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The role of Doxycyline as an MMP inhibitor in rabbit model of tuberculosis of the spine
Investigators
Title First Name Surname Institution Membership No.*
Department of
Orthopaedic and
Traumatology, Faculty
dr Otman Siregar 16826
of Medicine, University
of North Sumatera,
Medan, Indonesia
Department of
Orthopaedic and
Traumatology, Faculty
dr Benny Benny
of Medicine, University
of North Sumatera,
Medan, Indonesia
*At least 1 investigator must be a subscribing member of AOSpine
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Aims
To investigate the role of MMP inhibitor, Doxycyline, to decrease tissue destruction as
shown by the decrease in the serum level of MMP-9
Objectives
To investigate the relationship between MMP -9, TNF-α and (IL)-1 β and
tuberculosis infection of the spine in rabbit model
To investigate the relationship between the administration of oral doxycyline in
multiple doses and the serum level of MMP-9, TNF-α and (IL)-1 β in tuberculosis
of the spine in rabbit model
To investigate the relationship between the serum level of MMP -9 and the
expression of MMP-9 in the tissue
Background:
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Tuberculosis kills more than 1.5 million people per year, and standard treatment has
remained unchanged for more than 30 years, and multidrug - and C drug-resistant strains
are progressively emerging. Mortality rates remain high among patients even after they
have commenced TB treatment.
The only pharmacological agent licensed as an MMP inhibitor in the United States is
doxycycline, a tetracycline antibiotic, which is used in periodontal disease at
subantimicrobial doses to suppress collagenase activity, and has been proposed as an
agent to limit pathology in other infectious and inflammatory disease such as
atherosclerosis, arthritis, neurosistercosis and gram positive infection.
Methods:
In this experimental study, we will purchase 40 New Zealand white rabbits (aged 3 -4
weeks; body weights of 3.25±0.5 kg) and Mycobacterium tuberculosis strain H37Rv from
the institute of biologic product.
All animal experimental protocols will be processed by the Ethics Committee in Indonesia.
After numbering and weighing, the 40 rabbits will be randomly divided into 2 groups. A.
rabbits will be treated with M. tuberculosis (infection group; n = 30); and B. rabbits will be
treated with sterile normal saline (control group; n = 10). All rabbits will be examined
preoperatively by X-ray to exclude any with vertebral anomalies.
Preoperatively, peripheral blood samples will be collected from infection group and
control group. Sera will be separated by centrifugation at 1,000 g for 30 minutes and will
immediately stored at -80 °C. Measurement of TNF-α, IL-1β and MMP-9 will be performed
by ELISA as per manufacture’s instructions.
Under general anesthesia, a longitudinal incision will be made down from the 6 th lumbar
spinous process of the rabbits. At the upper part of the 6 th lumbar vertebra, an electric
drill will be used to bore a hole (0,3 mm diameter). This hole will be filled with medical
gelatin sponges. The, the gelatin sponges will be impregnated slowl y using a syringe with
0.2 ml of a TB suspension at a consentration of 0.5 mg/ml for the infection group or 0.2 ml
sterile normal saline for the control group. Rabbits then will be held individually in cages
for 8 weeks.
Then, the infection group then will be divided into three groups. The first group will
receive no treatment, the second group will receive doxycylin dosage 1, the third group
will receive dosage 2..
After 4 weeks of follow-up, serum TNF-α, IL-1β and MMP-9 will be measured by ELISA as
per manufacture’s instructions. Then, each rabbit will euthanized and
immunohistochemical examination will be revealed from the pathological vertebra. The
parameters measured objectively are serum TNF -α, IL-1β and MMP-9
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Administration of doxycyline in tuberculosis of the spine in rabbit model is expected to
decrease tissue destruction as shown by the decrease in the level of serum MMP -9 and
the decrease in the expression of MMP-9 in pathological tissue by immunohistochem istry
examination.
Budget:
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Thank you