Evaluation of stability and in vitro wound healing potential of melatonin loaded (lipid enriched) chitosan based microspheres

Marieta Duvnjak Romić; Anja Sušac; Jasmina Lovrić; Biserka Cetina-Čižmek; Jelena Filipović-Grčić; Anita Hafner

doi:10.2478/acph-2019-0049

Abstract

The aim of this study was to evaluate long-term stability and assess the wound healing potential of the innovative melatonin-loaded lipid-enriched hybrid system compared to conventional melatonin-loaded chitosan microspheres. The hybrid system contained nanostructured lipid carrier incorporated in the chitosan matrix, in order to modify melatonin release and alter physicochemical characteristics of the delivery system. Stability testing was performed during a six-month period under two conditions: refrigerated (5 ± 3 °C) and at room temperature (25 ± 2 °C/60 ± 5 % RH). Samples stored at both conditions were analyzed in terms of particle size, zeta potential, moisture content and thermal properties. At the end of testing, drug content was determined in all samples. Dressings wound healing potential was assessed by in vitro scratch test using human skin fibroblast cell line. Although both systems showed good stability characteristics, the addition of lipids in the system has improved its wound healing potential.

References

1. R. G. Frykberg and J. Banks, Challenges in the treatment of chronic wounds, Adv. Wound Care4 (2015) 560–582; https://doi.org/10.1089/wound.2015.063510.1089/wound.2015.0635
Search in Google Scholar Back to article
2. T. Shipperley, C. Martin and B. Healthcare, The physiology of wound healing : an emergency response, Surg. 35 (2011) 8–10; http://www.sciencedirect.com/science/article/pii/S0263931917301369
Search in Google Scholar Back to article
3. K. Kaplani, S. Koutsi, V. Armenis, F. G. Skondra, N. Karantzelis, S. Champeris Tsaniras and S. Taraviras, Wound healing related agents: Ongoing research and perspectives, Adv. Drug Deliv. Rev. (2018); https://doi.org/10.1016/j.addr.2018.02.00710.1016/j.addr.2018.02.007
Search in Google Scholar Back to article
4. Y.-K. Wu, N.-C. Cheng and C.-M. Cheng, Biofilms in chronic wounds: pathogenesis and diagnosis, Trends Biotechnol. 37 (2019) 505–517; https://doi.org/10.1016/j.tibtech.2018.10.01110.1016/j.tibtech.2018.10.011
Search in Google Scholar Back to article
5. R. J. Reiter, D.-X. Tan, L. Fuentes-Broto and M. Luciano, Chapter 8 – Melatonin: A multitasking molecule, Progress in Brain Res.181 (2010) 127–151; https://doi.org/10.1016/S0079-6123(08)81008-410.1016/S0079-6123(08)81008-4
Search in Google Scholar Back to article
6. R. Hardeland, A. T. Slominski, R. M. Slominski, R. J. Reiter, M. A. Zmijewski and R. Paus, Melatonin: A cutaneous perspective on its production, metabolism, and functions, J. Invest. Dermatol.138 (2018) 490–499; https://doi.org/10.1016/j.jid.2017.10.02510.1016/j.jid.2017.10.025582891029428440
Search in Google Scholar Back to article
7. A. Crooke, A. Guzman-Aranguez, A. Mediero, P. Alarma-Estrany, G. Carracedo, T. Pelaez, A. Peral and J. Pintor, Effect of melatonin and analogues on corneal wound healing: involvement of Mt₂ melatonin receptor, Curr. Eye Res.40 (2015) 56–65; https://doi.org/10.3109/02713683.2014.91454010.3109/02713683.2014.91454024892818
Search in Google Scholar Back to article
8. R. Song, L. Ren, H. Ma, R. Hu, H. Gao, L. Wang, X. Chen, Z. Zhao and J. Liu, Melatonin promotes diabetic wound healing in vitro by regulating keratinocyte activity, Am. J. Transl. Res.8 (2016) 4682–4693.
Search in Google Scholar Back to article
