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ID naloge: 143 Letnik: 2003 Predmet: stomatologija
PREŽIVETJE CELIC POZOBNICE PO IZBITJU ZOB
Avtor: Katarina Odar
Mentor: doc. dr. Nataša Ihan Hren
IZHODIŠCE: Izbitje zoba iz zobne jamice (ZJ) je ena najtežjih poškodb zob. Zdravljenje je ponovna vsaditev in ucvrstitev izbitega zoba v vsajenem položaju. V casu pred ponovno vsaditvijo poškodovanci shranjujejo zobe na razlicne nacine. Za uspeh zdravljenja je bistveno, da v tem casu preživi cim vec celic v pozobnici izbitih zob. Vecja kot je stopnja preživetja teh celic, manjša je verjetnost razgradnje zobne korenine in izgube vsajenega zoba. Pozobnica kirurško odstranjenih zob predstavlja enostavno dostopen model za preucevanje dogajanja v pozobnici izbitih zob.
NAMEN: Da bi prispevali k uspešnemu nacrtovanju zdravljenja izbitih zob, smo želeli na enostaven in objektiven nacin dolociti casovno odvisnost preživetja celic pozobnice po izbitju zob. To odvisnost smo želeli primerjati pri dveh nacinih shranjevanja izbitih zob pred ponovno vsaditvijo (v fiziološki raztopini ali brez nje) in preveriti vpliv nekaterih lastnosti bolnika ob izbitju zoba na stopnjo preživetja celic v pozobnici.
HIPOTEZA: Naše delovne hipoteze so bile, da delež mrtvih celic v pozobnici narašca s casom od odstranitve zob iz ZJ po doloceni krivulji, da nacin shranjevanja zob po njihovi odstranitvi vpliva na stopnjo preživetja celic pozobnice ter da ima nanjo pomemben vpliv tudi starost bolnika ob odstranitvi zoba.
METODE: Uporabili smo metodo, ki še ni bila uporabljena za dolocanje preživetja celic pozobnice. Kirurško odstranjene tretje kocnike smo shranjevali v zaprtih posodicah (5ml). Prvi skupini zob (n=64) smo dodali fiziološko raztopino, zobe druge skupine (n=41) pa smo pustili suhe. Iz zob smo ob razlicnih casih v razponu od 0 do 240 minut odvzemali vzorce pozobnice in preiskovali stopnjo preživetja njenih celic z barvanjem mrtvih celic s propidijevim jodidom in štetjem vseh celic s pretocno citometrijo (PC). Iz skupine 1 smo izlocili 13, iz skupine 2 pa 11 vzorcev, ki niso bili pripravljeni na standarden nacin in jih loceno obravnavali v podskupinah A in B. Za statisticno obdelavo podatkov smo uporabili multiplo in enostavno linearno regresijsko analizo, Chowov test in dvosmerni t test za neodvisne spremenljivke. Na 6 dodatnih zobeh (skupina 3) smo histološko preverili vsebino vzorca.
REZULTATI: Z multiplo linearno regresijsko analizo, v katero smo vkljucili le podatke za zobe, preiskane do 200 minut po odstranitvi iz ZJ, smo preverili vpliv casa in nacina shranjevanja zob ter vpliv starosti in spola bolnika na preživetje celic pozobnice. Le za cas smo ugotovili statisticno znacilen vpliv (p<0,001). Narašcanje odstotka mrtvih celic pozobnice je bilo hitrejše v drugi skupini (napoved z enostavno linearno regresijsko analizo je 0,187% na minuto) kot v prvi (0,094% na minuto), vendar razlika ni statisticno znacilna (Chowov test; p=0,186). Aritmeticna sredina zabeleženih odstotkov mrtvih celic je bila v vzorcih podskupine A statisticno znacilno nižja kot v ostalih vzorcih skupine 1 (t-test; p=0,001) in v vzorcih podskupine B statisticno znacilno nižja kot v ostalih vzorcih skupine 2 (p<0,001).
