Abstract
The major challenge with treatment of dermal wounds is accelerating healing process, while preventing the scar formation. Herein, we have fabricated layer-by-layer (LbL) polyelectrolyte multilayer films containing epidermal growth factor (EGF) and TGF-β siRNA to improve excisional wound healing and decrease scar formation. The chitosan and sodium alginate LbL thin films showed 13.0 MPa tensile strength and 2.22 N/cm2 skin adhesion strength. The LbL thin films were found to be cytocompatible, where A431 epidermal keratinocytes adhered to the film and showed 86.2 ± 0.8% cell growth compared with cells cultured in the absence of LbL thin film. In contrast, LbL thin film did not promote the Escherichia coli and Staphylococcus aureus bacterial colony formation. In a C57BL/6 mouse excisional wound model, application of LbL thin films containing TGF-β siRNA significantly (p < 0.05) reduced the TGF-β protein expression and collagen production. The LbL thin films containing EGF showed improved wound contraction (<9 days post excision). The co-delivery of TGF-β siRNA and EGF using LbL thin films resulted in accelerated wound healing and decreased collagen deposition. Furthermore, the LbL thin films with TGF-β siRNA and EGF combination showed greater reepithelialization. Taken together, we have successfully demonstrated the co-delivery of TGF-β siRNA and EGF peptide using LbL thin films to promote wound healing and decrease scar formation.
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References
Boateng JS, Matthews KH, Stevens HNE, Eccleston GM. Wound healing dressings and drug delivery systems: a review. J Pharm Sci. 2008;97:2892–923.
Guo S, DiPietro LA. Factors affecting wound healing. J Dent Res. 2010;89:219–29.
Shaw TJ, Martin P. Wound repair at a glance. J Cell Sci. 2009;122:3209–13.
Sgonc R, Gruber J. Age-related aspects of cutaneous wound healing: a mini-review. Gerontology. 2013;59:159–64.
Singh N, Armstrong DG, Lipsky BA. Preventing foot ulcers in patients with diabetes. JAMA. 2005;293:217–28.
Shah NJ, Hyder MN, Quadir MA, Courchesne NMD, Seeherman HJ, Nevins M, et al. Adaptive growth factor delivery from a polyelectrolyte coating promotes synergistic bone tissue repair and reconstruction. Proc Natl Acad Sci U S A. 2014;111:12847–52.
Choi JS, Leong KW, Yoo HS. In vivo wound healing of diabetic ulcers using electrospun nanofibers immobilized with human epidermal growth factor (EGF). Biomaterials. 2008;29:587–96.
Yang Y, Xia T, Zhi W, Wei L, Weng J, Zhang C, et al. Promotion of skin regeneration in diabetic rats by electrospun core-sheath fibers loaded with basic fibroblast growth factor. Biomaterials. 2011;32:4243–54.
Almquist BD, Castleberry SA, Sun JB, Lu AY, Hammond PT. Combination growth factor therapy via electrostatically assembled wound dressings improves diabetic ulcer healing in vivo. Adv Healthc Mater. 2015. doi:10.1002/adhm.201500403.
Occleston NL, O’Kane S, Goldspink N, Ferguson MWJ. New therapeutics for the prevention and reduction of scarring. Drug Discov Today. 2008;13:973–81.
Mustoe TA, Cooter RD, Gold MH, Hobbs FD, Ramelet AA, Shakespeare PG, et al. International advisory panel on scar management. International clinical recommendations on scar management. Plast Reconstr Surg. 2002;110:560–71.
Armour A, Scott PG, Tredget EE. Cellular and molecular pathology of HTS: basis for treatment. Wound Rep Reg. 2007;15:S6–17.
Gurtner GC, Werner S, Barrandon Y, Longaker MT. Wound repair and regeneration. Nature. 2008;453:314–21.
Burd A, Huang L. Hypertrophic response and keloid diathesis: two very different forms of scar. Plast Reconstr Surg. 2005;116:150e–7.
Lin RY, Sullivan KM, Argenta PA, Meuli M, Lorenz HP, Adzick NS. Exogenous transforming growth factor-beta amplifies its own expression and induces scar formation in a model of human fetal skin repair. Ann Surg. 1995;222:146–54.
Beanes SR, Dang C, Soo C, Ting K. Skin repair and scar formation: the central role of TGF-β. Expert Rev Mol Med. 2003;5:1–22.
Cutroneo KR. TGF-b–induced fibrosis and SMAD signaling: oligo decoys as natural therapeutics for inhibition of tissue fibrosis and scarring. Wound Rep Reg. 2007;15:S54–60.
Liu W, Wang DR, Cao YL. TGF-b: a fibrotic factor in wound scarring and a potential target for anti-scarring gene therapy. Curr Gene Ther. 2004;4:123–36.
Frederiksen K, Guy RH, Petersson K. Formulation considerations in the design of topical, polymeric film-forming systems for sustained drug delivery to the skin. Eur J Pharm Biopharm. 2015;91:9–15.
Follmann HDM, Martins AF, Gerola AP, Burgo TAL, Nakamura CV, Rubira AF, et al. Antiadhesive and antibacterial multilayer films via layer-by-layer assembly of TMC/heparin complexes. Biomacromolecules. 2012;13:3711–22.
Huang R, Li W, Lv X, Lei Z, Bian Y, Deng H, et al. Biomimetic LBL structured nanofibrous matrices assembled by chitosan/collagen for promoting wound healing. Biomaterials. 2015;53:58–75.
