Serbian Dental Journal, vol.

63, No4, 2016 159

ORIGINAL ARTICLE  DOI: 10.1515/sdj-2016-0016

ORIGINALNI RAD UDC: 616.314.2; 615.46:616.314-7

The effect of zinc oxide based sealer on bone defects

healing
Marija Nikolić, Jelena Popović, Jovanka Gašić, Radomir Barac
University of Niš, Medical faculty, Department of Restorative dentistry and Endodontics, Clinic of Dentistry, Niš, Serbia

SUMMARY
Introduction Obturation as the final phase of endodontic treatment aims to provide complete hermetic filling along the entire
length of the canal system from the coronal opening to the apical end. The aim of this study was to evaluate histological response
of bone tissue on the implantaton of zinc oxide based material in artificially prepared defect in the mandible of rats.
Material and method For the experiment, sixteen male Wistar rats were used. Using sterile steel burs a defect was made in man-
dible, between the midline and mental foramen. Zinc oxide based sealer was implanted in the defects of experimental group while
the defects of control group healed spontaneously. One half of animals in both groups were sacrificed after thirty days, and the
second half after ninety days. Microscopic preparations consisted of the defect with surrounding bone and after processing were
analysed using light microscopy.
Results The thirtieth day after implantation of the material, fibrovascular connective tissue was noted, with scant chronic inflam-
matory cell infiltrate. Away from the experimentally made defect, in the depth of the bone, lamellar bone with well-formed larger
osteons was noted as well as enlarged Volkmann and Haversian canals. Ninety days after implantation of the material, there was no
restitutio ad integrum, but intense focal remodelling of bone tissue was noted.
Conclusion Endomethasone N slowed down bone tissue healing process by showing the signs of prolonged inflammation in bone
tissue in which it has been implanted. Extension of the healing process is reflected in the slow replacement of fibrovascular con-
nective tissue with newly formed bone tissue.
Keywords: sealers; obturation; bone healing

INTRODUCTION are highly toxic, and should not be used as sealers [7]. The
toxicity is attributed mainly to the presence of eugenol
The aim of оbturation as the final phase of endodontic or formaldehyde in some formulations of these materi-
treatment is to provide complete hermetic filling along als [8, 9].
the entire length of the canal system from the coronal The aim of this study was to evaluate histological re-
opening to the apical end. It should break communica- sponse of bone tissue on the implantation of zinc oxide
tion between endodontic and periodontal tissues and based material in artificially prepared defects in the man-
thus prevent reinfection of periapical region [1]. Materi- dible of rats.
als used for obturation should have a number of physical,
chemical and biological properties and should not extend
beyond the end of the canal system [1]. When a bioma- MATERIALS AND METHOD
terial comes in contact with the tissues and fluids of the
human body there is always some form of interaction and For the experiment, 16 Wistar male rats were used, weight
therefore it is necessary for material to be biocompatible 160-180 g (approved by the Ethics Committeeof the Fac-
and harmless in the biological surrounding [2]. ulty of Medicine in Niš No. 01 3797). During the experi-
Overinstrumentation and consequently overfilling of mental procedure, animals were anaesthetized by intra-
the root canal significantly increase the risk of adverse im- peritoneal injection of ketamine hydrochloride (0.1 ml
pacts of filling material [3]. Overfilling causes inflamma- per 100 g of weight). After preparation procedure, be-
tion and slows down wound healing [4, 5]. The negative tween midline and mental foramen of the left mandible
effect is usually more pronounced until the material fully of all animals, a defect (diameter of 1.4mm and 1.6mm
sets and it is attributed to the components of the material depth) was made using sterile stainless steel fissure burs.
[6]. Slow release of harmful agents over long periods of Animals of the experimental group (n = 12) were di-
time depends on the solubility of materials and the degree vided into the two subgroups:
of material exposure to tissue fluids [2].  • The first subgroup (n = 6) was sacrificed after 30
Zinc oxide based sealers are most commonly used days;
in the clinical practice as well as comparative sealers in •T  he second subgroup (n = 6) was sacrificed after 90
studies that examine the effect of biomaterials [4, 7]. The days.
results of these studies, however, are not encouraging. Ac- In bone defects of experimental group animals, „Endo-
cording to some authors, certain materials from this group methasone N” (Septodont, France) prepared by the manu-

