Capillary and arteriolar pericytes attract innate leukocytes exiting through venules and 'instruct' them with pattern-recognition and motility programs.

Coordinated navigation within tissues is essential for cells of the innate immune system to reach the sites of inflammatory processes, but the signals involved are incompletely understood. Here we demonstrate that NG2(+) pericytes controlled the pattern and efficacy of the interstitial migration of leukocytes in vivo. In response to inflammatory mediators, pericytes upregulated expression of the adhesion molecule ICAM-1 and released the chemoattractant MIF. Arteriolar and capillary pericytes attracted and interacted with myeloid leukocytes after extravasating from postcapillary venules, 'instructing' them with pattern-recognition and motility programs. Inhibition of MIF neutralized the migratory cues provided to myeloid leukocytes by NG2(+) pericytes. Hence, our results identify a previously unknown role for NG2(+) pericytes as an active component of innate immune responses, which supports the immunosurveillance and effector function of extravasated neutrophils and macrophages.

Frequency of translocation of USP-6 in the aneurysmal bone cyst of the jaw.

BACKGROUND: This study aims to detect USP6 translocation in aneurysmal cysts located in the jaw and to give an overview of demographic data. METHODS: The present retrospective cohort study includes 10 patients who underwent surgery due to an aneurysmal cyst of the jaw in our hospital between 2002 and 2021. Unstained formalin-fixed and paraffin-embedded tissue sections cut at 4 µm thickness were subjected to USP6 FISH testing. RESULTS: All patients underwent surgical treatment. In four of ten patients (40%) USP-6-translocations have been found. CONCLUSION: Based on the study, it is hypothesized that the aneurysmal bone cyst of the jaw bone may be subject to a different pathomechanism than that of the long bones. Therefore, it seems conceivable that the primary cause of aneurysmal bone cysts in the jaw might differ.

Accuracy of maxillary positioning using computer-designed and manufactured occlusal splints or patient-specific implants in orthognathic surgery.

OBJECTIVE: To determine the accuracy of maxillary positioning using computer-designed and manufactured occlusal splints or patient-specific implants in orthognathic surgery. MATERIAL AND METHODS: A retrospective analysis of 28 patients that underwent virtually planned orthognathic surgery with maxillary Le Fort I osteotomy either using VSP-generated splints (n = 13) or patient-specific implants (PSI) (n = 15) was conducted. The accuracy and surgical outcome of both techniques were compared by superimposing preoperative surgical planning with postoperative CT scans and measurement of translational and rotational deviation for each patient. RESULTS: The 3D global geometric deviation between the planned position and the postoperative outcome was 0.60 mm (95%-CI 0.46-0.74, range 0.32-1.11 mm) for patients with PSI and 0.86 mm (95%-CI 0.44-1.28, range 0.09-2.60 mm) for patients with surgical splints. Postoperative differences for absolute and signed single linear deviations between planned and postoperative position were a little higher regarding the x-axis and pitch but lower regarding the y- and z-axis as well as yaw and roll for PSI compared to surgical splints. There were no significant differences regarding global geometric deviation, absolute and signed linear deviations in the x-, y-, and z-axis, and rotations (yaw, pitch, and roll) between both groups. CONCLUSIONS: Regarding accuracy for positioning of maxillary segments after Le Fort I osteotomy in orthognathic surgery patient-specific implants and surgical splints provide equivalent high accuracy. CLINICAL RELEVANCE: Patient-specific implants for maxillary positioning and fixation facilitate the concept of splintless orthognathic surgery and can be reliably used in clinical routines.

Cancer testis antigen (PRAME) as an independent marker for survival in oral squamous cell carcinoma (OSCC).

