Peer W. Kämmerer

Bisphosphonates: restrictions for vasculogenesis and angiogenesis: inhibition of cell function of endothelial progenitor cells and mature endothelial cells in vitro.

Bisphosphonate-associated osteonecrosis of the jaws (BP-ONJ) is one of the main side effects in patients treated with bisphosphonates for metastasis to the bone or osteoporosis. BP-ONJ usually occurs in patients treated with highly potent nitrogen-containing bisphosphonates. The exact mechanism of action and etiopathology is still unknown. In addition to inhibition of bone remodelling, an anti-angiogenetic effect has become the focus of research. The aim of these study was to investigate the effect of different bisphosphonates on human umbilicord vein endothelial cells (HUVEC) and endothelial progenitor cells (EPC), which play an important role in angiogenesis. Using varying concentrations, the impact of one non-nitrogen-containing bisphosphonate (clodronate) and three nitrogen-containing bisphosphonates (ibandronate, pamidronate and zoledronate) on HUVEC and EPC was analysed. The biologic behaviour of HUVEC after incubation with different bisphosphonates was measured in a Boyden migration assay as well as in a 3D angiogenesis assay. The number of apoptotic cells was measured by Tunnel assay. To underline the importance of neoangiogenesis in the context of BP-ONJ, we measured the EPC number after incubation with different bisphosphonates in vitro. HUVEC and EPC were significantly influenced by bisphosphonates at different concentrations compared with the non-treated control groups. The nitrogen-containing bisphosphonates pamidronate and zoledronate had the greatest impact on the cells, whereas clodronate followed by ibandronate was less distinct on cell function. These results underline the hypothesis that inhibited angiogenesis induced by bisphosphonates might be of relevance in the development and maintenance of BP-ONJ. The increased impact by highly potent bisphosphonates on HUVEC and EPC may explain the high prevalence of BP-ONJ in patients undergoing this treatment.

Ten-year retrospective follow-up study of the TiOblast dental implant.

PURPOSE Long-term results in the clinical outcome of different implant systems, including high patient numbers and a long follow-up time, are rare. This retrospective study evaluated the cumulative survival rate of a self-tapping, cylindrical implant system with a conical implant-abutment connection after 10 years of prosthetic loading. MATERIALS AND METHODS A total of 516 TiOblast implants (Astra Tech AB, Mölndal, Sweden) were placed in 108 patients. The patients were treated in the Department of Oral and Maxillofacial Surgery, Johannes Gutenberg University, Mainz, Germany, between September 1994 and May 2005. The main indications for implantation were the treatment of edentulous mandibles (74%) and partial edentulism (15%). Twenty-three implants were placed postradiation, and a further 64 implants were irradiated after insertion. In 153 implants, a bony augmentation was conducted prior to implantation. RESULTS The in situ rate was 89.7% after an average implantation time of 108 months. Eighty-three patients with 403 implants were available for investigation. Seventeen patients with 76 implants have died since 1994. Absence of osseointegration (n = 22), peri-implantitis (n = 18), fracture of the implants (n = 9), failing of primary stability (n = 2), and implants next to tumors (n = 2) were the reasons of explantation in 26 patients. Under analysis with different implant success-assessment criteria, the success rate showed results from 76 to 89%. CONCLUSION With respect to the critical patient selection including a high number of patients with minor and major augmentations, the 10-year clinical use of the studied implant system showed acceptable results.

Early implant healing: promotion of platelet activation and cytokine release by topographical, chemical and biomimetical titanium surface modifications in vitro

