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Introduction: The purpose of this study was to evaluate the influence of different surface topographies on the expression of bone cell-associated proteins, such as osteoprotegerin (OPG), osteocalcin and alkaline phosphatase (AP), and the production of the extracellular matrix (ECM) in vitro. Another aspect was the question as to whether a hydroxyapatite (HA) coating offers additional advantages. Vacuum plasma-sprayed (VPS) pure titanium was used to generate different surface topographies., Materials and Methods: The in vitro response of human bone marrow cells to VPS implants (porosity ranging from 25 to 50%, pore size ranging from 50 to 200 microm and roughness ranging from 0.191 to 0.547 mm) and cancellous structured titanium (cs-Ti) as a reference material (55% porosity, pore size of 500 microm, roughness 0.836 mm) were compared. The expression of bone cell-associated proteins, such as OPG, osteocalcin and alkaline phosphatase (AP), was evaluated. Scanning electron microscopy (SEM) was used to judge the production of ECM., Results: All implant materials induced the release of OPG, osteocalcin and AP. Significant differences were evident between the cs-Ti and the different VPS-Ti surface structures. There was no difference in the response between the VPS-Ti surfaces. SEM showed a dense and increased production of ECM on the VPS-Ti surfaces. An additional HA coating caused a faster production of ECM and higher levels of OPG., Conclusions: The in vitro data presented here demonstrate the superiority of VPS-Ti surfaces over cs-Ti, which is already in clinical use. Differences between the VPS-Ti surfaces were not evident. Presumably, VPS-Ti surfaces offer good prerequisites for a successful integration of the implant in the surrounding tissue. An additional HA coating could influence these events positively.

The objective of this study was to evaluate the bone ingrowth of a new vacuum plasma sprayed titanium surface (vps-ti) in comparison to cs-titanium implants in a göttinger minipig model. Fifteen göttinger minipigs each received the two implants, vacuum plasma sprayed titanium with a porosity of 50% and a pore size of 200 microm (vps-ti) and an implant with a similar porosity but a different pore size 500 microm (cs-ti), at the proximal femur metaphysis by press-fit technique. The pigs were euthanized at three different postsurgical periods: 4, 8 and 12 weeks. Each femur was harvested and qualitative (macroscopic and microscopic) and quantitative (histomorphometric) histological analysis was done on histological slides. The results indicated that there was a difference in bone ingrowth between the two implants, whereas the bone ingrowth of vps-ti was superior to cs-ti after 4 and 8 weeks healing time. 12 weeks post implantationem no statistiscal difference was evident. The pore size of 200 microm seemed superior to a pore size of 500 microm. Whether or not these effects lead to a better mechanical stability remains unanswered.

Aim: The purpose of the present study was to evaluate the influence of ultra-high-molecular-weight polyethylene (UHMW-PE), which is the major constituent of the material debris formed as a result of orthopaedic implant wear, on the cellular differentiation in a modified in vitro model., Methods: UHMW-PE particles (Ø < or = 7.5 microm) were suspended in soluble collagen type I and subsequently solidified in different concentrations (10(5), 10(6) and 10(7) particles per well) on the bottom of the wells. Human bone marrow cells in a concentration of 3 x 10(6) cells per well were seeded on the collagen-particle substrate and maintained for up to 72 h. The response of the cells to the particles was examined by light microscopy, scanning electron microscopy and FACS analysis compared to cells on control collagen surfaces without any particles., Results: Light and scanning microscopic evaluation revealed that the UHMW-PE particles, which had built large conglomerates (Ø 7.5 microm), were mainly surrounded by the cells and less phagocytosed. The results of the FACS analysis revealed significant differences in CD3/CD4 positive, CD14 positive and CD19 positive cells (p < 0.05). A significant elevation of CD3/CD4 positive and CD14 positive cells (p < 0.05) was observed after the period of culture (72 h) whereas a significant decrease could be detected in the case of CD19 positive cells., Conclusion: The results demonstrate that the particle-induced response by UHMW-PE limits itself not only to the particle macrophage contact but influences also the differentiation of the bone marrow. Moreover, the results confirm that the present method is useful to evaluate the in vitro effects of UHMW-PE wear particles with direct particle cell contact. Although the particles built large conglomerates, it could be shown that a change of the immune-competent cells also occurred.