ZAKLJUCKI: Potrdili smo našo prvo hipotezo. Casovni potek odmiranja celic pozobnice v prvih 200 minutah po odstranitvi zob iz telesa lahko približno ponazorimo z enacbo premice. Zavrnili smo hipotezo, da nacin shranjevanja zob vpliva na stopnjo preživetja celic pozobnice. Shranjevanje izbitih zob brez dodanega medija v tesno zaprtih majhnih posodicah, ne da bi pred tem s koreninske površine odstranili kri, ohranja preživetje celic pozobnice podobno dobro kot fiziološka raztopina, ce zobe shranjujemo do 200 minut po odstranitvi iz ZJ. Tudi našo tretjo hipotezo smo zavrnili. Starost poškodovancev ne vpliva na preživetje celic v pozobnici izbitih zob, ki jih shranjujemo na obravnavana nacina do 200 minut. Metoda, ki smo jo uporabili, se je izkazala kot ustrezna, vendar uporabna le pod dolocenimi pogoji. Vzorce lahko preiskujemo le v casu do 200 minut po odstranitvi zob iz ZJ. Biti morajo ustrezno pripravljeni, ker nacin odvzema vzorca lahko vpliva na odstotek mrtvih celic, ki ga zabeleži PC.
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[Abstract / English version]
PREŽIVETJE CELIC POZOBNICE PO IZBITJU ZOB
Author: Katarina Odar
Mentor: doc. dr. Nataša Ihan Hren
BACKGROUND: Tooth avulsion is one of the most severe injuries to the teeth. The treatment of choice is replantation and immobilisation of the avulsed tooth. Prior to replantation, the teeth are often being stored in various manners. Periodontal ligament (PDL) cell viability is one of the most crucial factors affecting the prognosis of avulsed teeth. The higher the survival rate of the PDL cells during the time delay between avulsion and replantation, the lower the risk of root resorption and tooth loss. The PDL of the surgically extracted teeth provides us an easily accessible experimental model to study the PDL of the avulsed teeth.
AIM: To objectively determine the time - dependence of PDL cell viability after the avulsion and to compare it between the two types of storage conditions (with and without the presence of physiologic saline solution). We also wanted to determine the impact of the patient's age on the PDL cell viability. We wanted to make a contribution for a more successful treatment planning.
HYPOTHESIS: The percentage of the nonviable PDL cells increases with time after the tooth removal (storage time) and follows a certain time-curve. The storage conditions after the avulsion can affect the PDL cell survival of the stored teeth. The age of the patient has an important impact on the PDL cell survival.
METHODS: The method we used has never been used for this purpose before. Surgically extracted human third molars were stored in tightly closed plastic containers (5 ml). To the first group of teeth (n=64), physiologic saline solution was added, the second group (n=41) was left dry. After various times ranging from 0 to 240 minutes, the PDL samples were obtained from the root surfaces and the cell viability was determined by using a nonvital cell staining with propidium iodide and total cell counting in a flow cytometer. 13 samples fom group 1 and 11 samples from group 2 that weren't prepared in a standard manner were studied separately in subgroups A and B. For the statistical analysis multiple and simple linear regression analysis, Chow's test and two - tailed independent t - test were used. Additional 6 samples (group 3) were examined histologically.
RESULTS: We tested the impact of storage time, storage conditions and age and sex of the patient on the PDL cell survival with multiple linear regression analysis in which the data for the teeth examined up to 200 minutes after the extraction was included. Only time showed a statistically significant impact (p<0,001). The increment of the nonviable cell percentage was greater in the second group of teeth (0,187% per minute according to the simple linear regression) than in the first group (0,094% per minute), but the difference was not statistically significant (Chow's test; p=0,186). The mean percentage of nonviable cells noted by the flow cytometry was significantly lower in subgroup A compared to the rest of the group 1(t-test; p=0,001) and in subgroup B compared to the rest of the group 2 (p<0,001).
CONCLUSIONS: Our first hypothesis was confirmed. The time course of the PDL cell viability loss can be approximated with simple linear equation for storage times up to 200 minutes. Our second hypothesis was not confirmed. The dry storage of avulsed teeth in tightly closed 5 ml containers with blood left on the root surfaces can maintain PDL cell viability as good as the physiologic saline solution storage during the storage times up to 200 minutes. Our third hypothesis was also not confirmed, age has no impact on the PDL cell survival .The method that we used in the study was proved to be useful only under certain conditions. Only teeth stored for up to 200 minutes can be meaningfuly investigated and the samples have to be obtained in an adequate manner, because some factors during the sample preparation can alter the percentage of nonviable cells noted by the flow cytometry.
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