Henning Hanken

Prognostic Relevance of Circulating Tumor Cells in Blood and Disseminated Tumor Cells in Bone Marrow of Patients with Squamous Cell Carcinoma of the Oral Cavity

Purpose: Current staging methods for squamous cell carcinomas (SCC) of the oral cavity (OSCC) need to be improved to predict the risk of individual patients. Because hematogenous tumor cell dissemination is a key event in tumor progression, we assessed the prognostic significance of disseminated tumor cells (DTC) in bone marrow and circulating tumor cells (CTC) in peripheral blood from patients with OSCC. Experimental Design: From 110 patients with OSCC, tumors were surgically resected (R0) without neoadjuvant therapy. The CellSearch system was used to enumerate CTCs. Bone marrow was aspirated from the iliac crest, and mononuclear cells (MNC) were enriched by Ficoll density gradient centrifugation. To detect DTCs, MNCs were immunostained with the pan-keratin antibody A45-B/B3. Results were correlated with clinicopathologic parameters and clinical outcome such as recurrence and death during follow-up time (mean 916 days). Results: Ten of 80 patients (12.5%) harbored CTCs in peripheral blood, whereas in 18 of 90 patients (20.0%) DTCs in bone marrow could be detected. Surprisingly, in only 2 patients (1.8%) CTCs and DTCs were detected simultaneously. Significant correlations could be found for CTCs and tumor size (P = 0.04), nodal status and DTCs (P = 0.02), and distant metastasis with CTCs (P = 0.004) and DTCs (P = 0.005). Univariate and multivariate analyses revealed that CTCs and DTCs were significant and independent predictors of recurrence-free survival (P < 0.001). Conclusions: Both DTCs and CTCs are independent prognostic markers in patients with OSCC, predicting relapse with higher sensitivity at various disease stages than routine staging procedures. Bone marrow might be an interesting target organ for future therapeutic interventions. Clin Cancer Res; 20(2); 425–33. ©2013 AACR.

The detection of oral pre- malignant lesions with an autofluorescence based imaging system (VELscopeTM) – a single blinded clinical evaluation

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Optimized in vitro procedure for assessing the cytocompatibility of magnesium-based biomaterials.

Magnesium (Mg) is a promising biomaterial for degradable implant applications that has been extensively studied in vitro and in vivo in recent years. In this study, we developed a procedure that allows an optimized and uniform in vitro assessment of the cytocompatibility of Mg-based materials while respecting the standard protocol DIN EN ISO 10993-5:2009. The mouse fibroblast line L-929 was chosen as the preferred assay cell line and MEM supplemented with 10% FCS, penicillin/streptomycin and 4mM l-glutamine as the favored assay medium. The procedure consists of (1) an indirect assessment of effects of soluble Mg corrosion products in material extracts and (2) a direct assessment of the surface compatibility in terms of cell attachment and cytotoxicity originating from active corrosion processes. The indirect assessment allows the quantification of cell-proliferation (BrdU-assay), viability (XTT-assay) as well as cytotoxicity (LDH-assay) of the mouse fibroblasts incubated with material extracts. Direct assessment visualizes cells attached to the test materials by means of live-dead staining. The colorimetric assays and the visual evaluation complement each other and the combination of both provides an optimized and simple procedure for assessing the cytocompatibility of Mg-based biomaterials in vitro.

Leech Therapy in Reconstructive Maxillofacial Surgery

PURPOSE Corrective plastic surgery is indicated after accidents, burns, cancer surgery, or postoperative wound healing disorders with large tissue defects. The range of reconstructive techniques includes local skin flaps, pedicle grafts, and microvascular anastomosed flaps in the case of large defects. Main complications such as arterial and venous insufficiency caused by a vessel collapse or a vascular spasm are reported regularly in the area of anastomosed vessels and are the concern of any surgeon. Today, leeches are used if wound healing is at risk because of hemodynamic imbalance or a venous insufficiency. PATIENTS AND METHODS A retrospective evaluation of 148 patients who underwent medical leech therapy in the case of local or pedicaled flaps and some patients who had undergone reconstruction with microvascular flaps from 2005 and 2010 was conducted. Our sample had the typical symptoms of venous congestions of their flaps, despite suture removal, relief of pressure on the flap, and the elimination of a hematoma beyond the flap after surgery. Medical leech therapy was used in these cases. RESULTS Our series has confirmed the excellent and predictable healing after medical leech therapy for local and microsurgical anastomosed flaps in the case of venous congestion. CONCLUSION Leech therapy should be considered as a reliable additional procedure and an advantage in maxillofacial and plastic reconstructive surgery to remedy complications resulting from a hemodynamic imbalance or venous insufficiency in the immediate postoperative period.