BACKGROUND: The objective was to assess the expression patterns of the cancer testis antigen PRAME, NY-ESO1, and SSX2 in oral squamous cell carcinoma (OSSC) and to correlate the expression with clinical and histopathological parameters including progression-free survival analysis. METHODS: The study variables of this retrospective cohort study (n = 83) included demographic data, histopathological data, and information on progression-free survival. PRAME expression patterns were rated based on immunohistochemistry on tissue microarrays (TMA). The survival rate was assessed by Kaplan-Meier method and Cox regression model. The primary predictor variable was defined as the expression of PRAME and the outcome variable was progression-free survival. RESULTS: Analysis of progression-free survival using Kaplan-Meier method showed that patients with positive expression of PRAME had lower probabilities of progression-free survival (p < 0.001). According to the Cox regression model, the level of PRAME expression had a considerable and significant independent influence on progression-free survival (positive PRAME expression increasing the hazards for a negative outcome by 285% in our sample; HR = 3.85, 95% CI: 1.45-10.2, p = 0.007). The expression of SSX2 (n = 1) and NY-ESO-1 (n = 5) in our samples was rare. CONCLUSION: PRAME is expressed in OSCC and appears to be a suitable marker of progression-free survival, correlates with severe course, and may allow identification of high-risk patients with aggressive progression.

Immunohistochemical expression of CD44 in oral squamous cell carcinoma in relation to histomorphological parameters and clinicopathological factors.

AIMS: Oral squamous cell carcinoma (OSCC) is characterised by its variable clinical course. In addition to the routinely used TNM and Union for International Cancer Control systems, patient-specific prognostic/predictive biomarkers are needed. Promising biomarkers include the determination of the cancer stem cell compartment, which can be identified by CD44 expression (among other things). The aim of this study was to evaluate the impact of CD44 in OSCC in terms of correlation with histomorphology, especially targeting features of EMT, and its influence on patient prognosis. METHODS AND RESULTS: A well-characterised cohort of 108 therapy-naive OSCCs with complete long-term follow-up and matched lymph node metastases were evaluated for CD44 expression by immunohistochemistry. CD44 expression was correlated with histomorphological characteristics (including tumour differentiation and tumour budding), clinicopathological parameters, and follow-up data. Overexpression of CD44 was detected in 37% of OSCCs within the tumour centre, in 39% of OSCCs at the invasive margin, and in 16% of lymph node metastases. CD44 overexpression at the invasive margin was significantly correlated with poor histopathological differentiation, and specifically with high tumour budding activity and single-cell invasion as signs of epithelial-mesenchymal transition (EMT). CD44 overexpression within the tumour core region and in lymph node metastases was identified as an independent prognostic factor for poor overall, disease-specific and disease-free survival in subsets of patients with advanced OSCC. CONCLUSION: Our study demonstrates the association of CD44 with tumour aggressiveness and EMT, as well as the independent prognostic impact of CD44 in a subset of OSCCs, which underlines the role of tumour cell stemness as a key factor in malignant behaviour in this disease.

What Factors Are Associated With Regional Recurrence After Operative Treatment of Oral Squamous Cell Carcinoma?

PURPOSE: A considerable proportion of oral squamous cell carcinoma (OSCC) recurrences involve the neck, which has a substantial impact on prognosis and is poorly understood. The purpose of this study was to analyze clinical and pathologic characteristics of regional recurrence of OSCC and identify possible risk factors. MATERIALS AND METHODS: A single-center retrospective cohort study was designed to address the study purpose. All patients who were treated surgically for primary OSCC with or without adjuvant therapy between 2010 and 2015 were considered for inclusion with respect to predefined criteria, and demographic, clinical, and pathologic variables were collected. The lymph node status after primary OSCC treatment (pN) was defined as the predictor variable, and the occurrence of regional recurrence served as the primary outcome variable. Further variables of special interest were the histologic differentiation (G status) of the primary OSCC and the values of the lymph node ratio and log odds of positive lymph nodes. Descriptive, inferential, and appropriate time-dependent (Kaplan-Meier analysis, Cox regression model) statistics were computed. The level of statistical significance was set at P ≤ .05. RESULTS: The study sample was composed of 171 patients (70 women and 101 men; average age, 62.4 years). Neck failure occurred in 18 patients (11%) either in combination with local recurrence (15 patients) or in isolation (3 patients). Poor histologic differentiation of the primary tumor was identified as an independent risk factor for regional recurrence. Most neck recurrences manifested in previously unaddressed levels (IV and V). CONCLUSIONS: Regional recurrence of OSCC might be associated with specific clinicopathologic parameters of the primary tumor. The importance of these parameters for OSCC prognosis assessment and recurrence prediction should be elucidated in further studies.

