Grading and quantification of dental fluorosis in zebrafish larva.
OBJECTIVE
The prevalence and severity of dental fluorosis in primary teeth are different from permanent teeth. Previous animal models of dental fluorosis mainly focus on juvenile rats, mice and zebrafish. Our experiment aims to set a dental fluorosis model using zebrafish larva and explore the characteristics of the first generation teeth by fluoride treatment.


METHODS
After the zebrafish eggs were laid, they were exposed to excess fluoride (19ppm, 38ppm and 76ppm) for five days. The morphological characteristics of first generation teeth were examined by H&E staining, whole-mount alizarin red and alcian blue staining, and scanning electron microscope (SEM) technique.


RESULTS
With whole-mount alizarin red and alcian blue staining, the tooth cusps presented red in normal control. 19ppm and 38ppmm fluoride resulted in extensive red staining from tooth cusps to the lower 1/3 of teeth. 76ppm fluoride caused malformed teeth with uneven red staining. H&E staining showed that excess fluoride caused cystic-like changes in 38ppm and 76ppm groups. SEM revealed the dose dependent pathological changes in zebrafish enameloid with fluoride treatment. Based on SEM findings, we set 0-4 dental fluorosis index (DFI) score to label the severity of dental fluorosis.


CONCLUSIONS
Excess fluoride presented a dose dependent fluorosis changes in the teeth of zebrafish larva. The DFI scores in our experiment reflect dose dependent fluorosis changes in a good way and will benefit the future research of dental fluorosis.
State Key Laboratory of Military Stomatology, National Clinical Research Center for Oral Diseases, Shaanxi Key Laboratory of Oral Diseases, Department of Oral Biology, Clinic of Oral Rare and Genetic Diseases, School of Stomatology, The Fourth Military Medical University, 145 Changle West Road, Shaanxi, Xi'an 710032, People's Republic of China; Department of Prosthodontics, School of Stomatology, The Fourth Military Medical University, 145 Changle West Road, Shaanxi, Xi'an 710032, People's Republic of China.
Yutao
Y
Zhang
State Key Laboratory of Military Stomatology, National Clinical Research Center for Oral Diseases, Shaanxi Key Laboratory of Oral Diseases, Department of Oral Biology, Clinic of Oral Rare and Genetic Diseases, School of Stomatology, The Fourth Military Medical University, 145 Changle West Road, Shaanxi, Xi'an 710032, People's Republic of China.
Yanli
Y
Zhang
State Key Laboratory of Military Stomatology, National Clinical Research Center for Oral Diseases, Shaanxi Key Laboratory of Oral Diseases, Department of Oral Biology, Clinic of Oral Rare and Genetic Diseases, School of Stomatology, The Fourth Military Medical University, 145 Changle West Road, Shaanxi, Xi'an 710032, People's Republic of China.
Xueni
X
Zheng
State Key Laboratory of Military Stomatology, National Clinical Research Center for Oral Diseases, Shaanxi Key Laboratory of Oral Diseases, Department of Oral Biology, Clinic of Oral Rare and Genetic Diseases, School of Stomatology, The Fourth Military Medical University, 145 Changle West Road, Shaanxi, Xi'an 710032, People's Republic of China.
Rongchen
R
Xu
Department of Prosthodontics, School of Stomatology, The Fourth Military Medical University, 145 Changle West Road, Shaanxi, Xi'an 710032, People's Republic of China.
Huiming
H
He
State Key Laboratory of Military Stomatology, National Clinical Research Center for Oral Diseases, Shaanxi Key Laboratory of Oral Diseases, Department of Oral Biology, Clinic of Oral Rare and Genetic Diseases, School of Stomatology, The Fourth Military Medical University, 145 Changle West Road, Shaanxi, Xi'an 710032, People's Republic of China. Electronic address: xhduan@fmmu.edu.cn.
Xiaohong
X
Duan
D000818:Animals; D002327:Cariostatic Agents / Q000008:administration & dosage / Q000633:toxicity; D003743:Dental Enamel / Q000737:chemistry / Q000187:drug effects / Q000473:pathology; D004195:Disease Models, Animal; D004305:Dose-Response Relationship, Drug; D005260:Female; D005459:Fluorides / Q000008:administration & dosage / Q000633:toxicity*; D009050:Fluorosis, Dental / Q000000981:diagnostic imaging / Q000378:metabolism / Q000473:pathology*; D007814:Larva; D008297:Male; D008855:Microscopy, Electron, Scanning; D010710:Phosphates / Q000008:administration & dosage / Q000633:toxicity; D014074:Tooth Calcification / Q000187:drug effects; D015027:Zebrafish
D002327:Cariostatic Agents; D010710:Phosphates; C012980:fluorophosphate; D005459:Fluorides
Dental fluorosis; Fluoride; Larva; Zebrafish