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Orofacial clefts have been associated with specific cleft candidate genes which encode regulatory proteins required for orofacial region development. Cleft candidate genes encode proteins involved with the cleft morphopathogenesis process, but their exact interactions and roles are relatively unclear in human cleft tissue. This study evaluates the presence and correlations of Sonic Hedgehog (SHH), SRY-Box Transcription Factor 3 (SOX3), Wingless-type Family Member 3A (WNT3A) and 9B (WNT9B) protein containing cells in different cleft tissue. Non-syndromic cleft-affected tissue was subdivided into three groups—unilateral cleft lip (UCL) (n = 36), bilateral cleft lip (BCL) (n = 13), cleft palate (CP) (n = 26). Control tissue was obtained from five individuals. Immunohistochemistry was implemented. The semi-quantitative method was used. Non-parametric statistical methods were applied. A significant decrease in SHH was found in BCL and CP tissue. SOX3, WNT3A and WNT9B had a significant decrease in all clefts. Statistically significant correlations were found. The significant decrease in SHH could be associated with BCL and CP pathogenesis. SOX3, WNT3A and WNT9B could have morphopathogenetic involvement in UCL, BCL, and CP. Similar correlations imply the presence of similar pathogenetic mechanisms in different cleft variations.

The zoonotic parasite Dirofilaria repens has spread toward north in Europe, and cases of autochthonous dirofilariosis caused by D. repens have emerged in the Baltic countries Estonia, Latvia, and Lithuania. We conducted a review on the emergence of dirofilariosis in humans and domestic dogs in these three countries in northeastern Europe. Based on the available literature and reports, the first finding in the Baltic countries was made in Latvia in 2008, followed by the first in Lithuania in 2010, and the first in Estonia in 2012. In all three countries, further findings were reported soon after the first reports. By the end of 2019, autochthonous human D. repens infections had been described from Latvia and Lithuania, and autochthonous canine D. repens infections had been described from all three Baltic countries. While no epidemiological studies estimating prevalence or incidence of the human infections have been published from the three countries, a substantial proportion of investigated dogs have tested positive for microfilariae in studies performed in Latvia and Lithuania. Dirofilariosis is an emerging zoonosis in northern Europe, and the summarized data confirm that D. repens has become established and endemic in the Baltic countries. The available data do not provide a good overview of the situation, and further epidemiological studies are needed. Awareness about the recently emerged zoonotic parasite should be increased among medical doctors, veterinarians, and the general public. Managing this zoonotic infection is a public health challenge that needs to be addressed using a One Health approach. Investigating the spread of D. repens in the Baltic countries could be useful for better preparedness for the anticipated further spread to the Nordic countries.

Introduction\ud \ud The Erasmus+O-Health-EDU project aims to gain a comprehensive view of oral health professional (OHP) education in Europe, through the development of web-based surveys and online toolkits. A glossary to facilitate a common language through which academic teams could cooperate and communicate more accurately was identified as a key need within the project. The aim of ARTICULATE was thus to create a shared language, with a European focus, for terms and concepts used in the field of OHP education.\ud \ud \ud \ud Methods\ud \ud The methodology was developed from those published for construction of other glossaries with a circular and iterative process: the creation of content and definitions by a group of experts in OHP education, the testing of “fitness for purpose” of the content, and stakeholder consultation. All creation steps were followed by refinements based on testing results and stakeholder comments. The final glossary was then launched as an online resource including a built-in mechanism for user feedback.\ud \ud \ud \ud Results\ud \ud The scope and structure of the glossary were mapped out at a workshop with 12 dental education experts from 7 European countries. A total of 328 terms were identified, of which 171 were finally included in ARTICULATE. After piloting with a close group of other colleagues, the glossary was opened for external input. Thirty European Deans or Heads of Education assessed the definition of each term as “clear” or “not clear.” A total of 86 definitions were described as “clear” by all individuals. Terms deemed unclear by at least one individual were revisited and changes made to 37 of the definitions. In conjunction with the launch of the glossary, a range of stakeholder organisations were informed and asked to participate in an open global consultation by providing feedback online. Since its launch in June 2021, the ARTICULATE website (https://o-health-edu.org/articulate) has had an average of 500 visits/month. To promote community ownership, forms embedded on the ARTICULATE webpage allow users to give feedback and suggest new terms. A standing taskforce will meet regularly to consider amendments and make changes to ensure that the glossary remains a relevant and up-to-date resource over time.\ud \ud \ud \ud Conclusion\ud \ud ARTICULATE is a unique, evolving, online glossary of terms relating to OHP education, created as a resource for all interested OHP educators. The glossary is a key output of the O-Health-Edu project, which relies on a comprehensive vision of OHP education to address the future oral health needs of the European population.

