Bone modelling in the implantation bed

We consider results from tissue culture studies and the comparative histology of mineralized tissues and other natural tissue interfaces which may have some relevance in understanding the abnormal biology of the immediate environment of an implant in bone. We discuss factors influencing setting, colonization, and migration on natural and artificial substrates by various cell types which may make or remove matrix near the implant. A knowledge of mechanisms of mineral and organic matrix destruction by osteoclasts and other cells must be important in addition to an understanding of the interaction of local and systemic hormones with bone cells. More studies of the role of the immune system in implant failure are urgently required.     

Local calibrated bone mineral density in the mandible presented using a color coding scheme.

Calibrated information on bone mineral density (BMD) may be used in dental implantology to measure "bone quality". It can be used to estimate the expected primary implant stability preoperatively and to guide the surgeon in selecting optimum implant types and operation techniques. Using a preoperative dental computed tomography (Dental-CT) scan, all of this information can be obtained without additional examinations and thus without additional X-ray exposure of the patient. In contrast to bone mineral determination in other body regions, local BMD values are important in the jaw bone. Therefore, a regimen where color-coded information on local bone mineralization is superimposed on Dental-CT images is proposed using the original CT volumes as well as reformatted views.     

Histological analysis of cells and matrix mineralization of new bone tissue induced in rabbit femur bones by Mg–Zr based biodegradable implants

The biological efficacy of bone inducing implant materials in situ can be assessed effectively by performing histological analysis. We studied the peri-implant bone regeneration around two types of biodegradable magnesium–zirconium alloys, Mg–5Zr and Mg–Zr–2Sr, using histological, histochemical and immunohistochemical methods in the femur of New Zealand White strain rabbits. Our study includes three animal groups: (a) Mg–5Zr, (b) Mg–Zr–2Sr and (c) control. In each group three animals were used and in groups ‘a’ and ‘b’ the respective alloys were implanted in cavities made at the distal ends of the femur; control animals were left without implants to observe natural bone healing. Qualitative assessment of the cellularity and matrix mineralization events of the newly formed bone tissue was done at three months after implantation by histological methods in methyl methacrylate embedded tissue without decalcifying the bone. Quantitative mineral content and density of the new bone (NB) were evaluated by the statistical analysis of dual energy X-ray absorptiometry (DXA) data obtained from three animals in each experimental group. Based on our analysis we conclude that Mg–Zr–2Sr alloy showed better osseointegration of the newly formed bone with the implant surface. Our methodology of studying peri-implant osteoinduction of degradable implants using low temperature methyl methacrylate embedding resin can be useful as a general method for determining the bio-efficacy of implant materials.     

Genetics of osteoporosis

Osteoporosis is a common disease with a strong genetic component. In recent years, some progress has been made in understanding the genetic basis of osteoporosis. Genetic factors contribute to osteoporosis by influencing not only bone mineral density but also bone size, bone quality, and bone turnover. Meta-analysis has been used to define the role of several candidate genes in osteoporosis. Some quantitative trait loci that regulate bone mass identified by linkage studies in humans and experimental animals have been replicated in multiple populations. Genes that cause monogenic bone diseases also contribute to regulation of bone mass in the normal population. Genome-wide association studies and functional genomics approaches have recently begun to apply to genetic studies of osteoporosis. In the future, not only single gene but also the entire gene networks involved in osteoporosis and regulation of bone mass will systematically be discovered through integrative genomics.     

Effect of mineral content of human bone on in vitro resorption

Summary Osteoclasts, mechanically isolated from chick long bones, were grown in vitro on slices of human rib and femur. Evidence of their activity was assessed by secondary electron and backscattered electron (BSE) imaging in the SEM. BSE imaging was also used to study the relative degree of mineralisation of the bone matrix in which resorption had taken place. All bone phases were resorbed, from osteoid through to densely mineralised interstitial bone and reversal (cement) lines. Resorbing osteoclasts crossed reversal lines between osteons of different mineral density and moved both from higher to lower and lower to higher density phases. Where single loci spanned reversal lines, and thus breached bone of two different mineral densities, depth of demineralisation was inversely related to mineral density. The presence of an annular zone around some resorption loci, which may be caused by demineralisation beneath the osteoclast clear zone, was confirmed. Also, BSE imaging of polished substrata showed that significantly more osteoclastic activity had occurred at their surfaces than was apparent from the amount of cavitation present.     

