Abstract
The aim of the present study was to determine the effects of UV irradiation on the conversion of 7-dehydrocholesterol (7-DHC), which has been coated onto a polystyrene surface, to cholecalciferol (D3), and the resulting effect on the formation of vitamin D (1,25-D3) by MC3T3-E1 cells. The changes in gene expression of the enzymes regulating its hydroxylation, Cyp27b1 and Cyp27a1, were monitored as well as the net effect of the UV-treated 7-DHC coating on cell viability and osteoblast differentiation. MC3T3-E1 cells were found to express the enzymes required for synthesizing active 1,25-D3, and we found a dose-dependent increase in the production of both 25-D3 and 1,25-D3 levels for UV-activated 7-DHC samples unlike UV-untreated ones. Cell viability revealed no cytotoxic effect for any of the treatments, but only for the highest dose of 7-DHC (20 nmol per well) that was UV-irradiated. Furthermore, osteoblast differentiation was increased in cells treated with some of the higher doses of 7-DHC when UV-irradiated, as shown by collagen-I, osterix and osteocalcin relative mRNA levels. The conversion of 7-DHC to preD3 exogenously by UV irradiation and later to 25-D3 by MC3T3-E1 cells was determined for the optimum 7-DHC dose (0.2 nmol per well), i.e. 8.6 ± 0.7% of UV-activated 7-DHC was converted to preD3 and 6.7 ± 2.8% of preD3 was finally converted to 25-D3 under the conditions studied. In conclusion, we demonstrate that an exogenous coating of 7-DHC, when UV-irradiated, can be used to endogenously produce active vitamin D. We hereby provide the scientific basis for UV-activated 7-DHC coating as a feasible approach for implant therapeutics focused on bone regeneration.
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Abbreviations
- 7-DHC:
-
7-Dehydrocholesterol
- D3:
-
Cholecalciferol
- 25-D3:
-
25-Hydroxyvitamin D3
- 1,25-D3:
-
1,25-Dihydroxyvitamin D3
- Cyp27a1:
-
Vitamin D3 25-hydroxylase
- Cyp27b1:
-
25-Hydroxyvitamin D3-1alpha-hydroxylase
References
G. J. Atkins, P. H. Anderson, D. M. Findlay, K. J. Welldon, C. Vincent, A. C. Zannettino, P. D. O’Loughlin and H. A. Morris, Metabolism of vitamin D3 in human osteoblasts: evidence for autocrine and paracrine activities of 1 alpha,25-dihydroxyvitamin D3, Bone, 2007, 40, 1517–1528.
R. St-Arnaud, The direct role of vitamin D on bone homeostasis, Arch. Biochem. Biophys., 2008, 473, 225–230.
B. E. C. Nordin and H. A. Morris, Osteoporosis and vitamin D, J. Cell. Biochem., 1992, 49, 19–25.
D. Ayyar, F. Floyd, D. Barwick, P. Hudgson and D. Weightman, Myopathy in chronic renal failure, Q. J. Med., 1974, XLIII, 509–524.
P. Irani, Electromyography in nutritional osteomalaic myopathy, J. Neurol. Neurosurg. Psychiatry, 1976, 686–693.
L. Ceglia, Vitamin D and skeletal muscle tissue and function, Mol. Aspects Med., 2008, 29, 407–414.
J. B. Eastwood, P. J. Bordier and E. M. Clarkson, The contrasting effects on bone histology of vitamin D and of calcium carbonate in the osteomalacia of chronic renal failure, Clin. Sci. Mol. Med., 1974, 47, 23–42.
J. B. Eastwood, T. C. B. Stamp, H. E. De Wardener, The effect of 25 hydroxy vitamin D3 in the osteomalacia of chronic renal failure, Clin. Sci. Mol. Med., 1977, 52, 499–508.
M. Samson, H. J. J. Verhaar, P. A. F. Jansen, P. L. de Vreede, J. W. Manten and S. A. Duursma, Muscle strength, functional mobility and vitamin D in older women, Aging Clin. Exp. Res., 2000, 12, 455–460.
M. Pfeifer, B. Begerow, H. W. Minne, C. Abrams, D. Nachtigall and C. Hansen, Effects of a short-term vitamin D and calcium supplementation on body sway and secondary hyperparathyroidism in elderly women, J. Bone Miner. Res., 2000, 15, 1113–1118.
