Amelogenin control over apatite crystal growth is affected by the pH and degree of ionic saturation.
OBJECTIVE
To study the mechanisms which promote the interactions of amelogenin proteins with the forming mineral to establish suitable conditions for the biomimetic synthesis of enamel in vitro.


METHODS
Saturated calcium phosphate solutions were used in conjunction with recombinant amelogenin proteins to induce mineral formation on glass-ceramics substrates containing oriented fluoroapatite crystals (FAP). The height of mineral layers formed on these substrates within 24 h was measured by atomic force microscopy (AFM).


METHODS
The effect of protein concentration, pH and degree of saturation (DS) on the growth of apatite mineral was evaluated. Mineralization experiments were performed at 0, 0.4 and 1.6 mg/ml amelogenin concentrations. Mineralization solutions were used at pH values of 6.5, 7.4, 8.0 and 8.8 and DS of calcium and phosphate between 9 and 13.


METHODS
Height and morphology of mineralized layer formed on glass-ceramic substrates as determined from AFM measurements.


RESULTS
Homogeneous nucleation and crystal growth of thin layers on the FAP were observed, when calcium and phosphate ions were added. The height of these layers grown on (001) planes of FAP was strongly dependent on the protein concentration and pH. At concentrations of 0 and 0.4 mg/ml crystal grew 5-15 nm on the FAP, while they grew approximately to 200 nm at 1.6 mg/ml. The enhanced crystal growth was observed only at pH 6.5, 7.4 and 8.0, while layers only 20 nm thick were obtained at pH 8.8. An increase in DS resulted in uncontrolled growth of calcium phosphate mineral covering large areas of the substrate.


CONCLUSIONS
Protein concentration, pH and the saturation of the mineralizing solution need to be considered carefully to provide suitable conditions for amelogenin-guided growth of apatite crystals.
Habelitz
S
S
Department of Preventive and Restorative Dental Sciences, University of California, San Francisco, 94143, USA. shabeli@itsa.ucsf.edu
Denbesten
P K
PK
Marshall
S J
SJ
Marshall
G W
GW
Li
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D000566:Amelogenesis; D053523:Amelogenin; D001031:Apatites / Q000737:chemistry*; D040761:Biomimetic Materials / Q000737:chemistry; D002130:Calcium Phosphates / Q000737:chemistry; D003460:Crystallization; D003746:Dental Enamel Proteins / Q000494:pharmacology* / Q000502:physiology*; D006801:Humans; D006863:Hydrogen-Ion Concentration; D018625:Microscopy, Atomic Force; D009994:Osmolar Concentration; D011994:Recombinant Proteins / Q000494:pharmacology
D053523:Amelogenin; D001031:Apatites; D002130:Calcium Phosphates; D003746:Dental Enamel Proteins; D011994:Recombinant Proteins; C020243:calcium phosphate; C025105:fluorapatite