The beneficial effect of cold atmospheric plasma on parameters of molecules and cell function involved in wound healing in human osteoblast-like cells in vitro.

The aim of this study was to analyse the effect of cold atmospheric plasma (CAP) on human osteoblast-like cells in vitro. Additionally, underlying intracellular mechanisms were to be studied. Human osteoblast-like (MG63) cells were exposed to CAP for 60 s. The effects of CAP on key molecules essential for the wound healing response were studied using real-time PCR, ELISA and immunocytochemistry. For studying intracellular signalling pathways, MAP kinase MEK 1/2 was blocked. Cell viability was analysed by an XTT assay and with an EVE automated cell counter. Cell migration was examined by an in vitro wound healing assay.CAP exposition on osteoblast-like cells caused a significant upregulation of interleukin (IL)-1ß, IL-6, IL-8, tumor necrosis factor (TNF)α, cyclooxygenase (COX)2, collagen (COL) 1α, matrix metalloproteinase (MMP)1, Ki67, proliferating-cell-nuclear-antigen (PCNA) and chemokine ligand (CCL)2 mRNA expression at 1 day. Interestingly, after blocking of MAP kinase, CAP-induced upregulation of Ki67 was inhibited by 57%. Moreover, CAP treatment improved significantly osteoblast-like cell viability as compared to untreated cells at 1 day. Beneficial effect of CAP treatment was shown by an in vitro wound healing assay, displaying a significant faster wound closure. Our findings provide evidence that CAP exposure effects gene and protein regulation in human osteoblast-like cells. Furthermore, CAP treatment has a positive impact on wound closure in an in vitro setting and might improve existing concepts of hard tissue regeneration in the future.

Interactions of human MSC with head and neck squamous cell carcinoma cell line PCI-13 reduce markers of epithelia-mesenchymal transition.

OBJECTIVES: Cancer progression is influenced by tumor microenvironment and communication of stromal cells and tumor cells. Interactions may enhance epithelial-mesenchymal transition (EMT) of tumor cells through signaling proteins such as Wnt/beta-catenin and matrix metalloproteinases (MMP), as well as loss of cellular integrity, which affects invasion, progression, and metastasis of head and neck squamous cell carcinoma (HNSCC). In this study, we are testing the hypothesis that interactions of human mesenchymal stromal cells (MSCs) with HNSCC might influence the expression of markers of EMT and tumor progression by co-culturing human MSC with the PCI-13 HNSCC line. MATERIALS AND METHODS: Pooled MSCs were derived from the iliac bone marrow of seven patients and co-cultured in transwell permeable membrane wells with tumor cells of the established HNSCC cell line PCI-13 (UICC: T3, N1, M0). MSCs were characterized through fluorescence-activated cell sorting (FACS) analysis. Expression of Wnt3, E-cadherin, beta-catenin, MMP14, cathepsin b, and ETS1 was assessed by quantitative RT-PCR. RESULTS: We were able to show that co-culture of MSCs and PCI-13 leads to a significantly reduced expression of Wnt3, MMP14, and beta-catenin compared to controls, whereas the expression of cathepsin b and ETS1 was not significantly different between co-cultures and controls. CONCLUSION: Our results suggest that the interaction between MSCs and PCI-13 may suppress EMT in cancer cells. CLINICAL RELEVANCE: The influence of MSCs can suppress the onset of EMT in HNSCC, affecting tumor progression and therapy.

Retrospective study on the pharyngeal airspace in the lateral cephalogram - A mathematical model to predict changes due to bimaxillary orthognathic surgery.

