Peter Proff

Functional 3-D analysis of patients with unilateral cleft of lip, alveolus and palate (UCLAP) following lip repair.

INTRODUCTIONnParticular importance is attached to lip repair cleft surgery, as numerous functional and aesthetic aspects have to be taken into account simultaneously. Spatial assessment of function and depiction of dynamic deviations is reasonable for describing surgical outcome in addition to long standing static analysis. This study aimed at 3D analysis of the oral area after reconstruction in patients with unilateral cleft lip, alveolus and palate.nnnPATIENTS AND METHODSnTwelve patients with unilateral cleft lip, alveolus and palate who underwent surgery according to Tennison-Randall were enrolled in this study. Soft tissue dynamics was analysed during passive stretching and active contraction of the lips, and photogrammetry was used for comparing relative changes of length and displacement vectors. The spatial coordinates of surgically significant and reproducible landmarks along the red-white lip junction were analyzed.nnnRESULTSnStatic analysis of the lips revealed a good result with far-reaching symmetry in all cases. Regarding dynamic behaviour, two groups could be distinguished showing clear differences of passive distension and contraction behaviour.nnnCONCLUSIONnDespite nominally identical surgical techniques and comparable static-morphological outcomes, dynamic analysis revealed differences pointing to a need for optimization.

In vivo study of apoptosis as a creative agent of embryonic development of the primary nasal duct in rats

INTRODUCTIONnThe first embryonic part of the nasal cavity is the primary nasal duct, beginning with the olfactory placode and ending with the oronasal membrane. Aim of this study was to investigate the cellular processes (apoptosis, proliferation) being responsible for development and opening of the primary nasal duct.nnnMATERIAL AND METHODSnIn this study developmental processes in at least three regions of the primary nasal duct (opening, middle, end) were examined by sectioning 38 rat fetuses on day 13.5 after conception. Apoptotic cells were detected by active caspase-3 antibodies and proliferating cells were examined by Ki-67 antibodies.nnnRESULTSnMultiple apoptotic events were diagnosed on the basis and proliferative cells on the top of this duct.nnnCONCLUSIONnApoptosis and proliferation play an important role in the process of opening the bottom of the primary nasal duct and for development of the nasal septum, philtrum as well as the primary palate. Mesenchymal proliferation seems to play a minor role in the process of opening the primary nasal duct.

Treatment of patients with cleft lip, alveolus and palate – a short outline of history and current interdisciplinary treatment approaches

Clefts of lip, alveolus and palate have been known for a long time. First tangible evidence of surgical therapy in terms of cheiloplasty, however, does not date further back than the 4th century after Christ. It was Werner Hagedorn from Magdeburg who laid the foundations of geometrical anatomical surgical lip repair in 1884. The procedures designed by LeMesurier, Tennison, Randall or Millard in the 1950s, and by Pfeifer in 1970 are part of todays cleft therapy applied by the different schools of surgery.

Apoptosis as a creative agent of embryonic development of bucca, mentum and nasolacrimal duct. An in vivo study in rats.

INTRODUCTIONnFor embryonal facial development several fusion processes between different facial prominences are necessary. If fusion fails to appear, various facial clefts may occur, known as median (e.g. lower median cleft lip), oblique (e.g. open nasolacrimal duct) or lateral facial clefts (macrostomia, lateral cleft).nnnMATERIAL AND METHODSnThe development of 3 different facial regions (bucca, mentum, and nasolacrimal duct) was examined in rats using serial histological sections on day 13.5 after conception. Common procedures were used (staining for active caspase-3 and for Ki-67) for histological assessment about the role of apoptotic and proliferative processes in the fusion zones of buccal, mental and nasolacrimal areas.nnnRESULTSnMultiple apoptotic events were detected in epithelial cells of the respective regions, the proliferative centers were located in the mesenchymal surroundings of fusion zones.nnnCONCLUSIONnA substantial precondition for fusion of facial prominences are proliferative and apoptotic processes in epithelial and mesenchymal cells. Apoptosis contributes to the development of bucca, mentum and the nasolacrimal duct. Absence of apoptoses may be responsible for facial clefts.