Address for correspondence: Address for correspondence: Marija NIKOLIĆ, Clinic of Dentistry, Bulevar dr Zorana Đinđića
52, 18000 Niš, Serbia; [email protected]
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160 Nikolić M. et al. The Effect of Zinc Oxide Based Sealer on Bone Defects Healing

Figure 1. Histological finding of the defect 30 days after implan- Figure 2. Histological finding of the defect 30 days after implanta-
tation of Endomethasone N. Fibrovascular connective tissue with tion of Endomethasone N. Thickening of cement lines and expan-
scarce inflammatory cell infiltrate is visible. Surrounding bone is sion of Volkmann and Haversian canals in the bone tissue away from
without osteon-type structure (HE, 800×). the edges of the defect are visible (HE, 400×).
Slika 1. Histološki nalaz defekta u kosti 30 dana nakon implanta- Slika 2. Histološki nalaz defekta u kosti 30 dana nakon implan-
cije endometazona N. Fibrovaskularno vezivno tkivo sa oskudnim tacije endometazona N. Zadebljanje cementnih linija i proširenje
inflamatornim ćelijskim infiltratom. Okolna kost bez građe po tipu Folkmanovih i Haversovih kanala u koštanom tkivu udaljenom od
osteona (HE, 800×). ivice defekta (HE, 400×).

Figure 3. Histological finding of the defect 90 days after implanta- Figure 4. Histological finding of the defect 30 days after implanta-
tion of Endomethasone N. Newly formed bone with diffuse distribu- tion of Endomethasone N. Bone shows normal micromorphology
tion of mature connective tissue is noted (HE, 400×). away from the defect (HE, 800×).
Slika 3. Histološki nalaz defekta u kosti 90 dana nakon implantacije Slika 4. Histološki nalaz defekta u kosti 90 dana nakon implantacije
endometazona N. Novoformirana kost sa difuzno raspoređenim endometazona N. Kost udaljena od defekta je normalne mikromor-
zrelim vezivnim tkivom (HE, 400×). fologije (HE, 800×).

facturer’s instructions was implanted. Bone defects in the RESULTS

control group (n = 4) were left to heal spontaneously with-
out any implants. Two animals of the control group were Experimental group
sacrificed after thirty days and two animals after ninety
days. After the estimated time the animals were sacrificed The thirtieth day after the implantation of material, fi-
by an overdose of anaesthetic (ketamine hydrochloride). brovascular connective tissue was noted, with scant focal
The tissue samples were made by resection of the man- chronic inflammatory cell infiltrate, while surrounding
dible consisting of the defect and surrounding bone. bone was hypocellular without the presence of Haversian
Tissue samples were fixed in 10% buffered formalin, system (in 5 out of 6 samples) (Figure 1). Small osteo-
demineralized in 10% formic acid, dehydrated in a series cytes with hyperhromatic nuclei were stored in slightly
of graded alcohols and embedded in paraffin wax. Cut- enlarged lacunae. Osteoblasts were few in number, while
ting was performed in the buccolingual direction on the the presence of osteoclasts was inconspicuous. In areas
microtome with 2μm thick glass knives. The slides were of non-resorbed material, which was lost during process-
stained with hematoxylin and eosin. Microscopic analysis ing of histopathological preparations, empty spaces were
was performed using light microscopy with digital camera noticed. Away from experimentally made defect, in the
Leica DFC 295. depth of the bone, lamellar bone with well-formed larger
osteons was noted as well as enlarged Volkmann and Ha-
versian canals (Figure 2). Expanded interstitial lamellae
were observed in osteons, osteocytes and lacunae were
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 Stomatološki glasnik Srbije. 2016;63(4):159-166 161