The aim of this study was the development of a statistical model for reliable prediction of Posterior Airway Space (PAS) changes in lateral cephalograms (LCR) of patients after bimaxillary orthognathic surgery. The LCRs of patients who underwent bimaxillary orthognathic surgery were retrospectively analyzed. The anteroposterior dimension of the PAS was measured at three levels in the pre-operative and postoperative LCR: On the nasopharyngeal (SPAS), oropharyngeal (MAS), and hypopharyngeal level (IAS). The data of 139 patients were collected. The following changes of the PAS were measured: in class II patients SPAS: 0.291 mm (SD = 2.570 mm); MAS: 2.444 mm (SD = 2.986 mm); IAS: 0.750 mm (SD = 3.017 mm); in class III patients SPAS: 1.377 mm (SD 3.212 mm); MAS: 0.962 (SD: = 3.135 mm); IAS: 0.370 mm (SD = 3.468 mm). Linear regression analysis showed for class II patients, a significant influence of mandibular movement on MAS (p = 0.049) and a significant effect of maxillary and mandibular movements on SPAS (p = 0.001) and MAS (p = 0.022) in class III patients. The other jaw displacements had no significant impact on the investigated PAS levels. While the presented method does not permit exact prediction of the dimension of the PAS, it is still an easily accessible method of orientation for the surgeon. The surgeon can initiate three-dimensional examinations to provide exact three-dimensional prediction based on this calculation.

A novel serum-free medium for the isolation, expansion and maintenance of stemness and tissue-specific markers of primary human periodontal ligament cells.

PURPOSE: Periodontal ligament (PDL) cell cultures are classically maintained in serum-containing media. However, unwanted side-effects of these conditions on cellular and molecular characteristics demand a serum-free alternative. Even though these limitations are well known and efforts for the development of adequate serum-free alternatives have been made, these approaches for replacement remained unsuccessful so far. This study aimed at developing a well-defined, serum-free formulation supporting both isolation from tissue samples and efficient expansion of PDL cells. Here, of particular focus was the perpetuation of tissue-characteristic markers detectable in primary tissues and of stemness features. BASIC PROCEDURES: Primary PDL cell cultures from generally healthy human donors (n = 3) were maintained in basal media N2B27 and E6 together with different concentrations of growth and attachment factors. Cell proliferation was recorded via microscopy and WST assay. Gene expression of RUNX2, Periostin, ALP, CD73, CD90, CD105, CD45, SOX10 and SOX2 was compared to primary PDL explants via qRT-PCR. Immunocytochemistry was performed for anti-CD105, SSEA-3, CD271, HNK1. Serum-containing sDMEM medium served as control. MAIN FINDINGS: N2B27 medium substituted with 25 ng/mL EGF, 25 ng/mL IGF1, 0.5 mg/mL Fetuin plus gelatine coating (designated N2B27-PDLsf) emerged as potent serum-free formulation ensuring adequate culture isolation and expansion. Here, PDL primary tissue signature markers RUNX2 and Periostin remained stable in N2B27-PDLsf compared to controls (229.0-fold ±101.0 and 83.2-fold ±9.6 increase). Additionally, stemness markers ALP and CD105 were significantly upregulated on transcriptional, and CD105 and SOX2 on protein level. PRINCIPAL CONCLUSIONS: This investigation identified a novel serum-free medium for the isolation, and expansion of primary human PDL cells with constantly high proliferation rates. Here, purity and stemness properties are maintained. Thus, N2B27-PDLsf represents a valid replacement for serum-containing media in PDL cultures.

Intermittent administration of the circulating form of human parathyroid hormone (hPTH-1-37) prevents bone loss in ovariectomized rats.

The circulating hormonal form of human parathyroid hormone (hPTH-1-37) has been assessed in vitro as well as in vivo in the ovariectomized rat, a model for postmenopausal osteoporosis. In vitro, hPTH-1-37 induces a dose-dependent cAMP formation and increases vitality as well as alkaline phosphatase activity in UMR106 osteosarcoma cells. Differentiation and proliferation of osteoclasts in rat bone marrow-derived stem cell preparations are decreased. Daily hPTH-1-37 s.c. administration in ovariectomized rats for 60 days results in augmented formation of new bone, in amplified femural bone density, and in thickening of the calvaria.

An analysis of quality of life in 130 families having small children with cleft lip/palate using the impact on family scale.