M. F. Holick, Vitamin D: its role in cancer prevention and treatment, Prog. Biophys. Mol. Biol., 2006, 92, 49–59.
J. S. Adams and M. Hewison, Unexpected actions of vitamin D: new perspectives on the regulation of innate and adaptive immunity, Nat. Clin. Pract. Endocrinol. Metab., 2008, 4, 80–90.
J. L. Omdahl, H. A. Morris and B. K. May, Hydroxylase enzymes of the vitamin D pathway: expression, function, and regulation, Annu. Rev. Nutr., 2002, 22, 139–166.
P. H. Anderson, S. Iida, J. H. T. Tyson, A. G. Turner and H. A. Morris, Bone CYP27B1 gene expression is increased with high dietary calcium and in mineralising osteoblasts, J. Steroid. Biochem. Mol. Biol., 2010, 121, 71–75.
I. Aiba, T. Yamasaki, T. Shinki, S. Izumi, K. Yamamoto, S. Yamada, H. Terato, H. Ide and Y. Ohyama, Characterization of rat and human CYP2J enzymes as vitamin D 25-hydroxylases, Steroids, 2006, 71, 849–856.
B. Lehmann and M. Meurer, Extrarenal sites of calcitriol synthesis: the particular role of the skin, Recent Results Cancer Res., 2003, 164, 135–145.
M. Schuessler, N. Astecker, G. Herzig, G. Vorisek and I. Schuster, Skin is an autonomous organ in synthesis, two-step activation and degradation of vitamin D3: CYP27 in epidermis completes the set of essential vitamin D3-hydroxylases, Steroids, 2001, 66, 399–408.
D. D. Bikle, Vitamin D regulated keratinocyte differentiation, J. Cell. Biochem., 2004, 92, 436–444.
B. W. Hollis, 25-Hydroxyvitamin D3-1a-hydroxylase in porcine hepatic tissue: subcellular localization to both mitochondria and microsomes, Proc. Natl. Acad. Sci. U. S. A., 1990, 87, 6009–6013.
D. Zehnder, R. Bland, M. C. Williams, R. W. McNinch, A. J. Howie, P. M. Stewart and M. Hewison, Extrarenal expression of 25-hydroxyvitamin D3-1a-hydroxylase, J. Clin. Endocrinol. Metab., 2001, 86, 888–894.
M. Hewison, L. Freeman, S. V. Hughes, K. N. Evans, R. Bland, A. G. Eliopoulos, M. D. Kilby, P. A. H. Moss and R. Chakraverty, Differential regulation of vitamin D receptor and its ligand in human monocyte-derived dendritic cells, J. Immunol., 2003, 170, 5382–5390.
S. Hansdottir, M. M. Monick, S. L. Hinde, N. Lovan, D. C. Look and G. W. Hunninghake, Respiratory epithelial cells convert inactive vitamin D to its active form: potential effects on host defense, J. Immunol., 2008, 181, 7090–7099.
G. A. Howard, R. T. Turner, D. J. Sherrard and D. J. Baylink, Human bone cells in culture metabolize 25-hydroxyvitamin D3 to 1,25-dihydroxyvitamin D3 and 24,25-dihydroxyvitamin D3, J. Biol. Chem., 1981, 256, 7738–7740.
F. Ichikawa, K. Sato, M. Nanjo, Y. Nishii, T. Shinki, N. Takahashi and T. Suda, Mouse primary osteoblasts express vitamin D3 25-hydroxylase mRNA and convert 1a-hydroxyvitamin D3 into 1a,25-dihydroxyvitamin D3, Bone, 1995, 16, 129–135.
D. K. Panda, S. A. Kawas, M. F. Seldin, G. N. Hendy and D. Goltzman, 25-Hydroxyvitamin D 1a-hydroxylase: structure of the mouse gene, chromosomal assignment, and developmental expression, J. Bone Miner. Res., 2001, 16, 46–56.
P. H. Anderson, P. D. O’Loughlin, B. K. May and H. A. Morris, Modulation of CYP27B1 and CYP24 mRNA expression in bone is independent of circulating 1,25(OH)2D3 levels, Bone, 2005, 36, 654–662.
M. Van Driel, H. A. P. Pols, J. P. T. M. Van Leeuwen, Osteoblast differentiation and control by vitamin D and vitamin D metabolites, Curr. Pharm. Des., 2004, 10, 2535–2555.