Evaluation of patient satisfaction after therapy of unilateral clefts of lip, alveolus and palate

INTRODUCTIONnCleft lip, alveolus and palate (CLAP) is a craniofacial abnormality and is one of the most frequent human developmental anomalies. Therapy of clefts does not only comprise surgical closure of the cleft, but rather aims at an aesthetically and functionally optimal result at adult age.nnnMATERIAL AND METHODSnThirty-three cleft patients with total clefts of lip, alveolus and palate were enrolled in this study. Osseous bridging of the alveolar cleft (osteoplasty) was performed in all patients followed by different types of subsequent treatment. All patients answered a questionnaire to assess their satisfaction with the treatment result and their facial appearance. Patient satisfaction was correlated to the type of alveolar cleft repair.nnnRESULTSnThe returned questionnaires revealed varying patient satisfaction with their appearance, occlusal conditions, and dental aesthetics depending on the type of dental treatment in the alveolar cleft area. Questionnaire analysis by gender revealed clear gender-dependent differences in self-rated satisfaction.nnnCONCLUSIONnAesthetics gain increasing importance for self-perception. Therefore, patient satisfaction with her facial appearance should move even more into focus of therapy of clefts.

Orthodontic forces add to nicotine-induced loss of periodontal bone

ObjectivesNicotine is considered an etiologic factor for chronic inflammatory phenomena within the periodontal ligament that may result in loss of periodontal attachment. Considering that smokers account for 26u2009% of adult and 12u2009% of adolescent patients in orthodontic practice, we performed in vivo and in vitro studies as to whether orthodontic forces may add to the nicotine-induced loss of periodontal bone.MethodsFourteen male rats (Fischerxa0344 inbred) were used. Seven of these served as controls, while the other seven received daily subcutaneous injections of 1.89xa0mg L-nicotine per kg body weight. Both groups were exposed to orthodontic mesialization of the first two upper left molars using a NiTi closed-coil spring, the contralateral side serving as control. Periodontal bone loss was assessed by cone-beam computed tomography (CBCT). Human periodontal fibroblasts were stressed by compression (2xa0g/cm2) and/or nicotine (3/5/7.5xa0µmol), and the expression of cyclooxygenase-2 (COX-2), prostaglandin E2 (PGE2), interleukin-6 (IL-6), osteoprotegerin (OPG), and receptor activator of nuclear factor κB ligand (RANKL) was determined at the transcriptional level by quantitative real-time polymerase chain reaction (qRT-PCR) and at the translational level by enzyme-linked immunosorbent assay (ELISA). In addition, differentiation of co-cultured murine RAW264.7 cells to osteoclast-like cells was quantified by tartrate-resistant acid phosphatase (TRAP) staining.ResultsOrthodontic force application in vivo led to a significant increase in nicotine-induced periodontal bone loss, and cell compression in vitro to increased COX-2, PGE2, IL-6, and RANKL expression, reduced OPG expression, and enhanced differentiation of RAW264.7 cells to osteoclast-like cells compared to nicotine alone.ConclusionAdditional loss of periodontal bone must be expected during orthodontic treatment of smokers. Clinicians should inform their patients of this increased risk and refrain from performing tooth movements before cessation of smoking.ZusammenfassungZielsetzungNikotin gilt als ein ätiologischer Faktor chronisch entzündlicher Prozesse im Zahnhalteapparat, die zum Verlust des parodontalen Attachments führen können. Raucher stellen mit 26u2009% der Erwachsenen und 12u2009% der Jugendlichen einen hohen Anteil von Patienten in kieferorthopädischer Behandlung. Wir prüften daher in vivo und in vitro die Hypothese, ob die Applikation kieferorthopädischer Kräfte zu einer Steigerung des nikotininduzierten parodontalen Knochenabbaus führt.Material und MethodenSieben männliche Fischer-344-Ratten erhielten tägliche Injektionen von 1,89xa0mg L-Nikotin pro kg Körpergewicht s.c., 7 weitere Tiere dienten als Kontrolle. In beiden Gruppen erfolgte eine kieferorthopädische Mesialisation der ersten beiden linken oberen Molaren mittels einer NiTi-Zugfeder, während die kontralaterale Oberkieferseite als Kontrolle diente. Der parodontale Knochenverlust wurde mittels digitaler Volumentomografie (DVT) bestimmt. Humane parodontale Fibroblasten wurden einem Druck von 2xa0g/cm2 und/oder 3/5/7,5xa0µM Nikotin ausgesetzt. Bestimmt wurde die Expression von COX-2, PGE2, IL-6, OPG und RANKL auf mRNA- (qRT-PCR) und Proteinebene (ELISA) und quantifiziert wurde die Differenzierung von kokultivierten RAW264.7-Zellen zu osteoklastenähnlichen Zellen mittels TRAP-Färbung.ErgebnisseDie kieferorthopädische Kraftapplikation führte in vivo zu einer signifikanten Zunahme des nikotininduzierten parodontalen Knochenverlustes sowie in vitro zu einer Steigerung der Expression von COX-2, PGE2, IL-6 und RANKL, einer Hemmung der OPG-Expression sowie einer vermehrten Differenzierung von osteoklastenähnlichen Zellen aus RAW264.7-Zellen gegenüber der alleinigen Wirkung von Nikotin.SchlussfolgerungenBei der kieferorthopädischen Behandlung von Rauchern ist mit einem vermehrten parodontalen Knochenverlust zu rechnen. Der praktizierende Kieferorthopäde sollte die Patienten über die erhöhten Risiken aufklären, und Zahnbewegungen sollten nur nach Einstellen des Nikotinabusus erfolgen.

