Abstract
Triiodothyronine (T3) is a thyroid hormone that can have varying effects on skin. In order to assess the effects of T3 on the human dermis, we prepared dermal equivalents using neonatal dermal cells via the process of self-assembly in the presence of differing concentrations of T3. These dermal equivalents were prepared in the absence of serum and a three dimensional matrix allowing for the direct assessment of different concentrations of T3 on dermal extracellular matrix formation. Three different concentrations of T3 were chosen, 20 pM, which is part of the base medium, 0.2 nM T3 and 2 nM T3. We find that self-assembled dermal equivalents formed under these conditions show a progressive “thinning” with increasing T3 concentrations. While we observed no change in total collagen content, inhibition of hyaluronate (HA) synthesis was observed in the 0.2- and 2-nM T3 constructs as compared to the 20-pM construct. Other glycosaminoglycan synthesis was not affected by increasing T3 concentrations. In order to identify the gene(s) responsible for inhibition of HA synthesis in the 2-nM T3 dermal equivalent, we conducted a differential gene array analysis. The results of these experiments demonstrate the differential expression of 40 genes, of these, 34 were upregulated and 6 genes were downregulated. The results from these experiments suggest that downregulation of HAS2 may be responsible for inhibition of hyaluronate synthesis in the self-assembled 2-nM T3 human dermal matrix.
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Benjamini Y.; Hochberg Y. Controlling the false discovery rate: a practical and powerful approach to multiple testing. J. R. Statist. Soc. B 57: 289–300; 1995.
Bolstad B. M.; Irizarry R. A.; Astrand M.; Speed T. P. A Comparison of Normalization Methods for High Density Oligonucleotide Array Data Based on Bias and Variance. Bioinformatics 19: 185–193; 2003.
Brown S.; Worsfold M.; Sharp C. Microplate Assay for the Measurement of Hydroxyproline in Acid-Hydrolyzed Tissue Samples. Biotechniques 30: 38–42; 2001.
Calabro A.; Benavides M.; Tammi M.; Hascall V. C.; Midura R. J. Microanalysis of enzyme digests of hyaluronan and chondroitin/dermatan sulfate by fluorophore assisted carbohydrate electrophoresis (FACE). Glycobiology 10: 273–81; 2000a.
Calabro A.; Hascall V. C.; Midura R. J. Adaptation of FACE methodology for microanalysis of total hyaluronan in chondroitin sulfate composition from cartilage. Glycobiology 10: 283–293; 2000b.
Contreras-Jurado C.; García-Serrano L.; Gomez-Ferrería M.; Costa C.; Paramio J. M.; Aranda A. The Thyroid Hormone Receptors as Modulators of Skin Proliferation and Inflammation. J. Biol. Chem 286: 24079–24088; 2011.
De Rycker C.; Vandelem J. L.; Hennen G. Effect of 3,5,3′-triiodothyronine on collagen synthesis by cultured human skin fibroblasts. FEBS Let 174: 34; 1984.
Fink C. W.; Ferguson J. L.; Smiley J. D. Effect of hyperthyroidism and hypothyroidism on collagen metabolism. J. Lab. Clin. Med 69: 950–959; 1967.
Fosang A. J.; Hey N. J.; Carney S. L.; Hardingham T. E. An ELISA plate-based assay for hyaluronan using biotinylated proteoglycan G1 domain. Matrix 10: 306–313; 1990.
Irizarry R. A.; Bolstad B. M.; Collin F.; Cope L. M.; Hobbs B.; Speed T. P. Summaries of Affymetrix GeneChip probe level data. Nucleic Acids Res 31: e15; 2003.
Itano N.; Kimata K. Mammalian Hyaluronan Synthases. IUBMB Life 54: 195–199; 2002.
Mehregan A. H.; Zamick P. The effect of triiodothyronine in healing of deep dermal burns and marginal scars of skin grafts. A histologic study. J. Cutan. Pathol 1: 113–116; 1974.
