دنیای نانو

دنیای نانو

سنتز و مشخصه‌یابی نانوکامپوزیت کراتین-سیلیکا مزوپروس با استفاده از کراتین استخراج‌شده از پر بلدرچین، ارزیابی فعالیت ضدباکتریایی و زیست‌سازگاری آن

نوع مقاله : مقاله پژوهشی

نویسندگان
فاطمه درویش زاده 1
آزاده حکمت 1
بنائی اقدس 2
1 گروه زیست شناسی، واحد علوم و تحقیقات، دانشگاه آزاد اسلامی، تهران، ایران
2 گروه نانو و بیوفیزیک، پژوهشکده علوم کاربردی جهاد دانشگاهی، دانشگاه شهید بهشتی، تهران، ایران
چکیده
با وجود پتانسیل بالای نانوپروتئین‌ها و نانوذرات سیلیکای مزوپوروس (MSNPs)، مطالعات محدودی به ترکیب هم‌زمان این دو ترکیب زیستی و معدنی، به‌ویژه با استفاده از کراتین طبیعی استخراج‌شده از پر پرندگان پرداخته‌اند. در این پژوهش، کراتین برای اولین بار از پر بلدرچین استخراج و با استفاده از روش اولتراسونیک به نانوکراتین (nK) تبدیل شد. نانوکراتین سپس بر روی MSNPs بارگذاری گردید. طیف‌سنجی مرئی-فرابنفش حضور پیوندهای پپتیدی و اسیدهای آمینه آروماتیک را در nK و نانوکامپوزیت (nK-MSNPs) تأیید کرد. طیف‌های مادون قرمز وجود گروه‌های عاملی مشخص کراتین مانند پیک آمید I، آمیدII و سیستئین (S-S) در نانوکراتین و باندهای Si-O-Si در محدوده cm⁻¹ 1100 و 800 مربوط به ساختار MSNPs در نانوکامپوزیت را تایید نمود. الگوی پراش پرتو ایکس ساختار آمورف را در nK-MSNPs نشان داد. میانگین اندازه nK و nK-MSNPs به‌ترتیب 4/245 و 4/211 نانومتر و شاخص پراکندگی آن‌ها به‌ترتیب 594/0 و 48/1 گزارش شد. پتانسیل زتای nK و nK-MSNPs به‌ترتیب 6/9- و 9/16- میلی‌ولت اندازه‌گیری گردید که نشان‌دهنده پایداری الکترواستاتیکی کامپوزیت بود. تصاویر میکروسکوپ الکترونی کاهش تجمع در nK-MSNPs را نسبت به nK تأیید کرد. نانوکامپوزیت در بازه‌ی ۹۰ روزه از نظر شیمیایی پایدار باقی ماند. نتایج آزمون سمیت سلولی (MTT) بر روی سلول‌های فیبروبلاست انسانی (hFF) نشان داد که nK-MSNPs حتی در غلظت‌های نسبتاً بالا فاقد اثر منفی معنادار بر بقاء سلولی است. بررسی فعالیت ضدباکتریایی نشان داد nK-MSNPs در باکتری‌های S. aureus وE. coli به‌ترتیب نواحی بازدارندگی 5/0±8/22  و 4/0±2/20 میلی‌متر ایجاد می‌کند که از نظر آماری معنادار (P<0.05) و قابل مقایسه با آنتی‌بیوتیک مرجع بود. بنابراین nK-MSNPs با ساختار متخلخل، پایداری طولانی‌مدت، زیست‌سازگاری و فعالیت ضدمیکروبی، به عنوان گزینه‌ای مناسب در مصارف زیست‌پزشکی می‌تواند باشد.
کلیدواژه‌ها
نانوکراتین
نانوذرات سیلیکا مزوپروس
نانوکامپوزیت
فعالیت ضدباکتریایی
پر بلدرچین
فیبروبلاست
موضوعات

