Interaction of titanium, zirconia and lithium disilicate with peri-implant soft tissue: study protocol for a randomized controlled trial.

BACKGROUND: Against the background of increasing use of dental implants, and thus an increasing prevalence of implant-associated complications, a deeper understanding of the biomolecular mechanisms in the peri-implant tissue is needed. Peri-implant soft tissue is in direct contact with transmucosal dental implant abutments. The aim of this trial is to distinguish the biomolecular and histological interactions of various dental abutment materials with peri-implant soft tissue. METHODS/DESIGN: The study is designed as a prospective, randomized, investigator-initiated clinical pilot trial with blinded assessment. We will ultimately include 24 eligible patients who opt for implant treatment to replace a single missing posterior tooth. Three months after implantation (submerged procedure), the study begins with the second-stage surgery. Each of the 24 patients will be given three different transmucosal abutments (zirconia, lithium disilicate, titanium) consecutively. The sequence in which the three materials are used is randomized. Peri-implant crevicular fluid is sampled weekly around the respective abutment for biomolecular analyses. After one month of wearing time, the stamping press from the second-stage surgery is used to gain a narrow gingival ring biopsy around the abutment for immunohistochemical analyses. The next abutment is then inserted. The same procedure is used for all three abutments. After sampling is completed, the patients will receive a definitive crown. The primary outcome measure of the trial is biomolecular detection of specific markers in the peri-implant crevicular fluid: matrix metalloproteinase 8, interleukin- 1ß, polymorphonuclear elastase, and myeloid-related protein MRP8/14 (calprotectin). Secondary outcome measures include immunohistochemical analyses and clinical parameters. DISCUSSION: The study design will allow us to perform correlation analyses between the clinical indices with biomarkers' expression in the interface of the transmucosal abutments and the peri-implant soft tissue. A deeper understanding of the three abutment materials' interactions with peri-implant soft tissue will help us understand the formation mechanisms of implant-associated complications and then develop prevention strategies. TRIAL REGISTRATION: The trial is registered at the German Clinical Trial Register and the International Clinical Trials Registry Platform by the WHO under DRKS00006555 (Registered on 27 October 2014).

Mechanism of high-power NIR laser bacteria inactivation.

Lasers are used in dentistry for a variety of indications. One of these is the disinfection of root canals or the sterilization of residual caries. Many studies have demonstrated the capacity to kill bacteria for lasers but the fundamental mechanism of the laser effect remains quite unclear. With our experiments we wanted to determine whether high-power NIR laser bacterial killing is caused by the light itself (photochemical effect) or by a photothermal process. In order to differentiate between mechanisms we heated bacteria suspensions of a nonpathogenic strain of E. coli by a water bath and by a diode laser (940 nm) with the same temporal temperature course. Furthermore, bacteria suspensions were irradiated while the temperature was fixed by ice water. Killing of bacteria was measured via fluorescence labelling. Comparison of killing rates between laser and water-based heating shows no significant differences. The most important parameter is the maximum temperature. Laser irradiation of bacteria at low temperatures does not result in killing. Our experiments show that at least for E. coli bacteria inactivation by high-power laser irradiation is solely based on a thermal process.

Additional Cyclin D(1) gene copies associated with chromosome 11 aberrations in cutaneous malignant melanoma.