Lymph nodal recurrence in levels IV and V in oral squamous cell carcinoma after neck dissection.

BACKGROUND: The aim of the study was to determine the risk of lymph node recurrence in levels IV and V after tumour resection and neck dissection of level I-III and level I-V. METHODS: Data from 228 patients suffering from OSCC were analysed retrospectively. Patients with level I-III neck dissection were compared to those with level I-V neck dissection in terms of number and location of nodal recurrence. The incidence of level IV-V recurrence in patients who had received level I-III neck dissection was compared with that of patients who had received level I-V neck dissection. The incidence of level IV-V recurrence was also compared between patients with pN0 and pN+ necks. RESULTS: Overall, 19 patients developed metastases. Only in two cases appeared nodal recurrence in levels IV or V. There was no statistically significant difference between both groups. CONCLUSIONS: Neck dissection of levels I-III seems to be sufficient treatment in cases of unsuspicious lymph nodes in levels IV and V, even in cases of positive nodes in levels I-III if adjuvant radiation therapy is applied. However, 5-year-disease free survival rate is lower in patients with nodal metastases in levels IV and V than in patients with metastases located in levels I-III.

Comparison of alternative N-staging models in patients with oral cancer. Is nodal yield or burden the most critical parameter?

BACKGROUND: There is increasing evidence in the literature that alternative N-classification systems offer a simpler and more precise risk stratification than the current N- classification in patients with oral squamous cell carcinoma. The purpose of this study is to compare three broadly proposed models incorporating lymph node ratio, log odds of positive lymph nodes and number of positive lymph nodes regarding disease-free and overall survival. METHODS: This is a retrospective study of patients treated in a single center between 2013 and 2019. Cox proportional hazard models, Kaplan Meier curves, the long rank test and the area under the curve were implemented to compare the risk-stratification ability of the three models. Moreover, a hazard ratio plot was calculated to investigate the association between nodal yield and disease-free survival. RESULTS: 231 patients were included. All three proposed N-models were significantly correlated to the patient's prognosis in the multivariate analysis. Pairwise comparisons between the groups showed that the current pN classification offers the worst stratification and that the model incorporating the number of positive lymph nodes had a better performance for predicting both endpoints. Finally, a decrease in hazard ratio was observed with each additional lymph node removed up to the number of 39 lymph nodes. CONCLUSIONS: The model incorporating the nodal burden offered a better predictive ability. The current N-classification cannot adequately identify patients with different prognosis. A non-linear relationship was found between nodal yield and survival, which could be the disadvantage of the lymph node ratio and log odds models.

Desmoglein-3 overexpression in oral squamous cell carcinoma is associated with metastasis formation and early recurrence: An immunohistochemical study.

The purpose of this study was to determine the expression patterns of specific desmosomal cadherins (desmogleins [DSG] 1/2/3) in oral squamous cell carcinoma (OSCC), and to examine possible associations with clinicopathological parameters and recurrence rates. Changes in desmosomal cadherin assembly may promote tumor metastasis formation. Patients with surgically treated OSCC with 36-60 months of follow-up (median 46 months) qualified for inclusion in this retrospective cohort study. Demographic, clinical and pathohistological data were collected. DSG-1/2/3 expression patterns were determined by an immunohistochemical approach on tissue microarrays. Descriptive and inferential statistics and survival analyses were computed (p ≤ 0.05). The study sample consisted of 88 patients (female: 38; male: 50; average age: 63.02 ± 17.5 years). DSG-3 overexpression was detected in 45 of 88 specimens. The expression rates for DSG-1 (28/88) and DSG-2 (14/88) were low and inconspicuous. DSG-3 overexpression was significantly associated with poor histologic differentiation (G3, p = 0.001), the presence of cervical node metastasis at primary diagnosis (N+ status, p = 0.001) and early recurrence (p = 0.001). Due to its possible relevance for lymph node metastasis formation and early OSCC recurrence, determination of DSG-3 expression in OSCC specimens may be a valuable tool for treatment planning and post-therapeutic risk assessment.