Bone repair after surgical intervention on cleft lip palate (CLP) depends on the coordinated action of multiple tissue regeneration factors. We determined the relative number and appearance of tissue factors: matrix metalloproteinase-2 (MMP-2), tissue inhibitor of metalloproteinase-2 (TIMP-2), bone morphogenetic protein 2/4 (BMP 2/4), transforming growth factor beta (TGF-ß), Wnt3a protein (Wnt3a), Runt-related transcription factor 2 (Runx2), basic fibroblast growth factor (bFGF) and osteoprotegerin in hard tissue of CLP patients during first time surgical intervention. Fourty-three CLP patients with 24 bone and 36 cartilage samples were involved. Immunohistochemistry was used to assess the levels of tissue factors and the semi-quantitative census method was used for quantification of immunological structures. The increased amount of MMP-2 and bFGF positive cells was detected in the CLP group in cartilage and bone (p < 0.05), compared to the controls. A statistically significant (p = 0.012) increased amount of BMP 2/4 positive cells was found in cartilage of CLP patients, in comparison to the control group. Increased appearance of MMP-2, bFGF in hard tissue of the CLP patients indicates the predominance of tissue degradation. Increased number of BMP2/4 positive chondrocytes suggests improved cartilage growth and better regeneration in CLP patients.

This consensus paper reports on the process of developing a renewed vision for Oral Health Professional (OHP) education across Europe, and forms part of a larger EU‐funded collaborative Erasmus+ project, “O‐Health‐Edu.” The vision aligns with the World Health Organisation milestones (2016) and resolutions (2021), and EU4Health programme (2020) objectives ‐ and projects 20 years into the future, to 2040. This longitudinal vision takes a multi‐stakeholder perspective to deliver OHP education that acts in the best interests of both students and patients, and sits within the context of a wider strategy for general health. Included, it is an infographic to help communicate the vision to various stakeholders of OHP education.

Craniofacial morphogenesis is highly complex, as is the anatomical region involved. Errors during this process, resulting in orofacial clefts, occur in more than 400 genetic syndromes. Some cases of cleft lip and/or palate (CLP) are caused by mutations in single genes; however, complex interactions between genetic and environmental factors are considered to be responsible for the majority of non-syndromic CLP development. The aim of the current study was to identify genetic risk factors in patients with isolated cleft palate (CP) by whole genome sequencing. Patients with isolated CP (n = 30) recruited from the Riga Cleft Lip and Palate Centre, Institute of Stomatology, Riga, were analyzed by whole genome sequencing. Pathogenic or likely pathogenic variants were discovered in genes associated with CP (TBX22, COL2A1, FBN1, PCGF2, and KMT2D) in five patients; hence, rare disease variants were identified in 17% of patients with non-syndromic isolated CP. Our results were relevant to routine genetic counselling practice and genetic testing recommendations. Based on our data, we propose that all newborns with orofacial clefts should be offered genetic testing, at least for a panel of known CLP genes. Only if the results are negative and there is no suggestive family history or additional clinical symptoms (which would support additional exome or genome-wide investigation), should multifactorial empiric recurrence risk prediction tools be applied for families.

