Trial Outcomes & Findings for Mechanisms of Ozone-Induced Alterations in Efferocytosis and Phagocytosis (NCT NCT03646877)

NCT ID: NCT03646877

Last Updated: 2026-06-15

Results Overview

Efferocytosis is defined by determining the number of apoptotic PMNs phagocytized by alveolar macrophages (BAL cytospins) versus the total number of macrophages ('efferocytic index'). The efferocytic index is a measure of how efficiently macrophages engulf and clear apoptotic (dying) cells. It can be determined by counting the number of macrophages that have a phagocytosed apoptotic body and dividing it by the total number of macrophages counted. A higher value is considered better.

Recruitment status

COMPLETED

Study phase

PHASE1

Target enrollment

35 participants

Primary outcome timeframe

Baseline, 21 days

Results posted on

2026-06-15

Participant Flow

Subjects were recruited from Duke University campus and surrounding Durham, NC, community using advertisement in newspapers, mailings, and the campus student newspaper. Subjects were screened to determine eligibility per protocol inclusion and exclusion criteria.

Participant milestones

Participant milestones
Measure
Ozone (O3), Then Filtered Air (FA)
The O3 level during the exposures will be 200 ppb, which has previously been used in human exposure studies without short or long term, untoward side effects (and comparable to peak levels attained during the summer in the Raleigh-Durham area of North Carolina). Ozone: Subjects will perform alternating 15 minutes rest with 15 minutes treadmill walk exercise periods for 135 minutes in while breathing Ozone (O3). Control will be treadmill walk with filtered room air in chamber. Filtered Air: Subjects will perform alternating 15 minutes rest with 15 minutes treadmill walk exercise periods for 135 minutes in while breathing filtered air.
Filtered Air (FA), Then Ozone (O3)
Control will be treadmill walk with filtered room air in chamber. Filtered Air: Subjects will perform alternating 15 minutes rest with 15 minutes treadmill walk exercise periods for 135 minutes in while breathing filtered air. The O3 level during the exposures will be 200 ppb, which has previously been used in human exposure studies without short or long term, untoward side effects (and comparable to peak levels attained during the summer in the Raleigh-Durham area of North Carolina). Ozone: Subjects will perform alternating 15 minutes rest with 15 minutes treadmill walk exercise periods for 135 minutes in while breathing Ozone (O3).
Overall Study
STARTED
17
18
Overall Study
COMPLETED
7
12
Overall Study
NOT COMPLETED
10
6

Reasons for withdrawal

Withdrawal data not reported

Baseline Characteristics

Mechanisms of Ozone-Induced Alterations in Efferocytosis and Phagocytosis

Baseline characteristics by cohort

Baseline characteristics by cohort
Measure
Ozone (O3) and Filtered Air (FA)
n=19 Participants
The O3 level during the exposures will be 200 ppb, which has previously been used in human exposure studies without short or long term, untoward side effects (and comparable to peak levels attained during the summer in the Raleigh-Durham area of North Carolina) (REF - Bromberg review). Ozone: Subjects will perform alternating 15 minutes rest with 15 minutes treadmill walk exercise periods for 135 minutes in while breathing Ozone (O3). Control will be treadmill walk with filtered room air in chamber. Filtered Air: Subjects will perform alternating 15 minutes rest with 15 minutes treadmill walk exercise periods for 135 minutes in while breathing filtered air.
Age, Continuous
23.75 years
STANDARD_DEVIATION 3.31 • n=20 Participants
Sex: Female, Male
Female
7 Participants
n=20 Participants
Sex: Female, Male
Male
12 Participants
n=20 Participants
Ethnicity (NIH/OMB)
Hispanic or Latino
5 Participants
n=20 Participants
Ethnicity (NIH/OMB)
Not Hispanic or Latino
14 Participants
n=20 Participants
Ethnicity (NIH/OMB)
Unknown or Not Reported
0 Participants
n=20 Participants
Race (NIH/OMB)
American Indian or Alaska Native
0 Participants
n=20 Participants
Race (NIH/OMB)
Asian
5 Participants
n=20 Participants
Race (NIH/OMB)
Native Hawaiian or Other Pacific Islander
0 Participants
n=20 Participants
Race (NIH/OMB)
Black or African American
3 Participants
n=20 Participants
Race (NIH/OMB)
White
11 Participants
n=20 Participants
Race (NIH/OMB)
More than one race
0 Participants
n=20 Participants
Race (NIH/OMB)
Unknown or Not Reported
0 Participants
n=20 Participants
Region of Enrollment
United States
19 Participants
n=20 Participants

PRIMARY outcome

Timeframe: Baseline, 21 days

Population: Participants with complete data sets for both O3 and FA.

Efferocytosis is defined by determining the number of apoptotic PMNs phagocytized by alveolar macrophages (BAL cytospins) versus the total number of macrophages ('efferocytic index'). The efferocytic index is a measure of how efficiently macrophages engulf and clear apoptotic (dying) cells. It can be determined by counting the number of macrophages that have a phagocytosed apoptotic body and dividing it by the total number of macrophages counted. A higher value is considered better.

