Microporous Hydroxyapatite-Based Ceramics Alter the Physiology of Endothelial Cells through Physical and Chemical Cues.

Incorporation of silicate ions in calcium phosphate ceramics (CPC) and modification of their multiscale architecture are two strategies for improving the vascularization of scaffolds for bone regenerative medicine. The response of endothelial cells, actors for vascularization, to the chemical and physical cues of biomaterial surfaces is little documented, although essential. We aimed to characterize in vitro the response of an endothelial cell line, C166, cultivated on the surface CPCs varying either in terms of their chemistry (pure versus silicon-doped HA) or their microstructure (dense versus microporous). Adhesion, metabolic activity, and proliferation were significantly altered on microporous ceramics, but the secretion of the pro-angiogenic VEGF-A increased from 262 to 386 pg/mL on porous compared to dense silicon-doped HA ceramics after 168 h. A tubulogenesis assay was set up directly on the ceramics. Two configurations were designed for discriminating the influence of the chemistry from that of the surface physical properties. The formation of tubule-like structures was qualitatively more frequent on dense ceramics. Microporous ceramics induced calcium depletion in the culture medium (from 2 down to 0.5 mmol/L), which is deleterious for C166. Importantly, this effect might be associated with the in vitro static cell culture. No influence of silicon doping of HA on C166 behavior was detected.

Dental and maxillofacial manifestations of Steinert's myotonic dystrophy: A retrospective case series and therapeutic investigation.

This retrospective study was performed to analyse the facial features and occlusal anomalies in 18 patients with Steinert's myotonic dystrophy (MD1). Medical and surgical management issues noted in this study may contribute to clinical decision-making. This series included 18 patients with MD1 who presented for maxillofacial consultations. For all patients, the following characteristics were assessed: sex, age, intellectual ability, oral condition, initial assessment of the occlusion and facial aspect. In total, 11 of 18 patients underwent surgery (10 achieved occlusion modification, whereas one did not). amongst patients who underwent surgery and achieved occlusion modification, six had stable class I results and four had unstable results or exhibited a slight degradation. Facial muscles play an important role in craniomaxillofacial development and facial aspects. A high prevalence of malocclusions is present in patients with MD1. Orthodontics and orthognathic surgery can improve the quality of life for affected patients. However, the long-term results of these treatments may be disappointing, and relapse can occur in patients with the most severe disease. Aspects of disease to consider while planning for surgery include oral health, risks of instability and relapse, and risks involving anaesthesia.

Pre-osteoblast cell colonization of porous silicon substituted hydroxyapatite bioceramics: Influence of microporosity and macropore design.

Silicate-substituted hydroxyapatite scaffolds containing multiscale porosity are manufactured. Model parts containing macropores of five cross-sectional geometries (circle, square, rhombus, star and triangle) and two sizes are shaped by microstereolithography. Three open microporosity contents (0.5, 23 or 37 vol%) are introduced in the ceramic. MC3T3-E1 pre-osteoblasts are seeded onto these scaffolds. Analysis of cell colonization inside the macropores after 7 and 14 days of cultivation shows that the cellular filling is proportional to the macropore size and strongly influenced by macropore shape. Straight edges and convex surfaces are detrimental. High aspect ratios, the absence of reentrant angles and the presence of acute angles, by creating concavities and minimizing flat surfaces, facilitate cell colonization. Rhombus and triangle cross-sections are thus particularly favorable, while square and star geometries are the least favored. An increase in the microporosity content strongly impairs cell growth in the macropores. The data are statistically analyzed using a principal components analysis that shows that macro- and microtopographical parameters of scaffolds must be collectively considered with correlated interactions to understand cell behavior. The results indicate the important cell sensing of topography during the initial step of cell adhesion and proliferation and evidence the need for an optimized scaffold design.

Functionalization of phosphocalcic bioceramics for bone repair applications.

