The macrophysical properties of granular biomaterials used to fill bone defects have rarely been considered. Granules of a given biomaterial occupy three-dimensional (3-D) space when packed together and create a macroporosity suitable for the invasion of vascular and bone cells. Granules of ?-tricalcium phosphate were prepared using polyurethane foam technology and increasing the amount of material powder in the slurry (10, 11, 15, 18, 21 and 25g). After sintering, granules of 1000-2000?m were prepared by sieving. They were analyzed morphologically by scanning electron microscopy and placed in polyethylene test tubes to produce 3-D scaffolds. Microcomputed tomography (microCT) was used to image the scaffolds and to determine porosity and fractal dimension in three dimensions. Two-dimensional sections of the microCT models were binarized and used to compute classical morphometric parameters describing porosity (interconnectivity index, strut analysis and star volumes) and fractal dimensions. In addition, two newly important fractal parameters (lacunarity and succolarity) were measured. Compression analysis of the stacks of granules was done. Porosity decreased as the amount of material in the slurry increased but non-linear relationships were observed between microarchitectural parameters describing the pores and porosity. Lacunarity increased in the series of granules but succolarity (reflecting the penetration of a fluid) was maximal in the 15-18g groups and decreased noticeably in the 25g group. The 3-D arrangement of biomaterial granules studied by these new fractal techniques allows the optimal formulation to be derived based on the lowest amount of material, suitable mechanical resistance during crushing and the creation of large interconnected pores.
Scaffolds of nonresorbable biomaterials can represent an interesting alternative for replacing large bone defects in some particular clinical cases with massive bone loss. Poly(styrene) microfibers were prepared by a dry spinning method. They were partially melted to provide 3D porous scaffolds. The quality of the material was assessed by Raman spectroscopy. Surface roughness was determined by atomic force microscopy and vertical interference microscopy. Saos-2 osteoblast-like cells were seeded on the surface of the fibers and left to proliferate. Cell morphology, evaluated by scanning electron microscopy, revealed that they can spread and elongate on the rough microfiber surface. Porous 3D scaffolds made of nonresorbable poly(styrene) fibers are cytocompatible biomaterials mimicking allogenic bone trabeculae and allowing the growth and development of osteoblast-like cells in vitro.
In Arabidopsis, lateral roots originate from pericycle cells deep within the primary root. New lateral root primordia (LRP) have to emerge through several overlaying tissues. Here, we report that auxin produced in new LRP is transported towards the outer tissues where it triggers cell separation by inducing both the auxin influx carrier LAX3 and cell-wall enzymes. LAX3 is expressed in just two cell files overlaying new LRP. To understand how this striking pattern of LAX3 expression is regulated, we developed a mathematical model that captures the network regulating its expression and auxin transport within realistic three-dimensional cell and tissue geometries. Our model revealed that, for the LAX3 spatial expression to be robust to natural variations in root tissue geometry, an efflux carrier is required--later identified to be PIN3. To prevent LAX3 from being transiently expressed in multiple cell files, PIN3 and LAX3 must be induced consecutively, which we later demonstrated to be the case. Our study exemplifies how mathematical models can be used to direct experiments to elucidate complex developmental processes.
Bone is composed of two phases. The organic phase is made of collagen fibrils assembled in broad fibers acting as a template for mineralization. The mineral phase comprises hydroxyapatite (HAP) crystals grown between and inside the collagen fibers. We have developed a biomimetic material using functionalized carbon nanotubes as scaffold to initiate in vitro mineralization. Biomimetic formation of HAP was performed on single-walled carbon nanotubes (SWCNTs) which have been grafted with carboxylic groups. Two types of nanotubes, HiPco(R) and Carbon Solutions(R), were oxidized via various acidic processes, leading to five different groups of carboxylated nanotubes, fully characterized by physical methods (thermogravimetric analysis, attenuated total reflectance infrared spectroscopy and X-ray photoelectron spectroscopy). All samples were dispersed in ultra-pure water and incubated for 2weeks in a synthetic body fluid, in order to induce the calcification of the SWCNTs. Scanning electron microscopy (SEM) and energy-dispersive X-ray analysis studies showed that Ca(2+) and PO(4)(3-) ions were deposited as round-shaped nodules (calcospherites) on the carboxylated SWCNTs. Fourier transform infrared and Raman spectroscopic studies confirmed the HAP formation, and image analysis made on SEM pictures showed that calcospherites and carboxylated SWCNTs were packed together. The size of calcospherites thus obtained in vitro from the HiPco(R) series was close to that issued from calcospherites observed in vivo. Functionalization of SWCNTs with carboxylic groups confers the capacity to induce calcification similar to woven bone.
Sinus lift elevation is an interesting method to restore bone mass at the maxilla in edentulated patients. We have investigated the histological effects of beta tricalcium phosphate (beta-TCP) combined with autograft bone for sinus lift elevation. A series of 14 patients who were candidate for dental implantation were grafted with beta-TCP granules and morcellized autograft bone harvested at the chin. beta-TCP was characterized by scanning electron microscopy and optical profilometry. Before implant placement, a small bone biopsy (2mm in diameter) was done. The amount of residual material and newly formed bone were determined by microcomputed tomography. Histological analysis was done on undecalcified sections stained by Goldners trichrome and osteoclast identification (TRAcP). beta-TCP served as a template for bone apposition by osteoblasts onto the granules surface. The material was simultaneously resorbed by TRAcP positive osteoclasts and macrophages. Fragments of the material remained buried in bone trabeculae as long as 12 months post-graft but the formed bone onto the granules surface had a lamellar texture. beta-TCP combined with autograft bone appears a suitable biomaterial for sinus lift augmentation before the placement of bone implants. The material favors the apposition of lamellar bone by osteoblasts and is simultaneous resorbed by two types of cells.
Aspergillus fumigatus is the most common agent of invasive aspergillosis, a feared complication in severely immunocompromised patients. Despite the recent commercialisation of new antifungal drugs, the prognosis for this infection remains uncertain. Thus, there is a real need to discover new targets for therapy. Particular attention has been paid to the biochemical composition and organisation of the fungal cell wall, because it mediates the host-fungus interplay. Conidia, which are responsible for infections, have melanin as one of the cell wall components. Melanin has been established as an important virulence factor, protecting the fungus against the hosts immune defences. We suggested that it might also have an indirect role in virulence, because it is required for correct assembly of the cell wall layers of the conidia.
Bone autograft remains a very useful and popular way for filling bone defects. In maxillofacial surgery or implantology, it is used to increase the volume of the maxilla or mandible before placing dental implants. Because there is a noticeable delay between harvesting the graft and its insertion in the receiver site, we evaluated the morphologic changes at the light and transmission electron microscopy levels. Five patients having an autograft (bone harvested from the chin) were enrolled in the study. A small fragment of the graft was immediately fixed after harvesting and a second one was similarly processed at the end of the grafting period when bone has been stored at room temperature for a 20 min +/- 33 s period in saline. A net increase in the number of osteocyte lacunae filled with cellular debris was observed (+41.5%). However no cytologic alteration could be observed in the remaining osteocytes. The viability of these cells is known to contribute to the success of autograft in association with other less well-identified factors.
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