M. Dard

Biomimetic coatings functionalized with adhesion peptides for dental implants.

A complete biological integration into the surrounding tissues (bone, gingiva) is a critical step for clinical success of a dental implant. In this work biomimetic coatings consisting either of collagen type I (for the gingiva region) and hydroxyapatite (HAP) or mineralized collagen (for the bone interface) have been developed as suitable surfaces regarding the interfaces. Additionally, using these biomimetic coatings as a matrix, adhesion peptides were bound to further increase the specificity of titanium implant surfaces. To enhance cell attachment in the gingiva region, a linear adhesion peptide developed from a laminin sequence (TWYKIAFQRNRK) was bound to collagen, whereas for the bone interface, a cyclic RGD peptide was bound to HAP and mineralized collagen using adequate anchor systems. The biological potential of these coatings deduced from cell attachment experiments with HaCaT human keratinocytes and MC3T3-E1 mouse osteoblasts showed the best results for collagen and laminin sequence coating for the gingiva region and mineralized collagen and RGD peptide coatings for regions with bone contact. Our concept opens promising approaches to improve the biological integration of dental implants.© 2001 Kluwer Academic Publishers

Tools for tissue engineering of mineralized oral structures

Abstract This paper presents a short review of three groups of tools which can be or are used for the tissue engineering of mineralized oral structures: growth factor delivery systems (GFDS) and surface bioactivation with covalent bound peptides or with nanomechanically linked proteins. According to the reported personal experience of the authors, GFDS have to face the following challenging issue before being used routinely in dentistry, e.g., as a tool for reparative dentinogenesis or bone healing: adaptation of the GFDS design to the tissue where it will be implanted in order to deliver the right dose of growth factor (GF) at the right time. The bioactivation of surfaces, for example of dental implants, with covalent bound peptides or nanomechanically linked proteins represents a second innovative way to improve dental health in the future. Here we report on the experimental use of cyclic RGD peptides grafted on polymethylmethacrylate to improve osteoblast adhesion. Furthermore, we show the potential advantage of immobilizing and incorporating collagen I on titanium implant surfaces. These techniques or a combination of them will help to create improvements, for example, of dental implants in the near future. They will also help to promote bone and dentin regeneration.

Evaluation of the effect of three calcium phosphate powders on osteoblast cells

The aim of the present study was to assess the effect of three calcium phosphate powders entering in the composition of bone substitute materials on osteoblast-cells activity. These powders were hydroxyapatite (HA) widely used as a biomaterial, nanocrystalline carbonate apatite (C A) very close to bone mineral crystals, and an experimental one: calcium phosphate cement-1 (CPC-1) composed of an amorphous Ca-P phase and brushite. The powders were physico-chemically characterized. The very reactive CPC-1 powder became transformed in cell culture medium: recrystallization of amorphous precursors and hydrolysis of brushite into poorly crystalline apatite occurred. Osteoblast-cells activity was evaluated: for low level of calcium phosphates (>100 μg/ml) CPC-1 enhanced proliferation and, to a lesser degree, differentiation on alkaline phosphatase activity. For 100 μg/ml of powders we observed a great alteration of biological activity of the osteoblasts: evaluation of proliferation indicated an inhibition for all samples, and a decrease of two differentiation markers: alkaline phosphatase activity and osteocalcin release were noticed, suggesting a down regulation due to the presence of large amount of mineral powder.

Adsorption of vascular endothelial growth factor to two different apatitic materials and its release

The aim of our study was to assess the ability of calcium phosphate powders to serve as growth factor carriers. Vascular endothelial growth factor (VEGF), in particular, is locally involved in the bone formation process throughout osteoblast differentiation. Two different apatitic substrates were tested: hydroxyapatite (HA), widely used as biomaterial, and nanocrystalline carbonated apatite (CA), which has a composition similar to bone mineral crystals. These materials have been compared for their VEGF adsorption and release properties. The adsorption of the growth factor was higher on CA than on HA probably due to differences of both the proteins and the powders involved. The specific activity of the VEGF released was also tested to determine the available activity for cells in contact with these materials. Interestingly, the bioactivity of the VEGF released from CA quantified on fetal bovine aortic endothelial cells (FBAE) by evaluating the proliferation activity, exhibited no marked difference compared to native VEGF. Qualitatively, VEGF adsorbed on CA material induced well-defined collagen type I immunostaining on osteoblast cells compared to the staining obtained after VEGF adsorption on HA.