Differential effect of Cyclosporin A and FK506 on SPARC mRNA expression by human gingival fibroblasts
Department of Human Morphology-LITA, University of Milan, Via Fratelli Cervi 93, 20090 Segrate, Milan, Italy. Biomedecine [?] Pharmacotherapy
(Impact Factor: 2.02).
07/2005; 59(5):249-52. DOI: 10.1016/j.biopha.2004.06.005
Secreted protein acidic and rich in cysteine (SPARC) is a glycoprotein that mediates cell-matrix interactions. In adults, its expression is mostly limited to tissue undergoing remodeling. During the development of Cyclosporin A (CsA)-induced gingival overgrowth (GO) a remodeling of the connective compartment occurs. By contrast, clinical trials showed that FK506 is not related to GO. SPARC expression and its involvement in GO is unknown. Our aim was, therefore, to analyze the effect of CsA and FK506 on SPARC gene expression.
Cultured human gingival fibroblasts were incubated with CsA, FK506 or with their vehicle (VH) for 24, 48 and 72 h. SPARC gene expression was determined by RT-PCR.
SPARC mRNA levels tended to increase 72 h after CsA treatment, whilst they are undetectable in FK506-treated fibroblasts, compared to VH.
This gene expression profile is consistent with the involvement of SPARC in the mechanisms leading to the development of CsA-induced GO. By contrast, the undetectable SPARC mRNA levels in FK506-treated fibroblasts suggest that FK506 may be associated with a role of ECM stabilization, that does not induce GO.
Available from: Nicoletta Gagliano
- "In adults, the expression of SPARC is limited mostly to tissue undergoing morphogenesis or remodeling due to wound healing, disease, or natural processes (Masson et al., 1998; Brekken and Sage, 2001; Gagliano et al., 2005). SPARC is expressed at high levels in bone tissue but is widely distributed in many other tissues, such as connective tissues, including the soft and mineralized tissue of teeth and their supporting periodontal tissues, according to the high turnover of the remodeling of healthy oral connective tissue (Bartold et al., 2000; Sodek et al., 2002). "
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ABSTRACT: Abstract Objective: We aimed to characterize the fibroblast phenotype of patients by analyzing gene and protein expression of cleft lip and/or cleft palate fibroblasts in relation to collagen turnover and extracellular matrix remodeling. Patients: Human palatal fibroblasts were obtained from three healthy subjects without cleft lip and/or cleft palate and from three subjects with nonsyndromic cleft lip and/or cleft palate. Collagen turnover-related gene and protein expression were analyzed by real-time polymerase chain reaction, Western and dot blots, and sodium dodecyl sulfate zymography. Results: Cleft lip and/or cleft palate fibroblasts, compared with controls, displayed a down-regulation of collagens type I and III messenger RNA (p < .0001 and p < .001, respectively) but an opposite tendency to increase protein levels. Cleft lip and/or cleft palate cells had higher lysyl hydroxylase-2b messenger RNA levels expressed in relation to collagen type I messenger RNA, down-regulated matrix metalloproteinase-1, tissue inhibitor of matrix metalloproteinase-1, and Secreted Protein Acidic and Rich in Cysteine messenger RNA (p < .0001 and p < .01, respectively). Pro-matrix metalloproteinase-1 tended to decrease, and pro-matrix metalloproteinase-2 and -9 were down-regulated (p < .01, p < .05, respectively), as was Secreted Protein Acidic and Rich in Cysteine protein expression (p < .05). Conclusions: Our results suggest that the cleft lip and/or cleft palate fibroblast phenotype is characterized by a tendency toward interstitial collagen deposition due to posttranslational modifications, such as decreased collagen degradation by matrix metalloproteinases and increased collagen cross-links. These findings may contribute to the knowledge of the cleft lip and/or cleft palate fibroblast phenotype and may be useful to the surgeon when considering the potential wound contraction and subsequent undesired scarring in cleft lip and/or cleft palate ocurring after the surgical closure of a cleft palate.