9. R. Murali, P. Thanikaivelan and K. Cheirmadurai, Melatonin in functionalized biomimetic constructs promotes rapid tissue regeneration in Wistar albino rats, J. Mater. Chem. B4 (2016) 5850; https://doi.org/10.1039/c6tb01221c.10.1039/C6TB01221C32263758
Search in Google Scholar Back to article
10. I. Garcia-Orue, J. L. Pedraz, R. M. Hernandez and M. Igartua, Nanotechnology-based delivery systems to release growth factors and other endogenous molecules for chronic wound healing, J. Drug Deliv. Sci. Technol.42 (2017) 2–17; https://doi.org/10.1016/j.jddst.2017.03.00210.1016/j.jddst.2017.03.002
Search in Google Scholar Back to article
11. J. Pardeike, A. Hommoss and R. H. Müller, Lipid nanoparticles (SLN, NLC) in cosmetic and pharmaceutical dermal products, Int. J. Pharm.366 (2009) 170–184; https://doi.org/10.1016/j.ijpharm.2008.10.00310.1016/j.ijpharm.2008.10.00318992314
Search in Google Scholar Back to article
12. G. Gainza, M. Pastor, J. J. Aguirre, S. Villullas, J. L. Pedraz, R. M. Hernandez and M. Igartua, A novel strategy for the treatment of chronic wounds based on the topical administration of rhEGF-loaded lipid nanoparticles: In vitro bioactivity and in vivo effectiveness in healing-impaired db/db mice, J. Control. Release185 (2014) 51–61; https://doi.org/10.1016/j.jconrel.2014.04.03210.1016/j.jconrel.2014.04.03224794895
Search in Google Scholar Back to article
13. N. Karimi, B. Ghanbarzadeh, H. Hamishehkar, B. Mehramuz and H. S. Kafil, Antioxidant, antimicrobial and physicochemical properties of turmeric extract-loaded nanostructured lipid carrier (NLC), Colloid. Interface Sci. Commun.22 (2018) 18–24; https://doi.org/10.1016/j.colcom.2017.11.00610.1016/j.colcom.2017.11.006
Search in Google Scholar Back to article
14. F. Saporito, G. Sandri, M. C. Bonferoni, S. Rossi, C. Boselli, A. I. Cornaglia, B. Mannucci, P. Grisoli, B. Vigani and F. Ferrari, Essential oil-loaded lipid nanoparticles for wound healing, Int. J. Nanomed.13 (2018) 175–186; https://doi.org/10.2147/IJN.S15252910.2147/IJN.S152529574796329343956
Search in Google Scholar Back to article
15. Z. Ma, A. Garrido-Maestu and K. C. Jeong, Application, mode of action, and in vivo activity of chitosan and its micro- and nanoparticles as antimicrobial agents: A review, Carbohydr. Polym.176 (2017) 257–265; https://doi.org/10.1016/j.carbpol.2017.08.08210.1016/j.carbpol.2017.08.08228927606
Search in Google Scholar Back to article
16. V. Patrulea, V. Ostafe, G. Borchard and O. Jordan, Chitosan as a starting material for wound healing applications, Eur. J. Pharm. Biopharm. (2015); https://doi.org/10.1016/j.ejpb.2015.08.00410.1016/j.ejpb.2015.08.00426614560
Search in Google Scholar Back to article
17. M. Duvnjak Romić, M. Š. Klarić, J. Lovrić, I. Pepić, B. Cetina-Čižmek, J. Filipović-Grčić and A. Hafner, Melatonin-loaded chitosan/Pluronic^® F127 microspheres as in situ forming hydrogel: An innovative antimicrobial wound dressing, Eur. J. Pharm. Biopharm.107 (2016) 67–79; https://doi.org/10.1016/j.ejpb.2016.06.01310.1016/j.ejpb.2016.06.01327329001
Search in Google Scholar Back to article
18. M. Duvnjak Romić, D. Špoljarić, M. Š. Klarić, B. Cetina-Čižmek, J. Filipović-Grčić and A. Hafner, Melatonin loaded lipid enriched chitosan microspheres – Hybrid dressing for moderate exuding wounds, J. Drug Deliv. Sci. Technol. (2019); https://doi.org/10.1016/j.jddst.2019.05.00410.1016/j.jddst.2019.05.004
Search in Google Scholar Back to article
19. B. N. Estevinho, F. Rocha, L. Santos and A. Alves, Microencapsulation with chitosan by spray drying for industry applications – A review, Trends Food Sci. Technol. 31 (2013) 138–155; https://doi.org/10.1016/j.tifs.2013.04.00110.1016/j.tifs.2013.04.001