عنوان مقاله English

Synthesis and Characterization of Keratin–Mesoporous Silica Nanocomposite Using Keratin Extracted from Quail Feathers, and Evaluation of Its Antibacterial Activity and Biocompatibility

نویسندگان English

Fatemeh Darvishzadeh 1
Azadeh Hekmat 1
Aghdas Banaei 2
1 Department of Biology, SR. C., Islamic Azad University, Tehran, Iran
2 Department of Nano and Biophysics, Research Institute for Applied Sciences (RIAS), ACECR, Tehran, Iran
چکیده English

Nanoproteins and mesoporous silica nanoparticles (MSNPs) have attracted considerable attention due to their unique biological and physicochemical properties; however, studies combining these biomacromolecules and inorganic carriers remain limited, particularly when employing natural keratin derived from avian feathers. In the present study, keratin was extracted for the first time from quail feathers and converted into nanokeratin (nK) through an ultrasonic process, followed by its loading onto MSNPs. UV–visible spectroscopy confirmed the presence of peptide bonds and aromatic amino acids in both nK and the nanocomposite (nK-MSNPs). Fourier-transform infrared spectra revealed characteristic keratin functional groups, including amide I, amide II, and cysteine (S-S), alongside Si-O-Si bands at 1100 and 800 cm⁻¹, indicative of MSNPs within the nanocomposite. X-ray diffraction confirmed an amorphous structure. The average particle sizes of nK and nK-MSNPs were 245.4 and 211.4 nm, respectively, with polydispersity indices of 0.594 and 0.148. Zeta potential values (-9.6 and -16.9 mV) demonstrated electrostatic stability, while electron microscopy images revealed reduced aggregation in the nanocomposite compared to nK alone. The nanocomposite also retained chemical stability over 90 days. Cytotoxicity evaluation using the MTT assay on human fibroblast cells (hFF) showed that nK-MSNPs were non-toxic even at relatively high concentrations. Moreover, antibacterial assays revealed significant inhibition zones of 22.8 ± 0.5 mm against Staphylococcus aureus and 20.2 ± 0.4 mm against Escherichia coli (P<0.05), comparable to a reference antibiotic. Collectively, these findings suggest that nK-MSNPs, owing to their porous structure, long-term stability, biocompatibility, and antibacterial activity, hold strong potential for biomedical applications.

کلیدواژه‌ها English

Nanokeratin
Mesoporous silica nanoparticles
Nanocomposite
Antibacterial activity
Quail feather
Fibroblast

  • Todros S, Todesco M, Bagno A. Biomaterials and their biomedical applications: From replacement to regeneration. Processes. 2021;9(11):1949. 10.3390/pr9111949.

  • Bhari R, Kaur M, Sarup Singh R. Chicken feather waste hydrolysate as a superior biofertilizer in agroindustry. Current Microbiology. 2021;78(6):2212-30. 10.1007/s00284-021-02491-z.

  • Martella E, Ferroni C, Guerrini A, Ballestri M, Columbaro M, S et al. Functionalized keratin as nanotechnology-based drug delivery system for the pharmacological treatment of osteosarcoma. International journal of molecular sciences. 2018;19(11):3670. 10.3390/ijms19113670.

  • Kamarudin NB, Sharma S, Gupta A, Kee CG, Chik SMSBT, Gupta R. Statistical investigation of extraction parameters of keratin from chicken feather using Design-Expert. 3 Biotech. 2017;7:1-9. 10.1007/s13205-017-0767-9.

  • Konop M, Czuwara J, Kłodzińska E, Laskowska AK, Sulejczak D, Damps T, et al. Evaluation of keratin biomaterial containing silver nanoparticles as a potential wound dressing in full-thickness skin wound model in diabetic mice. Journal of tissue engineering and regenerative medicine. 2020;14(2):334-46. 10.1002/term.2998.

  • Mousavi SZ, Manteghian M, Shojaosadati SA, Pahlavanzadeh H. Keratin nanoparticles: synthesis and application for Cu (II) removal. Advances in environmental technology. 2018;4(2):83-93. 10.22104/aet.2018.2948.1143.

