FORMAT FOR SUBMISSION OF PROJECTS INVOLVING RESEARCH IN

HUMAN SUBJECTS FOR CLEARANCE BY INSTITUTE ETHICS COMMITTEE

OF AIIMS

The research projects proposal submitted should be as follows:

1. Full Title of Study: Evaluation of inflammatory salivary biomarkers as a

surrogate indicator of systemic response to thermal
burn injuries: an observational study.
2. Name of Investigators / co- Signatures No. of projects
investigators (permanent already with
AIIMS Staff) with designation investigator
and departments

2.1 Dr. Shashank

Chauhan (PI)
Associate Professor
Department of Plastic, 2.1_____________________________________________
Reconstructive and Burns
Surgery
AIIMS, New Delhi-29

2.2 Dr. Maneesh Singhal

(Co-I)
Professor & Head
Department of Plastic,
Reconstructive and Burns 2.2_____________________________________________
Surgery
AIIMS, New Delhi-29

2.3 Dr. Shyam S Chauhan (Co-

I)
Professor & Head
Department of Biochemistry,
AIIMS, New Delhi 2.3_____________________________________________

2.4 Dr. Lokesh Kashyap (Co-I)

Professor & Head
Department of
Anaesthesiology, Crital Care
and Pain Medicine, AIIMS,
New Delhi 2.4_____________________________________________
2.5 Dr. Shailendra Kumar (Co-
I)
Associate Professor
Department of Plastic, 2.5_____________________________________________
Reconstructive and Burns
Surgery
AIIMS, New Delhi-29

2.6 Dr. Shivangi Saha (Co-I)

Assistant Professor 2.6_____________________________________________
Department of Plastic,
Reconstructive and Burns
Surgery
AIIMS, New Delhi-29

2.7 Dr. Suvashis Dash (Co-I)

Assistant Professor 2.7_____________________________________________
Department of Plastic,
Reconstructive and Burns
Surgery
AIIMS, New Delhi-29

Primary Objective: To investigate saliva as a surrogate

3. Objectives of the study
4. Justification for conduct of Blood has been chiefly utilised to assess biomarkers in
this study
biomarker for predicting outcomes by estimating the
salivary levels of inflammatory cytokines in patients with
thermal burn injury.
Secondary objectives:
1. To correlate the concentration of inflammatory
cytokines in saliva with the routine blood parameters of
inflammation.
2. To compare the levels of salivary biomarkers in different
types of thermal injuries such as scalds, flame and contact
burns.
burn patients. However, because of its invasiveness, it
cannot be used frequently to ascertain the prognosis,
especially in patients treated without needing a cannula or
in situations where blood collection is inappropriate.
Additionally, blood collection from children is challenging
in an outpatient setting. To avoid these limitations, there
have been substantial studies performed using saliva and
urine as diagnostic samples in pediatric patients; however,
 they are most commonly only to measure stress-related
markers. Only two studies— one by Carlton et al. (2022)
that examined salivary inflammatory cytokines and one by
Kulp et al. (2010) that evaluated urine inflammatory
cytokines—have been conducted using non-invasive
samples. However, in the study by Carlton et al. (2022),
the saliva
collection was performed 6-10 days after the burn injury,
and the salivary levels were not correlated with blood
picture. The most inflammatory cytokines would elevate
shortly after injury. Therefore, to understand the temporal
impact of burn injury on the salivary cytokine profiles,
samples should be taken at various intervals until healing.
In addition, the simultaneous collection of blood to
corroborate previous research findings and compare them
to the findings in saliva will potentially allow for the
development of a more robust non-invasive method of
evaluating burn injury.
5. Methodology i. Study Design:
Participants will be recruited in a consecutive
manner admitted to the Department of Plastic,
Reconstructive and Burns Surgery, Burns and
Plastic Surgery Block, All India Institute of
Medical Sciences, New Delhi, based on the
following inclusion criteria.
1. Flame burns, scald burns and thermal contact
burn injuries involving total body surface area
(TBSA) less than 20-50%.
2. Acute thermal burn injury (presenting within
24 hours of injury); and
3. All patients above 18 years of age.
The following criteria will be used to exclude the
participants in this study
1. Patients being managed on OPD basis
2. Patients with chemical burns, electric burns,
radiation burns or inhalational injuries
3. Patients with known history of chronic
inflammatory disease or systemic disease/
medications/ surgical procedure altering the
normal inflammatory/ immune mechanism
4. Patients with sepsis or on ventilator
ii. Sample Size:
The sample size was calculated according the
recent study by Carlton et al. 2022
(Carlton M, Voisey J, Jones L, Parker TJ,
Punyadeera C, Cuttle L. An Exploratory Study
Demonstrating That Salivary Cytokine Profiles
Are Altered in Children With Small Area Thermal
Injury. J Burn Care Res. 2022;43(3):613-624.)
with 95% confidence interval, 95% power and
5% significance level for detection of salivary IL-
7 in burn patients (Mean(SD): 18.60(8.5)) in
comparison with healthy controls (Mean(SD):
15.86(7.68)).
Accordingly, 217 patients were to be included in
the study. But it was inflated to a total of 250
patients to compensate for the possible drop
outs
and for convenience of distribution while
analyzing the results later on.
iii. Project Implementation Plan:
Participants will be recruited and examined in
the department of Plastic, Reconstructive and
Burns Surgery, AIIMS, New Delhi. The
Department of Biochemistry, AIIMS, New Delhi,
will perform the biochemical analysis. The
statistical analysis will be performed with the
help of the Department of Biostatistics, AIIMS,
New Delhi.
History and examination
A detailed history of burn injury and medical
history will be obtained from the caregivers at
the time of admission as a part of the normal
case sheet data recording. Variables that will be
recorded from each patient include
demographics such as age, gender, ethnicity,
geographic location and socio-economic status,
and injury data such as cause of burn injury, the
occurrence of adequate first aid, anatomical
location of the burn, percentage of TBSA burned,
depth of burn injury. Data will be collected as per
the performa.
Sample
Both saliva and blood will be collected at the
following time periods (Total 8 times): at the
time of admission or 0 hours (T0), 24 hours (T1),
48 hours (T2), 72 hours (T3), 7th day (T4), 15th
day (T5), 30th day (T6) and 90th day (T7). The
first week is divided into multiple time points
because of the dramatic differences anticipated
within this time period. The third-month sample
may serve as baseline data for comparison. Both
blood and saliva will be collected from each
patient per the proposed timeline.
Blood is routinely collected in all admitted burns
patients throughout the course of admission.
Saliva collection
The patient will be urged to hold off on eating,
drinking, and brushing for 15 minutes before
saliva collection. Next, the patient will be asked
to rinse their mouth with water. Then the patient
will be asked to drool through the lower lip into
a conical tube (Eppendorf, Hamburg, Germany)
by leaning their head forward and without any
oral
movements for 4-5 minutes. In the case the
patient cannot provide a sample by themselves, a
sterile, plastic transfer pipette will be used to
collect the saliva from their mouth and then
dispensed into the conical tube. The passive
drool sample of 5ml will be refrigerated at 4 o C
until transported to the laboratory for
processing.
Sample analysis
The saliva sample will be centrifuged within
seven days of collection (3000 rpm at 220C, for
10 min) and frozen at -800C until biochemical
analysis. The levels of IL-1β, IL-4, IL-7, IL-8, MCP-
1, and TNFα in all samples will be assessed using
ELISA technique according to manufacturer
recommendations within 4 months of sample
collection. The blood sample will be analysed
and reported as per the routine protocol.
iv. Ethics Review:
Ethical approval will be obtained before the start
of the study from the institutional ethical
committee, AIIMS, New Delhi
v. Data collection & statistical analysis plan:
All continuous variables will be presented as
mean ± standard deviation, and the frequencies
of categorical variables will be presented as
percentages. The concentration of salivary
biomarkers will be expressed in pg/mL or
ng/mL. Suitable statical tests will be used based
on the normality of the data for testing the
significance of levels of salivary biomarkers at
various time points. Grouping based on based on
the age, gender or %TBSA burn injuries may be
performed based on the characteristics of the
included sample. Linear regression models will
be used to determine differences in the mean
 cytokine concentration and to evaluate changes
in burn area and mechanism. Correlation
analysis will be used to determine relationships
between cytokine concentration, blood
parameters, and clinical outcomes.
6. Permission from Drug 1. Required 2.  Not required
Controller General of 3. Received 4. Applied when:
India(DCGI)
7. Permission from DGFT if 1. Required 2.  Not required
applicable 3. Received 4. Applied when:
8. a) Safety measures for Not applicable.
proposed interventions