O-phospho-L-serine: a modulator of bone healing in calcium-phosphate cements.

Abstract Bone substitution materials are seen as an alternative to autogenous bone transplants in the reconstruction of human bone structures. The aim of the present animal study was to evaluate the clinical handling and the conditions of bone healing after the application of a phosphoserine and collagen-I-modified calcium-phosphate cement (Biozement D). The application of phosphoserine is supposed to influence the texture of the extracellular matrix. Standardised bone defects were created in the lower jaw of 10 adult minipigs. These defects were reconstructed with a pasty calcium-phosphate cement mixture. After a healing time of 4 months, the animals were sacrificed. The mandibles of all animals were resected and non-decalcified histological sections of the areas of interest were prepared. The experiment was evaluated by means of qualitative histology and histomorphometry. The hydroxyapatite cement entirely hardened intraoperatively. Modelling and handling of the cement was facile and the margin fit to the host bone was excellent. Histology showed that resorption started in the periphery and proceeded exceptionally fast. The bony substitution, especially in phosphoserine-endowed cements, was very promising. After a healing period of 4 months, phosphoserine cements showed a bone regeneration of nearly two-thirds of the defect sizes. In the applied animal experiment, the newly developed hydroxyapatite collagen-I cement is well suited for bone substitution due to its easy handling, its excellent integration and good resorption characteristics. The addition of phosphoserine is very promising in terms of influencing resorption features and bone regeneration.

Prenatal diagnostics of cleft deformities and its significance for parent and infant care.

INTRODUCTIONnThis study aimed to demonstrate clefts of the secondary palate in embryos found to have cleft lip in order to evaluate the validity of prenatal ultrasound examination and, thus, to assess the significance of this diagnostic method for coordination and care of parents and infant.nnnPATIENTS AND METHODSnOver a period of 2.5 years, 7 fetuses with cleft deformities were examined sonographically during the 20th and 25th gestational week. The results were compared to postnatal clinical diagnosis. This study was made by two experienced examiners using 2D ultrasound devices (Acuson 128 XP-10/ C7; Toshiba Aplio XV). Postnatal clinical diagnosis was made by an orthodontist.nnnRESULTSnThree of the ultrasound-based diagnoses coincided with the postnatal result. In three of the examined cases the extent of the cleft was underestimated, whereas a greater extent suspected in one patient could not be confirmed clinically.nnnCONCLUSIONnThe results of the present study support the propositions of current literature: Diagnosis of a cleft of the lip and the alveolar process could correctly be made by an experienced ultrasound diagnostician. However, problems arise with clefts of the secondary palate.

Effects of mechanical and bacterial stressors on cytokine and growth-factor expression in periodontal ligament cells

