F. Z. Cui

Three‐dimensional nano‐HAp/collagen matrix loading with osteogenic cells in organ culture

Transplantation of osteogenic cells with a suitable matrix is one strategy for engineering bone tissue. Three-dimensional distribution and growth of cells within the porous scaffold are of clinical significance for the repair of large bony defects. A nano-HAp/collagen (nHAC) composite that mimics the natural bone both in composition and microstructure to some extent was employed as a matrix for the tissue engineering of bone. A porous nHAC composite was produced in sheet form and convolved to be a three-dimensional scaffold. Using organ culture techniques and the convolving method, we have developed three-dimensional osteogenic cells/nHAC constructs in vitro. Scanning electron microscopic and histological examination has demonstrated the development of the cells/material complex. Spindle-shaped cells migrating out of bone fragments continuously proliferated and migrated throughout the network of the coil. The porous nHAC scaffold provided a microenvironment resembling that seen in vivo, and cells within the composite eventually acquired a tridimensional polygonal shape. In addition, new bone matrix was synthesized at the interface of bone fragments and the composite.

Tissue response to nano-hydroxyapatite/collagen composite implants in marrow cavity.

The tissue response to a nano-hydroxyapatite/collagen composite implanted in a marrow cavity was investigated by histology and scanning electron microscopy. A Knoop microhardness test was performed to compare the mechanical behavior of the composite and bone. The ultrastructural features of the composite, especially the carbonate-substituted hydroxyapatite with low crystallinity and nanometer size, made it a bone-resembling material. It was bioactive, as well as biodegradable. At the interface of the implant and marrow tissue, solution-mediated dissolution and giant cell mediated resorption led to the degradation of the composite. Interfacial bone formation by osteoblasts was also evident. The process of implant degradation and bone substitution was reminiscent of bone remodeling. The composite can be incorporated into bone metabolism instead of being a permanent implant. For lack of the hierarchical organization similar to that of bone, the composite exhibited an isotropic mechanical behavior. However, the resistance of the composite to localized pressure could reach the lower limit of that of the femur compacta.

PREPARATION OF CALCIUM PHOSPHATE COATINGS ON TITANIUM IMPLANT MATERIALS BY SIMPLE CHEMISTRY

A two-step chemical treatment has been developed in our group to prepare commercially pure titanium (cpTi) surfaces that will allow calcium phosphate (Ca-P) precipitation during immersion in a supersaturated calcification solution (SCS) with ion concentrations of [Ca2+] = 3.10 mM and [HPO4(2-)] = 1.86 mM. It was observed that a precalcification (Pre-Ca) procedure prior to immersion could significantly accelerate the Ca-P deposition process. In this work, the bioactivity of chemically treated cpTi and Ti6Al4V was further verified by applying commercially available Hanks balanced salt solution (HBSS), an SCS with very low ion concentrations of [Ca2+] = 1.26 mM and [HPO4(2-)] = 0.779 mM, as the immersion solution. It was found that a uniform and very dense apatite coating magnesium impurities was formed if the Pre-Ca procedure was performed before immersion, as compared with the loose Ca-P layer obtained from the abovementioned high concentration of SCS. The formation of a microporous titanium dioxide thin surface layer on cpTi or Ti6Al4V by the two-step chemical treatment could be the main reason for the induction of apatite nucleation and growth from HBSS. Variations of pH values, Ca and P concentrations, and immersion time in HBSS were investigated to reveal the detailed process of Ca-P deposition. The described treatments provide a simple chemical method to prepare Ca-P coatings on both cpTi and Ti6Al4V.

PREPARATION OF BIOACTIVE MICROPOROUS TITANIUM SURFACE BY A NEW TWO-STEP CHEMICAL TREATMENT

Microporous oxide layers allowing fast deposition of calcium phosphate layers (CPLs) were formed on commercially pure titanium (c.p.Ti) after the application of a newly developed two-step chemical treatment. The micropores were of submicrometre size. The two-step treatment was carried out by etching c.p.Ti samples with HCl and H2SO4 first and then treating them in boiling 0.2 N NaOH solution at 140 °C for 5 h. Conformal CPLs, about 20 μm thick, were deposited on the two-step treated c.p.Ti surface by means of a two-day immersion in an in vitro supersaturated calcification solution. The CPL was characterized to be mainly composed of two sublayers, i.e. an outside loose octacalcium phosphate crystal sublayer and an inside dense carbonated apatite sublayer. A scratching test indicated that the apatite sublayer was strongly bonded to the c.p.Ti substrate. Moreover, it was observed that the untreated or single-step treated c.p.Ti surfaces are not only morphologically different from one another but significantly different from the two-step treated one, in that no precipitation was observed on them up to 14 d immersion in the same calcification solution. It is indicated that the two-step chemical treatment is a simple and easily controllable method to prepare bioactive titanium surfaces and subsequently to induce the rapid precipitation of conformal and adherent CPL from in vitro supersaturated calcification solutions.