M. Van Driel, M. Koedam, C. J. Buurman, M. Hewison, H. Chiba, A. G. Uitterlinden, H. A. P. Pols, J. P. T. M. Van Leeuwen, Evidence for auto/paracrine actions of vitamin D in bone: 1a-Hydroxylase expression and activity in human bone cells, FASEB J., 2006, 20, E1811–E1819.
V. J. Woeckel, R. D. A. M. Alves, S. M. A. Swagemakers, M. Eijken, H. Chiba, B. C. J. Van Der Eerden, J. P. T. M. Van Leeuwen, 1a,25-(OH)2D3 acts in the early phase of osteoblast differentiation to enhance mineralization via accelerated production of mature matrix vesicles, J. Cell. Physiol., 2010, 225, 593–600.
P. H. Anderson, S. Iida, J. H. T. Tyson, A. G. Turner and H. A. Morris, Bone CYP27B1 gene expression is increased with high dietary calcium and in mineralising osteoblasts, J. Steroid Biochem. Mol. Biol., 2010, 121, 71–75.
M. F. Holick, J. A. MacLaughlin and M. B. Clark, Photosynthesis of previtamin D3 in human skin and the physiologic consequences, Science, 1980, 210, 203–205.
M. K. Nemanic, J. Whitney and P. M. Elias, In vitro synthesis of vitamin D-3 by cultured human keratinocytes and fibroblasts: action spectrum and effect of AY-9944, Biochim. Biophys. Acta, Gen. Subj., 1985, 841, 267–277.
B. Lehmann, T. Genehr, P. Knuschke, J. Pietzsch and M. Meurer, UVB-induced conversion of 7-dehydrocholesterol to 1a,25- dihydroxyvitamin D3 in an in vitro human skin equivalent model, J. Invest. Dermatol., 2001, 117, 1179–1185.
J. A. MacLaughlin, R. R. Anderson and M. F. Holick, Spectral character of sunlight modulates photosynthesis of previtamin D3 and its photoisomers in human skin, Science, 1982, 216, 1001–1003.
L. Xu, Z. Korade and N. A. Porter, Oxysterols from free radical chain oxidation of 7-dehydrocholesterol: product and mechanistic studies, J. Am. Chem. Soc., 2010, 132, 2222–2232.
D. E. MacDonald, B. E. Rapuano, N. Deo, M. Stranick, P. Somasundaran and A. L. Boskey, Thermal and chemical modification of titanium–aluminum–vanadium implant materials: effects on surface properties, glycoprotein adsorption, and MG63 cell attachment, Biomaterials, 2004, 25, 3135–3146.
N. Håkan, Initial reactions of whole blood with hydrophilic and hydrophobic titanium surfaces, Colloids Surf., B, 1996, 6, 329–333.
I. Schuster, H. Egger, D. Bikle, G. Herzig, G. S. Reddy, A. Stuetz, P. Stuetz and G. Vorisek, Selective inhibition of vitamin D hydroxylases in human keratinocytes, Steroids, 2001, 66, 409–422.
P. Tarroni, I. Villa, E. Mrak, F. Zolezzi, M. Mattioli, C. Gattuso and A. Rubinacci, Microarray analysis of 1,25(OH)2D3 regulated gene expression in human primary osteoblasts, J. Cell. Biochem., 2011, 640–649.
G. S. Stein, J. B. Lian, J. L. Stein, A. J. Van Wijnen and M. Montecino, Transcriptional control of osteoblast growth and differentiation, Physiol. Rev., 1996, 76, 593–629.
A. Ulsamer, M. J. Ortuno, S. Ruiz, A. R. G. Susperregui, N. Osses, J. L. Rosa and F. Ventura, BMP-2 induces osterix expression through up-regulation of Dlx5 and its phosphorylation by p38, J. Biol. Chem., 2008, 283, 3816–3826.
T. L. Chen and D. Fry, Hormonal regulation of the osteoblastic phenotype expression in neonatal murine calvarial cells, Calcif. Tissue Int., 1999, 64, 304–309.
N. Kurihara, S. Ishizuka, M. Kiyoki, Y. Haketa, K. Ikeda and M. Kumegawa, Effects of 1,25-dihydroxyvitamin D3 on osteoblastic MC3T3-E1 cells, Endocrinology, 1986, 118, 940–947.