The use of buccal fat pad (BFP) as a pedicled graft in cleft palate surgery

The buccal fat pad (BFP) as pedicled graft was originally used in reconstructing medium sized intraoral defects. Promising results concerning the use of BFP in cleft palate surgery have been published recently. The aim of this article is to report on the use of BFP as a pedicled graft in cleft palate surgery and to discuss promising results for this reconstructive surgical concept. A retrospective evaluation of 24 patients who had BFP pedicled flaps used for the prevention and repair of Type III (Pittsburgh Fistula Classification) cleft palate fistulas, to obstruct the retromolar space of Ernst and in case of wide clefts, from 2005 to 2010, was conducted. In all cleft palate patients, the recipient area fully epithelialized within 4 weeks or less. No recurrence was seen and the donor site healed well without aesthetic or significant functional impairment. This series confirms the excellent and predictable healing of BFP intraorally and the minimal morbidity associated with the use of such grafts. The results of this study allow the authors to recommend that the BFP pedicled flap is considered as a reliable alternative procedure to expand the therapeutic options. The BFP graft provides an advantage in reconstructive cleft palate surgery.

Immunohistochemical analysis of p16 expression, HPV infection and its prognostic utility in oral squamous cell carcinoma.

OBJECTIVES Functional inactivation of p16 is an early and frequent event in the carcinogenesis of tumours of the head and neck region. However, the prognostic relevance of p16 protein expression for these tumours has been controversial. This study aims to examine immunohistochemical expression of p16 and HP virus in a large number of oral carcinomas, and possible correlation with clinical features of the tumours and survival of the patients. METHODS Two tissue microarrays composed of 222 oral carcinomas and 427 squamous cell carcinomas of the head and neck region were used for this study. Sections were stained immunohistochemically (anti-p16), and PCR analysis (HPV status) was carried out. Correlation of p16 expression/HPV status with features of tumours and with survival of the patients was analysed by means of Chi-squared test and using Kaplan-Meier analysis, respectively. RESULTS p16 expression was found immunohistochemically in 74% of tumours, but was not significantly correlated with features of the tumours, but recurrence-free survival of the patients (P = 0.009) if located predominantly nuclear. On the other hand neither intensity of p16 expression (P = 0.41) nor HPV status (P = 0.82) had any effect on these two aspects. CONCLUSION Immunohistochemical expression of p16 alone provides a limited tool for diagnosis and prognosis of carcinomas of the head and neck region. Immunohistochemical analysis of p16 depending on its intracellular location might serve as a surrogate marker for HPV infection.

Her2 expression and gene amplification is rarely detectable in patients with oral squamous cell carcinomas

PURPOSE Her2 (ErbB2) transforms cells when overexpressed and is an important therapeutic target in breast cancer. Contrary to breast cancer, studies on Her2 overexpression and gene amplification in squamous cell carcinomas of the head and neck region described largely different results. This study was undertaken to learn more on the prevalence and clinical significance of HER2 amplification and overexpression in squamous cell carcinomas of the head and neck. MATERIALS AND METHODS Her2 expression and gene amplification was analyzed by immunohistochemistry (IHC) and fluorescence in situ hybridization (FISH) on two tissue microarrays composed of 427 squamous cell carcinomas of the head and neck region and 222 oral squamous cell carcinomas. Results were compared with clinicopathological features. RESULTS Her2 expression and gene amplification was rarely detectable in squamous cell carcinomas of the head and neck region and unrelated to tumor phenotype or survival of the patients with oral squamous carcinoma. DISCUSSION Our results demonstrate that Her2 protein and gene amplification was only detectable in a small subset of squamous cell carcinomas of the head and neck region as well as oral squamous cell carcinomas. However, it can be speculated that those few patients with Her2 overexpressing and gene amplificated tumors may possibly benefit from an anti-Her2 therapy.

Face Transplantation: On the Verge of Becoming Clinical Routine?

