Biocompatible Aloe vera and Tetracycline Hydrochloride Loaded Hybrid Nanofibrous Scaffolds for Skin Tissue Engineering
Abstract
:1. Introduction
2. Results and Discussion
2.1. Characterization of Nanofibers
2.2. Drug Release
2.3. Cell Viability
2.4. Cell Proliferation
2.5. Expression of Collagen
2.6. Cell-scaffold Interactions
2.7. Expression of F-actin
2.8. Antimicrobial Activity
3. Materials and Methods
3.1. Materials
3.2. Fabrication of Electrospun Nanofibers
3.3. Characterization of Nanofibrous Scaffolds
3.4. Drug Release
3.5. Human Dermal Fibroblast (hDF)
3.6. MTS Assay
3.7. Cell-scaffold Interactions
3.8. CMFDA Staining
3.9. Sirius Red Staining
3.10. F-actin Staining
3.11. Disc Diffusion Assay
3.12. Statistical Analysis
4. Conclusions
Author Contributions
Funding
Conflicts of Interest
References
- Simpson, C.L.; Patel, D.M.; Green, K.J. Deconstructing the skin: Cytoarchitectural determinants of epidermal morphogenesis. Nat. Rev. Mol. Cell Biol. 2011, 12, 565–580. [Google Scholar] [CrossRef]
- Vig, K.; Chaudhari, A.; Tripathi, S.; Dixit, S.; Sahu, R.; Pillai, S.; Dennis, V.A.; Singh, S.R. Advances in skin regeneration using tissue engineering. Int. J. Mol. Sci. 2017, 18, 789. [Google Scholar] [CrossRef] [PubMed]
- Ozcelik, H.; Hindie, M.; Hasan, A.; Engin, N.; Cell, V.; Barthes, J.; Özçelik, H.; Hindié, M.; Ndreu-halili, A.; Vrana, N.E. Cell microenvironment engineering and monitoring for tissue engineering and regenerative medicine. Biomed Res. Int. 2014, 2014, 18. [Google Scholar]
- Olson, J.L.; Atala, A.; Yoo, J.J. Tissue Engineering: Current Strategies and Future Directions. Chonnam Med. J. 2011, 47, 1–13. [Google Scholar] [CrossRef] [PubMed]
- Inchingolo, F.; Tatullo, M.; Marrelli, M.; Inchingolo, A.M.; Inchingolo, A.D.; Dipalma, G.; Flace, P.; Girolamo, F.; Tarullo, A.; Laino, L.; et al. Regenerative surgery performed with platelet-rich plasma used in sinus lift elevation before dental implant surgery: An useful aid in healing and regeneration of bone tissue. Eur. Rev. Med. Pharmacol. Sci. 2012, 16, 1222–1226. [Google Scholar] [PubMed]
- Nayak, S.; Dey, S.; Kundu, S.C. Skin equivalent tissue-engineered construct: Co-cultured fibroblasts/ keratinocytes on 3D matrices of sericin hope cocoons. PLoS ONE 2013, 8, 1–17. [Google Scholar] [CrossRef] [PubMed]
- Chan, B.P.; Leong, K.W. Scaffolding in tissue engineering: General approaches and tissue-specific considerations. Eur. Spine J. 2008, 17, 467–479. [Google Scholar] [CrossRef]
- Paduano, F.; Marrelli, M.; Alom, N.; Amer, M.; White, L.J.; Shakesheff, K.M.; Tatullo, M. Decellularized bone extracellular matrix and human dental pulp stem cells as a construct for bone regeneration. J. Biomater. Sci. Polym. Ed. 2017, 28, 730–748. [Google Scholar] [CrossRef] [Green Version]
- Strong, A.L.; Neumeister, M.W.; Levi, B. Stem cells and tissue engineering: Regeneration of the skin and its contents. Clin Plast Surg. 2018, 44, 635–650. [Google Scholar] [CrossRef]
- Andreu, V.; Mendoza, G.; Arruebo, M.; Irusta, S. Smart dressings based on nanostructured fibers containing natural origin antimicrobial, anti-inflammatory, and regenerative compounds. Materials (Basel) 2015, 8, 5154–5193. [Google Scholar] [CrossRef]
- Harrison, K. Introduction to polymeric scaffolds for tissue engineering. In Biomedical Polymers, 1st ed.; Woodhead Publishing Limited: Cambridge, UK, 2007; pp. 1–32. [Google Scholar]
