PRACA POGLĄDOWA
Wykorzystanie komórek nabłonkowych z jamy ustnej w monitoringu biologicznym ludzi
 
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Instytut Ekologii Terenów Uprzemysłowionych w Katowicach Dyrektor: dr hab. inż. Jan Skowronek
AUTOR DO KORESPONDENCJI
Ewa Błaszczyk   

Instytut Ekologii Terenów Uprzemysłowionych ul. Kossutha 6; 40-844 Katowice tel. 32 254 60 31 wew. 247
 
Med Środow. 2012;15(4):129–138
SŁOWA KLUCZOWE
STRESZCZENIE ARTYKUŁU
Jedną z podstawowych metod określania stopnia zagrożenia środowiskowego ludzi jest oznaczenie substancji szkodliwych w poszczególnych elementach środowiska (powietrze, woda, gleba, żywność). W odróżnieniu od monitoringu środowiska – monitoring biologiczny ludzi (HBM) umożliwia oszacowanie wielkości dawki wchłoniętej, ogólnej bądź zlokalizowanej w określonym narządzie. HBM pozwala na ocenę narażenia na substancje, które wchłaniane są do organizmu wszystkimi drogami i z różnych nośników zanieczyszczeń. Polega na pomiarze wskaźników, tzw. biomarkerów w płynach ustrojowych (krew, mocz, ślina, itd.) lub w tkankach i narządach. Biomarkery dzielimy na markery ekspozycji, skutków oraz wrażliwości. Szczególnie przydatną metodą jest oznaczanie adduktów, czyli związków substancji rakotwórczych (lub ich metabolitów) z białkami lub DNA, będących markerem narażenia. Za biomarkery skutków biologicznych uznawane są różne zmiany cytogenetyczne, w tym także obecne w komórkach mikrojądra. Są to struktury pozajądrowe zawierające fragmenty chromatyny (powstałe w skutek pęknięć DNA) lub całe chromosomy (uszkodzenia wrzeciona podziałowego). Większość badań poziomu adduktów DNA i mikrojąder była dotychczas prowadzona w komórkach limfocytów krwi obwodowej. W chwili obecnej prowadzenie badań z wykorzystaniem krwi, zwłaszcza u dzieci jest obostrzone względami bioetycznymi, dlatego zainteresowaniem naukowców zaczęły się cieszyć testy wykorzystujące komórki nabłonkowe z jamy ustnej. Komórki nabłonkowe są głównym budulcem tkanki nabłonkowej, a także stanowią około 60% wszystkich komórek organizmu człowieka. Tkanka nabłonkowa pełni przede wszystkim funkcję okrywającą i wyściełającą zewnętrzne oraz wewnętrzne powierzchnie organizmu. Nabłonek w różnych częściach organizmu człowieka uległ wysokiej specjalizacji, co związane jest z jego budową i pełnioną funkcją. Jamę ustną człowieka jest pokrywa nabłonek wielowarstwowy płaski, który zbudowany jest z komórek określanych jako keratynocyty. Komórki nabłonkowe jamy ustnej mogą podlegać dwóm drogom różnicowania – w kierunku nabłonków rogowaciejących i nierogowaciejących. W niniejszej pracy w oparciu o nasze dotychczasowe doświadczenia oraz dostępną literaturę przedstawiono procedury badawcze zawierające opis poboru komórek nabłonkowych z jamy ustnej oraz przygotowanie go w ten sposób, aby mógł zostać wykorzystany zarówno do analizy adduktów DNA, jak i testu mikrojądrowego

One of the basic methods for determining the degree of environmental risk posed to humans is identification of harmful substances in various environmental elements (air, water, soil, food). In contrast to environmental monitoring human biological monitoring (HBM) enables the estimation of an absorbed dose, general or localized in a specific organ. HBM enables the assessment of exposure to substances which are absorbed by the body via different exposure pathways and with different contaminant carriers. It is based on the measurement of indicators, the so-called biomarkers, in body fluids (blood, urine, saliva, etc.) or in tissues and organs. Biomarkers can be divided into markers of exposure, effects and susceptibility. A particularly useful method is determination of adducts, i.e. carcinogenic compounds (or their metabolites) with proteins or DNA, which are markers of exposure. Biomarkers of biological effects are different cytogenetic changes, including micronuclei. These are extranuclear structures containing fragments of chromatin (arising as a result of DNA breaks) or whole chromosomes (damage to the spindle apparatus during mitosis). Up to now most studies on the DNA adduct levels and micronuclei have been conducted in peripheral lymphocytes. At present, studies using blood, especially in children to restricted to ethical aspects, and therefore tests using epithelial cells from the oral cavity have become more popular. Epithelial cells are the main building material of an epithelial tissue which makes up about 60% of all cells of the human body. The main function of the epithelial tissue is covering and lining of the outer and inner surfaces of the body. Epithelium underwent high specialisation in various parts of the human body, which is associated with its structure and function. Human oral cavity is covered by stratified squamous epithelium, which is comprised of cells called keratinocytes. Oral epithelial cells may differentiate in two directions: towards keratinized or nonkeratinized oral epithelia. In this study, based on our past experience and the available literature, research procedures for the collection of oral epithelial cells and their proper preparation for using them both for the analysis of DNA adducts and micronucleus assay are presented.
SKRÓTY
HBM – monitoring biologiczny ludzi; TP – temperatura pokojowa, PBS – zbuforowany roztwór soli fizjologicznej
 
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