(1) Background: Despite cleft lips and palates belonging to the most common orofacial congenital anomalies, their morphopathogenesis is not yet fully understood. The study aimed to determine the distribution and relation of cytokines interferon-γ (IFN-γ), tumor necrosis factor-alpha (TNF-α), interleukin (IL)-2, IL-7, IL-12, and IL-13 in the cleft affected mucosa of the lip. (2) Materials and Methods: Twenty cleft lip (CL) mucosal samples and seven control tissues of oral cavity mucosa were included in the study. Specimen were obtained during reconstruction surgeries and processed by hematoxylin and eosin staining and immunohistochemistry for IFN-γ, TNF-α, IL-2, IL-7, IL-12, and IL-13. (3) Results: The distribution of cytokines was higher overall in the cleft affected epithelium compared to the connective tissue, with TNF-a, IL-2, and IL-12 displaying the highest number of immunopositive cells. With the exception of IL-2, CL specimen showed higher immunoreactivity. IFN-γ displayed only minor immunoreactivity, with no expression in the control epithelium. Correlation analysis was strongest between CL epithelial IL-13 and IFN-γ (z = 0.71, p &lt, 0.0001). (4) Conclusions: The CLP affected epithelium displays high degrees of plasticity in expressing different cytokines, pointing towards the stimulation of a local adaptive immune response based on consistent inflammatory processes.

Cleft lip and palate are common congenital pathologies that affect the human population worldwide. The formation of cleft lip is associated with multiple genes and their coded proteins, which regulate the development of craniofacial region, but the exact role of these factors is not always clear. The use of morphological studies for evaluation of human cleft-affected tissue has been limited because of insufficiency of available pathological material. The aim of this study was to detect and compare the immunohistochemical expression of cleft candidate gene coded proteins (DLX4, MSX2, HOXB3, SHH, PAX7, SOX3, WNT3A, and FOXE1) in the non-syndromic unilateral cleft lip patient tissue and control group tissue. A semiquantitative counting method was used to evaluate the tissue in biotin-streptavidin-stained slides. Statistically significant differences between the patient and control groups were found for the number of immunoreactive structures for SHH (p = 0.019) and FOXE1 (p = 0.011) in the connective tissue and SOX3 (p = 0.012) in the epithelium. Multiple statistically significant very strong and strong correlations were found between the immunoreactives in cleft-affected tissue. These significant differences and various correlations indicate that multiple morphopathogenetic pathways are possibly involved in unilateral cleft lip pathogenesis. Therefore, we further discuss these possible interactions.

The prevalence and burden of untreated oral diseases throughout the life course remains high worldwide, and inequalities in oral health and dental care are increasing. This is a major public health issue that is not being enough addressed by the health care systems. To better manage populations' oral health, oral health professionals must be trained to adapt to population needs and societal and technological changes. Furthermore, dental institutions must fulfill their social responsibility by prioritizing educational and research activities that promote advancing individual and community health. In Europe, great variability exists between dental programs within the same country or between countries. This variability is an issue as European graduates can practice around the European Union through mutual recognition of their qualifications. This might lead to inequities in the availability, accessibility, acceptability and quality of health services. The convergence of competencies and quality standards at the international level must thus be better clearly identified, defined and improved. The purpose of the O-Health-Edu, EU funded project, “Advancing Oral Health: A vision for Dental Education” is thus to assess the current situation concerning the oral health professionals' education (OHP) and to identify educational priorities so that future graduating OHPs have competencies aligned with emerging population needs. The project is supported within the KA203 - “Cooperation for Innovation and the Exchange of Good Practices - Strategic Partnerships for higher education” Erasmus program. It began in October 2019 for three years, with eight European university partners and the ADEE (Association for Dental Education in Europe). The firsts steps consist of a scoping review to identify the available information followed by a questionnaire survey to gather more comprehensive data so that to build an opened data source, and a glossary of dental education terms. Key messages Few Information about the current situation of dental education in Europe. Graduates must be equipped with adequate competencies to face populations needs.