Figure 5. Histological finding of the defect after 30 days (no implan- Figure 6. Histological finding of the defect after 90 days (no im-
tation of material- control group). Fibrovascular connective tissue plantation of material- control group). Newly formed bone with
surrounding newly formed bone can be seen (HE, 400×). osteon- type organization and borders to the lamellar bone are
Slika 5. Histološki nalaz defekta kosti nakon 30 dana (kontrolna gru- visible (HE, 200×).
pa). Fibrovaskularno vezivno tkivo okružuje ostrvca novoizgrađene Slika 6. Histološki nalaz defekta kosti nakon 90 dana (kontrolna
kosti (HE, 400×). grupa). Novostvorena kost sa organizacijom po tipu osteona i gra-
nica prema lamelarnoj kosti (HE, 200×).

placed in oval contours. Border cement lines were baso- intaoseal) is considered to be superior test. Tested mate-
philic, enlarged, with amorphous to fine-grained appear- rials are commonly implanted into the tissues of rats [5,
ance. In one case (1 of 6) giant cell reaction to a foreign 8, 10], rabbits [11], guinea pigs [12]. Intraoseal models
body was observed. provide an adequate environment for in vivo testing of
Ninety days after material implantation, osteoplasia endodontic materials. In this experiment, the method of
and trabeculae of different widths surrounded by osteo- mandible bone implantation was chosen because of its
blasts were observed. Newly formed bone was young and specificity [13].
morphologically immature. There was no restitutio ad in- Eugenol (4-allyl-2-methoxyphenol) is clove oil extract
tegrum, but focal remodeling of bone tissue was intense and a component of zinc oxide eugenol (ZnOE). It causes
(Figure 3). Fibrous connective tissue was focally distrib- periapical toxicity and inhibits growth and proliferation
uted and reduced (in all samples). Bone tissue, away from of human osteoblastic U2OS cell line. As cell growth, at-
experimental area showed normal morphology (Figure 4). tachment, proliferation and synthesis of matrix play an
important role in wound healing and tissue regeneration,
it can be assumed that eugenol in tissue may disturb the
Control group processes of wound healing [14, 15].
Thirty days after the material (Endomethasone N)
Thirtieth day after preparation of experimental defect in implantation, its presence was macroscopically observed
all control samples, osteosynthesis activity and fibrovas- inside the experimental defects. During the process of
cular connective tissue were observed (Figure 5). Young making histological preparations, the material was in
bone tissue with osteocytes in enlarged oval lacunae was most cases partially or completely rinsed from the im-
noted. Thickened edges of osteons with increased baso- plantation site and experimental defects were represented
phil responses were present away from the experimental as empty spaces. After the experimental period of 90 days,
defect. No signs of inflammation in tissue were noted. microscopic analysis did not show tested material (Endo-
Ninety days after preparation of artificial defect, an ad methasone N) present.
integrum bone healing was noted in all samples of the Thirtieth day after implantation of Endomethasone
control group. Osteons of newly formed bone tissue had N, fibrovascular connective tissue by type of callus was
smaller diameter and smaller number of concentric la- observed as well as new bone formation. Inflammatory
mellae (Figure 6). response, in all samples, was slightly pronounced except in
one case where the response was more type of reaction to
the foreign body with granulation tissue. On the ninetieth
DISCUSSION day the absence of inflammatory reactions was recorded
and slower degree of fibrovascular proliferation in relation
Before introducing any material in the clinical practice to the period of 30 days. Connective tissue was mostly re-
it is necessary to conduct appropriate tests. Cytotoxicity placed with young bone tissue with osteon type structure.
testing of endodontic materials by in vitro tests provides In the control group after the period of 30 days an in-
control of the experiment conditions but does not cor- flammatory reaction was observed which disappeared
respond completely to clinical situation. For this reason, until the ninetieth day. Also, in the control group, the
implantation of materials (subcutaneous, intramuscular, presence of connective tissue in the second period (90
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162 Nikolić M. et al. The Effect of Zinc Oxide Based Sealer on Bone Defects Healing