Most parents are emotionally traumatized when confronted by the birth of a baby with an orofacial cleft (OFC). Affected families may have to compensate for increased financial, social and personal impacts before primary treatment is completed. This study was conducted to identify factors influencing the quality of life (QoL) of families having young children with OFC. A self-administered questionnaire containing the impact on family scale was applied in 130 consecutive families having children with OFC aged between 6 and 24 months. The results were related to the type of cleft and the time of initial diagnosis using non-parametric tests and multivariate correlation analysis (P<0.05). In families having children with isolated cleft lip, financial and social impacts were reduced, but problems in coping were increased when compared to families with children having cleft lip and palate or isolated cleft palate. Total impact was highest in families having children with isolated cleft palate, probably due to later surgery for reconstruction. Prenatal diagnosis of OFC did not reduce the general impact on affected families, but increased the social impact. The relation of certain impacts to distinct types of cleft might allow more tailored support of affected families and improve their QoL.

Reconstruction of orbital wall defects with calcium phosphate cement: clinical and histological findings in a sheep model.

The aim of this pilot study was to investigate the potential of calcium phosphate cement in the treatment of orbital wall defect fractures in an adult sheep model, and to compare this alloplastic material to autologous calvaria split-bone grafts. Clinical, volumetric and histological examinations were carried out of both reconstruction materials. The use of cement made intraoperative corrections easier to perform, and increased the precision of reconstruction of the orbital volume. This material also proved to be osseoconductive. The two materials were used successfully in combination. Regions of most intensive remodelling were the anterior orbital floor and the adjacent orbital rim. The preliminary results of this study demonstrate the potential of calcium phosphate cement as a useful biomaterial in the reconstruction of the anterior orbital region. Further animal and clinical trials are necessary to investigate its ability as a carrier for mediators where bone healing requires influence or support.

Influence of TGF-ß1 on tumor transition in oral cancer cell and BMSC co-cultures.

OBJECTIVES: TGF-ß1 signaling modulates epithelial mesenchymal transitions (EMT) of head and neck squamous cell carcinoma (HNSCC). Bone marrow mesenchymal stromal cells (BMSC) are able to exert a regulating influence on the expression of markers of EMT in HNSCC cells. It was thus the aim of this study to test the hypothesis that TGF-ß1 modulates the interactions of tumor transition between BMSCs and HNSCC, affecting the expression of E-cadherin, Vimentin, Snail, Twist, MMP14 and beta-catenin. Furthermore, we analyzed alterations in the AKT-signaling of tumor and stroma cells. MATERIALS AND METHODS: BMSCs were isolated from iliac bone marrow aspirates and co-cultured in trans-well permeable membrane wells with tumor cells of the established HNSCC cell line PCI-13. Following the induction with TGF-ß1 under serum free conditions the expression of Vimentin and E-Cadherin was assessed via immunofluorescence. A quantitative RT-PCR analysis of tumor transition markers E-cadherin, Vimentin, Snail, Twist, MMP14 and beta-catenin was performed. Changes in AKT-Signaling were identified via protein analysis. RESULTS: In non-induced co-cultures, BMSC were able to suppress Vimentin in PCI-13 as a marker of tumor transition. In TGF-ß1 induced co-cultures PCI-13 significantly increased the expression of Vimentin, Twist, Snail, MMP14, GSK3a, PRAS40, 4E-BP1, and AMPKa compared to monolayer controls. TGF-ß1 co-cultured BMSC demonstrated a significant increase of Snail, PRAS40, mTOR, GSK3a/b, Bad, PDK1 and 4E-BP1. CONCLUSIONS: TGF-ß1 was able to attenuate the modulating influence of BMSC in co-culture and drive the co-culture towards a progressive tumor transition, affecting the expression of markers of EMT, AKT-Signaling and proliferative checkpoints.

Ultrasonic surgery--an alternative way in orthognathic surgery of the mandible. A pilot study.

