بررسی زیست سازگاری داربست PLA پوشش داده شده با لاپونیت بر روی سلول های بنیادی مزانشیمی مغز استخوان انسان

نوع مقاله : مقاله پژوهشی

نویسندگان

1 گروه زیست شناسی، واحد علوم و تحقیقات، دانشگاه آزاد اسلامی، تهران، ایران

2 گروه بیومواد، پژوهشکده فناوری نانو و مواد پیشرفته، پژوهشگاه مواد و انرژی، کرج، ایران

3 پژوهشکده سیستم‌های پلیمری عامل، موسسه فرانهوفر برای تحقیقات پلیمری کاربردی، پوتسدام، آلمان

4 مرکز تحقیقات نانو تکنولوژی پزشکی و مهندسی بافت، پژوهشکده علوم تولید مثل یزد، دانشگاه علوم پزشکی و خدمات بهداشتی درمانی شهید صدوقی یزد، یزد، ایران

5 گروه نانوتکنولوژی پزشکی، واحد علوم و تحقیقات، دانشگاه آزاد اسلامی، تهران، ایران

چکیده

مهندسی بافت استخوان رویکردی امیدوارانه جهت توسعه درمان­های مناسب جدید برای رفع آسیب­های بافت استخوانی است. یکی از اهداف مهم در این رشته، ساخت داربست­هایی با تقلید از ماتریکس خارج سلولی است. هدف از این مطالعه تولید داربست پلی­لاکتیک اسید/لاپونیت (PLA/LAP) و بررسی رفتار سلول­های بنیادی مزانشیمی مغز استخوان انسانی (hBMSCs) بر روی آن بود. ابتدا داربست PLA به روش الکتروریسی ساخته شد و سپس LAP با غلظت 8/0 درصد وزنی (LAP0.8%) بر روی آن پوشش داده شد. مورفولوژی داربست توسط میکروسکوپ الکترونی روبشی (SEM) و طیف­سنجی پراش انرژی پرتو ایکس (EDX)، ساختار شیمیایی آن توسط طیف­سنجی ATR-FTIR و میزان آبدوستی داربست با آزمون زاویه تماس آب بررسی شد. نهایتاً زیست­سازگاری داربست و بقای سلولی توسط تست MTT، بر روی سلول­های hBMSC انجام شد. نتایج حاصل از مورفولوژی داربست نشان­دهنده پوشش­دهی موفق LAP0.8% بر روی داربست PLA بود. همچنین، سطح آبدوستی داربست PLA پس از پوشش­دهی با LAP بهبود یافت. زیست­سازگاری داربست تا 24 ساعت بعد از کشت سلولی و بقای سلول­های hBMSC تا 72 ساعت پس از کشت (001/0 ≥ p ) تایید شد. از نتایج بدست آمده در این تحقیق به نظر می­رسد که داربست  PLA/LAP0.8%بدلیل حفظ زیست­سازگاری و بقای سلولی که بواسطه حضور یون­های موجود در نانوذره LAP است، می­تواند کاندیدای مناسبی برای برنامه­های مهندسی بافت استخوان باشد.

کلیدواژه‌ها


عنوان مقاله [English]

Evaluation of Biocompatibility of PLA Scaffold Coated with Laponite on Human Bone Marrow Mesenchymal Stem Cells

نویسندگان [English]

  • Zahra Orafa 1
  • Shiva Irani 1
  • Ali Zamanian 2
  • Hadi Bakhshi 3
  • Habib Nikukar 4
  • Behafarid Ghalandari 5
1 Department of Biology, Science and Research Branch, Islamic Azad University, Tehran, Iran
2 Biomaterials Group, Research Institute of Nanotechnology and Advanced Materials, Materials and Energy Research Institute, Karaj, Iran
3 Operating Polymer Systems Research Institute, Fraunhofer Institute for Applied Polymer Research, Potsdam, Germany
4 Medical Nanotechnology and Tissue Engineering Research Center, Yazd Reproductive Sciences Research Institute, Shahid Sadoughi University of Medical Sciences and Health Services, Yazd, Iran
5 Department of Medical Nanotechnology, Science and Research Branch, Islamic Azad University, Tehran, Iran
چکیده [English]