Introduction\ud \ud The variability in oral health professional education is likely to impact on the management of oral health needs across Europe. This scoping review forms the initial part of a larger EU‐funded collaborative Erasmus+ project, ‘O‐Health‐Edu’. The aim of this scoping review is to investigate how oral health professional education in Europe is reported.\ud \ud \ud Methods\ud \ud The PRISMA and Arksey & O’Malley methodological frameworks for scoping reviews were used to guide reviewers in answering the research question “How is oral health professional education reported in Europe?”. The search strategy encompassed published literature searches, internet searches and further searching of relevant documents from educational organisations, regulators and professional bodies. Once the search strategy was developed it was sent to key stakeholders for consultation. Sources were reviewed by two authors (JD, JF), and included in the review if they reported on oral health professional education in Europe.\ud \ud \ud Results\ud \ud A total of 508 sources were retrieved from all of the searches. 405 sources were excluded as they did not report on the topic of interest, leaving 103 sources that reported on oral health professional education in Europe. Handsearching the references of published sources lead to a further 41 sources being screened, of which 15 were included. In total, 33 duplications were removed and the final number of included sources was 85. The average year of publication for the included sources was 2007, with sources most commonly published in journals dedicated to dental education. Surveys represented the most common form of reporting. From the data obtained, four broad themes of reporting were evident: Dental education at a Programme Level, Dental education at a Discipline Level, Other Oral Health Professional Education, and, Postgraduate Education and Continuous Professional Development.\ud \ud \ud Conclusion\ud \ud The reporting of dental and oral health professional education in Europe is limited. Whilst there are many useful documents that provide guidelines on dental education, there is limited knowledge on how education is implemented and delivered. There is a greater need for comprehensive educationally driven programme level data on oral health professional education across Europe.

Introduction. Facial morphogenesis occurs from the fourth to the twelfth gestation week, when the cells from nerve crest migrate to the region of face, forming the primary palate. The cleft palate is an abnormality in embryogenesis period, which is characterized by the absence of fusion of palatal shelves. The incidence of cleft lip and palate is one in 700 live births. In recent years the effect of different genes and signaling molecules, including Runx2, Wnt3 and OPG have been studied in the development of cleft palate, because these substances are considered to be regulators of pathogenesis responsible for formation of bone and cartilage tissue and particularly the bone. Aim of the Study. The aim of the work was to evaluate the expression of Runx2, Wnt3 and OPG in palate bone and nasal cartilage for children with cleft palate. Material and methods.Eleven bone and cartilage samples were obtained from 21 children of the lip, soft and hard palate correction surgery. All the patients were diagnosed with clefts of the lip, alveolar process of maxilla, and palate. In the tissue sections using the immunohistochemistry method (IMH), were determined Runx2 (code: AB192256, 1: 250, Abcam GB, rabbit), Wnt3 (code: AB1992, 1: 800, Abcam GB, rabbit), and OPG (code: A0611, 1: 100, The Orbit USA, rabbit) local expression. We used a semi-quantitative census method for quantifying the positive structures. Results. Runx2 expression was observed in five patient bone tissue samples and six patient cartilage tissue samples. Of the Runx2 positive bone tissue, in one case we observed occasional, in two cases- few, in one case- moderate to numerous and in one case numerous positive osteocytes while in tissue of cartilage in two cases we observed few, in one case- few to moderate, in two casesmoderate, and in one case numerous positive chondrocytes. A significant difference in Wnt3 expiation was observed between bone and cartilage tissues. Wnt3 expressing chondrocytes were observed in all samples, where in one case- occasional, in three cases few, in one case-moderate, and in six cases-numerous positive cartilage cells were observed. The expression of the gene in the bone was observed in nine cases, which contained mostly occasional or few positive structures, except in three cases where in one Wnt3 was marked by few to moderate and in two cases numerous positive osteocytes. OPG expression was observed in all samples, but in the cartilage, the expression was more pronounced. In the cartilage in seven cases, there were numerous positive chondrocytes, in one case- few to moderate, in two cases moderate to numerous and in one case few to moderate number of chondrocytes. OPG showed variable expression. In four cases, we observed occasional to few, in one case few to moderate, in one case- moderate, in one case moderate to numerous and in four cases numerous positive bone cells. Conclusion. Cartilage tissue expresses significantly more Runx2, Wnt3 genes and OPG proteins, indicating a greater compensatory tissue capacity. In the case of palate clefts, the high expression of Wnt3 and OPG and lower expression of Runx2 could indicate a significant tissue proliferation which predominates over mineralization and ossification processes.