The Cleft Palate-Craniofacial Journal 07/2010; 47(4):393-9. DOI:10.1597/07-196.1 · 1.20 Impact Factor
Available from: Denise Carleto Andia
- "In agreement with previous studies (Hernandez et al, 2000; James et al, 2000), the results of the present investigation show that tacrolimus administration for brief periods of treatment (60 and 120 days) does not induce gingival overgrowth. It has been suggested that in vitro tacrolimus does not affect the collagen type I gene and protein expression, transforming growth factor (TGF-b1) and tissue inhibitor of matrix metalloproteinase (MMPs) (TIMP-1) mRNA levels, although it significantly increases MMP-1 gene and protein expression and MMP-2 mRNA levels (Gagliano et al, 2005b). "
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ABSTRACT: Tacrolimus, an immunosuppressive drug used in organ transplantation, has been reported not to induce gingival overgrowth. However, prevalence studies are limited, and the methods used for assessing gingival overgrowth varies among studies.
The purpose of this study was to evaluate the effects of up to 240 days of tacrolimus therapy on gingival tissues of rats.
Rats were treated for 60, 120, 180 and 240 days with daily subcutaneous injections of 1 mg/kg body weight of tacrolimus. After histological processing, the oral and connective tissue, volume densities of fibroblasts (Vf), collagen fibers (Vcf) and other structures (Vo) were assessed in the region of the lower first molar.
After 60 and 120 days of treatment with tacrolimus, gingival overgrowth was not observed. The gingival epithelium, connective tissue, as well as the values for Vf, Vcf, and Vo were similar to those of the control rats (P>0.05). After 180 and 240 days of the treatment, gingival overgrowth was associated with a significant increase in the gingival epithelium and connective tissue as well as an increase in the Vf and Vcf (P<0.05).
Within the limits of the experimental study, it may be concluded that the deleterious side effects of tacrolimus on the gingival tissues of rats may be time-related.
Oral Diseases 01/2008; 14(1):67-72. DOI:10.1111/j.1601-0825.2006.01354.x · 2.43 Impact Factor
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ABSTRACT: Cyclosporin A is a powerful immunosuppressive drug with considerable impact on transplants and is able to modify extracellular matrix (ECM) composition. It has recently been demonstrated that cyclosporin A stimulates the production of the cytokine family. Cytokines such as interleukin, transforming growth factor beta(1), and bone morphogenetic protein induce the deposition of glycosaminoglycans (GAGs), proteoglycans, and collagen fibers in the connective ECM. ECM composition is very important for normal tissue development and function. In this work, we examine the effects caused by cyclosporin A on cultures of normal human palate fibroblasts in order to evaluate interleukin, transforming growth factor beta II, and bone morphogenetic protein II membrane receptor induction and extracellular GAG changes such as hyaluronic acid, heparin sulfate, and chondroitin sulfate.
Palate fibroblasts were maintained for 24 h in serum-free 199 medium containing 5 microg/mL (3)H glucosamine hydrochloride. After this time, TGF II and BMP II receptors were determined by microarray analysis and GAG classes by the biochemical method.
The results show that TGFbeta(1) II and BMP II membrane receptors are significantly inhibited in cyclosporin A-treated cultures as compared to controls, whereas IL-1R2 membrane receptors are stimulated. The behavior of total intra- and extracellular GAGs is significantly increased in cyclosporin A-treated cultures, whereas the ratio between non-sulfated/sulfated GAGs decreases (p <or=0.01) vis-à-vis controls.
Because they form a highly complicated macromolecular network in the ECM, which provides an indication of cell function and gene expression and modulates growth factor activities, GAG changes are related to modification of ECM functions. Our data show that cyclosporin A causes biochemical changes to ECM through alterations in cytokines and respective membrane receptor linkages.
Archives of Medical Research 10/2007; 38(7):717-22. DOI:10.1016/j.arcmed.2007.03.007 · 2.65 Impact Factor
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