Search in Google Scholar Back to article
20. B. R. P. Cabral, P. M. de Oliveira, G. M. Gelfuso, T. de S. C. Quintão, J. A. Chaker, M. G. de O. Karnikowski and E. F. Gris, Improving stability of antioxidant compounds from Plinia cauliflora (jabuticaba) fruit peel extract by encapsulation in chitosan microparticles, J. Food Eng.238 (2018) 195–201; https://doi.org/10.1016/j.jfoodeng.2018.06.00410.1016/j.jfoodeng.2018.06.004
Search in Google Scholar Back to article
21. D. P. Gaspar, M. M. Gaspar, C. V. Eleutério, A. Grenha, M. Blanco, L. M. D. Gonçalves, P. Taboada, A. J. Almeida and C. Remunán-López, Microencapsulated solid lipid nanoparticles as a hybrid platform for pulmonary antibiotic delivery, Mol. Pharmaceutics14 (2017) 2977–2990; https://doi.org/10.1021/acs.molpharmaceut.7b0016910.1021/acs.molpharmaceut.7b0016928809501
Search in Google Scholar Back to article
22. T. Wang, Q. Hu, M. Zhou, J. Xue and Y. Luo, Preparation of ultra-fine powders from polysaccharide-coated solid lipid nanoparticles and nanostructured lipid carriers by innovative nano spray drying technology, Int. J. Pharm.511 (2016) 219–222; https://doi.org/10.1016/j.ijpharm.2016.07.00510.1016/j.ijpharm.2016.07.00527395801
Search in Google Scholar Back to article
23. H. Salminen, J. Ankenbrand, B. Zeeb, G. Badolato Bönisch, C. Schäfer, R. Kohlus and J. Weiss, Influence of spray drying on the stability of food-grade solid lipid nanoparticles, Food Res. Int.119 (2018) 741–750; https://doi.org/10.1016/j.foodres.2018.10.05610.1016/j.foodres.2018.10.056
Search in Google Scholar Back to article
24. D. G. Sami, H. H. Heiba and A. Abdellatif, Wound healing models: A systematic review of animal and non-animal models, Wound Med.24 (2019) 8–17; https://doi.org/10.1016/j.wndm.2018.12.00110.1016/j.wndm.2018.12.001
Search in Google Scholar Back to article
25. A. Stunova and L. Vistejnova, Dermal fibroblasts – A heterogeneous population with regulatory function in wound healing, Cytokine Growth Factor Rev.39 (2018) 137–150; https://doi.org/10.1016/j.cytogfr.2018.01.00310.1016/j.cytogfr.2018.01.003
Search in Google Scholar Back to article
26. I. Rubelj, Telomere Q-PNA-FISH – reliable results from stochastic signals, PLoS One9 (2014) e92559; https://doi.org/10.1371/journal.pone.009255910.1371/journal.pone.0092559
Search in Google Scholar Back to article
27. F. Felice, Y. Zambito, E. Belardinelli, A. Fabiano, T. Santoni and R. Di Stefano, Effect of different chitosan derivatives on in vitro scratch wound assay: A comparative study, Int. J. Biol. Macromol.76 (2015) 236–241; https://doi.org/10.1016/j.ijbiomac.2015.02.04110.1016/j.ijbiomac.2015.02.041
Search in Google Scholar Back to article
28. A. D. Kulkarni, D. B. Bari, S. J. Surana and C. V Pardeshi, In vitro, ex vivo and in vivo performance of chitosan-based spray-dried nasal mucoadhesive microspheres of diltiazem hydrochloride, J. Drug Deliv. Sci. Technol.31 (2016) 108–117; https://doi.org/10.1016/j.jddst.2015.12.00410.1016/j.jddst.2015.12.004
Search in Google Scholar Back to article
29. S. Demarger-Andre and A. Domard, Chitosan carboxylic acid salts in solution and in the solid state, Carbohydr. Polym.23 (1994) 211–219; https://doi.org/10.1016/0144-8617(94)90104-X10.1016/0144-8617(94)90104-X
Search in Google Scholar Back to article
30. N. El Moussaoui and A. Bendriss, The influence of storage conditions on melatonin stability, Int. J. Eng. Res. Technol.3 (2014) 2243–2246.
Search in Google Scholar Back to article
