Acute rhinosinusitis (ARS) is an inflammation of the nasal passages and the lining of the
sinuses due to an infection. Clinical symptoms and signs of ARS include nasal
blockage/obstruction/congestion or runny nose, sneezing, headache, sinus pain (facial
pain), and loss of smell. According to statistics in the US, there are about 18-22
million ARS visits to the doctor each year that estimated cost of treatment of up to 11
billion dollars. Up to 28.9 million adults were diagnosed with rhinosinusitis, accounting
for 11.6% of the population. In a 5-year statistics at the National Otorhinolaryngology
Hospital of Vietnam, rhinosinusitis patients in the working age group from 16 to 50 years
old account for 87%. Acute otitis media (AOM) is inflammation of the middle ear mucosa
accompanied by effusion into the middle ear due to infection, which may be associated
with the presence or absence of perforation of the tympanic membrane. Symptoms of AOM
include fever, purulent discharge from the ear canal (ear discharge or otorrhea), earache
(rubbing or pulling on the earlobe, tossing or turning, difficulty sleeping or crying),
headache, and hearing loss temporary (poor response to sound), vomiting or diarrhea. AOM
is children's most common ear disease and a common cause of hearing loss. About 90% of
children under two years of age have had at least one episode, and about 80% of preschool
children have otitis media. The prevalence is now about 20% at 2 years and 8% at the age
of 8 years. Bacteria that cause acute rhinosinusitis and acute otitis media include S.
pneumoniae, H. influenzae, M. catarrhalis, and S. aureus... Rhinosinusitis and bacterial
otitis media usually require antibiotic treatment; however, inappropriate antibiotic
therapy will increase drug resistance and side effects, thereby increasing medical costs.
In recent years, the role of probiotics in respiratory diseases has also been mentioned
in convincing studies. However, these probiotic products are mainly used orally, with
very few products being prepared as nasal spray suspensions. Here, the investigators
conducted a study that the direct spray of probiotics into the nose effectively supports
treating acute rhinosinusitis and acute otitis media.
The objective was to investigate the symptomatic support treatment effects of the
probiotic product LiveSpo Navax® as a liquid-suspension form containing Bacillus spores
of safe B. subtilis ANA4 and B. clausii ANA39 strains in children and adults having ARS
and children having ARS accompanied by AOM caused by bacteria, investigators evaluation
of improved symptom efficacy of LiveSpo Navax® in patients; and measurement of changes in
bacterial concentrations, and major cytokine indicators in the nasopharyngeal and ear
samples before and after 3 days or 7 days using LiveSpo Navax®.
Methods: A randomized, blind, controlled clinical trial is conducted. The patients or
parents of patients must provide the following information: full name, sex, age,
address...After informed consent,120 eligible patients are divided into 2 groups (n =
60/group each) including the ARS group and the ARS accompanied by the AOM (ARS & AOM)
group. 60 patients with ARS will be randomized into two subgroups (n = 30/subgroup): the
control - acute rhinosinusitis subgroup (named Control - ARS subgroup) uses 0.9% NaCl
physiological saline and an experimental - acute rhinosinusitis subgroup (called the
Navax - ARS subgroup) use the probiotics LiveSpo Navax®. And similarly, 60 children with
ARS accompanied by AOM will be randomized into two subgroups (n = 30/subgroup): the
control - acute rhinosinusitis accompanied by acute otitis media group (named Control -
ARS & AOM group) uses 0.9% NaCl physiological saline and an experimental - acute
rhinosinusitis accompanied by acute otitis media group (called the Navax - ARS & AOM
group) use the probiotics LiveSpo Navax®. The patient is given a coded spray in the form
of a blind sample to ensure the study's objectivity. After the follow-up patient time,
nasopharyngeal samples on both ARS and ARS accompanied by AOM groups and ear samples on
the ARS accompanied by AOM group will be collected at day 0 and day 3 or/and 7 to
evaluate potential reductions in bacteria load and modulation of cytokine, IgA, and the
presence of probiotic spores in the patient's the nasopharyngeal and ear samples.
Real-time polymerase chain reaction (Real-time PCR) for detection of microorganisms in
nasopharyngeal and ear samples: semi-quantitative assays for measuring changes in
bacterial concentrations are conducted by the real-time PCR routine protocol, which has
been standardized under International Organization for Standardization 5189:2012 (ISO
15189:2012) criteria and used in Vietnam National Children's Hospital. Detection of B.
subtilis ANA4 and B. clausii ANA39 are also conducted by real-time PCR SYBR® Green (SYBR®
Green is a dsDNA-binding dye), standardized routinely in Spobiotic Research Center,
ANABIO R&D Ltd.
ELISA assays for cytokine levels: pro-inflammatory cytokines levels (pg/mL), including
interleukin (IL-6, IL-8) and TNF-alpha, and immunoglobin A (ng/mL) are quantified using
an enzyme-linked immunosorbent assay kit (ELISA) according to the manufacturer's
instructions. The patients are monitored for symptoms of ARS and AOM at days 0, 3, or/and
7, depending on the degree of compliance with the follow-up re-examination of the
outpatients. Doctors observe the patient's health conditions, and their patients of
information are filled into medical records. During this study, patients are asked to
abstain from the consumption of other probiotics, either via nasal spray or oral
administration.
Data collection and statistical analysis: individual medical records are collected, and
the patient's information is gathered and systematized. The efficacy of LiveSpo Navax® is
evaluated and compared to 0.9% Sodium Chloride (NaCl) physiological saline based on the
following clinical and sub-clinical criteria obtained in Navax and Control groups: (i)
effective the symptomatic-relieving; (ii) the reduction levels (2^△Ct) of bacteria
concentrations.△Ct for target genes is calculated as Ct (threshold cycle of real-time PCR
assay) at day 3 or/and 7 - Ct at day 0 while Ct of internal control is adjusted to be
equal among all samples; (iii) the reduction levels of IL-6, IL-8, and TNF-alpha
cytokines and IgA. The tabular analysis is performed on dichotomous variables using the
χ2 test or Fisher's exact test when the expected value of any cell is below five.
Continuous variables are compared using the Wilcoxon test, t-test, or the Mann-Whitney
test when data are not normally distributed. The correlations among the variables are
assessed by Spearman's correlation analysis. Statistical and graphical analyses are
performed on GraphPad Prism v8.4.3 software (GraphPad Software, CA, USA). The
significance level of all analyses is set at p < 0.05. P-values.