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Extra cellular matrix used in-vitro model system for hydroxyapatite formation

Emilia V. Pecheva 1Liliana Pramatarova 1Manfred F. Maitz 2Mihn T. Pham 2Alexey Kondyurin 3

1. Institute of Solid State Physics, Bulgarian Academy of Sciences (ISSP-BAS), 72, Tzarigradsko Chaussee blvd., Sofia 1784, Bulgaria
2. Forschungszentrum Rossendorf (FZR), Dresden 01314, Germany
3. Institute of Polymer Research, Dresden 01069, Germany


Many are the hypotheses related to the mechanisms of biomineralization process, particularly hydroxyapatite (HA) nucleation and growth. Surface topography is regarded as one of them, as well as supersaturated solutions, the presence of hydroxyl groups, chemistry of the nucleation points, organic components and etc.
The aim of this work is to investigate the influence of the organic components deposited on different solid surfaces upon HA formation. For this purpose the surfaces of stainless steel, silicon and silica glass substrates are coated with extra cellular matrix (ECM) to promote interactions with bone-forming cells. The ECM proteins are known as ligands in reactions with cell surface receptors involved in bone physiology. The osteoblast-like cell line SAOS-2 is allowed to synthesize and assemble its own ECM on the substrates under standard cell culture conditions. Cells are then selectively removed resulting in substrates coated with a thin film of ECM.
The so-modified samples are immersed in a solution, supersaturated with respect to Ca and P ions (so called simulated body fluid, which resembles the composition of human blood plasma) and a layer is grown. X-ray diffractometric techniques (XRD and EDX), spectroscopic (FTIR and Raman), microscopic techniques (SEM) are used to highlight the likeness of the grown artificial biomimetic HA/ECM protein structure with natural bone tissue.
A homogeneous HA layer is found on the so-modified samples, which consists of particles with regular sphere shape with mean diameter size of 5 micrometers. Well resolved IR and Raman peaks reveal the formation of crystalline HA layer, self organized inside the ECM matrix.


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Presentation: poster at E-MRS Fall Meeting 2003, Symposium E, by Emilia V. Pecheva
See On-line Journal of E-MRS Fall Meeting 2003

Submitted: 2003-05-12 09:09
Revised:   2009-06-08 12:55