Nodal Disease and Survival in Oral Cancer: Is Occult Metastasis a Burden Factor Compared to Preoperatively Nodal Positive Neck?

The impact of neck involvement and occult metastasis (OM) in patients with oral squamous cell carcinoma (OSCC) favors an elective neck dissection. However, there are barely any existing data on survival for patients with OM compared with patients with positive lymph nodes detected preoperatively. This study aims to compare survival curves of patients suffering from lymph nodal metastases in a preoperatively N+ neck with those suffering from OM. In addition, clinical characteristics of the primary tumor were analyzed to predict occult nodal disease. This retrospective cohort study includes patients with an OSCC treated surgically with R0 resection with or without adjuvant chemoradiotherapy between 2010 and 2016. Minimum follow-up was 60 months. Kaplan-Meier analysis was used to compare the survival between patients with and without occult metastases and patients with N+ neck to those with occult metastases. Logistic regression was used to detect potential risk factors for occult metastases. The patient cohort consisted of 226 patients. Occult metastases occurred in 16 of 226 patients. In 53 of 226 patients, neck lymph nodes were described as suspect on CT imaging but had a pN0 neck. Higher tumor grading increased the chance of occurrence of occult metastasis 2.7-fold (OR = 2.68, 95% CI: 1.07-6.7). After 12, 24, 48 and 60 months, 82.3%, 73.8%, 69% and 67% of the N0 patients, respectively, were progression free. In the group with OM occurrence, for the same periods 66.6%, 50%, 33.3% and 33.3% of the patients, respectively, were free of disease. For the same periods, respectively, 81%, 63%, 47% and 43% of the patients in the N+ group but without OM remained disease free. The predictors for progression-free survival were a positive N status (HR = 1.44, 95% CI: 1.08-1.93) and the occurrence of OM (HR = 2.33, 95% CI: 1.17-4.64). The presence of occult metastasis could lead to decreased survival and could be a burdening factor requiring treatment escalation and a more aggressive follow-up than nodal disease detected in the preoperative diagnostic imaging.

Digital planning and individual implants for secondary reconstruction of midfacial deformities: A pilot study.

Objective: To evaluate the feasibility and accuracy of implementing three-dimensional virtual surgical planning (VSP) and subsequent transfer by additive manufactured tools in the secondary reconstruction of residual post-traumatic deformities in the midface. Methods: Patients after secondary reconstruction of post-traumatic midfacial deformities were included in this case series. The metrical deviation between the virtually planned and postoperative position of patient-specific implants (PSI) and bone segments was measured at corresponding reference points. Further information collected included demographic data, post-traumatic symptoms, and type of transfer tools. Results: Eight consecutive patients were enrolled in the study. In five patients, VSP with subsequent manufacturing of combined predrilling/osteotomy guides and PSI was performed. In three patients, osteotomy guides, repositioning guides, and individually prebent plates were used following VSP. The median distances between the virtually planned and the postoperative position of the PSI were 2.01 mm (n = 18) compared to a median distance concerning the bone segments of 3.05 mm (n = 12). In patients where PSI were used, the median displacement of the bone segments was lower (n = 7, median 2.77 mm) than in the group with prebent plates (n = 5, 3.28 mm). Conclusion: This study demonstrated the feasibility of VSP and transfer by additive manufactured tools for the secondary reconstruction of complex residual post-traumatic deformities in the midface. However, the median deviations observed in this case series were unexpectedly high. The use of navigational systems may further improve the level of accuracy.

Accuracy of free-hand positioned patient specific implants (PSI) in primary reconstruction after inferior and/or medial orbital wall fractures.