  • Posati T, Listwan A, Sotgiu G, Torreggiani A, Zamboni R, Aluigi A. Keratin/hydrotalcites hybrid sponges as promising adsorbents for cationic and anionic dyes. Frontiers in bioengineering and biotechnology. 2020;8:68. 10.3389/fbioe.2020.00068.

  • Babalola R, Ayeni AO, Joshua PS, Ayoola AA, Isaac UO, Aniediong U, et al. Synthesis of thermal insulator using chicken feather fibre in starch-clay nanocomposites. Heliyon. 2020;6(11). 10.1016/j.heliyon.2020.e05384.

  • Khodaei D, Álvarez C, Mullen AM. Biodegradable packaging materials from animal processing co-products and wastes: An overview. Polymers. 2021;13(15):2561. 10.3390/polym13152561.

  • Avancini G, Guerrini A, Ferroni C, Tedesco D, Ballestri M, Columbaro M, et al. Keratin nanoparticles and photodynamic therapy enhance the anticancer stem cells activity of salinomycin. Materials Science and Engineering: C. 2021;122:111899. 10.1016/j.msec.2021.111899.

  • Guglielmelli A, Rosa P, Contardi M, Prato M, Mangino G, Miglietta S, et al. Biomimetic keratin gold nanoparticle-mediated in vitro photothermal therapy on glioblastoma multiforme. Nanomedicine. 2021;16(2):121-38. 10.2217/nnm-2020-0349.

  • Donato RK, Mija A. Keratin associations with synthetic, biosynthetic and natural polymers: An extensive review. Polymers. 2019;12(1):32. 10.3390/polym12010032.

  • Wang X, Shi Z, Zhao Q, Yun Y. Study on the structure and properties of biofunctional keratin from rabbit hair. Materials. 2021;14(2):379. 10.3390/ma14020379.

  • Sarma A. Biological importance and pharmaceutical significance of keratin: a review. International Journal of Biological Macromolecules. 2022;219:395-413. 10.1016/j.ijbiomac.2022.08.002.

  • Han X, Wang L, Du J, Dou J, Yuan J, Shen J. Keratin-dopamine conjugate nanoparticles as pH/GSH dual responsive drug carriers. Journal of Biomaterials Science, Polymer Edition. 2020;31(18):2318-30. 10.1080/09205063.2020.1803182.

  • de Q. Souza GE, Burin GR, de Muniz GI, Alves HJ. Valorization of feather waste in Brazil: structure, methods of extraction, and applications of feather keratin. Environmental Science and Pollution Research. 2023;30(14):39558-67. 10.1007/s11356-023-25788-x.

  • Yilma KT, Tesfaye T, Enawgaw H, Ayele M, Limeneh DY, Gibril M, et al. Optimizing the Extraction of Keratin from Cattle Hoof Using Central Composite Design. Advances in Materials Science and Engineering. 2022;2022(1):4866467. 10.1155/2022/4866467.

  • Low CT. Human hair keratin and its interaction with metal ions. 2020. 10.32657/10356/142466.

  • Banasaz S, Ferraro V. Keratin from animal by-products: structure, characterization, extraction and application- a review. Polymers. 2024;16(14):1999. 10.3390/polym16141999.

  • Giteru SG, Ramsey DH, Hou Y, Cong L, Mohan A, Bekhit AEDA. Wool keratin as a novel alternative protein: A comprehensive review of extraction, purification, nutrition, safety, and food applications. Comprehensive reviews in food science and food safety. 2023;22(1):643-87. 10.1111/1541-4337.13087.

  • Zhang Y, Zhang N, Wang Q, Wang P, Yu Y. A facile and eco-friendly approach for preparation of microkeratin and nanokeratin by ultrasound-assisted enzymatic hydrolysis. Ultrasonics Sonochemistry. 2020;68:105201. 10.1016/j.ultsonch.2020.105201.