days) was not observed, and the experimental defect was Other authors reported different results from ours. Za-
completely replaced by newly formed bone tissue. The falon et al. found moderate to severe inflammatory reac-
presence of connective tissue in the experimental group tion after 15 days of implantation, while after 30, 60, 90
after 30 days and its reduction after 90 days suggest that days, there was no reaction [10]. The time period was the
after longer time it may be completely replaced by newly same so the difference can be attributed to the experimen-
formed bone. tal procedure and the difference in material composition
Results of the current study were similar to other of Endomethasone and Endomethasone-N.
studies [13, 14]. Tassery et al. describe mild to moder- The most ingredients of Endomethasone are released
ate infiltration of inflammatory cells after 4 weeks when during setting reaction due to its high solubility [21]. Seal-
material Super EBA (based on zinc oxide) was implanted er components diffuse easier in the freshly mixed state,
in the mandible of guinea pigs. This reaction decreased although they could be released by hydrolysis even after
over time and it was not observed after 12 weeks. This material is set [15]. Continuous release of toxic compo-
study did not find healing disruption due to increased in- nents such as paraformaldehyde and eugenol may explain
flammatory reaction [13]. The initial inflammatory effect persistent inflammation over longer period of time.
was attributed to eugenol and etoxybenzoic acid (Super
EBA ingredient) that were released [16], but the setting
of the material reduced their amount [17]. High degree CONCLUSION
of binding of these substances to serum and extracellu-
lar proteins may contribute to reduced toxicity and mild Endomethasone N slows down bone tissue healing pro-
inflammation [18]. cess by showing the signs of prolonged inflammation in
Triches et al. examined inflammatory reaction of sub- bone tissue in which it was implanted. Prolonged healing
cutaneous tissue in rats using histomorphological param- is reflected in slow replacement of fibrovascular connec-
eters such as - quantity of inflammatory cells, presence of tive tissue with newly formed bone tissue. However, per-
blood vessels, area of necrosis and the thickness of fibrous manent disturbance of morphofunctional relations in the
capsule. A mild to moderate tissue reaction to Endometh- bone tissue was not found.
asone N was described after 7 days with predominance
of chronic inflammatory cells, while after 42 days tissue
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Received: 19.07.2016 • Accepted: 01.11.2016

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164 Nikolić M. et al. The Effect of Zinc Oxide Based Sealer on Bone Defects Healing

Uticaj materijala za opturaciju na bazi cink-oksida na zarastanje

koštanih defekata
Marija Nikolić, Jelena Popović, Jovanka Gašić, Radomir Barac
Univerzitet u Nišu, Medicinski fakultet, Odeljenje za bolesti zuba i endodonciju, Klinika za stomatologiju, Niš, Srbija

KRATAK SADRŽAJ
Uvod Cilj opturacije kao završne faze endodontskog tretmana je da se obezbedi kompletno hermetično zatvaranje duž cele dužine
kanala od koronarnog otvora da apeksne granice.
Cilj studije bio je da se proveri histološki odgovor koštanog tkiva na implantaciju materijala na bazi cink-oksida u arteficijalno
preparisan defekt u mandibuli pacova.
Metode rada U eksperimentu je upotrebljeno 16 pacova Wistar soja, muškog pola. Upotrebom sterilnih, čeličnih fisurnih borera
formiran je defekt između medijalne linije i mentalnog otvora u mandibuli pacova. Siler baziran na cink-oksidu je implantiran u
defekte eksperimentalne grupe, dok su defekti kod kontrolne grupe ostavljeni da spontano zarastaju. Polovina životinja iz obe grupe
je žrtvovana posle trideset, a druga polovina posle devedeset dana. Mikroskopski preparati su se sastojali od defekta i okolne kosti
i posle pripreme su analizirani svetlosnom mikroskopijom.
Rezultati Trideset dana posle implantacije je uočeno fibrovaskularno vezivno tkivo sa oskudnim hroničnim inflamatornim infil-
tratom. Dalje od eksperimentalnog defekta, u dubini kosti, zapažena je lamelarna kost sa dobro formiranim većim osteonima i
proširenim Haversovim i Folkmanovim kanalima. Devedeset dana posle implantacije materijala nije došlo do restitutio ad integrum,
ali je fokalno remodelovanje kosti bilo intenzivno.
Zaključak Endometazon N usporava proces zarastanja koštanog tkiva jer pokazuje znake produženog zapaljenja u koštanom tkivu
u koje je implantiran. Produženje procesa zarastanja ogleda se i u usporenoj zameni fibrovaskularnog vezivnog tkiva novonastalim
koštanim tkivom.
Ključne reči: sileri; opturacija; zarastanje kosti