Analysis of the risk of transmitting bovine spongiform encephalopathy through bone grafts derived from bovine bone

Bone substitutes of bovine origin are widely used for treatment of bone defects in dental and orthopedic surgery. Due to the occurrence of BSE and the new variant of Creutzfeldt Jakob Disease risks of transmitting diseases through the use of such materials need to be carefully evaluated. Risk analysis can either be based on theoretical assessments or experimental evidence. Here we present a comparative study on two bovine bone substitutes (Bio-Oss and Osteograf/N) which is based on theoretical values. Furthermore, for one of these materials, i.e. Bio-Oss, the prion inactivation capacity of one of the production steps was experimentally evaluated. Theoretical and experimental data indicate that the use of these materials does not carry a risk of transmitting BSE to patients.     

Art of replacing craniofacial bone defects

In the history of medicine, many surgeons have been tried to reconstruct lost tissue and correct deformity, attempts to use implant materials have probably paralleled those involving autogenous tissue. Recently there has been an acceleration in the understanding of the requirements and potentials of implant materials caused by collaboration between material scientists, biomaterials engineers, clinicians, and clinical investigators. Alloplastic materials have become an essential part of reconstructing the function and contour of the craniofacial skeleton. Bone is a specialized form of connective tissue, which provides support, and protects vital and delicate organs. Bone is embryologically derived from mesenchymal tissue through membranous and endochondral ossification. In the clinical field, the need for bone graft has been increased due to trauma, tumor, craniosynostosis, and pure esthetic bone surgery. Various types of bone grafts have been used to repair craniofacial bone defects over many years, but the autogenous graft has many disadvantages, such as, limited donor sites, donor morbidity, pain, growth deformity and resorption. Many surgeons working in a number of centers around the world have created substitutes and simpler methods for bone replacement. As the alloplatic bone substitute has been advanced, many synthetic substitutes are replaced by bone in vivo over time. The ideal material should be cost effective, non-toxic, non-antigenic, non-carcinogenic, and inert in the body fluids, be easily shaped at the operating table, and maintain its desired form and consistency in situ. This article reviews several of the more commonly used materials for craniofacial reconstruction and summarizes their mechanical properties and clinical aspects.     

In vitro characterizations of mesoporous hydroxyapatite as a controlled release delivery device for VEGF in orthopedic applications

Following bone implant surgery, prolonged ischemic conditions at the implant site often result in postsurgical complications like failure of osseointegration at the bone-implant interface which can lead to implant failure. Thus, restoration of the vascular supply is paramount to the proper development of the bone. High surface area mesostructured materials have been shown to be attractive candidates for bone regeneration to enhance cell adhesion and cell proliferation. This study uses hydroxyapatite, a naturally occurring mineral in the bone, fabricated to a range of suitable pore sizes, infused with vascular endothelial growth factor (VEGF), to be progressively released to stimulate revascularization. In this study, several characterizations including nitrogen adsorption analysis, Fourier-transformed infrared spectroscopy, X-ray diffraction, field emission scanning electron microscope, and transmission electron microscope were used to evaluate the synthesized mesoporous hydroxyapatite (MHA). The results showed that MHA can gradually release VEGF for enhancing revascularization, which is beneficial for orthopedic applications. ? 2012 Wiley Periodicals, Inc. J Biomed Mater Res Part A: 100A:3143-3150, 2012.     