This work is devoted to the processing of bone morphogenetic protein (BMP-2) functionalized silicate substituted hydroxyapatite (SiHA) ceramic spheres. The motivation behind it is to develop injectable hydrogel/bioceramic composites for bone reconstruction applications. SiHA microspheres were shaped by spray drying and thoroughly characterized. The silicate substitution was used to provide preferred chemical sites at the ceramic surface for the covalent immobilization of BMP-2. In order to control the density and the release of the immobilized BMP-2, its grafting was performed via ethoxysilanes and polyethylene glycols. A method based on Kaiser's test was used to quantify the free amino groups of grafted organosilanes available at the ceramic surface for BMP-2 immobilization. The SiHA surface modification was investigated by means of X-ray photoelectron spectroscopy, Fourier transformed infrared spectroscopy and thermogravimetry coupled with mass spectrometry. The BMP-2 bioactivity was assessed, in vitro, by measuring the luciferase expression of a stably transfected C3H10 cell line (C3H10-BRE/Luc cells). The results provided evidence that the BMP-2 grafted onto SiHA spheres remained bioactive.

Quantitative analysis of vascular colonisation and angio-conduction in porous silicon-substituted hydroxyapatite with various pore shapes in a chick chorioallantoic membrane (CAM) model.

UNLABELLED: The development of scaffolds for bone filling of large defects requires an understanding of angiogenesis and vascular guidance, which are crucial processes for bone formation and healing. There are few investigations on the ability of a scaffold to support blood vessel guidance and it this is of great importance because it relates to the quality and dispersion of the blood vessel network. This work reports an analysis of vascularisation of porous silicon-substituted hydroxyapatite (SiHA) bioceramics and the effects of pore shape on vascular guidance using an expedient ex ovo model, the chick embryo chorioallantoic membrane (CAM) assay. Image analysis of vascularised implants assessed the vascular density, fractal dimension and diameter of blood vessels at two different scales (the whole ceramic and pores alone) and was performed on model SiHA ceramics harbouring pores of various cross-sectional geometries (circles, square, rhombus, triangles and stars). SiHA is a biocompatible material which allows the conduction of blood vessels on its surface. The presence of pores did not influence angiogenesis related-parameters (arborisation, fractal dimension) but pore geometry affected the blood vessel guidance and angio-conductive potential (diameter and number of the blood vessels converging toward the pores). The measured angles of pore cross-section modulated the number and diameter of blood vessels converging to pores, with triangular pores appearing of particular interest. This result will be used for shaping ceramic scaffolds with specific porous architecture to promote vascular colonisation and osteointegration. STATEMENT OF SIGNIFICANCE: An expedient and efficient method, using chick embryo chorioallantoic membrane (CAM) assays, has been set up to characterise quantitatively the angiogenesis and the vascular conduction in scaffolds. This approach complements the usual cell culture assays and could replace to a certain extent in vivo experiments. It was applied to silicon-substituted hydroxyapatite porous bioceramics with various pore shapes. The material was found to be biocompatible, allowing the conduction of blood vessels on its surface. The presence of pores does not influence the angiogenesis but the pore shape affects the blood vessel guidance and angio-conductive potential. Pores with triangular cross-section appear particularly attractive for the further design of scaffolds in order to promote their vascular colonisation and osteointegration and improve their performances.

A new custom made bioceramic implant for the repair of large and complex craniofacial bone defects.

INTRODUCTION: Neurosurgery and Maxillofacial Surgery Departments of Limoges University Hospital Centre have developed a new concept of a custom made ceramic implant in hydroxyapatite (HA) for the reconstruction of large and complex craniofacial bone defects (more than 25 cm(2)). MATERIALS AND METHODS: The manufacturing process of the implants used a stereolithography technique that produces implants with three-dimensional shapes derived directly from the scan file of the patient's skull without moulding or machining. Eight patients received 8 implants between 2005 and 2008. RESULTS: The surgical procedure is simple and fast. The post-operative follow-up was 12 months. No major complications (infection or fracture of the implant) were observed. The cosmetic result was considered satisfactory by both patients and surgeons. CONCLUSIONS: These new implants are well suited for reconstruction of large craniofacial bone defects (greater than 25 cm(2)) in adults and children over 8 years.