  • Du J, Wang L, Han X, Dou J, Yuan J, Shen J. Keratin-tannic acid complex nanoparticles as pH/GSH dual responsive drug carriers for doxorubicin. Journal of Biomaterials Science, Polymer Edition. 2021;32(9):1125-39. 10.1080/09205063.2021.1906074.

  • Pakdel M, Moosavi-Nejad Z, Kermanshahi RK, Hosano N, Qamsari EM, Hosano H. Keratin nanoparticles derived from feather waste for novel antibacterial delivery. International Journal of Biological Macromolecules. 2025;298:139676. 10.1016/j.ijbiomac.2025.139676.

  • Biradar AI, Sarvalkar PD, Teli SB, Pawar C, Patil P, Prasad NR. Photocatalytic degradation of dyes using one-step synthesized silica nanoparticles. Materials Today: Proceedings. 2021;43:2832-8. 10.1016/j.matpr.2020.11.946.

  • Hekmat A, Haertlé T, Leblanc RM, Khan HY, Khan RH, Saboury AA. A Review on Interaction of Nanomaterials of Group-XIV (G14) Elements of the Periodic Table with Proteins and DNA: Applications in Biotechnology and Pharmacy. BioNanoScience. 2024;14(2):1978-2003. 10.1007/s12668-024-01423-y.

  • Nairi V, Medda S, Piludu M, Casula MF, Vallet-Regì M, Monduzzi M, et al. Interactions between bovine serum albumin and mesoporous silica nanoparticles functionalized with biopolymers. Chemical Engineering Journal. 2018;340:42-50. 10.1016/j.cej.2018.01.011.

  • Maurya SD, Singh A. Extraction and Characterization of Keratin from Waste Broiler Chicken Feathers. Environmental Protection Research. 2023:372-81. 10.37256/epr.3220233579.

  • Subroto E, Lembong E, Filianty F, Indiarto R, Primalia G, Putri M, et al. The analysis techniques of amino acid and protein in food and agricultural products. Int J Sci Technol Res. 2020;9(10):29-36.

  • Miranda MP. Comparison of the effect of sodium chloride concentration on protein determination: Bradford and Biuret methods. Analytical Biochemistry. 2024;687:115450. 10.1016/j.ab.2023.115450.

  • Ferroni C, Varchi G. Keratin-based nanoparticles as drug delivery carriers. Applied Sciences. 2021;11(20):9417. 10.3390/app11209417.

  • Khan MUA, Razaq SIA, Mehboob H, Rehman S, Al-Arjan WS, Amin R. Antibacterial and hemocompatible pH-responsive hydrogel for skin wound healing application: In vitro drug release. Polymers. 2021;13(21):3703. 10.3390/polym13213703.

  • Rai S, Dash D, Agarwal N. Introducing the new face of CLSI M100 in 2023: An explanatory review. Indian Journal of Medical Microbiology. 2023;46:100432. 10.1016/j.ijmm.2023.100432.

  • Kummari R, Puja R, Bose K. Protein quantitation and detection. Textbook on cloning, expression and purification of recombinant proteins: Springer; 2022. p. 279-99. 10.1007/978-981-16-4987-5_11.

  • Hekmat A, Hatamie S, Saboury AA. The effects of synthesized silver nanowires on the structure and esterase-like activity of human serum albumin and their impacts on human endometrial stem cells. Inorganic and Nano-Metal Chemistry. 2024;54(5):399-412. 10.1080/24701556.2022.2034859.

  • Ge N, Zhang Y, Zhang H, Cheng L, Shi X, editors. Extraction and characterization of keratin and keratin hydrogels from wasted rabbit hair. Journal of Physics: Conference Series; 2021: IOP Publishing. 10.1088/1742-6596/1790/1/012008.

  • Banerjee S, Vishakha K, Das S, Sangma PD, Mondal S, Ganguli A. Antibacterial and antibiofilm activity of facile synthesized biocompatible keratin-copper nanoparticles against multidrug resistant uro-pathogens. World Journal of Microbiology and Biotechnology. 2022;38(2):20. 10.1007/s11274-021-03187-z.