Introduction: Repair of large skeletal defects using bone allografts has become a routine procedure in orthopaedic and trauma surgery. Different procedures of sterilisation (82.5 degrees C disinfection; 121 degrees C autoclaving; PES; Tutoplast; 25 kGy gamma irradiation) are available to inactivate bacteria and fungi, including their spores, as well as viruses in human bone allografts. The efficiency of these procedures has been proven. However, the effects on the cellular response are rarely investigated. This present in vitro study investigates the immunological answer of human bone marrow cells to human allogenous and autologous bone platelets which were sterilised by different methods., Materials and Methods: Human bone marrow cells and the bone platelets were harvested from patients undergoing a total hip replacement. All patients provided informed consent. Human bone platelets, 10 mm in diameter, 3 mm in height, were produced from femoral heads which were removed within the scope of total hip replacements. They were sterilised by different procedures or were disinfected (gamma radiotherapy, PES/ethanol treatment, Tutoplast procedure, 121 degrees C autoclaving, > 82.5 degrees C thermodisinfection). In addition, an autologous in vitro bone donation was simulated and compared with the allogenous bone grafts. Endobon was evaluated as a bovine hydroxyapatite ceramic. As control a human bone marrow cell culture without bone platelets was used. Over a period of four weeks the changes of the immunogenic cell populations were analysed in vitro (FACS analysis). Light and scanning microscopy were done to reveal morphological differences. As a vitality test the trypan-blue staining was performed., Results: Light and scanning microscopy demonstrated large differences between the various sterilisation and disinfection methods. After 4 weeks the autologous bone platelets were completely covered with homogenously distributed human osteoblast like cells. The heat-sterilised/disinfected transplants demonstrated similar effects compared to the autologous bone grafts while the irradiated bone platelets demonstrated less cell coverage. 2/3 of the cells were vital on average after four weeks, with the exception of the irradiated bone platelets. The FACS analysis revealed in comparison to the control group provable differences in the immunological answer for the autologous bone donation as well as for the differently sterilised or disinfected allogenous bone grafts. The heat sterilisation or, respectively, disinfection methods compared to the autologous bone donation demonstrated almost similar in vitro effects. By far the worst results, characterised by an excessively increased portion of cytotoxic T-cells and a decreased amount of viable cells, were seen in the 25 kGy gamma irradiation samples., Conclusions: The results demonstrate the influence of the different sterilisation and disinfection procedures on the differentiation of human marrow cells (host). Similar in vitro effects were seen for the autologous and heat-treated bone platelets. The treatment of allogenous bone grafts with PES/ethanol and the Tutoplast procedures showed, just as Endobon, only low differences in comparison with the control cultures. The worse results in the case of the irradiated bone platelets may be explained by the production of free radicals which led to an excessive cell death.

There is considerable evidence that orthopaedic wear debris plays a crucial role in the pathology of aseptic loosening of joint prostheses. The purpose of the present study was to evaluate the influence of ultra-high-molecular-weight polyethylene (UHMW-PE) on the cytokine response in a modified in vitro model. UHMW-PE particles (psi < 7.5 microm) were suspended in soluble collagen type I and subsequently solidified in different concentrations (105,106 and 107 particles per well) on the bottom of the wells. Human bone marrow cells in a concentration of 3 x 106 cells per well were seeded on the collagen-particle substrata and maintained for up to 12 days. The cytokine response (IL-1&#95;, IL-6 and TNF-&#95;) of the cells to the particles were examined by ELISA compared to cells on control collagen surfaces without any particles. Assays for viability using LDH activity were done immediately. Light and scanning microscopic evaluation revealed that the UHMWPE particles, which have built large conglomerates (psi7.5&#95;m), were mainly surrounded by the cells and less phagocytosed. The results of the cytokine release revealed significant differences in interleukin (IL)6, tumor necrosis factor (TNF)- &#95; and IL-1beta. The cell viability was not affected by the UHMW-PE particles. The results demonstrate that the particle induced cytokine response by UHMW-PE is mainly by the release of Interleukin 6 and TNF- &#95;. Moreover the results confirm that the present method is useful to evaluate the in vitro effects of UHMW-PE wear particles with direct particle cell contact.

Introduction: This study deals with the ingrowth behaviour of biomaterials (hydroxyapatite, cp-titanium, cobalt-chromium-molybdenum and PAEK) in relationship to the immunological competence in an animal model. Measured were the production of extracellular matrix (ECM) after implantation in non-immunocompetent naked mice and immunocompetent wild mice. Intention of the trial was to find out if either the immunological competence or the duration of implantation influences the quantity of produced ECM. In addition, the ingrowth behaviour was investigated under these conditions by using four different biomaterials., Material and Methods: Biomaterials (hydroxyapatite, cp-titanium, cobalt-chromium-molybdenum and PAEK) were implanted for 14 or 60 days, respectively. CLSM, SEM and SEM-EDX were used for analysis of the ECM and for measuring the distance between ECM and the biomaterials. CLSM was also used for the detection of collagen I and III as a parameter of the quality of osteointegration., Results: In all cases a matrix grew on the surface of the biomaterials. The CLSM detected a co-localisation of collagen I and III. In the case of hydroxyapatite collagen I and III were found at a distance of 1 micro m over the surface. The largest space between the surface of the implant and the ECM was found in the case of PAEK. The smallest space was in the case of hydroxyapatite. In all investigated biomaterials the proportion of collagen I to collagen III varied through the duration of implantation., Discussion: As is known from the literature we found different ingrowth behaviours on using different biomaterials. Furthermore, we found a statistically significant influence of the immunological competence of the host with regard to ECM production. We draw the conclusion that immunological competence improves the ingrowth behaviour of biomaterials.

The validity of isovolumic parameters of left ventricular contractility in assessing ventricular function at rest has been questioned repeatedly, and with some justification. By contrast, auxotonic parameters such as left ventricular ejection fraction provide a clinically useful index of functional impairment even under resting conditions. The correlation between isovolumic and auxotonic parameters during exercise remains unclear, however. To evaluate this relation, isovolumic m (max dP/dt, Vpm) and auxotonic (EF, Vcf) parameters of contractility were determined simultaneously at rest and after both leg raising and exercise using a manometer-equipped angiography catheter in 45 patients. Under resting conditions, there was no significant correlation between the parameters (Vpm: Vcf,r = 0,226). After leg raising (r = 0.543) and exercise (r = 0.801), the correlation became markedly closer. The presence of hypo- or akinetic regions did not affect the correlation. In patients with cardiomyopathy and uniform functional impairment, the correlation did not improve in comparison with the group of patients characterized by asynergy. The specificity of the various parameters was evaluated in patients with and without significant coronary-artery stenosis. The Vcf values during exercise yielded virtually optimal separation of the two groups, whereas isovolumic parameters were far less specific.