The aim of this report is to present preliminary results and experiences using an ultrasonic bone-cutting device in bilateral sagittal split osteotomies of the mandible (BSSRO) with particular attention to possible damages to the inferior alveolar nerve (IAN). Seven patients with class II or class III malocclusion were treated by BSSRO with a conventional combined orthognathic and surgical approach. The osteotomy was carried out using an ultrasonic bone-cutting device. Subjective neurosensory deficits of the inferior alveolar nerve were assessed on 14 sides. Compared to the conventional techniques using saws, chisels and burs, the use of the ultrasonic device was more time-consuming, but the osteotomies were carried out at a high level of precision. In addition, this procedure offered the advantage of a blood-free surgical field and thus provided good control of the surgical procedure. Subjective neurosensory disturbances of the IAN showed a continuous decrease from 57.1% (eight sides) 2 months after the surgical procedure to 14.3% (2 sides) after 5 months and to 7.1% 7 months after BSSRO. Within the seven patients of this pilot study associated neurosensory disturbances were low. A possible advantage in terms of nerve protection is subject to a prospective study.

Effect of platelet-rich-plasma on cranial distraction osteogenesis in sheep: preliminary clinical and radiographic results.

The purpose of this study was to investigate the effect of platelet-rich-plasma (PRP) on cranial distraction osteogenesis. Standardized calvaria critical size defects (6 cm x 5 cm) were created in 16 adult female sheep. Bifocal cranial transport distraction osteogenesis with autogenous free calvaria bone grafts (2 cm x 4 cm) was performed at a rate of 1mm once daily to a total of 30 mm. The 16 sheep involved in the experiment were randomly divided into four groups, four animals in each: Group 1 (no PRP, latency 5 days); Group 2 (no PRP, latency 0 day), Group 3 (PRP, latency 5 days) and Group 4 (PRP, latency 0 day). After a consolidation phase of 6 weeks, the animals were sacrificed and specimens harvested for conventional radiological and 3D quantitative computer tomographic (3D-QCT) assessment. New bone was generated in the distraction zone in all groups. There were significantly (P < 0.05) higher densities in the proximal region of the distraction regenerate in Group 4 (PRP, latency 0 day) compared to Group 2. However, no significant differences in mean density of the total distraction regenerate were found, neither in volume of the bony regenerate between the experimental groups. This study showed that PRP only had an effect on bone regeneration if active distraction was started immediately after application of PRP in the distraction gap.

Dental implants in patients with orofacial clefts: a long-term follow-up study.

Aim of this study was to determine the success of implants that were inserted in patients with cleft of the lip, alveolus and palate (CLAP) and to identify prognosis-relevant factors. In a prospective evaluation, 75 implants inserted in combination with bone grafting at cleft sites of 45 patients were examined. The observation period extended 5.5 years in average (minimum 1.5, maximum 11.3 years). Implant success was evaluated clinically and radiographically and was compared to age- and gender-matched control groups. Statistic assessment included Kaplan-Meier survival analysis, Log rank tests and Cox regression analysis. In total, 10 implants were lost in 8 patients, resulting in an implant survival rate of 82.2% at the end of the observation period. Patient-related parameters of age, gender or type of cleft had no significant influence. The length of an implant was significantly related to an improved survival rate (P<0.01). Implant survival was less in CLAP patients when compared to implant insertions in a non-cleft control group, but improved when compared to patients with bone grafting for other indications. It is concluded that implants combined with bone grafting can offer a reliable alternative in patients with CLAP.

Can piezoelectric ultrasound osteotomies result in serious noise trauma?

The use of ultrasound to cut bone in oral and craniofacial surgery has increased. There is concern that the application of ultrasound to the craniofacial skeleton might represent a potential hazard to the inner ear because of sound transmission by bone conduction resulting in hearing trauma. Conventional and ultrasound osteotomies were performed on human specimens of temporal bone containing an intact middle and inner ear. The equivalent sound pressure was measured with a microphone at the round window, which had been calibrated with a bone conduction audiometer. Conventional osteotomy with a rose burr resulted in maximum sound pressures of 125dB(A) consisting of major frequency components at 2100, 7600, and 9300Hz. Ultrasound osteotomy resulted in maximum sound pressures of 122dB(A) and exhibited major frequency components at around 10kHz, 20kHz, and 26.5kHz. Ultrasound osteotomies have no acoustic advantage over conventional osteotomies. Both osteotomy techniques can produce noise-induced hearing trauma, especially when applied over longer durations of time. This appears to be more relevant for ultrasound osteotomies, because the bone cutting efficiency is usually poorer than in conventional osteotomies. Surgeons should consider the risk of noise-induced potential damage to the inner ear when selecting the method of osteotomy.