- Kim, P.H.; Cho, J.Y. Myocardial tissue engineering using electrospun nanofiber composites. BMB Rep. 2016, 49, 26–36. [Google Scholar] [CrossRef] [PubMed] [Green Version]
- Agarwal, S.; Wendorff, J.H.; Greiner, A. Progress in the field of electrospinning for tissue engineering applications. Adv. Mater. 2009, 21, 3343–3351. [Google Scholar] [CrossRef] [PubMed]
- Ulery, B.D.; Nair, L.S.; Laurencin, C.T. Biomedical applications of biodegradable polymers. J. Polym. Sci. Part B Polym. Phys. 2011, 49, 832–864. [Google Scholar] [CrossRef] [PubMed] [Green Version]
- Khosravi, A.; Ghasemi-Mobarakeh, L.; Mollahosseini, H.; Ajalloueian, F.; Masoudi Rad, M.; Norouzi, M.R.; Sami Jokandan, M.; Khoddami, A.; Chronakis, I.S. Immobilization of silk fibroin on the surface of PCL nanofibrous scaffolds for tissue engineering applications. J. Appl. Polym. Sci. 2018, 135, 1–8. [Google Scholar] [CrossRef]
- Amar, S.; Resham, V.; Saple, D.G. Aloe Vera: A Short Review. Indian J. Dermatol. 2008, 53, 163–166. [Google Scholar]
- Rahman, S.; Carter, P.; Bhattarai, N. Aloe Vera for Tissue Engineering Applications. J. Funct. Biomater. 2017, 8, 6. [Google Scholar] [CrossRef]
- Kant, V.; Gopal, A.; Pathak, N.N.; Kumar, P.; Tandan, S.K.; Kumar, D. Antioxidant and anti-inflammatory potential of curcumin accelerated the cutaneous wound healing in streptozotocin-induced diabetic rats. Int. Immunopharmacol. 2014, 20, 322–330. [Google Scholar] [CrossRef]
- Zorofchian Moghadamtousi, S.; Abdul Kadir, H.; Hassandarvish, P.; Tajik, H.; Abubakar, S.; Zandi, K. A review on antibacterial, antiviral, and antifungal activity of curcumin. Biomed Res. Int. 2014, 2014, 1–12. [Google Scholar] [CrossRef]
- Garrido-Mesa, N.; Zarzuelo, A.; Gálvez, J. Minocycline: Far beyond an antibiotic. Br. J. Pharmacol. 2013, 169, 337–352. [Google Scholar] [CrossRef]
- Soares, P.B.F.; de Menezes, H.H.M.H.; de Naves, M.M.; Taga, E.M.; de Magalhães, D. Effect of absorbent tetracycline-loaded membrane used in the reduction of periodontal pockets: An in vivo study. Braz. Dent. J. 2009, 20, 414–418. [Google Scholar] [CrossRef]
- Agnes Mary, S.; Giri Dev, V.R. Electrospun herbal nanofibrous wound dressings for skin tissue engineering. J. Text. Inst. 2015, 106, 886–895. [Google Scholar] [CrossRef]
- Lee, D.E.; Ayoub, N.; Agrawal, D.K. Mesenchymal stem cells and cutaneous wound healing: Novel methods to increase cell delivery and therapeutic efficacy. Stem Cell Res. Ther. 2016, 7, 1–8. [Google Scholar] [CrossRef] [PubMed]
- Li, P.; Guo, X. A review: Therapeutic potential of adipose-derived stem cells in cutaneous wound healing and regeneration. Stem Cell Res. Ther. 2018, 9, 1–7. [Google Scholar] [CrossRef] [PubMed]
- Parisi, L.; Toffoli, A.; Ghiacci, G.; Macaluso, G.M. Tailoring the interface of biomaterials to design effective scaffolds. J. Funct. Biomater. 2018, 9, 1–31. [Google Scholar] [CrossRef]
- Karuppuswamy, P.; Venugopal, J.R.; Navaneethan, B.; Laiva, A.L.; Sridhar, S.; Ramakrishna, S. Functionalized hybrid nanofibers to mimic native ECM for tissue engineering applications. Appl. Surf. Sci. 2014, 322, 162–168. [Google Scholar] [CrossRef]
- Shababdoust, A.; Ehsani, M.; Shokrollahi, P.; Zandi, M. Fabrication of curcumin-loaded electrospun nanofiberous polyurethanes with anti-bacterial activity. Prog. Biomater. 2017, 7, 23–33. [Google Scholar] [CrossRef] [Green Version]
