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Biochemical and morphological studies of human diploid and fluoride-resistant fibroblasts in vitro
1Department of Preventive and Community Dentistry, Nippon Dental University, Tokyo, Japan.
Abstract:
Fluoride-resistant (FR 30) lines were produced from human diploid fibroblast (NDU-1) cells by progressively increasing the concentration of fluoride (F) in the growth medium up to 1.58 mM. This concentration completely inhibited the growth of the original cells. The resistant cells had decreased incorporation of [14C]-leucine and an increased rate of the cell division. The activities of acid phosphatase, alkaline phosphatase, lactate dehydrogenase, leucine aminopeptidase, glutamic oxaloacetic transaminase, glutamic pyruvic transaminase and alpha-hydroxybutyrate dehydrogenase in the FR 30 cells were lower than in the NDU-1 cells. The FR 30 cells had irregular shapes and sizes; the amount of undeveloped rough endoplasmic reticulum and the number of lysosomes were increased. These biochemical and morphological changes in FR 30 cells suggest that their metabolic activities were depressed by F, although they have some degree of F resistance.
Insights
Fluoride-resistant cells (FR 30) were developed from human diploid fibroblast (NDU-1) cells. These resistant cells exhibited altered metabolic activity and morphology, indicating fluoride
Area of Science:
- Cell Biology
- Biochemistry
- Toxicology
Background:
- Human diploid fibroblast (NDU-1) cells were used to study cellular responses to fluoride.
- Fluoride exposure can inhibit cell growth and alter cellular functions.
Purpose of the Study:
- To develop and characterize fluoride-resistant (FR 30) cell lines from human diploid fibroblasts.
- To investigate the biochemical and morphological changes associated with fluoride resistance.
Main Methods:
- Progressively increasing fluoride concentration in the growth medium to select for resistant cells.
- Assessing [14C]-leucine incorporation, cell division rates, enzyme activities, and cell morphology.
Main Results:
- FR 30 cells demonstrated resistance to 1.58 mM fluoride, a concentration lethal to parent NDU-1 cells.
- Resistant cells showed decreased [14C]-leucine incorporation and increased cell division rates.
- Enzyme activities (acid phosphatase, alkaline phosphatase, LDH, etc.) were lower in FR 30 cells.
- FR 30 cells exhibited irregular shapes/sizes, increased undeveloped rough endoplasmic reticulum, and more lysosomes.
Conclusions:
- Fluoride exposure depresses metabolic activities in human diploid fibroblasts.
- FR 30 cells possess a degree of fluoride resistance despite significant biochemical and morphological alterations.
- These changes suggest complex adaptive mechanisms in response to chronic fluoride exposure.