Abstract
We have previously reported that interferon-alpha-2b (IFN-α-2b) can be encapsulated in liposomes without compromising its anti-fibrogenic effects on dermal fibroblasts in vitro. This study was conducted to determine whether this preparation applied topically to guinea pig wounds can affect their healing. The rationale for this approach is that systemic administration of IFN-α-2b by injection for treatment of dermal fibrosis is uncomfortable, requires a large quantity of the cytokine and cannot be easily used in children. Liposomes are potentially useful vehicles for the topical delivery of drugs. Empty sonicated liposome vesicles were mixed with various concentrations of IFN-α-2b and then dried and rehydrated. An enzyme-linked immunosorbent assay (ELISA) was used to determine the efficiency of encapsulation and the stability of the preparation under experimental conditions. A total of 36 full thickness skin wounds (6/animal, 3 on each side) were made with an 8 mm disposable punch. Each wound on the right side received cream (100 mg/wound) containing 3000 units of liposome-encapsulated IFN-α-2b, while wounds on the left side received cream containing empty liposomes. There was a significant reduction in rate of contraction of wounds treated with IFN-α-2b as early as 5 days after wounding. This reduction remained significant up to 10 days. Northern analysis, used to evaluate the expression of mRNAs for type I and type III collagens in response to IFN-α-2b showed a marked reduction in abundance of the transcripts for the pro-α1(I) chain of type 1 collagen on days 11 and 14 after wounding. Similarly, the level of mRNA for type III procollagen was markedly reduced as early as day 7 and remained depressed up to day 14. These findings were consistent with results obtained for the total collagen content in tissue samples. Cellularity of the IFN-α-2b-treated wounds, assessed by vimentin content, was also markedly reduced at day 7 and remained depressed up to day 14. Liposome associated IFN-α-2b applied 5 days after completion of epithelialization reduced mRNA for the pro-α1(I) chain of type 1 collagen., confirming its transepidermal penetration and effectiveness. The activity of liposome-associated IFN-α-2b in vivo supports the concept of the topical use of this anti-fibrogenic agent for treatment of fibroproliferative disorders.
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References
Scott PG, Ghahary A, Chambers M, Tredget EE: Biological basis of hypertrophic scarring. In: Advances in Structural Biology, vol. 3. 1994, pp 157–201
Rudolph R: Wide spread scars, hypertrophic scars and keloids. Clin Plast Surg 14: 253–260, 1987
Tredget EE, Shankowsky HA, Pannu R, Nedelec B, Iwashina T, Ghahary A, Taerum TV, Scott PG: Transforming growth factor-β in thermally injured patients with hypertrophic scars: Effects of interferon-α-2b. Plast Reconstr Surg 102: 1317–1328, 1998
Weiner N, Williams N, Birch G, Ramachandran C, Shipman C Jr, Flynn G: Topical delivery of liposomally encapsulated interferon evaluated in a cutaneous herpes guinea pig model. Antimicrob Agents Chemother 33: 1217–1221, 1989
Sen GC, Lengyel P: The interferon system. A bird's eye view of its biochemistry. J Biol Chem 267: 5017–5020, 1992
Tsutsumi Y, Kakumu S, Yoshioka K, Arao M, Inoue M, Wakita T: Effects of various cytokines on collagen synthesis by rat normal hepatocytes in primary cultures and fibroblasts. Digestion 44: 191–199, 1989
Tredget EE, Shen YJ, Liu G, Forsyth N, Smith C, Harrop RA, Scott PG, Ghahary A: Regulation of collagen synthesis and mRNA levels in normal and hypertrophic scar fibroblasts in vitro by interferon alpha-2b. Wound Rep Reg 1: 156–165, 1993
Harrop AR, Ghahary A, Scott PG, Forsyth N, Friedland A, Tredget EE: Regulation of collagen synthesis and mRNA expression in normal and hypertrophic scar fibroblasts in vitro by interferon-g. J Surg Res 58: 471–477, 1995
Rosenbloom J, Feldman G, Freundlich B, Jimenez SA: Transcriptional control of human diploid fibroblast collagen synthesis by gamma interferon. Biochem Biophys Res Commun 123: 365–372, 1984