Background and Aims: Morphogenesis of the upper lip and palate is a complex process involving highly regulated interactions between epithelial and mesenchymal cells. Genetic evidence in humans and mice indicates the involvement of matrix metalloproteinases (MMPs) and their endogenous tissue inhibitors (TIMPs) in cleft lip palate (CLP) aetiology. This study investigated whether expression of MMP-2, MMP-8, MMP-9, TIMP-2, and TIMP-4, which are essential for the upper lip and palate fusion, is dysregulated in children with CLP. Methods: Oral mucosa tissue samples were obtained from patients with complete unilateral (CU) CLP (n = 25) and complete bilateral (CB) CLP (n = 19) during corrective plastic surgery and in unaffected control subjects (n = 10). MMPs and TIMPs expression was assessed by immunohistochemistry, and the data were analyzed using the Kruskal - Wallis test with the Bonferroni correction. Results: In CLP patients, MMP-2, TIMP-2 immunoreactivity in the oral mucosa was seen to have a few to abundant structures, but the overall number of MMP-2, TIMP-2-positive structures was greater than that in controls (P < 0.01). The total number of TIMP-4, MMP-9-positive cells showed a significant decrease in the CBCLP compared with that of CUCLP (P < 0.001). MMP-8 expression trends in the CLP group were similar to those of the control group. Conclusions: The results suggest that TIMP-4 and MMP-9 are the main ECM remodeling regulatory proteins expressed in CUCLP affected tissues of the oral mucosa. The increased expression of MMP-2 and TIMP-2 in CLP tissues implicates these factors in the regulation of cell migration during ECM turnover independently of different types of clefts. Investigation of MMP and TIMP expression in tissue samples from patients with CLP appears to be a promising approach to the etiopathogenesis of CLP.

Background and objectives: Cleft lip palate takes the second place among all anomalies. The complex appearance of cytokines and proliferation markers has still not been clarified despite their possible crucial role in cleft tissue. Therefore, the aim of work was the detection of appearance of pro- and anti-inflammatory cytokines and proliferation marker Ki67, and their inter-correlations in cleft affected lip (CAL). Materials and Methods: The lip material was obtained from 16 children aged before primary dentition during plastic surgery. Control was obtained from 7 non-CAL oral tissue. Tissues were stained for IL-1, IL-4, IL-6, IL-8, IL-10 and Ki67 immunohistochemically. Non-parametric statistic, Mann&ndash, Whitney and Spearman&rsquo, s coefficient were used. Results: All cytokines positive cells were observed more into the epithelium. Statistically significant difference was seen between epithelial IL-1, IL-10, IL-8 and Ki67 positive cells and IL-10-, IL-4-containing connective tissue cells in comparison to the control. Strong positive correlation was detected in CAL epithelium between IL-10 and IL-8, IL-10 and IL-4, IL-10 and IL-1, IL-1 and IL-8, IL-1 and IL-4, IL-4 and IL-8, IL-8 and Ki67, IL-10 and Ki67, but moderate&mdash, in connective tissue between IL-1 and IL-10, IL-1 and IL-4. Conclusion: The CAL epithelium is the main source for the interleukins. Rich similar expression of IL-1 and IL-10 suggests the balance between pro-and anti-inflammatory tissue response on basis of dysregulated tissue homeostasis (increase of IL-8). The correlations between the different ILs -1, -4, -8, -10 in CAL epithelium seem to indicate the self-protection compensatory mechanism for intensification of local inflammatory-immune response without involvement of IL-6. The correlations between Ki67 and cytokines indicate the involvement of IL-8 and IL-10 in stimulation of cellular proliferation. IL-4 and IL-10 expression from CAL connective tissue simultaneously to IL-1, IL-4 and IL-10 inter-correlations there suggests the intensification of local immune response regulated probably by main pro-inflammatory cytokine&mdash, IL-1.