- Širc, J.; Hobzová, R.; Kostina, N.; Munzarová, M.; Juklíčková, M.; Lhotka, M.; Kubinová, Š.; Zajícová, A.; Michálek, J. Morphological characterization of nanofibers: Methods and application in practice. J. Nanomater. 2012, 2012, 14. [Google Scholar] [CrossRef]
- Balaji, A.; Jaganathan, S.K.; Ismail, A.F.; Rajasekar, R. Fabrication and hemocompatibility assessment of novel polyurethane-based bio-nanofibrous dressing loaded with honey and Carica papaya extract for the management of burn injuries. Int. J. Nanomedicine 2016, 11, 4339–4355. [Google Scholar]
- Shah, S.A.A.; Imran, M.; Lian, Q.; Shehzad, F.K.; Athir, N.; Zhang, J.; Cheng, J. Curcumin incorporated polyurethane urea elastomers with tunable thermo-mechanical properties. React. Funct. Polym. 2018, 128, 97–103. [Google Scholar] [CrossRef]
- Trang Mai, T.T.; Thuy Nguyen, T.T.; Le, Q.D.; Nguyen, T.N.; Ba, T.C.; Nguyen, H.B.; Hoa Phan, T.B.; Tran, D.L.; Nguyen, X.P.; Park, J.S. A novel nanofiber Cur-loaded polylactic acid constructed by electrospinning. Adv. Nat. Sci. Nanosci. Nanotechnol. 2012, 3, 4. [Google Scholar] [CrossRef]
- Liao, N.; Unnithan, A.R.; Joshi, M.K.; Tiwari, A.P.; Hong, S.T.; Park, C.H.; Kim, C.S. Electrospun bioactive poly (ε-caprolactone)–cellulose acetate–dextran antibacterial composite mats for wound dressing applications. Colloids Surfaces A Physicochem. Eng. Asp. 2015, 469, 194–201. [Google Scholar] [CrossRef]
- Suganya, S.; Venugopal, J.; Agnes Mary, S.; Ramakrishna, S.; Lakshmi, B.S.; Giri Dev, V.R. Aloe vera incorporated biomimetic nanofibrous scaffold: A regenerative approach for skin tissue engineering. Iran. Polym. J. 2014, 23, 237–248. [Google Scholar] [CrossRef]
- Lee, H.; Yamaguchi, K.; Nagaishi, T.; Murai, M.; Kim, M.; Wei, K.; Zhang, K.Q.; Kim, I.S. Enhancement of mechanical properties of polymeric nanofibers by controlling crystallization behavior using a simple freezing/thawing process. RSC Adv. 2017, 7, 43994–44000. [Google Scholar] [CrossRef] [Green Version]
- Xiang, C.; Frey, M.W. Increasing mechanical properties of 2-D-structured electrospun nylon 6 non-woven fiber mats. Materials (Basel) 2016, 9, 270. [Google Scholar] [CrossRef] [PubMed]
- Wongkanya, R.; Chuysinuan, P.; Pengsuk, C.; Techasakul, S.; Lirdprapamongkol, K.; Svasti, J.; Nooeaid, P. Electrospinning of alginate/soy protein isolated nanofibers and their release characteristics for biomedical applications. J. Sci. Adv. Mater. Devices 2017, 2, 309–316. [Google Scholar] [CrossRef]
- Chen, S.C.; Huang, X.B.; Cai, X.M.; Lu, J.; Yuan, J.; Shen, J. The influence of fiber diameter of electrospun poly(lactic acid) on drug delivery. Fibers Polym. 2012, 13, 1120–1125. [Google Scholar] [CrossRef]
- Kumar, S.; Yadav, A.; Yadav, M.; Yadav, J.P. Effect of climate change on phytochemical diversity, total phenolic content and in vitro antioxidant activity of Aloe vera (L.) Burm.f. BMC Res. Notes 2017, 10, 1–12. [Google Scholar] [CrossRef]
- Xue, M.; Jackson, C.J. Extracellular matrix reorganization during wound healing and its impact on abnormal scarring. Adv. Wound Care 2015, 4, 119–136. [Google Scholar] [CrossRef]