Monitoring and analysis of bovine spongiform encephalopathy (BSE) testing in Denmark using statistical models

The evolution of monitoring and surveillance for bovine spongiform encephalopathy (BSE) from the phase of passive surveillance that began in the United Kingdom in 1988 until the present is described. Currently, surveillance for BSE in Europe consists of mass testing of cattle slaughtered for human consumption and cattle from certain groups considered to be at higher risk of having clinical or detectable BSE. The results of the ongoing BSE testing in Denmark have been analyzed using two statistical approaches: the "classical" frequentist and the Bayesian that is widely used in quantitative risk analysis. The analyses were intended to provide information for decision-makers, the media and the public as well as to provide inputs for future BSE surveillance models. The results to date suggest that the total number of BSE cases that will be found in Denmark in 2001 will not exceed 16.     

Function of matrix IGF-1 in coupling bone resorption and formation

Balancing bone resorption and formation is the quintessential component for the prevention of osteoporosis. Signals that determine the recruitment, replication, differentiation, function, and apoptosis of osteoblasts and osteoclasts direct bone remodeling and determine whether bone tissue is gained, lost, or balanced. Therefore, understanding the signaling pathways involved in the coupling process will help develop further targets for osteoporosis therapy, by blocking bone resorption or enhancing bone formation in a space- and time-dependent manner. Insulin-like growth factor type 1 (IGF-1) has long been known to play a role in bone strength. It is one of the most abundant substances in the bone matrix, circulates systemically and is secreted locally, and has a direct relationship with bone mineral density. Recent data has helped further our understanding of the direct role of IGF-1 signaling in coupling bone remodeling which will be discussed in this review. The bone marrow microenvironment plays a critical role in the fate of mesenchymal stem cells and hematopoietic stem cells and thus how IGF-1 interacts with other factors in the microenvironment are equally important. While previous clinical trials with IGF-1 administration have been unsuccessful at enhancing bone formation, advances in basic science studies have provided insight into further mechanisms that should be considered for future trials. Additional basic science studies dissecting the regulation and the function of matrix IGF-1 in modeling and remodeling will continue to provide further insight for future directions for anabolic therapies for osteoporosis.     

Balancing osteoblast functions and bacterial adhesion on functionalized titanium surfaces

The demand for orthopedic and dental implants will continue to grow, and for these applications, titanium and its alloys have been used extensively. While these implants have achieved high success rates, two major complications may be encountered: the lack of bone tissue integration and implant-centered infection. The surface of the implant, through its interactions with proteins, bacteria and tissue cells, plays a determining role in the success or failure of the implant. Ideally, to enhance the success of implants, their surfaces should inhibit bacterial colonization and concomitantly promote osteoblast functions. In this article, we discuss strategies for tailoring implant surfaces by exploiting the differences in the response of bacteria and osteoblasts to proteins and surface structures. Nevertheless, limitations still exist in the quest for an ideal implant surface. Further advances in this field will require concurrent development in surface modification techniques and a better understanding of the complex and highly inter-related events occurring at the implant surface after implantation.     

Bone reaction to nano hydroxyapatite modified titanium implants placed in a gap-healing model

Nanohydroxyapatite materials show similar chemistry to the bone apatite and depending on the underlying topography and the method of preparation, the nanohydroxyapatite may simulate the specific arrangement of the crystals in bone. Hydroxyapatite (HA) and other CaP materials have been indicated in cases in which the optimal surgical fit is not achievable during surgery, and the HA surface properties may enhance bone filling of the defect area. In this study, very smooth electropolished titanium implants were used as substrata for nano-HA surface modification and as control. One of each implant (control and nano HA) was placed in the rabbit tibia in a surgical site 0.7 mm wider than the implant diameter, resulting in a gap of 0.35 mm on each implant side. Implant stability was ensured by a fixating plate fastened with two side screws. Topographical evaluation performed with an optical interferometer revealed the absence of microstructures on both implants and higher resolution evaluation with AFM showed similar nanoroughness parameters. Surface pores detected on the AFM measurements had similar diameter, depth, and surface porosity (%). Histological evaluation demonstrated similar bone formation for the nano HA and electropolished implants after 4 weeks of healing. These results do not support that nano-HA chemistry and nanotopography will enhance bone formation when placed in a gap-healing model. The very smooth surface may have prevented optimal activity of the material and future studies may evaluate the synergic effects of the surface chemistry, micro, and nanotopography, establishing the optimal parameters for each of them.     

