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Accessing the Cytotoxicity and Cell Response to Biomaterials

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Human osteoblastic cells discriminate between 20-kDa amelogenin isoforms.

Elisabeth A Riksen¹, Christiane Petzold, Steven Brookes

¹Department of Biomaterials, Faculty of Dentistry, University of Oslo, Oslo, Norway.

European Journal of Oral Sciences

|January 17, 2012

Summary

Enamel matrix derivative (EMD) fractions A1 and A2 differentially affect human osteoblasts. A1 enhances proliferation and RUNX2 expression, while A2 boosts ALP activity and OPG/RANKL secretion, suggesting distinct roles in bone healing.

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Area of Science:

Biomaterials Science
Cell Biology
Periodontology

Background:

Enamel matrix derivative (EMD) promotes alveolar bone healing after periodontitis.
The specific roles of EMD's constituent fractions in osteoblast activity remain unclear.

Purpose of the Study:

To compare the effects of two EMD fractions, A1 and A2, on primary human osteoblasts.
To elucidate the distinct contributions of EMD components to osteoblast function and differentiation.

Main Methods:

Osteoblasts were cultured with EMD fractions A1, A2, or whole EMD (50 μg/ml).
Analysis included SDS-PAGE for protein composition, FTIR for secondary structure, and MALDI-MS for peptide identification.
Osteoblast proliferation (BrdU incorporation), gene expression (RUNX2, CD44, DMP1 mRNA), and protein secretion (IL-6, ALP, OPG, RANKL, osteocalcin) were measured.

Main Results:

Both A1 and A2 fractions contain amelogenins and exhibit distinct secondary structures and peptide masses.
Fraction A1 enhanced osteoblast proliferation, RUNX2 mRNA expression, and IL-6 secretion.
Fraction A2 increased alkaline phosphatase (ALP) activity, CD44 mRNA expression, OPG and RANKL secretion, and DMP1 mRNA expression.

Conclusions:

EMD fractions A1 and A2 exhibit differential effects on human osteoblasts, influencing proliferation, gene expression, and protein secretion.
These distinct cellular responses suggest that A1 and A2 may facilitate the differentiation of different osteoblast phenotypes, contributing uniquely to EMD's therapeutic effects in bone regeneration.