Ortho- and heterotopic bone grafts in bifocal transport osteogenesis for craniofacial reconstruction--an experimental study in sheep.

Bifocal transport osteogenesis (BTO) is a promising technique for the reconstruction of extended osseous craniofacial defects. Aim of this study was to determine the potential of this technique related to various donor sites of the transport segment. In 10 adult sheep critical size defects of the calvaria were treated by gradual movement of a transport segment consisting either of autogenous regional free calvarial bone grafts (n=5) or autogenous illiac free bone grafts (n=5). Latency period was 5 days; the rate of distraction was 1mm per day and extended approximately 40 days. The consolidation period was 28 days. After harvesting, specimens were investigated by conventional radiography, CT-scans, histologically and by fluorescence. In both groups transport osteogenesis resulted in a complete closure of the critical size defect. Membranous bone formation and remodeling occurred during the entire period of distraction and consolidation. The volumes and thickness of newly formed bone at the defect site were increased significantly when calvarial bone grafts were used (P<0.05). Iliac bone grafts became progrediently smaller during distraction, while the volume of calvarial grafts remained relatively constant (P<0.05). In conclusion, transport segments consisting of calvarial and iliac bone resulted in a reliable closure of craniofacial critical size defects in adult organisms; the application of calvarial bone grafts resulted in an increased extend of bone formation.

[Interdisciplinary treatment of carotid cavernous fistulas via the superior ophthalmic vein]. / Interdisziplinäre Behandlung spontaner A.-carotis-Sinus-cavernosus-Fisteln über die offene Punktion der V.ophthalmica superior.

BACKGROUND: The endovascular occlusion of symptomatic carotid cavernous fistulae (CCF) via the transfemoral approach is safe and effective. Due to anatomical variations or after unsuccessful transarterial therapy, a direct surgical approach to the superior ophthalmic vein (SOV) may be necessary. CASE REPORTS: In two patients with acute ophthalmologic symptoms coil occlusion of the CCF was performed after palpebral incision and cannulation of the SOV. RESULTS: In both patients preparation of the SOV was performed successfully and without complications. After coil embolization of the CCF both patients had complete resolution of symptoms within several weeks. During a follow-up of 12 months there were no recurrences, but both patients exhibited moderate blepharoptosis. CONCLUSION: Embolization of CCF via a surgically created approach is an effective procedure in selected cases when standard interventional treatment is not possible.

[In vivo administration of purified supernatant of fetal and postnatal osteoblast cultures: initial results]. / In-vivo-Applikation aufgereinigter Uberstände fetaler und postnataler Osteoblastenkulturen: Erste Ergebnisse.

With increasing age of an organism, the osseous regenerative potential is reduced. In osteoblast-like cell cultures, cells of older donors that would not proliferate under standard conditions start to proliferate and express a differentiated phenotype when supplemented with the supernatant of fetal osteoblast-like cultures. The aim of this study was an evaluation of the fetal supernatant actions in vivo. Rat calvaria-derived osteoblasts of fetal and 21-day-old donors were isolated enzymatically and cultured. Their osteoblastic phenotype was confirmed by the expression of typical osteoblast synthesis products. The supernatant of the cultures of each age group was collected and pooled. The supernatant was purified by HPLC and concentrated approximately ten times. Additionally, several age-dependent differences within the purified protein fractions were documented by MALDI. After resuspension, the purified supernatant proteins were transferred on a collagen carrier and implanted into critical size defects of the calvaria of adult Wistar rats. In a control group, the collagen carriers were implanted containing isotonic salt solution. After 2, 4, and 8 weeks, a radiographic and histologic analysis of the regeneration process was performed which revealed differences in the progress of mineralization. The methods used in this study might help to identify age-dependent differences regarding the osteoblastic synthesis of osteoanabolic peptides and their impact on the regeneration of osseous defects.