Effect of nickel-titanium shape memory metal alloy on bone formation

The aim of this study was to determine the biocompatibility of NiTi alloy on bone formation in vivo. For this purpose we used ectopic bone formation assay which goes through all the events of bone formation and calcification. Comparisons were made between Nitinol (NiTi), stainless steel (Stst) and titanium-aluminium (6%)-vanadium (4%) alloy (Ti-6Al-4V), which were implanted for 8 weeks under the fascia of the latissimus dorsi muscle in 3-month-old rats. A light-microscopic examination showed no chronic inflammatory or other pathological findings in the induced ossicle or its capsule. New bone replaced part of the decalcified matrix with mineralized new cartilage and bone. The mineral density was measured with peripheral quantitative computed tomography (pQCT). The total bone mineral density (BMD) values were nearly equal between the control and the NiTi samples, the Stst samples and the Ti-6Al-4V samples had lower BMDs. Digital image analysis was used to measure the combined area of new fibrotic tissue and original implanted bone matrix powder around the implants. There were no significant differences between the implanted materials, although Ti-6Al-4V showed the largest matrix powder areas. The same method was used for measurements of proportional cartilage and new bone areas in the ossicles. NiTi showed the largest cartilage area (p < or = 0.05). Between implant groups the new bone area was largest in NiTi. We conclude that NiTi has good biocompatibility, as its effects on ectopic bone formation are similar to those of Stst, and that the ectopic bone formation assay developed here can be used for biocompatibility studies.     

Tantalumpentoxide as a radiopacifier in injectable calcium phosphate cements for bone substitution

The chemical resemblance of calcium phosphate (CaP) cements and the mineral phase of bone is a problem in distinguishing CaP cement from bone tissue by means of common, noninvasive techniques (e.g., X-ray imaging and microcomputed tomography [μCT]). In this study, the feasibility of using tantalumpentoxide (Ta(2)O(5)) powder as radiopacifier in CaP cements was analyzed. A distal femoral condyle model in male adult Wistar rats was used. After 6 weeks of implantation time, the results were analyzed by means of μCT and histology. Unambiguous distinction of CaP cement from native bone tissue and volumetric measurements of the materials appeared to be possible by means of μCT scanning. Furthermore, there was no evidence of either inflammation or fibrous tissue around the implant materials or at the bone-material interface. In conclusion, the addition of Ta(2)O(5) as a radiopacifying additive to CaP cements allows discrimination between bone substitute and surrounding bone tissue. Consequently, Ta(2)O(5) represents an effective and biocompatible additive in CaP cements for in vivo monitoring purposes.     

Design optimization of functionally graded dental implant

The continuous increase of man's life span, and the growing confidence in using artificial materials inside the human body necessities introducing more effective prosthesis and implant materials. However, no artificial implant has biomechanical properties equivalent to the original tissue. Recently, titanium and bioceramic materials, such as hydroxyapatite are extensively used as fabrication materials for dental implant due to their high compatibility with hard tissue and living bone. Titanium has reasonable stiffness and strength while hydroxyapatite has low stiffness, low strength and high ability to reach full integration with living bone. In order to obtain good dental implantation of the biomaterial; full integration of the implant with living bone should be satisfied. Minimum stresses in the implant and the bone must be achieved to increase the life of the implant and prevent bone resorption. Therefore, the aim of the current investigation is to design an implant made from functionally graded material (FGM) to achieve the above advantages. The finite element method and optimization technique are used to reach the required implant design. The optimal materials of the FGM dental implant are found to be hydroxyapatite/titanium. The investigations have shown that the maximum stress in the bone for the hydroxyapatite/titanium FGM implant has been reduced by about 22% and 28% compared to currently used titanium and stainless steel dental implants, respectively.     