Cyclin D1 (CCND1, PRAD1, bcl-1) is associated with progression through G1 and entry into S phase of the cell cycle, as a partner of cyclin-dependent kinases (CDKs). The aim of this study was to evaluate cytogenetic aberrations of the gene locus at chromosome 11q13 in cutaneous malignant melanoma. Fluorescence in situ hybridisation (FISH) was applied on metaphase spreads of short-term primary cell cultures as well as on 5 microm paraffin tissue sections of primary melanomas and melanoma metastases, and on melanoma cell lines (C32, WM 266-4, HT 144, RPMI 7951, SK MEL 28). For FISH analysis DNA probes specific for Cyclin D1, centromere 11 and painting probes for whole chromosome 11 were used. FISH revealed additional Cyclin D1 gene copies in relation to centromere 11 copy numbers in 9/19 (47%) primary melanomas and in 7/20 (35%) melanoma metastases. The melanoma cell line SK MEL 28 also showed extra Cyclin D1 gene copies. A 24-colour FISH (mFISH) investigation revealed translocated chromosome 11 material to chromosomes 12, 14, 15, 20, 21, 22 and Y in these cases, respectively. Positivity for Cyclin D1 protein was detected by immunostaining in 17/19 (89%) primary melanomas (10 weak, 7 strong expression) and in 9/12 (75%) melanoma metastases (5 weak and 4 strong). All primary melanomas and 3/4 examined metastases with additional Cyclin D1 gene copies concomitantly revealed positive protein staining. Based on these results Cyclin D1 could well be involved in the pathogenesis of a subset of malignant melanoma. Additional studies will have to elucidate the role of further genes located at chromosome 11q13.

Numerical abnormalities of the Cyclin D1 gene locus on chromosome 11q13 in non-melanoma skin cancer.

Deregulation of the cell-cycle G1-restriction point control via abnormalities of Rb-pathway components is a frequent event in the formation of cancer. The aim of this study was to evaluate numerical aberrations of the Cyclin D1 (CCND1, PRAD1, bcl-1) gene locus at chromosome 11q13 in basal cell carcinomas (BCCs) and squamous cell carcinomas (SCCs) of the skin and to compare it with the Cyclin D1 protein expression. Fluorescence in situ hybridization with DNA-probes specific for the Cyclin D1 gene locus and the centromere of chromosome 11 as well as immunostaining for Cyclin D1 protein was applied on 5 microm serial paraffin sections. Six of the 30 (20%) SCCs showed additional Cyclin D1 gene copies and 2/30 (6.6%) cases had a loss of the Cyclin D1 gene locus in relation to the centromere 11 number. In contrast, only one of the 14 BCCs (7%) showed one additional Cyclin D1 gene copy in relation to the centromere 11 number. None of the BCCs demonstrated aneusomy for chromosome 11 in contrast to SCCs, where it was found in 21/30 (70%) cases. Twenty-six of the 30 (86.6%) cutaneous SCCs and 13/14 (93%) BCCs expressed Cyclin D1 protein. All SCCs and the BCC with additional Cyclin D1 gene copies showed positivity for Cyclin D1 protein. Both SCCs with less Cyclin D1 gene copies than centromere 11 signals showed a weak protein expression. Our findings suggest that numerical abnormalities of the Cyclin D1 gene locus could result in an altered gene-dose effect, possibly leading to an aberrant expression in affected tumor cells. This might result in deregulation of cell cycle control, eventually leading to uncontrolled cell cycle progression.

Expression of c-myc and bcl-2 in primary and advanced cutaneous melanoma.

Apoptosis is an important co-factor in the pathogenesis of a plethora of malignancies. Enhanced c-myc activation can result either in proliferation or apoptosis. Coexpression with antiapoptotic bcl-2, which abrogates the apoptotic function of c-myc might lead to an enormous growth advantage of cells. In order to elucidate the role of c-myc and bcl-2 as well as the coexpression of both genes in human melanoma, their expression was studied in four samples of normal skin (SK), 15 surgical margins (SM), 20 benign melanocytic nevi (MN), 20 primary melanomas (MM), and 30 melanoma metastases (MMET) by RT-PCR. These results were compared with immunohistochemistry (IH) in 7 SK, 7 SM, 26 MN, 50 MM, and 34 MMET. Similar results were found with both methods. However, MMET expressed c-myc (PCR 28/30, IH 23/34) as well as bcl-2 (PCR 27/30, IH 24/34) more frequently. Primary melanomas showed a similar expression pattern as SM and nevi. Moreover, in contrast to SK, SM, MN, and MM coexpression of bcl-2 and c-myc was found more frequently in MMET (PCR 25/30, p < 0.01, IH 19/34, p < 0.01). These results indicate that coexpression of c-myc and bcl-2 appears to be associated with advanced melanoma and contributes to the malignant phenotype.