[Determination of the toxicity of industrial waste by means of a bacterial test (author's transl)]. / Bestimmung der Toxizität von Industriemüll mit einem Bakterientest

The decision on whether industrial waste should be deposited on general or special dumps depends upon the toxicity of the waste and thus upon the risk of ground-water pollution and the killing off of micro-organisms responsible for breaking down waste products. Since chemical analyses that are to provide information on toxicity are frequently very costly, an attempt was made to determine the toxicity of solid or semi-solid waste with the aid of a bacterial test. As in the perforated-plate test, here, too, the formation and size of inhibition zones were used to determine the toxicity after diffusion of substances contained in industrial waste. About 30 different types of industrial waste were examined with this method. For almost all substances a positive correlation of the (known) constituents of waste to the formation of inhibition zones was found to exist. The influence exercised by solubility and diffusibility and other problems are dealt with.

[Endosseous implants for functional masticatory rehabilitation in the extremely atrophied edentulous maxilla]. / Enossale Implantate zur kaufunktionellen Rehabilitation des extrem atrophierten zahnlosen Oberkiefers.

PURPOSE: The aim of this retrospective study was to evaluate the long-term survival rate of dental implants in edentulous patients suffering from severe atrophy of the alveolar ridges in the upper jaw. PATIENTS AND METHODS: In total, 964 implants were inserted in 140 patients. A total of 481 implants were combined with an osteoplastic augmentation of the maxilla, and 483 implants were inserted directly in the atrophic bone. The success rate was determined using survival analysis, log rank tests and a Cox regression analysis. RESULTS: The overall survival rate for all implants was 42.2% during an observation period of 11 years. Between implants combined with an osteoplasty and implants inserted in local bone tissue there were no significant differences in the survival rate. The survival rate of implants combined with an osteoplasty was significantly reduced in women and in the case of repeated insertion or augmentation. Interestingly, a few of the patients treated with an osteoplasty demonstrated high numbers of individual implant failures. Those patients were postmenopausal women exclusively. Among them there is probably a certain group with a very high risk of implant failures. CONCLUSION: This study shows that oral rehabilitation with osteointegrated implants in patients with severely atrophic alveolar ridges in the upper jaw is still problematic.

[Johann Friedrich Dieffenbach (1792-1847) as the founder of plastic surgery. His contribution to maxillofacial surgery]. / Johann Friedrich Dieffenbach (1792-1847) als Begründer der plastischen Chirurgie. Sein Beitrag zur Mund-, Kiefer- und Gesichtschirurgie.

Johann Friedrich Dieffenbach (1792-1847) is generally known as the founder of modern plastic surgery. One focus of his work, apart from the physiology of transplantation and operative techniques, was reconstructive oral and maxillofacial surgery. This special aspect of plastic surgery as well as Dieffenbach's biography is presented in this historic article.

[Growth of a squamous epithelial carcinoma in an odontogenic cyst]. / Entstehung eines Plattenepithelkarzinoms auf dem Boden einer odontogenen Zyste.

Malignant change in the epithelium of an odontogenic cyst and growth of an invasive squamous cell carcinoma is rare. The diagnosis of malignant disease is mostly only established by histological evaluation of excised tissue because the initial clinical appearance does not differ much from that of odontogenic cysts. The clinical course of one patient after removal of an impacted canine and adjacent cystic tissue is presented. Histological evaluation of the specimen led to diagnosis of a squamous cell carcinoma arising from the cystic epithelium. Partial resection of the maxilla was performed subsequently and 1 year postoperatively the patient was free from recurrence or metastasis. This case report underlines the importance of submitting any soft tissue excised in the treatment of odontogenic cysts to histologic evaluation.

[The Stromeyer hook. Life and work of the man behind the eponym]. / Stromeyer-Haken. Leben und Werk des Mannes hinter dem Eponym.

Georg Friedrich Stromeyer (1804-1876) is generally known as one of the founders of orthopedics and orthopedic surgery and also made many contributions to modern military medicine. Furthermore, every oral and maxillofacial surgeon in Germany knows him because of the "Stromeyer hook", which is used for elevation of zygomatic arch fractures. This special aspect as well as Stromeyer's biography is presented in this article from the history of medicine.