UVOD CILJ RADA

Opturacija kao završna faza endodontskog tretmana ima
za cilj obezbeđivanje kompletnog hermetičnog punjenja duž Cilj istraživanja bio je da se proveri histološki odgovor koštanog
celokupne dužine kanalskog sistema od koronarnog otvora do tkiva na implantaciju materijala na bazi cink-oksida u artefici-
apeksne granice. Uloga opturacije je da se spreči komunikacija jalno preparisan defekt u mandibuli pacova.
između endodonta i parodonta i na taj način onemogući rein-
fekcija periapeksne regije [1]. Materijali koji se upotrebljavaju bi
trebalo da poseduju niz fizičkih, hemijskih i bioloških osobina METOD RADA
i po pravilu ne bi trebalo da se prostiru izvan granica kanal-
skog sistema [1]. Kada biomaterijal dođe u kontakt sa tkivima U eksperimentu je korišćeno 16 Wistar pacova muškog pola,
i tečnostima ljudskog organizma, uvek postoji neka forma in- telesne mase 160–180 g (odobreno od strane Etičkog komiteta
terakcije i zato je potrebno da materijal bude biokompatibilan, Medicinskog fakulteta u Nišu, broj 01 3797). Tokom eksperi-
neškodljiv u biološkoj sredini, a da je pacijent zaštićen od bilo mentne procedure životinje su anestezirane intraperitoneal-
kakvih negativnih uticaja [2]. nom injekcijom ketamin-hidrohlorida (0,1 ml na 100 ml telesne
Preterana instrumentacija i sledstveno prepunjavanje ka- mase). Nakon postupka pripreme, između središnje linije i men-
nala korena značajno povećavaju rizik od nepovoljnog uticaja talnog otvora sa leve strane donje vilice svih životinja, naprav-
materijala za punjenje [3]. Prepunjavanje utiče na povećanu ljen je defekt (prečnika 1,4 mm i dubine 1,6 mm) korišćenjem
inflamaciju i usporeno zarastanje [4, 5]. Negativno dejstvo je sterilnog fisurnog čeličnog borera. Životinje eksperimentalne
obično izraženije dok materijal još nije potpuno vezan i uglav- grupe podeljene su u dve podgrupe:
nom se pripisuje sastavnim komponentama materijala [6]. • Prva podgrupa (n = 6) žrtvovana nakon 30 dana
Sporo oslobađanje negativnih agenasa tokom dužih perioda • Druga podgrupa (n = 6) žrtvovana nakon 90 dana
vremena zavisi od rastvorljivosti materijala, odnosno stepena
izloženosti materijala tkivnim tečnostima [2]. U formirane defekte kod eksperimentalne grupe životinja
Materijali na bazi cink-oksida sa eugenolom su najčešće implantiran je endometazon N (Septodont, France) pripremljen
upotrebljavani i obično se koriste kao komparativni sileri u stu- po uputstvu proizvođača. Preparisan defekt kod životinja kon-
dijama koje ispituju biološke efekte materijala [4, 7]. Rezultati trolne grupe (n = 4) ostavljen je da spontano zarasta bez ika-
ovih studija, međutim, nisu ohrabrujući. S obzirom na to da su kvog implantata. Dve životinje iz kontrolne grupe su žrtvovane
prema nekim autorima pojedini materijali iz ove grupe veoma posle 30, a dve posle 90 dana. Nakon predviđenog vremena
toksični, oni ih ne preporučuju za upotrebu [7]. Toksičnost se životinje su žrtvovane predoziranjem anestetikom (ketamin-
uglavnom pripisuje prisustvu eugenola ili formaldehida, koji hidrohlorid). Uzorci tkiva su napravljeni resekcijom mandibule,
neki od ovih materijala sadrže [8, 9]. a sastojali su se od defekta i okolne kosti. Uzorci tkiva su fik-
sirani u 10% puferisanom formalinu, demineralizovani u 10%
mravljoj kiselini, dehidrirani u seriji alkohola, a zatim kalupljeni
u parafinskom vosku. Sečenje je izvedeno u bukolingvalnom
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 Stomatološki glasnik Srbije. 2016;63(4):159-166 165