ObjectivesThe goal of the study was to examine the effects of a mechanical (orthodontic force simulation by static compressive loading) and a bacterial (endotoxins from a heat-inactivated gram-negative periodontal pathogen) stressor on the expression patterns of factors that are key to regulating osteoclastogenesis and bone remodeling.Materials and methodsThree experimental groups were formed with fifth-passage periodontal ligament (PDL) fibroblasts treated by the static application of compressive force (2xa0g/cm2), heat-inactivated aggregatibacter actinomycetemcomitans (1u2009×u2009107 cells), or both of these stressors combined. Real-time polymerase chain reaction (RT-PCR) was used to study gene expression of IL-6, IL-8, COX-2, IGF-1, VEGF, and MMP-13 in the 3xa0groups. Protein levels of COX-2, prostaglandin E2 (PGE2), and IL-8 production were quantified using immunoblotting and enzyme-linked immunosorbent assay (ELISA).ResultsThe mechanical stressor upregulated the genes of COX-2, IL-8, IGF-1, and MMP-13 in PDL fibroblasts and the bacterial stressor upregulated IL-6, IL-8, COX-2 and MMP-13. Both stressors in combination upregulated VEGF and caused COX-2 gene expression to increase further; the latter effect was also detected at the protein level and indirectly via the enhanced production of PGE2. We noted that the posttranscriptional regulation of IL-8 was induced by the mechanical stressor and influenced by PGE2.ConclusionWhile mechanical-stressor application increased the gene expression of COX-2, IL-8, and VEGF in the presence of the bacterial stressor, IL-8 production was posttranscriptionally regulated by the mechanical stressor, whereas COX-2 expression correlated with enhanced production of the inflammatory tissue hormone PGE2, which exerted a suppressive effect on endotoxin-induced IL-8 production.ZusammenfassungZielsetzungZiel der Studie war es, die Regulation von Faktoren, die einen entscheidenden Einfluss auf die Steuerung der Osteoklastogenese und Knochenremodellierung haben, in Abhängigkeit von biomechanischer Stressapplikation und Endotoxinen eines parodontal-pathogenen Erregers zu untersuchen.Material und MethodikFür die Untersuchung wurden humane Parodontalligamentfibroblasten in der 5. Passage verwendet, die entweder mit statischem Kompressionsdruck (2xa0g/cm2) oder mit hitzeinaktivierten A. actinomycetemcomitans (1u2009×u2009107 Zellen) behandelt wurden. Eine weitere Gruppe wurde zusätzlich mit Kraftapplikation und hitzeinaktivierten parodontal-pathogenen Bakterien behandelt. Von den 3 Untersuchungsgruppen erfolgte die Bestimmung der Genexpression von den Genen IL-6, IL-8, COX-2, IGF-1, VEGF und MMP-13 mittels RT(„real time“)-PCR. In einer weiteren proteinbiochemischen Untersuchung wurde die Bildung von COX-2, Prostaglandin E2 (PGE2) und IL-8 mittels Immunoblot und ELISA quantifiziert.ErgebnisseDie Simulation kieferorthopädischer Kraftapplikationen führte zu einer Hochregulation der Gene von COX-2, IL-8, IGF-1 und MMP-13 in Parodontalligamentfibroblasten. Die Anwesenheit von hitzeinaktivierten gramnegativen Bakterien bewirkte eine Hochregulation von IL-6, IL-8, COX-2 und MMP-13. Die kombinierte Anwendung beider Stressfaktoren ergab eine verstärkte Genexpression von VEGF und eine Potenzierung der Genexpression von COX-2, die auch auf Proteinebene und indirekt durch eine erhöhte Bildung von PGE2 nachweisbar war. IL-8 wurde durch die Anwendung von Kompressionskräften posttranskriptionell reguliert vorgefunden. Ein Einfluss von PGE2 auf die posttranskriptionelle Regulation von IL-8 konnte nachgewiesen werden.SchlussfolgerungDie zellexperimentelle Studie zeigt, dass die Anwendung biomechanischer Kräfte in Anwesenheit von bakteriellen Endotoxinen gram-negativer Bakterien eine Verstärkung der Genexpression von COX-2, IL-8 und VEGF bewirkt. Die Bildung von IL-8 wurde allerdings durch Kompressionskräfte posttranskriptionell reguliert, während die erhöhte COX-2-Expression mit einer verstärkten Bildung des inflammatorischen Gewebehormons PGE2 korreliert. Letzteres hatte einen supprimierenden Effekt auf die endotoxininduzierte Bildung von IL-8.

Influence of Cyclosporin A on human gingival keratinocytes in vitro

PURPOSEnGingival hyperplasia is a well known side effect of Cyclosporine A therapy. The aetiology of this is not totally understood and there is debate whether it is hyperplasia of the gingival epithelium or of the submucosal connective tissue, or both, and what roles play factors like age and gender of the patients, duration and dosage of the drug.nnnMATERIAL AND METHODSnThe influence of different Cyclosporine A concentrations (10(-6) g/ml; 5 x 10(-7) g/ml; 10(-9) g/ml) and of no medication (controls) on growth and proliferation of cultured human gingival keratinocytes was investigated after a culture period of 3, 6 and 9 days. Cell proliferation was assessed by counting anti Ki-67 stained nuclei, cell growth by counting total number of nuclei and by the EZ4U-assay.nnnRESULTSnThere was no significant correlation of the cell proliferation rate and cellular growth with either gender (p > 0.568) or duration of medication (p > 0.876); but Cyclosporine A concentration showed a highly significant influence on cellular growth (p = 0.0001). Inhibition of cell growth was dependent on drug dosage, but a low concentration of 10(-9) g/ml even stimulated cell growth.nnnCONCLUSIONSnThere is evidence that Cyclosporine A in low concentrations (10(-9) g/ml as applied in long-term therapy) stimulates gingival keratinocyte growth and therefore might be related to hyperplasia of the gingiva. However, high Cyclosporine A concentrations may inhibit cell growths and factors like gender of the patient did not show any influence in-vitro.