OBJECTIVES Platelet releasate has been shown to promote osteogenetic cell proliferation and differentiation. Topography and chemistry of biomaterials have high impact on platelet activation. More specifically, the bioactive cell adhesive peptide sequence Arg-Gly-Asp (RGD) triggers platelet activation mediated by the α(IIb) β(3) integrin receptor. Accordingly, topographical, chemical and biomimetical (immobilized RGD peptide) modifications of titanium (Ti) surfaces may enhance early platelet activation and bony healing of implants. Therefore, the aim of the study was to evaluate platelet activation with subsequent platelet-derived cytokine release by accordingly modified Ti surfaces. MATERIALS AND METHODS Pre-treated (PT; mean roughness [R(a)]=0.04 μm, contact angle [CA]=91°), acid-etched (A, R(a) =0.83 μm, CA=106°), large grit-sandblasted, acid-etched (SLA, R(a) =3.2 μm, CA=109°) as well as hydrophilically modified acid-etched (modA, R(a) =0.83 μm, CA=0) and modified large grit-sandblasted, acid-etched (modSLA, R(a) =3.2 μm; CA=0°) titanium surfaces were investigated. Additionally, RGD peptides were chemically immobilized on PT, A and SLA surfaces (PT-RGD [CA=18°], A-RGD [CA=0°], SLA-RGD [CA=0°]). The different Ti surfaces were incubated with platelet concentrate of three healthy volunteers at room temperature for 15 min and for 30 min. High thrombogenous collagen served as the control group. Out of the supernatant, platelet consumption was assessed via platelet count (PC). Cytokine release was quantified via the level of platelet-derived growth factor (PDGF) and vascular endothelial growth factor (VEGF). RESULTS After 15 min, especially the rough SLA surface showed a strong decrease in PC and a strong increase in VEGF and PDGF levels. After 30 min, high platelet consumption as well as high levels of VEGF and PDGF were measured for unspecifically modified (modA) and especially for biomimetic, specifically modified (PT-RGD, A-RGD) surfaces, indicating a delayed effect of the surface modifications on platelet activation. DISCUSSION Modifications of surface roughness modifications appear to influence early platelet activation and cytokine release after 15 min whereas surface chemistry modifications with increased hydrophilic properties and surface modifications via RGD peptide on plainer surfaces lead to a further, more specific promotion of platelet activation and degranulation after 30 min. The observed effect could be valuable for critical clinical situations like compromised bone sites.

Single nucleotide polymorphisms of the vascular endothelial growth factor gene associated with incidence of oral squamous cell carcinoma

BACKGROUND   Vascular endothelial growth factor (VEGF) plays an important role in promoting angiogenesis and is overexpressed in several malignancies. Polymorphisms of the VEGF gene can alter VEGF protein expression, which may be biologically significant and account for heterogeneity in disease risk and outcome. The aim of this case-control study was to evaluate potential associations between single nucleotide polymorphisms (SNP) of the VEGF gene with susceptibility of oral squamous cell carcinoma (OSCC). PATIENTS AND METHODS Five VEGF SNP (-1154 G/A, +405 G/C, +936 C/T, -2578 C/A and -460 C/T) were determined in peripheral blood isolated from 80 patients with OSCC and from 40 age- and gender-matched healthy volunteers (RT-PCR). RESULTS The +936 T allele and the -2578 C/A SNP were expressed significantly more often in the OSCC-group (P=0.002; P<0.0001) where three associations between two SNPs (+936 and +405, -2578 and -1154, -460 and -2578) were found. CONCLUSION Our findings provide support that +936 T allele and -2578 C/A SNP of the VEGF gene alone or in combination with other SNP are associated with OSCC. The SNPs may be used as biomarker for the development of specialized anti-VEGF drugs. Further studies must confirm the value of preoperative genetic analysis for prognosis.

Association of T-cell regulatory gene polymorphisms with oral squamous cell carcinoma

Costimulatory molecules have complementary effects on T-cell activation and their balance may control the development of oral cancer. The aim of this study was to determine the relevance of cytotoxic T-lymphocyte antigen 4 (CTLA-4), CD28 and inducible costimulator (ICOS) polymorphisms in oral squamous cell carcinoma (OSCC). Genotyping for CTLA-4 (-1661 A/G and +49 A/G), CD28 (0 C/G and +3160 G/T) and ICOS (+637 A/C and +1599 C/T) was performed in the 83 patients with OSCC, compared to the 40 unrelated healthy volunteers as controls. The genotype CTLA-4 -1661 was significantly different between the patient group and the control group. The allele CTLA-4 -1661 G was significantly found more frequent in patients with OSCC (p=0.001). In bivariate analysis, noticeable differences between OSCC and controls were seen. The combinations CTLA-4 -1661 G/G and CTLA-4 +49 A/G, ICOS +1559 C/T and ICOS +1559 C/C each with CTLA-4 -1661 G/G, ICOS +637 C/C and ICOS +637 A/C each with CTLA-4 -1661, CTLA-4 -1661 A/G and ICOS +637 C/C, CD28 +3160 G/T and CTLA-4 -1661 A/A and CD28 +3160 G/T and CTLA-4 -1661 A/G were seen in the patient group only. Especially the polymorphisms of the CTLA-4 -1661-genotype - alone and in combination with other T cell regulator polymorphisms - seem to be possible predisposing factors for OSCC. Therefore, they might be future targets for a primary prophylaxis or an individualised therapy.