Cleft lip and palate are amongst the most common congenital malformations worldwide presenting with variable manifestations. Previous research has been primarily focused on the genetical aspects of its complex and multifactorial etiology. In the present study, we investigated the role of cytokines as mediators of epithelial–mesenchymal crosstalk and local site inflammation in cleft affected infants. Lip material was obtained from 12 children aged before primary dentition who suffered from orofacial clefting. The quantification of 12 cytokines (Interleukin-2,4,5,6,10,12,13,17A, Tumor Necrosis Factor-α, Interferon-γ, Transforming Growth Factor beta-1 and Granulocyte-Colony Stimulating Factor) was done using ELISA. Nonparametric Spearman Rho was used to ascertain the correlation between the expression levels of different cytokines. A significantly strong positive correlation was found between IL-2 and IFN-γ coupled with an IL4/IFN-γ ratio favoring IFN-γ. These findings indicate a shift towards the preferential activation of the Th1 differentiation pathway. Further, a pathological reduction in TGFβ-1 levels was noted, which may contribute to mucosal damage. IL-6 was more highly correlated to IFN-γ and IL-12 indicating its potential proinflammatory role in cleft affected tissues. This preferential activation of Th1 cell differentiation and consistent expression of IL-2,6,13 and TNF-α in cleft patients may indicate certain underlying mechanisms for inflammation mediation at the site of clefting.

AimsTo provide evidence-based and peer-reviewed recommendations for the development of dental continuing professional development (CPD) learning e-modules.MethodsThe present recommendations are consensus recommendations of the DentCPD project team and were informed by a literature research, consultations from e-learning and IT expert, discussions amongst the participants attending a special interest group during the 2012 ADEE meeting, and feedback from the evaluation procedures of the exemplar e-module (as described in a companion paper within this Supplement). The main focus of these recommendations is on the courses and modules organised and offered by dental schools.Results and discussionE-modules for dental CPD, as well as for other health professionals’ continuing education, have been implemented and evaluated for a number of years. Research shows that the development of e-modules is a team process, undertaken by academics, subject experts, pedagogists, IT and web designers, learning technologists and librarians. The e-module must have clear learning objectives (outcomes), addressing the learners’ individual needs, and must be visually attractive, relevant, interactive, promoting critical thinking and providing feedback. The text, graphics and animations must support the objectives and enable the learning process by creating an attractive, easy to navigate and interactive electronic environment. Technology is usually a concern for learners and tutors; therefore, it must be kept simple and interoperable within different systems and software. The pedagogical and technological proficiency of educators is of paramount importance, yet remains a challenge in many instances.ConclusionsThe development of e-courses and modules for dental CPD is an endeavour undertaken by a group of professionals. It must be underpinned by sound pedagogical and e-learning principles and must incorporate elements for effective visual learning and visual design and a simple, consistent technology.