- Wakuda, Y.; Nishimoto, S.; Suye, S.I.; Fujita, S. Native collagen hydrogel nanofibres with anisotropic structure using core-shell electrospinning. Sci. Rep. 2018, 8, 1–10. [Google Scholar] [CrossRef]
- Brett, D. A review of collagen and collagen-based wound dressings. Wounds 2008, 20, 347–356. [Google Scholar]
- Jin, G.; Prabhakaran, M.P.; Kai, D.; Annamalai, S.K.; Arunachalam, K.D.; Ramakrishna, S. Tissue engineered plant extracts as nanofibrous wound dressing. Biomaterials 2013, 34, 724–734. [Google Scholar] [CrossRef] [PubMed]
- Abreu-Blanco, M.T.; Watts, J.J.; Verboon, J.M.; Parkhurst, S.M. Cytoskeleton responses in wound repair. Cell. Mol. Life Sci. 2012, 69, 2469–2483. [Google Scholar] [Green Version]
- Delaine-Smith, R.M.; Green, N.H.; Matcher, S.J.; MacNeil, S.; Reilly, G.C. Monitoring fibrous scaffold guidance of three-dimensional collagen organisation using minimally-invasive second harmonic generation. PLoS ONE 2014, 9, e89761. [Google Scholar] [CrossRef] [PubMed]
- Ramalingam, R.; Dhand, C.; Leung, C.M.; Ezhilarasu, H.; Prasannan, P.; Ong, S.T.; Subramanian, S.; Kamruddin, M.; Lakshminarayanan, R.; Ramakrishna, S.; et al. Poly-ε-caprolactone/gelatin hybrid electrospun composite nanofibrous mats containing ultrasound assisted herbal extract: Antimicrobial and cell proliferation study. Nanomaterials 2019, 9, 462. [Google Scholar] [CrossRef] [PubMed]
Nanofiber Construct | Fiber Diameter (nm) | Water Contact Angle (°) |
---|---|---|
PCL | 770 ± 98 | 128.3 ± 6 |
PCL/AV | 561 ± 49 **** | 47.3 ± 2.5 |
PCL/CUR | 695 ± 57 ** | 94.3 ± 3.7 |
PCL/AV/CUR | 665 ± 64 **** | 79 ± 1.6 |
PCL/AV/TCH | 360 ± 87 **** | 57.3 ± 5 |
Nanofiber Construct | Ultimate Tensile Stress (MPa) | Ultimate Tensile Strain (%) | Young’s Modulus (MPa) |
---|---|---|---|
PCL | 9.2 | 164.5 | 19.8 |
PCL/AV | 14.9 | 53.1 | 72.3 |
PCL/CUR | 11.1 | 64.2 | 79.7 |
PCL/AV/CUR | 14.4 | 65.2 | 25.5 |
PCL/AV/TCH | 20.0 | 40.1 | 92.6 |
Samples | Concentration (%) | Voltage (kV) | Flow Rate (ml/h) | Needle Size |
---|---|---|---|---|
PCL | 13 | 14 | 1.5 | 24G |
PCL/AV | 10:3 | 14 | 1.5 | 24G |
PCL/CUR | 10:3 | 14 | 1.5 | 24G |
PCL/AV/CUR | 9:3:1 | 14 | 1.5 | 24G |
PCL/AV/TCH | 9:3:1 | 15 | 1.5 | 24G |
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Ezhilarasu, H.; Ramalingam, R.; Dhand, C.; Lakshminarayanan, R.; Sadiq, A.; Gandhimathi, C.; Ramakrishna, S.; Bay, B.H.; Venugopal, J.R.; Srinivasan, D.K. Biocompatible Aloe vera and Tetracycline Hydrochloride Loaded Hybrid Nanofibrous Scaffolds for Skin Tissue Engineering. Int. J. Mol. Sci. 2019, 20, 5174. https://doi.org/10.3390/ijms20205174
Ezhilarasu H, Ramalingam R, Dhand C, Lakshminarayanan R, Sadiq A, Gandhimathi C, Ramakrishna S, Bay BH, Venugopal JR, Srinivasan DK. Biocompatible Aloe vera and Tetracycline Hydrochloride Loaded Hybrid Nanofibrous Scaffolds for Skin Tissue Engineering. International Journal of Molecular Sciences. 2019; 20(20):5174. https://doi.org/10.3390/ijms20205174
Chicago/Turabian StyleEzhilarasu, Hariharan, Raghavendra Ramalingam, Chetna Dhand, Rajamani Lakshminarayanan, Asif Sadiq, Chinnasamy Gandhimathi, Seeram Ramakrishna, Boon Huat Bay, Jayarama Reddy Venugopal, and Dinesh Kumar Srinivasan. 2019. "Biocompatible Aloe vera and Tetracycline Hydrochloride Loaded Hybrid Nanofibrous Scaffolds for Skin Tissue Engineering" International Journal of Molecular Sciences 20, no. 20: 5174. https://doi.org/10.3390/ijms20205174