31. M. Friciu, T. Savji, S. Zaraa and G. Leclair, Evaluation of stability of melatonin in extemporaneously compounded oral suspensions, J. Pharm. Pract. Res.46 (2016) 28–33; https://doi.org/10.1002/jppr.117110.1002/jppr.1171
Search in Google Scholar Back to article
32. P. Linse and M. Malmsten, Temperature-dependent micellization in aqueous block copolymer solutions, Macromolecules25 (1992) 5434–5439; https://doi.org/10.1021/ma00046a04810.1021/ma00046a048
Search in Google Scholar Back to article
33. A. Garcês, M. H. Amaral, J. M. Sousa Lobo and A. C. Silva, Formulations based on solid lipid nanoparticles (SLN) and nanostructured lipid carriers (NLC) for cutaneous use: A review, Eur. J. Pharm. Sci.112 (2018) 159–167; https://doi.org/10.1016/j.ejps.2017.11.02310.1016/j.ejps.2017.11.02329183800
Search in Google Scholar Back to article
34. B. M. Dulmovits and I. M. Herman, Microvascular remodeling and wound healing: A role for pericytes, Int. J. Biochem. Cell Biol.44 (2012) 1800–1812; https://doi.org/10.1016/j.biocel.2012.06.03110.1016/j.biocel.2012.06.031345511622750474
Search in Google Scholar Back to article
35. S. L. Percival, S. McCarty, J. A. Hunt, E. J. Woods, The effects of pH on wound healing, biofilms, and antimicrobial efficacy, Wound Rep. Reg.22 (2014) 174–186; https://doi.org/10.1111/wrr.1212510.1111/wrr.1212524611980
Search in Google Scholar Back to article
36. D. B. Friedman, D. L. Stauff, G. Pishchany, C. W. Whitwell, V. J. Torres and E. P. Skaar, Staphylococcus aureus redirects central metabolism to increase iron availability, PLoS Pathog.2 (2006) 0777–0789; https://doi.org/10.1371/journal.ppat.002008710.1371/journal.ppat.0020087155783216933993
Search in Google Scholar Back to article
37. X. Chen, L. H. Peng, Y. H. Shan, N. Li, W. Wei, L. Yu, Q. M. Li, W. Q. Liang and J. Q. Gao, Astragaloside IV-loaded nanoparticle-enriched hydrogel induces wound healing and anti-scar activity through topical delivery, Int. J. Pharm.447 (2013) 171–181; https://doi.org/10.1016/j.ijpharm.2013.02.05410.1016/j.ijpharm.2013.02.05423500766
Search in Google Scholar Back to article
38. V. Kant, A. Gopal, D. Kumar, A. Gopalkrishnan, N. N. Pathak, N. P. Kurade, S. K. Tandan and D. Kumar, Topical pluronic F-127 gel application enhances cutaneous wound healing in rats, Acta Histochem.116 (2014) 5–13; https://doi.org/10.1016/j.acthis.2013.04.01010.1016/j.acthis.2013.04.01023706531
Search in Google Scholar Back to article
39. G. Soybir, C. Topuzlu, Ö. Odabaş, K. Dolay, A. Bilir and F. Köksoy, The effects of melatonin on angiogenesis and wound healing, Surg. Today. 33 (2003) 896–901; https://doi.org/10.1007/s00595-003-2621-310.1007/s00595-003-2621-314669079
Search in Google Scholar Back to article
40. I. Garcia-Orue, G. Gainza, P. Garcia-Garcia, F. B. Gutierrez, J. J. Aguirre, R. M. Hernandez, A. Delgado and M. Igartua, Composite nanofibrous membranes of PLGA/Aloe vera containing lipid nanoparticles for wound dressing applications, Int. J. Pharm. (2019); https://doi.org/10.1016/j.ijpharm.2018.12.010.10.1016/j.ijpharm.2018.12.01030553008
Search in Google Scholar Back to article
41. M. Ghodrati, M. R. Farahpour and H. Hamishehkar, Encapsulation of Peppermint essential oil in nanostructured lipid carriers: In-vitro antibacterial activity and accelerative effect on infected wound healing, Colloids Surfaces A Physicochem. Eng. Asp. (2019); https://doi.org/10.1016/j.colsurfa.2018.12.04310.1016/j.colsurfa.2018.12.043
Search in Google Scholar Back to article

Evaluation of stability and in vitro wound healing potential of melatonin loaded (lipid enriched) chitosan based microspheres

Abstract

Paradigm

My account