                1-2#                                                        None
                1-3#                                                        None
                2-1#                                                        EO25
                2-2#                                                        EO25
                2-3#                                                        EO25
                3-1#        EO50      EO75
                3-2#        EO50      EO75
                3-3#        EO50      EO75
                4-1#        EO100   None
                4-2#        EO100  None
                4-3#        EO100   None
3.     Applications of test agents:  Test agents were topically applied to the rabbit ear at the dosage of 0.1 ml daily for 5 days each week for three consecutive weeks.  The site of application was a 2x2x2cm2 area at the external ear canal notch.
4. Evaluation methods:
(1)   Observational examination of the rabbit’s ear was performed at the end of the second and third week of application by visual scoring of comedogenicity.  The following scale was used for scoring.
Grading Scale:
0= No change
1= Slight follicular hyperkeratosis in some follicles;
2= Slight follicular hyperkeratosis;
4= Moderate follicular hyperkeratosis with some plugging of follicles; and,
5= Confluent plugging of follicles
(2)   Histological examination was performed at the end of the test.  A portion of the ear from the treated and control sites were biopsied.  Vertical step sections were evaluated for histologic evidence of follicular plugging as defined by the American Academy of Dermatology Comedogenicity Consensus Group recommendations as follows:
Grading Scale:
0= No comedogenesis
1= The presence of a relatively small amount of compact horny material occupying the infundibulum
2= The presence of a moderate amount of compact horny material that widens the infundibulum and extends into the sebaceous ducts
3= The presence of dense horny material that distends the follicle, resembling a human open comedo.
Test Results
1. EO25 treatment results

An observational score of 1 or less is considered to indicate a substance that is unlikely to produce a comedogenic response during human use and is therefore considered to be a negative result.
A histologic score of 1 of less is considered to indicate a substance that is unlikely to produce a comedogenic response during human use and is therefore considered to be a negative result.
4. Final Assessment:
On the basis of observational and histologic testing for comedogenicity, the Emu oil used in this test, from 25% to 100%, is non-comedogenic.      

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