Bone tissue engineering is a promising approach to develop new appropriate treatments for bone tissue damage. One of the important goals in this field is to fabricate the scaffolds by mimicking the extracellular matrix. The aim of this study was to study the fabrication of polylactic acid/Laponite (PLA/LAP) scaffold and to investigate the behavior of human bone marrow mesenchymal stem cells (hBMSCs) on it. First, PLA scaffold was fabricated by electrospinning technique, and then LAP (0.8 wt%) was coated on it. The morphology of the scaffold was examined by scanning electron microscopy (SEM) and Energy-dispersive X-ray (EDX) spectroscopy. The chemical structure of the scaffold was evaluated by ATR-FTIR spectroscopy and its hydrophilicity was tested by measuring the water contact angle. Finally, the biocompatibility of the scaffold and cell viability tested with MTT assay was performed on hBMSCs. The results of scaffold morphology showed a successful coat of LAP 0.8% on the surface of PLA scaffold. Furthermore, the hydrophilicity of PLA scaffold improved after coating with LAP 0.8%. The Biocompatibility of scaffold up to 24 hours and hMSCs viability up to 72 hours after cell culture were confirmed (p≤0.001). Based on the results of this study, it seems that PLA/LAP of 0.8% scaffold can be a promising candidate for bone tissue engineering applications by maintaining biocompatibility and cell viability due to the presence of ions in LAP nanoparticles.

کلیدواژه‌ها [English]