b) Results of relevant
laboratory tests

c) Result of studies in human

9. Plans to withdraw standard Yes ✔No

therapy during conduct of
research

10. Plan for provision of Not applicable.

coverage for medical risk (s)
during the study period

11. How you will maintain The records associated with this work will be kept
Confidentiality of subject? confidential.

12. Total Budget - Rs. 1. Patient 2. ✔ Project 3. Exempted

76,92,122 /- 4.
Other Agencies (Name) – Indian Council of Medical
Research (ICMR)
Who will bear the cost of
investigation / implants Not applicable.
drugs / contrasts?
13. Participant Information
Sheet ✔
(mark √ if yes)
14. Participant Informed ✔
Consent Form
(mark √ if yes)
1. Dr. Shashank Chauhan Yes ✔No
15. Conflict of interest for any
other investigator(s) (if yes, 2. Prof. Maneesh Singhal Yes ✔No
please explain in brief
 3. Prof. Lokesh Kashyap Yes ✔No

4. Prof. Shyam S Chauhan Yes ✔No

5. Dr. Shailendra Kumar Yes ✔No

6. Dr. Shivangi Saha Yes ✔No

7. Dr. Suvashis Dash Yes ✔No

16. Whether any work on this X(mark √ if yes, X if no)

project has started or not? (Please enclose a separate certificate to this effect).

17.Attached documents 17.1 Covering letter, through proper channel.

(If any) 17.2 Copy of the detailed protocol is mandatory.
17.3 Brief CV of Investigators (including No. of projects
with Principal Investigator)
17.4 Investigator’s Brochure: Record data
17.5 Undertaking that the study shall be done in
accordance with DST and GCP guidelines
17.6 In case of multi-centric study, IEC clearance of other
centres must be provided
17.7 Definite undertaking as to who will bear the
expenditure of injury related to the project
17.8 In case an insurance cover is intended, Insurance
certificate must be provided (as per DST guidelines)
17.9 Permission as mentioned in column 5.9
17.10 Certificate/undertaking as mentioned in column 16
17.11 In case of Clinical trials, proof of registration of
Clinical trial with DST needs to be submitted. Applied
17.12 Others