BACKGROUND: To assess the accuracy with which CAD/CAM-fabricated patient-specific titanium implants (PSI) are positioned for inferior and/or medial orbital wall reconstruction without the use of intraoperative navigation. METHODS: Patients who underwent a primary reconstruction of the orbital walls with PSI due to fractures were enrolled in this retrospective cohort analysis. The primary outcome variables were the mean surface distances (MSD) between virtually planned and postoperative PSI position and single linear deviations in the x-, y- and z-axis at corresponding reference points. Secondary outcome variables included demographic data, classification of orbital wall defects and clinical outcomes. RESULTS: A total of 33 PSI (orbital floor n = 22; medial wall, n = 11) were examined in 27 patients. MSD was on a comparable level for the orbital floor and medial wall (median 0.39 mm, range 0.22-1.53 mm vs. median 0.42 mm, range 0.21-0.98 mm; p = 0.56). Single linear deviations were lower for reconstructions of the orbital floor compared to the medial wall (median 0.45 vs. 0.79 mm; p < 0.05). There was no association between the occurrence of diplopia and the accuracy level (p = 0.418). CONCLUSIONS: Free-hand positioning of PSI reaches a clinically appropriate level of accuracy, limiting the necessity of navigational systems to selected cases.

Autofluorescence-guided surgery for the treatment of medication-related osteonecrosis of the jaw (MRONJ): a retrospective single-center study.

OBJECTIVE: Medication-related osteonecrosis of the jaw (MRONJ) has become a serious concern for patients under antiresorptive treatment, especially in the oncological setting. Different approaches have been described in the management of MRONJ, including innovative autofluorescence-guided surgery. However, until now, there has been a lack of data regarding the outcome. In this study, we evaluated the efficacy of minimally invasive autofluorescence-guided resection in MRONJ. STUDY DESIGN: Seventy-five patients with 82 lesions were included in this retrospective, single-center study. All included patients were diagnosed with MRONJ according to the American Association of Oral and Maxillofacial Surgeons guidelines and underwent autofluorescence-guided surgery with a minimum follow-up of 3 months. The primary outcome was complete integrity of the mucosa and absence of bone exposure. RESULTS: The MRONJ stages were stage 0 (3.7%), stage 1 (3.7%), stage 2 (75.6%), and stage 3 (17%). Overall, complete mucosal healing of all lesions after the first surgery was 81.7% (67 of 82), whereas it was 90.2% (74 of 82) after revision surgery. CONCLUSIONS: The study showed that autofluorescence-guided surgery is a safe and successful treatment option that can be considered for all stages of MRONJ.

Fatty Acid Receptor CD36 Functions as a Surrogate Parameter for Lymph Node Metastasis in Oral Squamous Cell Carcinoma.

PURPOSE: To investigate the expression pattern of CD36 in a patient population with oral squamous cell carcinoma (OSCC) and to correlate CD36 expression with clinical and histopathological parameters. The hypothesis was that CD36 expression correlates with the occurrence of lymph node metastasis. METHODS: To address the study objectives, a retrospective cohort study was conducted. Study variables included demographic, histopathological and survival data. CD36 expression patterns were assessed by immunohistochemistry on tissue microarrays (TMA). Logistic regression analysis, survival analysis and Cox proportional hazards model were performed. RESULTS: High CD36 expression correlated significantly with a higher T-status, grading and occurrence of lymph node metastasis. The logistic regression with binary N status as a dependent variable showed that high CD36 expression increased the chance for lymph node metastasis 45-fold (OR = 44.7, 95% CI: 10.0-316). Patients with high CD36 expression had lower probabilities of progression-free survival. CD36 had a small and non-significant independent influence on progression-free survival. CONCLUSIONS: CD36 is expressed in OSCC and correlates with tumor grading, T-status, and especially the occurrence of lymph node metastasis. CD36 may be useful for risk stratification regarding lymph node metastasis in OSCC.

Comparative analysis of bone regeneration behavior using recombinant human BMP-2 versus plasmid DNA of BMP-2.