Y. Maehata, S. Takamizawa, S. Ozawa, Y. Kato, S. Sato, E. Kubota and R. I. Hata, Both direct and collagen-mediated signals are required for active vitamin D3-elicited differentiation of human osteoblastic cells: roles of osterix, an osteoblast-related transcription factor, Matrix Biol., 2006, 25, 47–58.
T. Matsumoto, C. Igarashi, Y. Takeuchi, S. Harada, T. Kikuchi, H. Yamato and E. Ogata, Stimulation by 1,25-dihydroxyvitamin D3 of in vitro mineralization induced by osteoblast-like MC3T3-E1 cells, Bone, 1991, 12, 27–32.
R. L. Jilka, G. Hangoc, G. Girasole, G. Passeri, D. C. Williams, J. S. Abrams, B. Boyce, H. Broxmeyer and S. C. Manolagas, Increased osteoclast development after estrogen loss: mediation by interleukin-6, Science, 1992, 257, 88–91.
O. Kozawa, H. Tokuda, T. Kaida, H. Matsuno and T. Uematsu, Effect of vitamin D3 on interleukin-6 synthesis induced by prostaglandins in osteoblasts, Prostaglandins, Leukotrienes Essent. Fatty Acids, 1998, 58, 119–123.
R. Gruber, G. Nothegger, G. M. Ho, M. Willheim and M. Peterlik, Differential stimulation by PGE2 and calcemic hormones of IL-6 in stromal/osteoblastic cells, Biochem. Biophys. Res. Commun., 2000, 270, 1080–1085.
J. M. Tran, C. R. Kleeman and J. Green, Production of interleukin-6 by osteoblastic cells is independent of medium inorganic phosphate, Biochem. Mol. Med., 1995, 55, 90–95.
M. Giner, M. A. Rios, M. A. Montoya, M. A. Vazquez, L. Naji, R. Perez-Cano, RANKL/OPG in primary cultures of osteoblasts from post-menopausal women. Differences between osteoporotic hip fractures and osteoarthritis, J. Steroid Biochem. Mol. Biol., 2009, 113, 46–51.
N. J. Horwood, J. Elliott, T. J. Martin and M. T. Gillespie, Osteotropic agents regulate the expression of osteoclast differentiation factor and osteoprotegerin in osteoblastic stromal cells, Endocrinology, 1998, 139, 4743–4746.
T. Suda, E. Jimi, I. Nakamura and N. Takahashi, Role of 1 alpha,25-dihydroxyvitamin D3 in osteoclast differentiation and function, Methods Enzymol., 1997, 282, 223–235.
T. Suda, N. Takahashi, N. Udagawa, E. Jimi, M. T. Gillespie and T. J. Martin, Modulation of osteoclast differentiation and function by the new members of the tumor necrosis factor receptor and ligand families, Endocr. Rev., 1999, 20, 345–357.
G. P. Thomas, S. U. K. Baker, J. A. Eisman and E. M. Gardiner, Changing RANKL/OPG mRNA expression in differentiating murine primary osteoblasts, J. Endocrinol., 2001, 170, 451–460.
D. Zhang, Y. Q. Yang, X. T. Li and M. K. Fu, The expression of osteoprotegerin and the receptor activator of nuclear factor kappa B ligand in human periodontal ligament cells cultured with and without 1alpha,25-dihydroxyvitamin D3, Arch. Oral Biol., 2004, 49, 71–76.
P. A. Baldock, G. P. Thomas, J. M. Hodge, S. U. Baker, U. Dressel, P. D. O’Loughlin, G. C. Nicholson, K. H. Briffa, J. A. Eisman and E. M. Gardiner, Vitamin D action and regulation of bone remodeling: suppression of osteoclastogenesis by the mature osteoblast, J. Bone Miner. Res., 2006, 21, 1618–1626.
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Satué, M., Córdoba, A., Ramis, J.M. et al. UV-irradiated 7-dehydrocholesterol coating on polystyrene surfaces is converted to active vitamin D by osteoblastic MC3T3-E1 cells. Photochem Photobiol Sci 12, 1025–1035 (2013). https://doi.org/10.1039/c3pp50025j
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DOI: https://doi.org/10.1039/c3pp50025j