الفهرس 
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Abstract
Considering the Diabetes mellitus (DM) as one of the oldest conditions
characterized in humans, it has been recognized since antiquity and considered as a
major health problem at present. There is a need for efficient treatment. The aim of
the present investigation was to find out whether stem cells versus statins can
alleviate the well documented side effects of streptozotocin induced diabetes
mellitus on the periodontal ligament and alveolar bone of adult male Albino rats.
Materials and Methods
One hundred adult male albino rats with body weight ranging from180-200gr
were used in this investigation. They were divided into the following groups:-
Group1.1: consisted of 20 animals and served as controls.
Group2.1: consisted of 20 animals, they were subjected to a single
intraperitoneal injection of streptozotocin in a dose of 60mg. /kg body weight for
induction of diabetes mellitus
group 3.1: consisted of 20 animals. They were subjected to induction of
diabetes mellitus in the same way as group 2-1 animals. One week later, the
animals were treated with simvastatin in a daily intraperitoneal dose of 5mg/kg
body weight
group 4.1: consisted of 40 animals, they were subdivided as follows:
Subgroup. 4.1: consisted of 20 animals, they were subjected to induction of
diabetes mellitus, the same way as group 2.1 animals. One week later they were
subjected to single intravenous infusion of mesenchymal bone marrow stem cells
Summary
132
)1.5× 103 -106(cells per rat in 0.2ml.phosphate buffer saline, slowly injected into
the tail vein over two minutes period using a 22 gauge needle.
Subgroup. 4.2: consisted of 20 rats. They were used for isolation and culture of
bone marrow stem cells, at the Biochemistry Department, Faculty of
Medicine,Cairo University.
At the end of the experiment which lasted for three months the jaws of the
animals of all the groups were dissected out, fixed, decalcified and processed to be
sectioned.
The sections from periodontal ligaments& alveolar bone of rats were stained by
hematoxylin and eosin stain for histological examination and detection of any
structural changes, Masson᾿s trichrome stain for collagen fibers and
immunohistochemical stain for localization and detection of any changes in VEGF
and IL-1β. Other small specimens one cubic mm were taken and processed for
ultrastructural examination by TEM.
Results
Histological results:
In this study histological examination of the diabetic albino rats’ periodontal
ligaments& alveolar bone using H& E stain revealed several histological changes.
These changes included degenerative changes in the form of disorientation,
detachments and dissociation of the principle fibers, infiltration by inflammatory
cells, marked dilatation of blood vessels, a lot of howship,s lacunae and osteoclasts
on the surface of the bone, widening of the marrow cavities .The attachment
epithelium of the animals of this group showed an outstanding feature, which was
their extreme apical migration.
Summary
133
While histological examination of the diabetic albino rats’ periodontal
ligament& alveolar bone treated with statins (group 3.1) revealed that partial
improvements, mostly in the arrangement and association of the fibers of PDL.
Compared to diabetic group, the fibers mostly regained their arrangement and
association together in bundles, as dissociation of fibers and disorientation were at
minimum in this group of animals . Dilatation of the blood vessels was still
observed, however in lesser degree compared to diabetic animals . A lot of empty
howships lacunae were observed on the bone side, while on the root side
cementum showed new cementoid tissue formation. There was minimal apical
migration of the attached epithelium.
While histological examination of the diabetic albino rats’ periodontal
ligament& alveolar bone treated with stem cell using H& E stain (group 4.1)
revealed that marked improvement of the collagen fibers of the PDL with
increased density and grouped in thick bundles. chronic inflammatoty cells
infiltrates were at minimum. The attachment epithelium showed no noticeable
apical migration. Marked regeneration and attachment of the collagen fibers to the
bone& cementum were noticed.
Masson’s trichrome stain results
Sections stained with Masson’s trichrome stain of Periodontal ligament and
alveolar bone of diabetic animals revealed weakly positive reaction of collagen
fibers of the PDL& alveolar bone matrix to Masson’s trichrome stain indicating
weak reactivity of collagen fibers.
While Periodontal ligament and alveolar bone of sections of simvastatin treated
animals revealed moderately positive reaction of collagen fibers of the PDL&
alveolar bone matrix to Masson’s trichrome stain.
Summary
134
While Periodontal ligament and alveolar bone of sections of stem cells treated
diabetic animals revealed strongly positive reaction of collagen fibers of the PDL&
alveolar bone matrix to Masson’s trichrome stain.
Immunohistochemical results
Immunohistochemical localization of the VEGF in the periodontal ligament&
alveolar bone revealed improvement in the diabetic albino rats treated with statins
where they revealed moderately positive reactivity, while rats treated with stem
cells showed strongly positive staining reaction almost as the control group
compared with diabetic albino rats’ periodontal ligament & alveolar bone which
showed weaker staining reactivity.
While Immunohistochemical localization of the Interlukin 1β in the periodontal
ligament& alveolar bone showed also improvment in the diabetic albino rats
treated with statins where they revealed moderately positive reactivity, while rats
treated with stem cell showed weakly positive staining reaction almost as the
control group compared with diabetic albino rats’ periodontal ligament & alveolar
bone which showed strongly positive staining reactivity.
Ultrastructural results:
The electron microscopic examination of the periodontal ligament& alveolar
bone 2 months after the induction of diabetes with STZ has revealed that, marked
destruction and reduction in the density of the collagen fibers leaving a lot of
debris. The fibroblast cells showed definite signs of degeneration as they were
markedly shrunken with pyknotic nuclei, degenerated organelles and vacuolated
cytoplasm.
The ultrastructural examination of the periodontal ligament& alveolar bone of
the diabetic albino rats’ periodontal ligament& alveolar bone treated with statins
Summary (group3.1) showed moderate regenerative changes in the fibrous and cellular
elements. While periodontal ligament& alveolar bone of the diabetic albino rats
treated with stem cells (group4.1) showed marked regenerative changes in the
density of collagen fibers. where the collagen fibers were arranged in large and
thick bundles overlapping each other. There were minimal inflammatory cells,
normal fibroblasts. The alveolar bone repaired its normal ultrastructure with no
signs of osteoclastic activity.