Introduction. Face transplantation (FT) is an innovative achievement of modern reconstructive surgery and is on the verge of becoming a common surgical opportunity. This review article was compiled to provide an update on this surgical field, especially regarding clinical outcomes, benefits, and complications implied. Methods. We performed an extensive research on all English-language Medline articles, case reports, and reviews published online until September 15, 2013. Used search terms were “face transplantation,” “face transplant,” “facial transplantation,” “facial transplant,” “face allograft,” and “facial allograft.” Results. To date 27 FTs have been performed worldwide. 19 of these cases have been published in the Medline database. Long-term follow-up reports of FT cases are rare. Three deaths associated with the procedure have occurred to date. The clinical outcomes of FT are satisfying. Reinnervation of sensation has been faster than motor recovery. Extensive functional improvements have been observed. Due to strict immunosuppression protocols, no case of hyperacute or chronic rejection and no graft-versus-host disease have occurred to date. Conclusions. As studies on long-term outcomes are missing, particularly regarding immunosuppression-related complications, FT will stay experimental for the next years. Nevertheless, for a small group of patients, FT already is a feasible reconstructive option.

Intraosseous heat generation during sonic, ultrasonic and conventional osteotomy.

OBJECTIVES To assess heat generation in osteotomies during application of sonic and ultrasonic saws compared to conventional bur. METHODS Two glass-fiber isolated nickel-chromium thermocouples, connected to a recording device, were inserted into fresh bovine rib bone blocks and kept in 20 ± 0.5 °C water at determined depths of 1.5 mm (cortical layer) and 7 mm (cancellous layer) and 1.0 mm away from the planned osteotomy site. Handpieces, angulated 24-32°, were mounted in a vertical drill stand, and standardized weights were attached to their tops to exert loads of 5, 8, 15 and 20 N. Irrigation volumes of 20, 50 and 80 ml/min were used for each load. Ten repetitions were conducted using new tips each time for each test condition. The Mann-Whitney-U test was used for statistical analysis (p < 0.05). RESULTS Both ultrasonic and sonic osteotomies were associated with significantly lower heat generation than conventional osteotomy (p < 0.01). Sonic osteotomy showed non-significantly lower heat generation than ultrasonic osteotomy. Generated heat never exceeded the critical limit of 47 °C in any system. Variation of load had no effect on heat generation in both bone layers for all tested systems. An increased irrigation volume resulted in lower temperatures in both cortical and cancellous bone layers during all tested osteotomies. CONCLUSION Although none of the systems under the conditions of the present study resulted in critical heat generation, the application of ultrasonic and sonic osteotomy systems was associated with lower heat generation compared to the conventional saw osteotomy. Copious irrigation seems to play a critical role in preventing heat generation in the osteotomy site.

Outcome of microvascular free flaps in a high-volume training centre

PURPOSE Microvascular free tissue transfer allows major ablative defects following oncologic surgical and traumatic reasons to be reliably reconstructed in the head and neck region. A retrospective analysis of the microvascular flap procedures which were performed within one year in a high volume training centre was performed. PATIENTS AND METHODS The microvascular free flap procedures of the year 2011 were reviewed and followed up until the 31st December 2012. The type and indication of the reconstructive procedure, operation time, operating team, experience and level of training of the surgeons involved, postoperative IMC (intermediate care unit) and/or ICU (intensive care unit) time, inpatient time, flap revisions, further postoperative complications, preoperative and postoperative radiation of the patients, the placement of dental implants were studied. RESULTS From 1st of January 2011 to 31st of December, 2011, the data of 101 patients with 103 microvascular free flap procedures were analysed of which 72% (84 flaps) were harvested by residents. The patients ranged in age from 14 to 89 years (mean age 59 years, 71 males and 40 females). The mean operation time was 591 min with the longest operation times for scapular flaps (744 min) and the shortest operation times for ALT flaps (455 min). Mean inpatient time was 34.2 days with a minimal time for the fibular flaps of 27.2 days and a maximum of 45.7 days for the latissimus dorsi flaps. 24 flaps (23.3%) in total had to be revised with bleeding being the main cause of immediate revisions (41.7% of all revisions). 5 flaps (4.85% of all flaps) were lost despite a revision procedure meaning a successful revision rate in 79.2% of all revisions. CONCLUSION Microvascular reconstruction procedures are safe and should be considered as standard procedures for reconstruction of large defects especially in high volume training centres. Intensive flap monitoring and early revisions maximize the flap outcome.