pravcu na mikrotomu staklenim noževima 2 μm debljine. Pre- 10], zečeva [11], zamoraca [12]. Intraosealni modeli obezbe-
parati su bojeni hematoksilin-eozin metodom i mikroskopski đuju adekvatno okruženje za in vivo testiranje endodontskih
analizirani svetlosnim mikroskopom VH50 (Olimpys, Japan) materijala. U ovom eksperimentu je korišćena metoda koštane
opremljenim digitalnom kamerom Leica DFC 295. implantacije, a odabrana je mandibula zbog specifičnosti svoje
građe [13].
Eugenol (4 alil-2-metoksifenol) jeste ekstrakt ulja karanfilića
REZULTATI i predstavlja sastojak cinkoksid-eugenola (ZnOE). Izaziva peri-
apeksnu toksičnost i inhibiše rast i proliferaciju U2OS humane
Eksperimentalna grupa osteoblasne ćelijske linije. S obzirom na to da ćelijski rast, veziva-
nje, proliferacija i sinteza matriksa igraju važnu ulogu u zarasta-
Tridesetog dana posle implantacije materijala uočava se fibrova- nju rana i tkivnoj regeneraciji, može se pretpostaviti da oslobo-
skularno vezivno tkivo, sa oskudnim hroničnim inflamativnim đeni eugenol može uzrokovati poremećaje u zarastanju [14, 15].
ćelijskim infiltratom u vidu fokusa, dok je okolna kost hipo- U eksperimentalnom periodu od 30 dana unutar ekspe-
celularna i bez prisustva Haversovih sistema (kod pet od šest rimentalnih defekata makroskopski je zapaženo prisustvo ili
uzoraka) (Slika 1). Osteociti malih hiperhromatičnih jedara su manji ostaci materijala (endometazon N). Tokom tehnike obra-
smešteni u lako proširene lakune. Osteoblasti su malobrojni, de uzorkovane kosti i izrade histoloških preparata opturacioni
dok je prisustvo osteoklasta neupadljivo. Na mestima neresor- materijal je u najvećem broju slučajeva potpuno ili delimično
bovanog materijala, ispalog tokom obrade histopatoloških pre- ispao sa mesta implantacije, a eksperimentalni defekti svetlo-
parata, zapažaju se prazni prostori. Dalje od eksperimentalno sno-mikroskopski su bili predstavljeni kao prazni prostori.
načinjenog defekta, prema dubini kosti, uočava se lamelarna U eksperimentalnom periodu od 90 dana makroskopski i sve-
kost sa dobro formiranim većim osteonima, proširenih Haver- tlosno-mikroskopski primenjeni opturacioni materijal (endo-
sovih i Folkmanovih kanala (Slika 2). U osteonima se zapažaju metazon N) nije uočen.
proširene intersticijalne lamele, a osteociti su smešteni u lakune Tridesetog dana od implantacije materijala endometazona
ovalnih kontura. Granične cementne linije su bazofilne, prošire- N uočeni su fibrovaskularno vezivno tkivo po tipu kalusa i stva-
ne, sitnozrnastog do amorfnog izgleda. U jednom slučaju (jedan ranje nove kosti. Inflamativni odgovor je kod svih uzoraka bio
od šest) uočena je džinovsko-ćelijska reakcija na strano telo. blago izražen osim u jednom slučaju gde je uočen odgovor po
Devedeset dana nakon implantacije materijala u preparisan tipu reakcije na strano telo sa granulacionim tkivom i okolnom
defekt u koštanom tkivu se zapaža osteoplazija i formiranje tra- reaktivno izmenjenom kosti. Devedesetog dana zabeleženi su
bekula različite širine, koje su okružene osteoblastima. Novofor- odsustvo inflamativne reakcije u tkivu i manji stepen fibrova-
mirana kost je mlada, morfološki nezrela. Nije došlo do restitutio skularne proliferacije u odnosu na period od 30 dana. Vezivno