Hsu BB, Hagerman SR, Jamieson K, Castleberry SA, Wang W, Holler E, et al. Multifunctional self-assembled films for rapid hemostat and sustained anti-infective delivery. ACS Biomater Sci Eng. 2015;1:148–56.
Borges J, Mano JF. Molecular interactions driving the layer-by-layer assembly of multilayers. Chem Rev. 2014;114:8883–942.
Mandapalli PK, Labala S, Bojja J, Venuganti VK. Effect of pirfenidone delivered using layer-by-layer thin film on excisional wound healing. Eur J Pharm Sci. 2016;83:166–74.
Neuman RE, Logan MA. The determination of hydroxyproline. J Biol Chem. 1950;184:299–306.
Zhong SP, Zhang YZ, Lim CT. Tissue scaffolds for skin wound healing and dermal reconstruction. WIRE’s Nanomed Nanobiotechnol. 2010;2:510–25.
Chong EJ, Phan TT, Lim IJ, Zhang YZ, Bay BH, Ramakrishna S, et al. Evaluation of electrospun PCL/gelatin nanofibrous scaffold for wound healing and layered dermal reconstitution. Acta Biomater. 2007;3:321–30.
Jayakumar R, Divya Rani VV, Shalumon KT, Kumar PT, Nair SV, Furuike T, et al. Bioactive and osteoblast cell attachment studies of novel alpha- and beta-chitin membranes for tissue-engineering applications. Int J Biol Macromol. 2009;45:260–4.
Sudheesh Kumar PT, Lakshmanan VK, Anilkumar TV, Ramya C, Reshmi P, Unnikrishnan AG, et al. Flexible and microporous chitosan hydrogel/nano ZnO composite bandages for wound dressing: in vitro and in vivo evaluation. ACS Appl Mater Interfaces. 2012;4:2618–29.
Sudheesh Kumar PT, Praveen G, Raj M, Chennazhi KP, Jayakumar R. Flexible, micro-porous chitosan-gelatin hydrogel/nano fibrin composite bandages for treating burn wounds. RSC Adv. 2014;4:65081–7.
Khan TA, Peh KK, Ching HS. Mechanical, bioadhesive strength and biological evaluations of chitosan films for wound dressing. J Pharm Pharmaceut Sci. 2000;3:303–11.
Zahedi P, Rezaeian I, Ranaei-Siadat SO, Jafari SH, Supapho P. A review on wound dressings with an emphasis on electrospun nanofibrous polymeric bandages. Polym Adv Technol. 2010;21:77–95.
Khademhosseini A, Langer R, Borenstein J, Vacanti JP. Microscale technologies for tissue engineering and biology. Proc Natl Acad Sci U S A. 2006;103:2480–7.
Mina BM, Lee G, Kim SH, Nam YS, Lee TS, Park WH. Electrospinning of silk fibroin nanofibers and its effect on the adhesion and spreading of normal human keratinocytes and fibroblasts in vitro. Biomaterials. 2004;25:1289–97.
Kempf M, Miyamur Y, Liu PY, Chen ACH, Nakamura H, Shimizu H, et al. A denatured collagen microfiber scaffold seeded with human fibroblasts and keratinocytes for skin grafting. Biomaterials. 2011;32:4782–92.
Jewell CM, Lynn DM. Multilayered polyelectrolyte assemblies as platforms for the delivery of DNA and other nucleic acid-based therapeutics. Adv Drug Deliv Rev. 2008;60:979–99.
Taylor JM, Mitchell WM, Cohen S. Epidermal growth factor physical and chemical properties. J Biol Chem. 1972;247:5928–34.
Jones V, Grey JE, Harding KG. ABC of wound healing wound dressings. Br Med J. 2006;332:777–80.
Moura LIF, Dias AMA, Carvalho E, de Sousa HC. Recent advances on the development of wound dressings for diabetic foot ulcer treatment—a review. Acta Biomater. 2013;9:7093–114.
Mohan VK. Recombinant human epidermal growth factor (REGEN-D 150): effect on healing of diabetic foot ulcers. Diabetes Res Clin Pract. 2007;78:405–11.
Yu FSX, Yin J, Xu K, Huang J. Growth factors and corneal epithelial wound healing. Brain Res Bull. 2010;81:229–35.
Wang YW, Liou NH, Cherng JH, Chang SJ, Ma KH, Fu E, et al. siRNA-targeting transforming growth factor-b type I receptor reduces wound scarring and extracellular matrix deposition of scar tissue. J Investig Dermatol. 2014;134:2016–25.
Acknowledgments
This work was financially supported by a grant from the Department of Science and Technology, Government of India under SERB-Young Scientist award (SERB/F/1260/2012-13). Texture analyzer and multimode plate reader were procured using a grant from Department of Science and Technology—fund for improvement of science and technology infrastructure (DST FIST). PK Mandapalli received senior research fellowship from the Council of Scientific and Industrial Research to pursue doctoral studies.
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Mandapalli, P.K., Labala, S., Jose, A. et al. Layer-by-Layer Thin Films for Co-Delivery of TGF-β siRNA and Epidermal Growth Factor to Improve Excisional Wound Healing. AAPS PharmSciTech 18, 809–820 (2017). https://doi.org/10.1208/s12249-016-0571-6
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DOI: https://doi.org/10.1208/s12249-016-0571-6