  • Polylactic Acid
  • Laponite
  • Electrospinning
  • Bone tissue engineering
  1.  Agrawal S.K., Sanabria-DeLong N., Tew G.N., Bhatia S.R. 2008. Nanoparticle-reinforced associative network hydrogels. Langmuir, 24(22): 13148-13154.
  2. Ai J., Lotfi bakhshaiesh N., Irani S., Ebrahimi B.S. 2019. Evaluation of adhession and viability of endometrial stem cells-derived osteoblast-like cells cultured on PLGA/HA scaffold. Journal of Developmental Biology, 11(1):1-14. [In Persian]
  3. Aliakbari Ghavimi S., Solati Hashjin M., Ebrahimzadeh M.H. 2011. An introduction on tissue engineering. Iranian Journal of Orthopaedic Surgery, 9(4): 185-190. [In Persian]
  4. Arab‐Ahmadi S., Irani S., Bakhshi H., Atyabi F., Ghalandari B. 2021. Immobilization of carboxymethyl chitosan/laponite on polycaprolactone nanofibers as osteoinductive bone scaffolds. Polymers for Advanced Technologies, 32(2):755-765.
  5. Arya A., Sharma A.L. 2017. Insights into the use of polyethylene oxide in energy storage/conversion devices: a critical review. Journal of Physics D: Applied Physics, 50(44): 443002.
  6. Ataie M., Solouk A., Bagheri F., Seyed Jafari E. 2017. Regeneration of musculoskeletal injuries using mesenchymal stem cells loaded scaffolds. Tehran University Medical Journal TUMS Publications, 75(4): 241-250. [In Persian]
  7. Atrian M., Kharaziha M., Emadi R., Alihosseini F. 2019. Silk-LAPONITE® fibrous membranes for bone tissue engineering. Applied Clay Science, 174: 90-99.
  8. Bagheri Marandi G., Baharloui M. 2012. Synthesis of hydrogel nanocomposites of acrylamide-itaconic acid using laponite and study of crystal violet dye adsorption. Iran. Journal of Polymer Science and Technology, 24(6): 505-514. [In Persian]
  9. Becher T.B., Braga C.B., Bertuzzi D.L., Ramos M.D., Hassan A., Crespilho F.N., Ornelas C. 2019. The structure–property relationship in LAPONITE® materials: from Wigner glasses to strong self-healing hydrogels formed by non-covalent interactions. Soft Matter, 15(6): 1278-1289.
  10.  Castro-Aguirre E., Auras R., Selke S., Rubino M., Marsh T. 2018. Impact of nanoclays on the biodegradation of poly (lactic acid) nanocomposites. Polymers, 10(2): 202.
  11.  Fattahi F.S., Khoddami A., Avinc O. 2019. Poly (lactic acid)(PLA) nanofibers for bone tissue engineering. Journal of Textiles and Polymers, 7 (2): 47-64.
  12.  Gaharwar A.K., Kishore V., Rivera C., Bullock W., Wu C.J., Akkus O., Schmidt G. 2012. Physically crosslinked nanocomposites from silicate‐crosslinked PEO: mechanical properties and osteogenic differentiation of human mesenchymal stem cells. Macromolecular Bioscience, 12(6): 779-793.
  13.  Gao A., Liu F., Xue L., 2014. Preparation and evaluation of heparin-immobilized poly (lactic acid)(PLA) membrane for hemodialysis. Journal of Membrane Science, 452: 390-399.
  14.  Gao Y., Shao W., Qian W., He J., Zhou Y., Qi K., Wang L., Cui S., Wang R. 2018. Biomineralized poly (l-lactic-co-glycolic acid)-tussah silk fibroin nanofiber fabric with hierarchical architecture as a scaffold for bone tissue engineering. Materials Science and Engineering,84: 195-207.
  15.  Ghorbani S., Tirahi T., Soleimani M., Pour Beiranvand S. 2017. Wet electrospinning of 3D nanofiber poly (lactic) acid scaffolds for tissue engineering applications: Fabrication and characterization. Pathobiology Research, 20(2): 49-61. [In Persian]
  16.  Gonçalves M., Figueira P., Maciel D., Rodrigues J., Qu X., Liu C., Tomás H., Li Y. 2014. pH-sensitive Laponite/ doxorubicin/alginate nanohybrids with improved anticancer efficacy. Acta Biomaterialia, 10(1): 300-307.
  17.  Gouma P.I., Ramachandran K. 2008. Electrospinning for bone tissue engineering. Recent patents on nanotechnology, 2(1): 1-7.
  18.  Grémare A., Guduric V., Bareille R., Heroguez V., Latour S., L'heureux N., Fricain J.C., Catros S., Le Nihouannen D., 2018. Characterization of printed PLA scaffolds for bone tissue engineering. Journal of Biomedical Materials Research Part A, 106(4): 887-894.
  19.  Hegazy D.E., Mahmoud G.A. 2014. Radiation synthesis and characterization of polyethylene oxide/chitosan-silver nanocomposite for biomedical applications. Arabian Journal of Nuclear Science and Application, 47(2): 1-14.
  20.  Ilie C., Stinga G., Iovescu A., Purcar V., Anghel D.F., Donescu D. 2010. The influence of nonionic surfactants on the carbopol-peg interpolymer complexes. Revume Roumaine Chimie, 55(7): 409-417.
  21.  Imani S.M., Rabiee S.M., Moazami Goudarzi A., Dardel M. 2017. Investigation of the mechanical properties of the porous scaffolds used in bone tissue engineering by means of micromechanical modeling. Modares Mechanical Engineering, 17(9): 397-408. [In Persian]
  22.  Jalali Jahromi, A., Mirhosseini, M., Molla Hoseini, H., Nikukar, H. 2020. A Review on Commonly Used Scaffolds in Tissue Engineering for Bone Tissue Regeneration. SSU Journals, 28(1): 2235-2254. [In Persian]
  23.  Kao C.T., Lin C.C., Chen Y.W., Yeh C.H., Fang H.Y., Shie M.Y. 2015. Poly (dopamine) coating of 3D printed poly (lactic acid) scaffolds for bone tissue engineering. Materials Science and Engineering: C, 56: 165-173.