ad integrum, ali je fokalno remodelovanje koštanog tkiva inten- tkivo je najvećim delom zamenjeno mladim koštanim tkivom
zivno (Slika 3). Fibrozno vezivno tkivo je fokalno raspoređeno i sa građom po tipu osteona.
redukovano (kod svih uzoraka). Koštano tkivo dalje od eksperi- U eksperimentalnoj grupi u ovom istraživanju je u ranijem
mentalnog područja pokazuje normalnu morfologiju (Slika 4). periodu (30 dana) uočena zapaljenska reakcija, koja se do de-
vedesetog dana potpuno povukla. Takođe, kod kontrolne grupe,
za razliku od eksperimentalne, nije uočeno prisustvo vezivnog
Kontrolna grupa tkiva u drugom vremenskom periodu (90 dana), već je ekspe-
rimentalni defekt potpuno zamenjen novonastalim koštanim
Tridesetog dana od preparacije eksperimentalnog defekta kod tkivom. Prisustvo vezivnog tkiva kod eksperimentalne grupe
svih uzoraka kontrolne grupe zapaža se osteosintetska aktivnost nije bilo izraženo i primećeni su znaci njegovog smanjivanja
i fibrovaskularno vezivno tkivo (Slika 5). Prisutno je mlado ko- u odnosu na raniji period, pa bi se moglo pretpostaviti da će s
štano tkivo čiji osteociti imaju proširene ovalne lakune. Dalje od vremenom potpuno biti zamenjeno novostvorenom kosti.
eksperimentalnog defekta se uočavaju zadebljali rubovi osteona Reakcija tkiva slična dobijenoj u ovoj studiji sreće se i u istra-
pojačano bazofilne reakcije (Slika 6). U tkivu nema znakova živanjima drugih autora [13, 14]. Kod implantacije materijala
zapaljenja. Super EBA (na bazi cink-oksida) u mandibule zamoraca, Tassery
Devedeset dana od preparacije arteficijalnog defekta kod i sar. opisuju blagu do umerenu infiltraciju inflamativnim će-
kontrolne grupe (svi uzorci) uočava se potpuno zarastanje ad lijama posle četiri nedelje. Ova reakcija se smanjivala vreme-
integrum. Osteoni novonastalog koštanog tkiva pokazuju manji nom i nije zapažena posle 12. nedelje. Pomenuto istraživanje
dijametar i manji broj koncentričnih lamela (Slika 6). ne ukazuje na ometanje zarastanja niti na pojačanu reakciju
inflamacije [13]. Inicijalni inflamativni efekat se pripisuje eu-
genolu i etoksibenzoevoj kiselini (sastojak Super EBA), koji
DISKUSIJA se oslobađaju [16] i čije se otpuštanje smanjuje sa vezivanjem
materijala [17]. Visok stepen vezivanja ovih supstanci za serum
Pre preporuke za kliničku upotrebu nekog materijala neophod- i ekstracelularne proteine može doprineti redukovanoj toksič-
no je sprovesti odgovarajuće testove. Testiranje citotoksičnosti nosti i pratećoj blagoj inflamaciji [18].
endodontskih materijala in vitro testovima omogućava kontro- Inflamativna reakcija subkutanog tkiva pacova je dobijena
lisanje uslova eksperimenta, ali ne odgovara u dovoljnoj meri i u studiji Triches i sar., gde su ispitivani histomorfološki para-
kliničkoj situaciji. Iz tog razloga se tehnike implantacije (subku- metri kao što su kvantitet inflamativnih ćelija, prisustvo krv-
tane, intramuskularne, intraosealne) smatraju superiornijim. nih sudova, zona nekroze i debljina fibrozne kapsule. Opisana
Materijali koji se testiraju implantiraju se u tkiva pacova [5, 8, je blaga do umerena tkivna reakcija na endometazon N posle
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166 Nikolić M. et al. The Effect of Zinc Oxide Based Sealer on Bone Defects Healing