Outcome measures

Outcome measures
Measure
Ozone (O3)
n=8 Participants
The O3 level during the exposures will be 200 ppb, which has previously been used in human exposure studies without short or long term, untoward side effects (and comparable to peak levels attained during the summer in the Raleigh-Durham area of North Carolina). Ozone: Subjects will perform alternating 15 minutes rest with 15 minutes treadmill walk exercise periods for 135 minutes in while breathing Ozone (O3).
Filtered Air (FA)
n=8 Participants
Control will be treadmill walk with filtered room air in chamber. Filtered Air: Subjects will perform alternating 15 minutes rest with 15 minutes treadmill walk exercise periods for 135 minutes in while breathing filtered air.
Change in Ozone Induced Efferocytosis of Lung Macrophages (Ability to Clear Dead or Dying Cells)
0.478 ratio
Standard Deviation 0.089
0.470 ratio
Standard Deviation 0.090

PRIMARY outcome

Timeframe: Baseline, 21 days

Population: Data not collected.

Bronchoalveolar macrophages are incubated with pHrodo™ dye-labeled particles. Macrophage phagocytosis will be defined by determining the number of macrophages that contain fluorescent particles versus the total macrophages. Cell free hemoglobin and soluble CD163 - both are measured by commercial ELISA kit.

Outcome measures

Outcome data not reported

SECONDARY outcome

Timeframe: Baseline, 21 days

Population: Participants with complete data sets for both O3 and FA.

Measured by spectroscopy.

Outcome measures

Outcome measures
Measure
Ozone (O3)
n=19 Participants
The O3 level during the exposures will be 200 ppb, which has previously been used in human exposure studies without short or long term, untoward side effects (and comparable to peak levels attained during the summer in the Raleigh-Durham area of North Carolina). Ozone: Subjects will perform alternating 15 minutes rest with 15 minutes treadmill walk exercise periods for 135 minutes in while breathing Ozone (O3).
Filtered Air (FA)
n=19 Participants
Control will be treadmill walk with filtered room air in chamber. Filtered Air: Subjects will perform alternating 15 minutes rest with 15 minutes treadmill walk exercise periods for 135 minutes in while breathing filtered air.
Change in Ozone Induced Increase in Cell Free Hemoglobin in Bronchoalveolar Lavage Fluid
0.0000921391 micromolar (uM)
Standard Deviation 0.0000261954
0.000112267 micromolar (uM)
Standard Deviation 0.0000273103

SECONDARY outcome

Timeframe: Baseline, 21 days

Population: Participants with complete data sets for both O3 and FA.

Measured by enzyme-linked immunosorbent assay (ELISA)

Outcome measures

Outcome measures
Measure
Ozone (O3)
n=19 Participants
The O3 level during the exposures will be 200 ppb, which has previously been used in human exposure studies without short or long term, untoward side effects (and comparable to peak levels attained during the summer in the Raleigh-Durham area of North Carolina). Ozone: Subjects will perform alternating 15 minutes rest with 15 minutes treadmill walk exercise periods for 135 minutes in while breathing Ozone (O3).
Filtered Air (FA)
n=19 Participants
Control will be treadmill walk with filtered room air in chamber. Filtered Air: Subjects will perform alternating 15 minutes rest with 15 minutes treadmill walk exercise periods for 135 minutes in while breathing filtered air.
Change in Ozone Induced Increase in Soluble CD163 in Bronchoalveolar Lavage Fluid
96.03 ng/mL
Standard Deviation 57.97
97.16 ng/mL
Standard Deviation 47.92

Adverse Events

Ozone (O3)

Serious events: 0 serious events
Other events: 1 other events
Deaths: 0 deaths

Filtered Air (FA)

Serious events: 0 serious events
Other events: 0 other events
Deaths: 0 deaths

Serious adverse events

Adverse event data not reported

Other adverse events

Other adverse events
Measure
Ozone (O3)
n=35 participants at risk
The O3 level during the exposures will be 200 ppb, which has previously been used in human exposure studies without short or long term, untoward side effects (and comparable to peak levels attained during the summer in the Raleigh-Durham area of North Carolina) (REF - Bromberg review). Ozone: Subjects will perform alternating 15 minutes rest with 15 minutes treadmill walk exercise periods for 135 minutes in while breathing Ozone (O3).
Filtered Air (FA)
n=35 participants at risk
Control will be treadmill walk with filtered room air in chamber. Filtered Air: Subjects will perform alternating 15 minutes rest with 15 minutes treadmill walk exercise periods for 135 minutes in while breathing filtered air.
Nervous system disorders
Near syncope
2.9%
1/35 • Up to 21 days
All participants received both interventions.
0.00%
0/35 • Up to 21 days
All participants received both interventions.

Additional Information

Robert Tighe, MD

Duke University

Phone: 919-684-4894

Results disclosure agreements

  • Principal investigator is a sponsor employee
  • Publication restrictions are in place