Berman B, Duncan MR: Short-term keloid treatment in vivo with human interferon alfa-2b results in a selective and persistent normalization of keloidal fibroblast collagen, glycosaminoglycan and collagenase production in vitro. J Am Acad Dermatol 21: 694–702, 1989
Ghahary A, Shen YJ, Scott PG, Tredget EE: Expression of fibronectin mRNA in hypertrophic and normal dermal tissues and in vitro regulation by interferon alpha-2b. Wound Rep Reg 1: 166–174, 1993
Ghahary A, Shen YJ, Nedelec B, Scott PG, Tredget EE: Interferon gamma and alpha-2b differentially regulate the expression of collagenase and TIMP-1 mRNA in human hypertrophic and normal dermal fibroblasts. Wound Rep Reg 3: 13–21, 1995
Czaja MJ, Weiner FR, Eghbali M, Giambrone MA, Eghbali M, Zern MA: Differential effects of γ-interferon on collagen and fibronectin gene expression. J Biol Chem 262: 13348–13351, 1987
Veys EM, Mialants H, Verbruggen G, Grosclaude JP, Meyer W, Galazka A, Schindler J: Interferon gamma in rheumatoid arthritis — a double blind study comparing human recombinant interferon gamma with placebo.J Rheumatol 15:570–574, 1988
Jonas MM, Ragin L, Silva MO: Membranous glomerulonephritis and chronic persistent hepatitis B in a child: Treatment with recombinant interferon alpha. J Pediatr 119: 818–820, 1991
Castilla A, Prieto J, Fausto N: Transforming growth factors beta 1 and alpha in chronic liver disease. New Engl J Med 324: 933–940, 1991
Larrabee WF, East CA, Jaffe HF, Stephenson RN, Peterson KE: Intralesional interferon gamma treatment for keloid and hypertrophic scars. Arch Otolaryng Head Neck Surg 116: 1159–1162, 1990
Granstein RD, Rook A, Flotte TJ, Hass A, Gallo RL, Jaffe HF, Amento EP: A controlled trial of intralesional recombinant interferon-gamma in the treatment of keloidal scarring. Arch Dermatol 126: 1295–1302, 1990
Ghahary A, Shen Q, Rogers JA, Wang R, Fathi-Afshar A, Scott PG, Tredget EE: Liposome-associated interferon-α-2b functions as an antifibrogenic factor for human dermal fibroblasts. J Invest Dermatol 109: 55–60, 1997
Eppstein DA, Stewart WE: Binding and capture of human interferon alpha by reverse evaporation vesicles, multilamellar vesicles, and small unilamellar vesicles. J Interferon Res 1: 495–504, 1981
Traub P, Scherbarth A, Willingate-Theune J, Traub U: Large scale coisolation of vimentin and nuclear lamins from Ehrlich ascites tumor cells cultured in vitro. Prep Biochem 18: 381–404, 1988
Mechanic GL: Collagen biochemistry. In: D.J. Simmons, A.S. Kunin (eds). Skeletal Research. Academic Press, New York, 1979, pp 227–241.
Chomczynski P, Sacchi N: Single-step method of RNA isolation by acid guanidinium thiocyanate-phenol-chloroform extraction. Anal Biochem 162: 156–159, 1987
Stegemann H, Stalder K: Determination of hydroxyproline. Clin Chim Acta 18: 267–273, 1967
Nedelec B, Shen YJ, Ghahary A, Scott PG, Tredget EE: The effect of interferon alpha-2b on expression of cytoskeletal proteins in an in vitro model of wound contraction. J Lab Clin Med 126: 474–484, 1995
Dans MJ, Isseroff R: Inhibition of collagen lattice contraction by pentoxifylline and interferon-alpha-beta and-gamma. J Invest Dermatol 102: 118–121, 1994
Catty RHC: Healing and contraction of experimental full thickness wounds in the human. Brit J Surg 52: 542–548, 1965
Eppstein DA, Stewart WE: Altered pharmacological properties of liposome-associated human interferon-alpha. J Virol 41: 575–582, 1982
Giometto B, Miotto D, Botteri M, Alessio L, Scanarini M, An SF, Tavolato B: Follicula-Stellate cells of human pituitary adenomas: Immunohistochemical study of the monocyte/macrophage phenotype expression. Neuroendocrinology 65: 47–52, 1997
Mezei M, Gulasekharam V: Liposomes — a selective drug delivery system for the topical route of administration: Lotion dosage form. Life Sci 26: 1473–1477, 1980
Lasch J, Wohlrab W: Liposome-bound cortisol: A new approach to cutaneous therapy. Biomed Biochem Acta 45: 1295–1299, 1986
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Ghahary, A., Tredget, E.E., Shen, Q. et al. Liposome associated interferon-alpha-2b functions as an anti-fibrogenic factor in dermal wounds in the guinea pig. Mol Cell Biochem 208, 129–137 (2000). https://doi.org/10.1023/A:1007054424400
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DOI: https://doi.org/10.1023/A:1007054424400