Retrospective analysis of survival rates and marginal bone loss on short implants in the mandible

OBJECTIVES Short implants have become an interesting alternative to bone augmentation in dental implantology. Design of shorter implants and longer surveillance times are a current research issue. The goal of this study was to show the survival rates of short implants below 9 mm in the partly edentulous mandibular premolar and molar regions with fixed prosthetics. Marginal vertical and 2D bone loss was evaluated additionally. Different implant designs are orientationally evaluated. MATERIAL AND METHODS A total of 247 dental implants with fixed prosthetics (crowns and bridges) in the premolar and molar region of the mandible were evaluated; 47 implants were 9 mm or shorter. Patient data were evaluated to acquire implant survival rates, implant diameter, gender and age. Panoramic X-rays were analysed for marginal bone loss. RESULTS Average surveillance time was 1327 days. Cumulative survival rate (CSR) of short implants was 98% (1 implants lost) compared to 94% in the longer implants group without significance. Thirty-five of the short implants were Astratech (0 losses) and 12 were Camlog Screw Line Promote Plus (1 loss). Early vertical and two-dimensional marginal bone loss was not significantly different in short and regular length implant group with an average of 0.6 mm and 0.7 mm(2) in short implants over the observation period. CONCLUSIONS Within the limitations of this study, we conclude that short implants with a length of 9 mm or less have equal survival rates compared with longer implants over the observation period of 1-3 years.

Modulation of platelet activation and initial cytokine release by alloplastic bone substitute materials

OBJECTIVES Platelet-derived cytokines play a crucial role in tissue regeneration. In regenerative dental medicine, bone substitute materials (BSM) are widely used. However, initial interactions of BSM and platelets are still unknown. The aim of this study was to evaluate the potential of platelet activation and subsequent initial cytokine release by different commercial alloplastic BSM. MATERIAL AND METHODS Eight commercial BSM of different origins and chemical compositions (tricalcium phosphate, hydroxyapatite, bioactive glass: SiO(2) and mixtures) were incubated with a platelet concentrate (platelet-rich plasma, PRP) of three healthy volunteers at room temperature for 15 min. Platelet count, aggregation, degranulation (activated surface receptor CD62p) and cytokine release (Platelet-derived growth factor, Vascular endothelial growth factor) into the supernatant were quantified. Highly thrombogenic collagen served as a reference. RESULTS The investigated PRP samples revealed different activation patterns when incubated with different BSM. In general, SiO(2)-containing BSM resulted in high platelet activation and cytokine release. In detail, pure bioactive glass promoted platelet activation most significantly, followed by hybrid BSM containing lower ratios of SiO(2). Additionally, we found indications of cytokine retention by BSM of large specific surfaces. CONCLUSIONS Platelet activation as well as consecutive storage and slow release of platelet-derived cytokines are desirable attributes of modern BSM. Within the limits of the study, SiO(2)-containing BSM were identified as promising biomaterials. Further investigations on cytokine adsorption and cytokine release kinetics by the respective BSM have to be conducted.

Influence of a collagen membrane and recombinant platelet‐derived growth factor on vertical bone augmentation in implant‐fixed deproteinized bovine bone – animal pilot study

OBJECTIVES Combinations of bone substitute block materials with membrane techniques as well as with growth factors are possible options to enhance the prognosis of vertical bone augmentation. Therefore, the aim of the pilot study was to compare the influence of a collagen membrane and a signal protein (rhPDGF-BB) on vertical bone augmentation with a stable fixed block material (deproteinized bovine bone [DBB]). MATERIALS AND METHODS In 12 rabbits, a DBB-block was implant-fixed on the tibia in a split-leg-design. Included were: DBB only (control), DBB + collagen membrane (test), DBB + rhPDGF-BB (test) and DBB + rhPDGF-BB + collagen membrane (test). 24 samples were examined after 3 (n = 12) and 6 weeks (n = 12). Calculated parameters were new bone area (NBA;%), new vertical bone height (VBH; mm). Due to the pilot character of this study, single values are shown descriptively only. RESULTS After 3 weeks, there were constant higher NBA values in the rhPDGF-BB-group without membrane (NBA (%) DBB: 30/16/4; DBB + membrane: 25/17/7, DBB + rhPDGF-BB: 40/33/34, DBB + rhPDGF-BB + membrane: 0/30/16; VBH (mm) DBB: 1.2/1.2/1, DBB + membrane: 0.7/0.9/1, DBB + rhPDGF-BB: 0.7/0.9/1, DBB + rhPDGF-BB + membrane: 0/1.1/1). After 6 weeks, both membrane groups showed a constant higher NBA and VBH independent to the use of rhPDGF-BB (NBA DBB: 3/0/5, DBB + membrane: 20/35/31, DBB + rhPDGF-BB: 5/8/4, DBB + rhPDGF-BB + membrane: 31/35/40; VBH DBB: 0.3/0.3/0.6, DBB + membrane: 1.6/2.4/2.1, DBB + rhPDGF-BB: 0.4/0.7/0.8, DBB + rhPDGF-BB + membrane: 1.8/2/1.8). CONCLUSIONS For vertical augmentation, the addition of rhPDGF-BB to DBB-blocks may increase early bone growth. In the later phase, the use of a collagen membrane enhances new bone volume and height to a significant greater extend. Even if the results are higher than those in the non-membrane groups, the low gain of bone after the short time periods still needs improvement.