AimFree movement of dental professionals across the European Union calls for more uniform continuing education in dentistry to ensure up-to-date, high-quality patient care and patient safety. This article provides guidelines for the management and delivery of high-quality continuing professional development (CPD) by European dental schools and other CPD providers.MethodThe guidelines are based on an extensive literature inventory, a survey of existing practices (both available as separate publications), discussions during meetings of the Association for Dental Education in Europe in 2011 and 2012 and debate amongst the members of the DentCPD project team representing six dental schools.ResultsOn the basis of the literature review, survey and discussions, we recommend that (i) every dentist should be given the opportunity for CPD, (ii) providers should be quality-approved and impartial, (iii) educators should be approved, impartial, suitably trained, and with educational expertise, (iv) the mode of CPD delivery should suit the educational activity, with clear learning objectives or outcomes, (v) effort should be made to assess the learning, (vi) participant feedback should be collected and analysed to inform future developments and (vii) uniform use of the pan-European system of learning credit points (ECTS) should be implemented.ConclusionImplementation of these guidelines should make dental CPD more transparent to all relevant parties and facilitate the transferability of earned credits across the European Union. It will also enable better quality control within dentistry, resulting in enhanced dental care and ultimately the improvement in patient safety.

IntroductionIn the context of free movement, EU-citizens need assurance that dental practitioners providing their care have a degree/license to practice that meets EU-standards and that they maintain their knowledge and skills through ongoing education.AimOne aim of the ‘DentCPD’ project (HYPERLINK ‘http://www.dentcpd.org’ www.dentcpd.org) was to identify and agree essential CPD requirements for EU dentists. This paper reports the consensus process and outcomes.MethodsAgreement on core components of CPD was achieved through a three stage process: an online survey of dental educators' (n = 143) views on compulsory topics; a paper-based questionnaire to practitioners (n = 411); leading to a proposal discussed at the Association for Dental Education (ADEE) 2011 Lifelong Learning special interest group (SIG).ResultsFrom the online survey and practitioner questionnaire, high levels of agreement were achieved for medical emergencies (89%), infection control (79%) and the medically compromised patient (71%). The SIG (34 attendees from 16 countries) concluded that these three CPD topics plus radiation protection should be core-compulsory and three CPD topics should be core-recommended (health and safety, pain management, and safeguarding children & vulnerable adults). They also agreed that the teaching of all topics should be underpinned by evidence-based dentistry.ConclusionBuilding four core topics into CPD requirements and making quality-approved education and training available will ensure that all dentists have up-to-date knowledge and skills in topic areas of direct relevance to patient safety. In turn, this will contribute to patients having access to comparably high standards of oral health care across Europe.

Formation of mammalian lip and palate involves multiple developmental steps that finally lead to the fusion of the two bilateral palatal shelves along the facial midline. Recent studies confirm that on the basis successful orofacial formation is coordinated interactions of several factors including genes, growth factors, enzymes and their natural inhibitors. The aim of our study was to investigate cleft disordered tissue in children with cleft lip and palate for detection of local expression of key regulators of palatogenesis. The study involved 10 patients with unilateral cleft lip and palate at the age of three months to four years and eight months. Tissue samples were collected during the surgical procedure from the borders of the cleft region. Material was proceed for detection of PAX9, RYK, TGFβ-3, MMP-8, MMP-9 and TIMP-2 with biotin-streptavidin immunohistochemistry. Distribution of immunoreactive structures was detected semiquantitatively. Expression of PAX9 and RYK was more pronounced in cleft lip and palate disordered epithelium. The relative number of PAX9 positive epitheliocytes was very variable, while RYK mainly stained few cells localized into middle layers. Expression of TGFβ-3 was detected in the tissue of all patients. We saw numerous positive epitheliocytes in eight cases, but underlying connective tissue mainly demonstrated few or moderate number immunoreactive cells. Expression of MMP-8 was found only in epithelium of eight patients and it was slight, while MMP-9 and TIMP-2 marked variable number of positive epithelial and connective tissue cells. Conclusions. Expression of Pax-9 is more characteristic for cleft lip and palate affected tissue and probably facilitates tissue reconstruction due to its mitogenic effect. Rich expression of TGFβ-3 in cleft lip and palate disordered tissue may play a role in successful tissue remodelling and scar-free healing. Cleft lip and palate disordered tissue are characterizied by more pronounced expression of MMP-9, it slightly predominate expression of TIMP-2, giving evidence to the involvement of mentioned factors in the postnatal tissue remodeling.