Moeller L. C.; Dumitrescu A. M.; Walker R. L.; Meltzer P. S.; Refetoff S. Thyroid Hormone Responsive Genes in Cultured Human Fibroblasts. J. Clin. Endocrinol. Metab 90: 936–943; 2005.
Murata Y.; Ceccarelli P.; Refetoff S.; Horwitz A. L.; Matsui N. Thyroid hormone inhibits fibronectin synthesis by cultured human skin fibroblasts. J. Clin. Endocrinol. Metab 64: 334–339; 1987.
Noble P. W. Hyaluronan and its catabolic products in tissue injury and repair. Matrix Biol 21: 25–9; 2002.
Paus R. Exploring the “Thyroid–Skin Connection”: Concepts, Questions, and Clinical Relevance. J. Invest. Dermatol 130: 7–10; 2010.
Pouyani T.; Ronfard V.; Scott P. G.; Dodd C. M.; Ahmed A.; Gallo R.; Parenteau N. De Novo Synthesis of Human Dermis In Vitro in the Absence of a Three-Dimensional Scaffold. In Vitro Cell Dev. Biol.-Anim 45: 430–441; 2009.
Pouyani T.; Schaffer L.; Papp S. Tissue Engineered Fetal Dermal Matrices. In Vitro Cell Dev. Biol.-Anim 8: 493–506; 2012.
Safer J. D.; Crawford T. M.; Holick M. F. A Role for Thyroid Hormone in Wound Healing through Keratin Gene Expression. Endocrinology 145: 2357–2361; 2004.
Safer J. D.; Crawford T. M.; Holick M. F. Topical thyroid hormone accelerates wound healing in mice. Endocrinology 146: 4425–4430; 2005.
Safer J. D.; Fraser L. M.; Ray S.; Holick M. F. Topical triiodothyronine stimulates epidermal proliferation, dermal thickening, and hair growth in mice and rats. Thyroid 11: 717–724; 2001.
Smith T. J.; Bahn R. S.; Gorman C. A. Connective tissue, glycosaminoglycans and diseases of the thyroid. Endocrine Rev 10: 366–391; 1989.
Smith T. J.; Murata Y.; Horwitz A. L.; Philipson L.; Refetoff S. Regulation of glycosaminoglycan synthesis by thyroid hormone in vitro. J. Clin. Invest 70: 1066–1073; 1982.
Smyth G. K. Linear models and empirical Bayes methods for assessing expression in microarray experiments. Stat. Appl. Genet. Molec. Biol 3(1); 2004. Article 3.
Stern R. Devising a pathway for hyaluronan catabolism: are we there yet? Glycobiology 13: 105R–15R; 2003.
Stern R. Hyaluronan metabolism: a major paradox in cancer biology. Pathol Biol 53: 372–382; 2005.
van Beek N.; Bodó E.; Kromminga A.; Gáspár E.; Meyer K.; Zmijewski M. A.; Slominski A.; Wenzel B. E.; Paus R. Thyroid hormones directly alter human hair follicle functions: anagen prolongation and stimulation of both hair matrix keratinocyte proliferation and hair pigmentation. J. Clin. Endocrinol. Metab 93: 4381–4388; 2008.
Zamick P.; Mehregan A. H. Effect of l-tri-iodothyronine on marginal scars of skin grafted burns in rats. Plas. Reconstr. Surg 51: 71–75; 1973.
Acknowledgments
We thank Maria Ericsson and Louise Trakimas (Harvard Medical School) for their valuable assistance with Transmission Electron Microscopy. We are grateful to Gilberto Hernandez (Scripps Research Institute) for his assistance with RNA analysis. This research was supported by a grant from the Charles H. Hood Foundation to TP.
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Pouyani, T., Sadaka, B.H., Papp, S. et al. Triiodothyronine (T3) inhibits hyaluronate synthesis in a human dermal equivalent by downregulation of HAS2. In Vitro Cell.Dev.Biol.-Animal 49, 178–188 (2013). https://doi.org/10.1007/s11626-013-9583-7
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DOI: https://doi.org/10.1007/s11626-013-9583-7