Clinical, therapeutic and prognostic features of osteosarcoma of the jaws – Experience of 36 cases

INTRODUCTION Osteosarcoma of the jaws (OSJ) differs from osteosarcoma of other skeletal regions due to later development, a high mortality associated with the local disease, fewer incidences of metastases and its extreme rarity. In regard to clinical and pathological parameters as well as therapeutic approaches and prognosis, OSJ has not been specifically examined to date. In order to achieve a better understanding of this special malignancy, an evaluation of incidence, treatment and prognosis of patients with OSJ in our department over the past 38 years was conducted. MATERIALS AND METHODS A retrospective analysis of patients with OSJ between 1972 and 2010 was performed. Information regarding patient characteristics, site of the lesion, main presenting symptoms, latency of initial diagnosis, treatment, histology, local recurrence, development of metastatic disease, duration of follow up and survival was obtained. The data were compared to the literature. RESULTS Thirty-six patients (2-81 years, mean: 33.9, standard deviation: 21.3) were diagnosed and treated for OSJ (maxillar:mandibular nearly 1:2). Initial symptoms were local swelling (81%) and pain (47%). The latency period between fist symptoms and clinical presentation was 3.7 months (1-24). A radical resection alone was conducted in 15 patients. In nine patients, resection and radiotherapy was used. Resection with chemotherapy was the treatment of choice in seven patients. Five patients received a triple combination of resection, chemo- and radiation therapy. The osteoblastic subtype of osteosarcoma was most frequent (42%). In 15 cases (42%) local recurrences, in two cases (5%) metastasis were seen. Of these patients, 13 died within the observation period. One other patient (3%) died as a result of progressive pulmonary metastasis. A mean total survival rate of 61% could be seen whereas the highest survival rate (80%) was found in patients who were treated with neoadjuvant chemotherapy, radical resection and adjuvant radiation. Positive prognostic factors were a younger age and tumour-free resection margins. DISCUSSION OSJ is a highly lethal tumour entity. According to the data at hand, therapy should possibly include chemotherapy, radical resection and irradiation. Nevertheless, due to the rarity of OSJ, information remains limited and the treatment of choice should be within the focus of clinical multi-centre studies.

Vertical Osteoconductive Characteristics of Titanium Implants with Calcium‐Phosphate‐Coated Surfaces – A Pilot Study in Rabbits

INTRODUCTION Osteoconductive characteristics of different implant surface coatings are in the focus of current interest. The aim of the present study was to compare the vertical osteoconductivity at the implant shoulder of supracrestal inserted calcium-phosphate coated implants (SLA-CaP) with conventional sand-blasted/acid-etched (SLA) implants in a rabbit model. MATERIALS AND METHODS SLA-CaP and SLA implants were inserted bilaterally in the mandible of four rabbits in a split-mouth design. The implants were placed 2 mm supracrestal. After 3 weeks, at the left and right implant shoulder, the percentage of linear bone fill (PLF) as well as bone-implant contact (BIC-D) were determined. RESULTS After 3 weeks, newly formed woven bone could be found at the shoulder of the most of both surface-treated implants (75%). PLF was significantly higher in SLA-CaP implants (11.2% vs. 46.5%; n = 8, p = .008). BIC-D was significantly increased in the SLA-CaP implants (13.0% vs. 71.4%; n = 8, p < .001) as well. CONCLUSION The results of this study show for the first time that calcium-phosphate coated surfaces on supracrestal inserted implants have vertical osteoconductive characteristics and increase the bone-implant contact at the implant shoulder significantly in a rabbit model. In clinical long-term settings, these implants may contribute to a better vertical bone height.