Bone regeneration and the osteoinductive capacity of implants are challenging issues in clinical medicine. Currently, recombinant growth factors and nonviral gene transfer are the most frequently investigated methods for bone growth enhancement, although the more favorable method remains unclear. There is a lack of knowledge in literature about the in vivo comparison of these methods for bone regeneration. BMP-2, which is the most commonly used growth factor for osteogenesis, was applied at its most efficient dose as a recombinant growth factor (rhBMP-2) and as a growth-factor-encoding copolymer protected gene vector (pBMP-2) in a critical size bone defect (CSD) model to determine the most suitable method for bone regeneration. CSDs were induced bilaterally in 32 Sprague-Dawley rats. RhBMP-2 (62.5 µg) or pBMP-2 (2.5 µg) was embedded in poly(d,l-)lactide-coated titanium discs. Survival times were set at 14, 28, 56, and 112 days. After euthanasia, samples were analyzed via micro-computed tomography, polychrome sequential fluorescent labeling, and immunohistochemistry. Whereas defects in both groups were bridged with new bone after 56 days, rhBMP-2 initially induced ectopic new bone formation that was later remodeled in an unorganized hypodense manner. In contrast, pBMP-2 led to slower but steady bone regeneration with physiological tissue morphology, as confirmed by high osteoblast activity shown by osteocalcin staining. CD68 and TRAP staining verified high osteoclast activity for the rhBMP-2 group. pBMP-2 successfully induced locally controlled physiological bone regeneration, whereas rhBMP-2 triggered rapid and ectopic but insufficient bone formation. Thus, nonviral gene transfer appears to be more favorable for clinical applications. © 2018 Wiley Periodicals, Inc. J Biomed Mater Res Part A: 107A: 163-173, 2019.

Functional analysis of bioactivated and antiinfective PDLLA - coated surfaces.

Common scaffold surfaces such as titanium can have side effects; for example, infections, cytotoxicity, impaired osseointegration, or low regeneration rates for bone tissue. These effects lead to poor implant integration or even implant loss. Therefore, bioactive implants are promising instruments in tissue regeneration. Osteoinductive elements-such as growth factors and anti-infectives-support wound healing and bone growth and thereby enable faster osseointegration, even in elderly patients. In this study, titanium surfaces were coated with a poly-(d,l-lactide) (PDLLA) layer containing different concentrations of copolymer-protected gene vectors (COPROGs) to locally provide bone morphogenetic protein-2 (BMP-2) or activated anti-infective agents, such as chlorhexidine gluconate, triclosan, and metronidazole, to prevent peri-implantitis. The coated titanium implants were then loaded with osteoblasts, NIH 3T3 fibroblasts, and human mesenchymal stem cells in 96-well plates. When shielded by COPROGs as a protective layer and resuspended in PDLLA, BMP-2-encoding pDNA at relatively low doses (5.63 µg/implant) induced the local expression of BMP-2. A linear dose dependence, which is common for recombinant growth factors, was not found, probably due to the retention property of the PDLLA surface. PDLLA, in general, successfully retains additional elements, such as osteoconductive growth factors (BMP-2) and anti-infective agents, which was demonstrated using metronidazole, and thus prevents the systemic application of excessive doses. These bioactive implant surfaces that provide the local release of therapeutic gene vectors or anti-infective agents allow the controlled stimulation of the implant and scaffold osseointegration. © 2017 Wiley Periodicals, Inc. J Biomed Mater Res Part A: 105A: 1672-1683, 2017.

Monocytes, neutrophils, and platelets cooperate to initiate and propagate venous thrombosis in mice in vivo.

Deep vein thrombosis (DVT) is a major cause of cardiovascular death. The sequence of events that promote DVT remains obscure, largely as a result of the lack of an appropriate rodent model. We describe a novel mouse model of DVT which reproduces a frequent trigger and resembles the time course, histological features, and clinical presentation of DVT in humans. We demonstrate by intravital two-photon and epifluorescence microscopy that blood monocytes and neutrophils crawling along and adhering to the venous endothelium provide the initiating stimulus for DVT development. Using conditional mutants and bone marrow chimeras, we show that intravascular activation of the extrinsic pathway of coagulation via tissue factor (TF) derived from myeloid leukocytes causes the extensive intraluminal fibrin formation characteristic of DVT. We demonstrate that thrombus-resident neutrophils are indispensable for subsequent DVT propagation by binding factor XII (FXII) and by supporting its activation through the release of neutrophil extracellular traps (NETs). Correspondingly, neutropenia, genetic ablation of FXII, or disintegration of NETs each confers protection against DVT amplification. Platelets associate with innate immune cells via glycoprotein Ibα and contribute to DVT progression by promoting leukocyte recruitment and stimulating neutrophil-dependent coagulation. Hence, we identified a cross talk between monocytes, neutrophils, and platelets responsible for the initiation and amplification of DVT and for inducing its unique clinical features.