  • Zhang C, Xia L, Zhang J, Liu X, Xu W. Utilization of waste wool fibers for fabrication of wool powders and keratin: A review. Journal of Leather Science and Engineering. 2020;2:1-15. 10.1186/s42825-020-00030-3.

  • Pourjavaheri F, Pour SO, Jones OA, Smooker PM, Brkljača R, Sherkat F, et al. Extraction of keratin from waste chicken feathers using sodium sulfide and l-cysteine. Process Biochemistry. 2019;82:205-14. 10.1016/j.procbio.2019.04.010.

  • Sharma I, Pranaw K, Soni H, Rawat HK, Kango N. Parametrically optimized feather degradation by Bacillus velezensis NCIM 5802 and delineation of keratin hydrolysis by multi-scale analysis for poultry waste management. Scientific Reports. 2022;12(1):17118. 10.1038/s41598-022-21351-9.

  • Elizondo-Villarreal N, Gandara-Martínez E, García-Méndez M, Gracia-Pinilla M, Guzmán-Hernández AM, Castaño VM, et al. Synthesis and characterization of SiO2 nanoparticles for application as nanoadsorbent to clean wastewater. Coatings. 2024;14(7):919. 10.3390/coatings14070919.

  • Yildiz SN, Arslan TS, Arslan YE. Organic-inorganic biohybrid films from wool-keratin/jellyfish-collagen/silica/boron via sol-gel reactions for soft tissue engineering applications. Biomedical Materials. 2024;19(2):025032. 10.1088/1748-605X/ad2557.

  • Sellappan LK, Manoharan S. Fabrication of bioinspired keratin/sodium alginate based biopolymeric mat loaded with herbal drug and green synthesized zinc oxide nanoparticles as a dual drug antimicrobial wound dressing. International Journal of Biological Macromolecules. 2024;259:129162. 10.1016/j.ijbiomac.2023.129162.

  • Pakdel M, Moosavi-Nejad Z, Kermanshahi RK, Hosano H. Self-assembled uniform keratin nanoparticles as building blocks for nanofibrils and nanolayers derived from industrial feather waste. Journal of Cleaner Production. 2022;335:130331. 10.1016/j.jclepro.2021.130331.

  • Bhattacharjee S. DLS and zeta potential–what they are and what they are not? Journal of controlled release. 2016;235:337-51. 10.1016/j.jconrel.2016.06.017.

  • Rawle A. Basic of principles of particle-size analysis. Surface coatings international Part A, Coatings journal. 2003;86(2):58-65.

  • Wang X, Zhang Z, Xu H, Li X, Hao X. Preparation of sheep bone collagen peptide–calcium chelate using enzymolysis-fermentation methodology and its structural characterization and stability analysis. RSC advances. 2020;10(20):11624-33. 10.1039/D0RA00425A.

  • Shankar S, Rhim J-W. Eco-friendly antimicrobial nanoparticles of keratin-metal ion complex. Materials Science and Engineering: C. 2019;105:110068. 10.1016/j.msec.2019.110068.

  • Godoy-Gallardo M, Eckhard U, Delgado LM, de Roo Puente YJ, Hoyos-Nogués M, Gil FJ, et al. Antibacterial approaches in tissue engineering using metal ions and nanoparticles: From mechanisms to applications. Bioactive Materials. 2021;6(12):4470-90. 10.1016/j.bioactmat.2021.04.033.

  • Du J, Chen G, Yuan X, Yuan J, Li L. Multi-stimuli responsive Cu-MOFs@Keratin drug delivery system for chemodynamic therapy. Frontiers in Bioengineering and Biotechnology. 2023;11:1125348. 10.3389/fbioe.2023.1125348.

دنیای نانو
دوره 21، شماره 80
پاییز 1404
صفحه 40-59

  • تاریخ دریافت 21 مرداد 1404
  • تاریخ بازنگری 03 مهر 1404
  • تاریخ پذیرش 21 مهر 1404