Nanostructured materials for inhibition of bacterial adhesion in orthopedic implants: a minireview

Orthopedic implants may fail owing to different reasons: poor osseointegration at the tissue-implant interface, generation of wear debris, stress and strain imbalance between implant and surrounding tissues, and infections. To ensure success in orthopedics, implant materials must not evoke an undesirable inflammatory response, they must be habitable by bone-forming cells (favoring adhesion of osteoblasts), hinder formation of soft connective tissue (hindering adhesion of fibroblasts), and be anti-infective (discouraging bacterial adhesion). Recent studies have suggested that nanophase materials have a better efficacy as bone implants in favoring osseointegration compared to conventional orthopedic implant materials. This minireview discusses studies on nanophase materials as bone implants, focusing on the effect of these materials in inhibiting bacterial adhesion for the prevention of implant infections.     

Determining relevance of a weight-bearing ovine model for bone ingrowth assessment

A weight-bearing ovine model was used to quantify cancellous bone ingrowth and remodeling in porous-coated implants over 6, 12, and 24 weeks in situ. The null hypothesis for the investigation was that there would be no significant difference between the amount of cancellous bone ingrowth and rate of remodeling in this ovine model compared to a reported human bilateral implant model. Bone ingrowth progressed from 20.1 +/- 8.2% at 6 weeks in situ to 23.8 +/- 7.9% at 12 weeks, and 30 +/- 5.1% at 24 weeks. Fluorochrome analysis demonstrated a mineral apposition rate of 1.07 +/- 0.28 microm/day for bone at the porous-coating interface, whereas host bone remodeling at 0.89 +/- 0.23 microm/day. Histological analysis showed no adverse tissue or inflammatory response. The null hypothesis was supported in that regression analysis demonstrated that the amount of cancellous bone ingrowth over time (p = 0.545) and mineral apposition rate over time (p = 0.089) in this ovine model was not significantly different than reported human bilateral knee data. The results of this study appear to validate the ovine model for use in understanding skeletal attachment of porous-coated implants to cancellous bone in humans.     

Novel in vivo method for evaluation of healing around implants in bone

A material implanted in bone is always inserted into coagulating blood. Protein and cell interactions during this initial implantation time will govern later healing. Many studies have focused on the tissue surrounding implants. We have developed a method for evaluation of healing around implants in bone by studying cells adhering to the implant surface. Hydrophilic titanium discs were inserted into rat tibiae. Samples were retrieved after 1, 2, 4, and 8 days of implantation and were analyzed by fluorescence microscopy techniques and scanning electron microscopy. Both proliferating and apoptotic cells were found on the surface. Generally, cells closest to the implant surface were nonviable whereas cells in the fibrin network a distance from the surface were viable. Bone morphogenetic protein-2 (BMP-2) is an osteogenic substance. An increase in BMP-2-positive cells was seen during the implantation period, and a population of large BMP-2-positive cells appeared on the surface after 4 days of implantation. The method developed here is a suitable tool for rapid evaluation of the initial healing around implant material.     

Integration of metallic endoprotheses in dog femur studied by near-infrared Fourier-transform Raman microscopy.

The integration of hydroxyapatite-coated implants in dog femur was studied by near-infrared Fourier-transform Raman microscopy. Raman spectra were taken in lateral scans in step widths of 10-40 microm from the implant surface up to a distance of 320 microm into the bone tissue. The spectra were subjected to a component analysis for the quantitative determination of the protein and the inorganic components. This quantitative analysis is shown to be more reliable than conventional band fitting procedures and allows, for the first time, the quantitative distinction between the hydroxyapatite form of mature bone tissue and synthetic hydroxyapatite introduced by the implant coating. It is demonstrated that full mineralization of the ongrowing bone is not achieved after 6 months. In contrast, after a residence time of 18 months in the body, the Raman spectra reveal a complete calcification of the new bone tissue as indicated by content of biological hydroxyapatite that is the same as in mature bone tissue throughout the whole implant/bone interface. On the other hand, the content of synthetic hydroxyapatite is strongly reduced in the sample prepared after eighteen months implantation whereas for the shorter implantation time. substantial contributions of synthetic hydroxyapatite are found even at positions beyond the thickness of the implant coating. These results indicate that the coating material is actively involved in the mineralization of ongrowing bone. Possible mechanisms for the underlying transport processes in the implant/bone interface are discussed.