MMPs and TIMPs expression in facial tissue of children with cleft lip and palate

Abstract

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.

Ultrastructural analysis of collagen fibril diameter distribution in cleft lip

“…Studies elsewhere have shown that small and large fibrils have distinct roles in stiff state, and only the small fibrils have a role in the disease states and hence lending to changing properties (Goh & Holmes, ), which could be the case in our medial tissues. Nonetheless, the possible role of factors apart from CF‐ED or a consequence of CF‐ED cannot be ruled out for compromised extracellular matrix of the medial tissue, based on previously described ability of metalloproteinases and their inhibitors to participate in matrix remodeling during lip fusion (Smane‐Filipova et al, ). We postulate that remodeling of the extracellular matrix components (collagen fibers and/or proteoglycans) that form the migration substrate for the cranial neural crest cells mesenchyme (Henderson & Copp, ; Jiang et al, ) could be different across the two sides.…”

“…A fine modulation in the extracellular matrix in the orofacial region during development is essential for the cells to interact and respond to the remodeling or change in mechanical properties of the extracellular matrix (Gagliano et al, ; McDaniel et al, ), which is likely critical to cellular migration, differentiation (Badylak, ), and etiology of orofacial cleft (Mansell et al, ; Smane‐Filipova, Pilmane, & Akota., ). Studies in experimental models of fibroblast cells obtained from CL/P cases have suggested molecular mechanisms involved in phenotypic variation among fibroblasts (Bodo et al, ; Bosi et al, ).…”

“…Additionally, our group has recently found a variant in this gene to be associated with non‐syndromic cleft palate (Khan, Little, Mossey, Steegers‐Theunissen, Bonsi, et al, ). The morphogenesis of lip and palate fusion involves highly regulated sharing of signaling molecules, and so it is possible that lip fusion promotes palate fusion (Smane‐Filipova et al, ). Therefore, improper fusion of the lip may secondarily affect palate fusion (Meng, Bian, Torensma, & Von den Hoff, ).…”

Extracellular Matrix Remodeling During Palate Development

“… IHC 100 , 102 , 106 NR NR MMP-9/Gelatinase B Extensively expressed in palatal mesenchyme, transiently upregulated while palatal shelves elevating and fusing. qRT-PCR, IHC, ISH Human and mouse: 24 , 102 , 104 Cleaves laminin NR MMP13 Before palate shelf elevation: in the palatal mesenchyme and basement membrane, intensified gradually in the nasal-medial part. During the palatal shelf elevation and the palatal shelf fusion: intensified gradually in the tip and nasal-medial part and MES After the palatal shelf fusion: still strong in the nasal-medial mesenchyme and MES but decreased in other area.…”

“… IHC 100 NR NR TIMP-3 Palatal epithelium, transiently expressed in mid-oral and ventral-medial mesenchyme. IHC 100 NR NR TIMP-4 Oral mucosa IHC 104 NR NR ADAMTS ADAMTS9 Palatal capillary endothelium ISH 12 Versican Haploinsufficiendy of Adamts9 in Adamts20 mutant mice ( Adamts9 ± ;Adamts20 bt/bt ) Complete cleft palate 12 ADAMTS20 Palatal mesenchyme ISH 12 NR, not reported. IHC, immunohistochemical staining.…”