Expression of growth factors and growth factor receptors in human cleft-affected tissue

OBJECTIVE. To investigate cleft disordered tissue in children with cleft palate and cleft lip with or without alveolar clefting for detection of local tissue growth factors and growth factor receptors and compare findings. Design. Morphological analysis of human tissue. Patients. Three groups were studied: 14 patients with cleft palate at the age from eight months to 18 years and two months, 12 patients with cleft lip with or without alveolar clefting in the age from four months to 15 years and four months and 11 control patients. RESULTS. In general, cleft palate disordered tissue showed more prominent expression of BMP2/4 (z=3.574; p=0.0004) and TGFβ (z=2.127; p=0.033), while expression of TGFBR3 significantly higher was only in connective tissue (z=3.822; p=0.0001). Cleft lip affected tissue showed significantly pronounced expression of FGFR1 in general as well as separately in epithelium. CONCLUSIONS. The marked and statistically significant expression of BMP 2/4 in cleft palate disordered soft tissue probably is delayed, but still proliferation and differentiation as well as tissue, especially, bone remodeling contributing signal. Cleft palate affected tissue show more prominent expression of TGFβ, still the weak regional expression of TGFβ type III receptors prove the disordered tissue growth and changed TGFβ signalling pathway in postnatal pathogenesis. In general, expression of TGFβ, BMP 2/4 and FGFR1 is significantly different, giving evidence to the involvement of these mentioned factors in the cleft severity morphopathogenesis.publishersversionPeer reviewe

Barx1, growth factors and apoptosis in facial tissue of children with clefts

OBJECTIVE: Clefts of lip and palate belong to the most common birth defects worldwide. Growth factors and genes play an important role in tissue growth, differentiation and induction and upregulation of growth factors, apoptosis and matrix metalloproteinases might be involved in pathogenesis of facial clefts. The aim of this study was investigation of palate tissue in children with unilateral cleft lip palate for detection of local tissue growth factors, barx1 and apoptosis. MATERIALS AND METHODS:We investigated soft and hard palate tissue from 36 children with complete unilateral cleft lip and palate from cleft area.14 children were in age before and primary dentition, but 22 children were in mixed dentition period. We examined the localization of barx1, FGFR1, NGFR, TGFbeta, BMP2/4, MMP2, PGP 9,5 by immunohistochemistry. TUNEL method was performed for detection of apoptotic cells. RESULTS: Abundance of FGFR1 positive cells was seen almost in all cases. FGFR richly stained cells of soft and hard palate tissue. Abundance of NGFR positive cells was detected in basal epithelium, hair follicles, nerve fibers in wall of blood vessels and subepithelium, and was more often seen in children before mixed dentition. TGFbeta has showed intensive expression in epithelium, cartilage and bone in both dentition ages. Chondrocytes, fibroblasts and macrophages expressed MMP2 predominant before mixed dentition. Regional expression of barx1 was observed in epithelium before the mixed dentition, while during mixed dentition gene appeared in hyaline cartilage. TUNEL discovered apoptosis in both dentition ages. CONCLUSIONS: FGFR1 and TGFbeta are main tissue stimulating growth factors in both dentition ages. Expression of barx1 appears in cleft lip palate affected structures mainly in mixed dentition ages. NGFR and neuropeptides-containing structures are mainly characteristic in cleft tissue before mixed dentition. Distribution of genes, GF and apoptosis seem to correlate rather with dentition age than to type of CLP.publishersversionPeer reviewe

Expression of interferon regulatory factor 6, muscle segment homeobox 1, paired box gene 9, homeo box B3, and related to tyrosine kinases in human cleft-affected tissue

Publisher Copyright: © 2016 Journal of Orofacial Sciences | Published by Wolters Kluwer - Medknow.Background and Aim: Recent studies demonstrate direct roles of different genes during formation of secondary palate, but there are no still data about local expression and distribution of gene products in cleft palate affected human tissue. Thus, the aim of our study was to investigate cleft disordered cartilage and bone for detection of local expression of key regulators of palatogenesis and its correlations. Materials and Methods: The study involved 16 patients with unilateral cleft lip and palate. Tissue samples were proceeded for detection of interferon regulatory factor 6 (IRF6), muscle segment homeobox 1 (MSX1), paired box gene 9 (PAX9), homeo box B3 (HOXB3), and related to tyrosine kinases with biotin-streptavidin immunohistochemistry. Distribution of immunoreactive structures was detected semiquantitatively. Statistical analysis included the Mann-Whitney test and Pearson′s correlation test. Results: Statistically significant differences were found between expression of IGFR6, MSX1, and HOXB3 in the cartilage and bone. We also detected statistically significant correlation between the expressions of PAX9 and MSX1 in the bone tissue. Conclusions: Cleft lip and palate disordered cartilage is characterized by more pronounced expression of IRF6, MSX1, and PAX9. Expression of HOXB3 is more characteristic for cleft lip and palate affected bone. Considered as a whole, our results suggest that the cleft lip and palate affected cartilage seems more plastic in tissue remodeling what can probably result in qualitative postoperative tissue reconstruction.Peer reviewe

Lūpas un aukslēju šķeltņu zonas audu funkcionālā morfoloģija operāciju materiālā. Promocijas darba kopsavilkums

The Doctoral Thesis was developed at the Department of Morphology of the Institute of Anatomy and Anthropology of the Rīga Stradiņš University. Defence: on the 28th June, 2010 in the Hippocrates Lecture Room of the Rīga Stradiņš University, Dzirciema Street 16, Riga.The Promotion work has been elaborated with ESF project support “Support for doctoral and post-doctoral investigations Riga Stradins University

Functional Morphology of the Cleft Lip and Palate Affected Tissue in the Material of Operations. Summary of the Doctoral Thesis

Promocijas darbs veikts Rīgas Stradiņa Universitātes Anatomijas un antropoloģijas institūtā Morfoloģijas katedrā. Aizstāvēšana: 2010. gada 28. jūnijā plkst. 15.00 Rīgas Stradiņa universitātes promocijas padomes atklātajā sēdē Hipokrāta auditorijā, Rīgā, Dzirciema ielā 16.Projekts veikts ar ESF projekta „Atbalsts doktorantiem studiju programmas apguvei un zinātniskā grāda ieguvei Rīgas Stradiņa Universitātē” atbalst

Functional Morphology of the Cleft Lip and Palate Affected Tissue in the Material of Operations. Doctoral Thesis

Promocijas darbs veikts Rīgas Stradiņa Universitātes Anatomijas un antropoloģijas institūtā Morfoloģijas katedrā. Aizstāvēšana: 2010. gada 28. jūnijā plkst. 15.00 Rīgas Stradiņa universitātes promocijas padomes atklātajā sēdē Hipokrāta auditorijā, Rīgā, Dzirciema ielā 16.Projekts veikts ar ESF projekta „Atbalsts doktorantiem studiju programmas apguvei un zinātniskā grāda ieguvei Rīgas Stradiņa Universitātē” atbalst

Expression of TGFβ-3, PAX9, RYK, matrix metalloproteinases and their tissue inhibitors in human cleft-affected tissue

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.publishersversionPeer reviewe