Effect of Keratin Preparations on Cementoblast OCCM- 30 and Fibroblast L929 Cells

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Author(s)
Love, Robert
Sajeev, Jeeson
Griffith University Author(s)
Year published
2020
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Aim: To assess the in vitro effect of keratin preparations on the viability and proliferation of cementoblast OCCM-30 and fibroblast L929 cells and on the mineralization capability of the OCCM-30 cells.
Methodology: Cells of L929 and OCCM-30 were assessed for viability in growth media supplemented with 10, 1 and 0.1 mg/ml of keratin using a LIVE/ DEAD assay and confocal laser scanning microscopy. Cell proliferation was tested with similar keratin concentrations using an alamarBlue® proliferation assay at 0, 24, 48 and 72 h. Alkaline phosphatase assay was performed for OCCM-30 cells with keratin concentrations of 1 mg/ml, 0.1 ...
View more >Aim: To assess the in vitro effect of keratin preparations on the viability and proliferation of cementoblast OCCM-30 and fibroblast L929 cells and on the mineralization capability of the OCCM-30 cells. Methodology: Cells of L929 and OCCM-30 were assessed for viability in growth media supplemented with 10, 1 and 0.1 mg/ml of keratin using a LIVE/ DEAD assay and confocal laser scanning microscopy. Cell proliferation was tested with similar keratin concentrations using an alamarBlue® proliferation assay at 0, 24, 48 and 72 h. Alkaline phosphatase assay was performed for OCCM-30 cells with keratin concentrations of 1 mg/ml, 0.1 mg/ml at 3, 6, and 10 days and an alizarin red assay was performed at 14 days. Results: The viability assay showed concentrations at 1 mg/ml or greater of keratin proved toxic to L929 cells. The proliferation assay for OCCM-30 cells showed keratin concentration at 10 mg/ml prevented its proliferation and was significantly lower than the other keratin groups (P < 0.0001). There was significantly higher OCCM-30 cell ALP activity with the control and 0.1 mg/ml groups compared with the 1 mg/ml group at 6 d and 12 days (P < 0.0001). The alizarin red assay was consistent with the ALP activity of the OCCM-30 cells. Conclusion: Low concentration keratin preparations were shown to allow normal cell proliferation of L929 and OCCM-30 cells and normal mineralization of the cementoblasts.
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View more >Aim: To assess the in vitro effect of keratin preparations on the viability and proliferation of cementoblast OCCM-30 and fibroblast L929 cells and on the mineralization capability of the OCCM-30 cells. Methodology: Cells of L929 and OCCM-30 were assessed for viability in growth media supplemented with 10, 1 and 0.1 mg/ml of keratin using a LIVE/ DEAD assay and confocal laser scanning microscopy. Cell proliferation was tested with similar keratin concentrations using an alamarBlue® proliferation assay at 0, 24, 48 and 72 h. Alkaline phosphatase assay was performed for OCCM-30 cells with keratin concentrations of 1 mg/ml, 0.1 mg/ml at 3, 6, and 10 days and an alizarin red assay was performed at 14 days. Results: The viability assay showed concentrations at 1 mg/ml or greater of keratin proved toxic to L929 cells. The proliferation assay for OCCM-30 cells showed keratin concentration at 10 mg/ml prevented its proliferation and was significantly lower than the other keratin groups (P < 0.0001). There was significantly higher OCCM-30 cell ALP activity with the control and 0.1 mg/ml groups compared with the 1 mg/ml group at 6 d and 12 days (P < 0.0001). The alizarin red assay was consistent with the ALP activity of the OCCM-30 cells. Conclusion: Low concentration keratin preparations were shown to allow normal cell proliferation of L929 and OCCM-30 cells and normal mineralization of the cementoblasts.
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Journal Title
Austin Journal of Biotechnology & Bioengineering
Volume
7
Issue
1
Copyright Statement
© Love et al. 2020. This is an Open Access article distributed under the terms of the Creative Commons Attribution 4.0 International (http://creativecommons.org/licenses/by/4.0/) which permits unrestricted use, distribution, and reproduction in any medium, provided the original work is properly cited.
Subject
Endodontics