Synthesis and biocompatibility of porous nano-hydroxyapatite/collagen/alginate composite

Porous nano-hydroxyapatite/collagen/alginate (nHAC/Alginate) composite containing nHAC and Ca-crosslinked alginate is synthesized biomimetically. This composite shows a significant improvement in mechanical properties over nHAC material. Mechanical test results show that the compressive modulus and yield strength of this composite are in direct proportion to the percentage of Ca-crosslinked alginate in the composite. Primary biocompatibility experiments in vitro including fibroblasts and osteoblasts co-culture with nHAC/alginate composite indicated the high biocompatibility of this composite. Therefore the composite can be a promising candidate of scaffold material for bone tissue engineering.

Morphological behaviour of osteoblasts on diamond-like carbon coating and amorphous C–N film in organ culture

Similar to diamond-like carbon (DLC) coating, amorphous carbon nitride (C-N) films can be extremely hard and wear-resistant. They may serve as candidates for the solution to the problem of aseptic loosening of total hip replacements. Morphological behaviour of osteoblasts on silicon, DLC-coated silicon and amorphous C-N film-deposited silicon in organ culture was investigated by scanning electron microscopy. Cells on the silicon wafers were able to attach, but were unable to follow this attachment with spreading. In contrast, the cells attached, spread and proliferated on the DLC coatings and amorphous C-N films without apparent impairment of cell physiology. The morphological development of cells on the coatings and films was similar to that of cells in the control. The preliminary results support the biocompatibility of DLC coating and are encouraging for the potential biomedical applications of amorphous C-N film.

Preparation of bioactive Ti6Al4V surfaces by a simple method.

Boiling diluted alkali incubation was found to be an effective way to prepare bioactive Ti6Al4V surfaces, whether polished or not, as indicated in vitro after immersion in two different supersaturated calcification solutions (SCSs). The induction of calcium phosphate (Ca-P) precipitation from the SCSs is most probably made possible by the formation of a new TiO2 surface layer and a large number of submicron-scaled etched pits therein. The morphologies and composition of the Ca-P deposited from different SCSs are entirely different from each other. The processes on Ti6Al4V surfaces during treatment and immersion were investigated in detail by means of scanning electron microscopy combined with energy dispersive X-ray analysis, X-ray photoelectron spectroscopy and X-ray diffraction.

Observations of damage morphologies in nacre during deformation and fracture

The deformation, fracture and toughening mechanisms of nacre from a kind of fresh-water bivalve mollusc (Cristaria plicata) were studied by SEM, TEM and microindentation tests. Experimental results revealed a strong anisotropy of the damage behaviour reflecting the microstructural character of nacre. The fractured surface parallel to the cross-sectional surface of nacre was much more tortuous than that parallel to the platelet surface. The crack line on the cross-sectional surface was step-like, while that on the platelet surface was polygonal. Sliding of aragonite layer combined with the plastic deformation of organic matrix is the main plastic deformation mechanism of nacre. Three main toughening mechanisms have been found acting in concert: crack deflection, fibre pull-out and organic matrix bridging.

Bone growth in biomimetic apatite coated porous Polyactive® 1000PEGT70PBT30 implants

We recently, developed a simple one-day one-step incubation method to obtain bone-like apatite coating on flexible and biodegradable Polyactive 1000PEGT70PBT30. The present study reports a preliminary biological evaluation on the coated polymer after implantation in rabbit femurs. The porous cylindrical implants were produced from a block fabricated by injection molding and salt leaching. This technique provided the block necessary mechanical integrity to make small cylinders (diameter 3.5 x 5 mm2) that were suitable for implantation in rabbits. The coating continuously covered the surface of the polymer, preserving the porous architecture of outer contour of the cylinders. Two defects with a diameter of 3.5 or 4 mm were drilled in the proximal and distal part of femur diaphysis. The implants were inserted as press-fit or undersized into the cortex as well as in the marrow cavity. The polymer swelled after implantation due to hydration, leading to a tight contact with the surrounding bone in both defects. The adherence of the coating on the polymer proved to be sufficient to endure a steam sterilization process as well as the 15% swelling of the polymer in vivo. The coated Polyactive 1000PEGT70PBT30 has a good osteoconductive property, as manifested by abundant bone growth into marrow cavity along the implant surface during 4-week implantation. A favorable bioactive effect of the coating with an intimate bone contact and extensive bone bonding with this polymer was qualitatively confirmed. Concerning the bone ingrowth into the porous implant in the defect of 4 mm diameter, only marginal bone formation was observed up to 8 weeks with a maximal penetration depth of about 1 mm. The pore interconnectivity is important not only for producing a coating inside the porous structure but also for bone ingrowth into this biodegradable material. This preliminary study provided promising evidence for a further study using a bigger animal model.

Hierarchical structure of ivory: from nanometer to centimeter

Abstract Hierarchical structure of ivory has been investigated in terms of the organization of collagen fibrils in three dimensions, the spatial relationship of apatite crystals with collagen fibrils as well as the morphology and sizes of the crystals. Ivory can be characterized as two sets of radially-distributed layers of collagen fibril bundles interweaving to form a network. Within one layer, the collagen fibril bundles with diameter of about 2 μm lie nearly parallel to one another. They rotate by about 90° from one layer to its neighbor. The collagen fibril bundles are composed of densely-packed mineralized collagen fibrils with a diameter of 60–200 nm. Plate-shaped apatite crystals deposit mainly within the gap zone of the collagen fibrils. The average sizes of the crystals are measured to be 31 nm in length, 20 nm in width and 3 nm in thickness.