sedam dana sa predominacijom hroničnih inflamativnih ćelija, kada se prebaci preko apeksa korena zuba. Neslaganje rezultata
dok je 42. dana tkivo ostalo blago inflamirano, sa prisustvom može se pripisati različitim životinjskim modelima i eksperi-
limfocita, fibroblasta i dobro organizovanih kolagenih vlakana mentalnom dizajnu.
[14]. Autori su došli do zaključka da su svi testirani sileri (endo- Podaci iz literature, kod ispitivanja endometazona in vivo
metazon N, endofil (na bazi ZnO), Sealer 26 (na bazi CaOH2)) subkutanim testom ukazuju na rezultate različite od dobijenih
iritansi, čija se toksičnost vremenom smanjuje. u ovoj studiji. Zafalon i sar. su ustanovili umerenu do jaku in-
Kada su u pitanju dugi vremenski periodi, rezultati dobijeni flamativnu reakciju nakon 15 dana od implantacije, a posle 30,
u ovoj studiji su saglasni sa rezultatima drugih autora koji po- 60, 90 dana reakcije nije bilo [10]. S obzirom na to da se radi o
tvrđuju da endometazon N usporava procese reparacije i izaziva istom vremenskom periodu, razlike se mogu pripisati eksperi-
kasnu zapaljensku reakciju [14, 19, 20]. mentalnoj proceduri i razlici u sastavu materijala endometazo-
U istraživanju koje su sproveli Suzuki & Souza, kod endo- na i endometazona N.
metazona nije zapaženo idealno zarastanje ni posle 90 dana. Najviše sastojaka endometazon oslobađa tokom reakcije ve-
Eksperiment je sproveden na psima čiji su kanali korena zuba zivanja, što se može objasniti velikom rastvorljivošću materijala
prepunjavani ovim materijalom, a zatim patohistološki analizi- [21]. Komponente silera lakše difunduju u sveže zamešanom
rana reakcija periradiksnog tkiva. Hronični zapaljenski ćelijski stanju, iako se eugenol može osloboditi hidrolizom i iz vezanog
infiltrat različitog stepena javio se kod svih uzoraka, dok su materijala [15]. Kontinuirano ispuštanje toksičnih komponena-
gigantske ćelije bile prisutne kod osam od deset uzoraka [20]. ta poput eugenola i paraformaldehida može objasniti perzisti-
Dobijeni rezultati u ovoj studiji takođe ukazuju na hroničnu ranje inflamacije tokom dužeg vremenskog perioda.
inflamativnu reakciju koštanog tkiva na endometazon u perio-
du posle 30 dana, koja je slabog intenziteta, što je u saglasnosti
sa autorima koji ukazuju na blagu inflamaciju sa znacima re- ZAKLJUČAK
generacije posle 42. dana od subkutane implantacije endome-
tazona kod pacova [14]. Na osnovu dobijenih rezultata može se zaključiti da endometa-
Devedesetog dana od implantacije nisu uočeni znaci zapalje- zon N usporava proces zarastanja koštanog tkiva jer pokazuje
nja, već preovladavanje regenerativnih procesa. Ovi se rezultati znake produženog zapaljenja u koštanom tkivu u koje je im-
ne slažu sa nalazima autora koji hronični zapaljenski infiltrat plantiran. Produženje procesa zarastanja ogleda se i u usporenoj
registruju i posle perioda dužih od 90 dana (šest meseci) kod zameni fibrovaskularnog vezivnog tkiva novonastalim koštanim
prepunjenih kanala endometazonom kod majmuna [19]. Do- tkivom, ali materijal ne dovodi do trajnog narušavanja morfo-
bijene rezultate objašnjavaju iritacijom koju materijal izaziva funkcionalnih odnosa u tkivu.

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