  24.  Kerativitayanan P., Tatullo M., Khariton M., Joshi P., Perniconi B., Gaharwar A.K. 2017. Nanoengineered osteoinductive and elastomeric scaffolds for bone tissue engineering. ACS Biomaterials Science and Engineering, 3(4): 590-600.
  25.  Khakestani M., Jafari S.H., Zahedi P., Bagheri R., Hajiaghaee R. 2017. Physical, morphological, and biological studies on PLA/n HA composite nanofibrous webs containing E quisetum arvense herbal extract for bone tissue engineering. Journal of Applied Polymer Science, 134(39): p.45343.
  26.  Kung F.C., Kuo Y.L., Gunduz O., Lin C.C. 2019. Dual RGD-immobilized poly (L-lactic acid) by atmospheric pressure plasma jet for bone tissue engineering. Colloids and Surfaces B: Biointerfaces, 178: 358-364.
  27.  Li H.L., Zhou G.X., Shan Y.K., Yuan M.L. 2013. The Mechanical Properties and Hydrophilicity of Poly (L-Lactide)/Laponite Composite Film. In Advanced Materials Research, 706: 340-343.
  28.  Li T., Liu Z.L., Xiao M., Yang Z.Z., Peng M.Z. Di Li C., Zhou X.J., Wang J.W. 2018. Impact of bone marrow mesenchymal stem cell immunomodulation on the osteogenic effects of laponite. Stem Cell Research and Therapy, 9(1): 100.
  29.  Li X., Wang L., Fan Y., Feng Q., Cui F.Z., Watari F. 2013. Nanostructured scaffolds for bone tissue engineering. Journal of Biomedical Materials Research Part A, 101(8): 2424-2435.
  30.  Loiola L.M., Más B.A., Duek E.A., Felisberti M.I. 2015. Amphiphilic multiblock copolymers of PLLA, PEO and PPO blocks: synthesis, properties and cell affinity. European Polymer Journal, 68: 618-629.
  31.  Miri V., Mansourizadeh F., Sagha M., Asadi A., Golmohammadi M.G. 2015. Fabrication and evaluation of the morphology, biodegradability, and chemical characteristics of the nano-fibrous scaffold poly-l-lactic-acid (plla) and its application in neural tissue engineering. Studies in Medical Sciences, 25(11): 988-997. [In Persian]
  32.  Monnier A., Al Tawil E., Nguyen Q.T., Valleton J.M., Fatyeyeva K., Deschrevel B. 2018. Functionalization of poly (lactic acid) scaffold surface by aminolysis and hyaluronan immobilization: How it affects mesenchymal stem cell proliferation. European Polymer Journal, 107: 202-217.
  33.  Motamedian F.S.T.S.R., Khosraviani F.G.K., Khojasteh A. 2012. Craniomaxillofacial bone engineering by scaffolds loaded with stem cells: a systematic review. Journal Dental School, 30(2):115-131.
  34. Nair B.P., Sindhu M., Nair P.D. 2016. Polycaprolactone-laponite composite scaffold releasing strontium ranelate for bone tissue engineering applications. Colloids and Surfaces B: Biointerfaces, 143: 423-430.
  35.  Oliveira J.E., Moraes E.A., Marconcini J.M., C. Mattoso L.H., Glenn G.M., Medeiros E.S. 2013. Properties of poly (lactic acid) and poly (ethylene oxide) solvent polymer mixtures and nanofibers made by solution blow spinning. Journal of Applied Polymer Science, 129(6): 3672-3681.
  36.  Ordikhani F., Dehghani M., Simchi A. 2015. Antibiotic-loaded chitosan–Laponite films for local drug delivery by titanium implants: cell proliferation and drug release studies. Journal of Materials Science: Materials in Medicine, 26(12): 1-12.
  37.  Peng Q., Xu, P., Xiao S. 2018. Porous Laponite/Poly (L-lactic acid) Membrane with Controlled Release of TCH and Efficient Antibacterial Performance. Fibers and Polymers, 19(3): 477-488.
  38.  Puppi D., Chiellini F., Piras A.M., Chiellini E. 2010. Polymeric materials for bone and cartilage repair. Progress in polymer Science, 35(4): 403-440.
  39.  Ramesh S., Yuen T.F., Shen C.J. 2008. Conductivity and FTIR studies on PEO–LiX [X: CF3SO3−, SO42−] polymer electrolytes. Spectrochimica Acta Part A: Molecular and Biomolecular Spectroscopy, 69(2): 670-675.
  40.  Ranjbar M.M., Shaki H., Kargozar S. 2019. Fabrication of Nanofibrous Hybrid Scaffolds from Polylactic Acid-Graphene and Gelatin for Application in Bone Tissue Engineering. Iranian Journal of Polymer Science and Technology, 31 (6): 566-576. [In Persian]
  41.  Salerno A., Fernández-Gutiérrez M., del Barrio J.S.R., Domingo C. 2015. Bio-safe fabrication of PLA scaffolds for bone tissue engineering by combining phase separation, porogen leaching and scCO2 drying. The Journal of Supercritical Fluids, 97: 238-246.
  42.  Sharifi F.F., Irani S., Zandi M., Soleimani M. 2014. Synthesis and surface modification of polycaprolactone nanofibers for tissue engineering. Journal of Ardabil University of Medical Sciences (Jaums), 14 (3): 217-228. [In Persian]
  43.  Shen R., Xu W., Xue Y., Chen L., Ye H., Zhong E., Ye Z., Gao J., Yan Y. 2018. The use of chitosan/PLA nano-fibers by emulsion eletrospinning for periodontal tissue engineering. Artificial Cells, Nanomedicine and Biotechnology, 46(sup2): 419-430.
  44.  Silva T.N., Gonçalves R.P., Rocha C.L., Archanjo B.S., Barboza C.A.G., Pierre M.B.R., Reynaud F., de Souza Picciani P.H. 2019. Controlling burst effect with PLA/PVA coaxial electrospun scaffolds loaded with BMP-2 for bone guided regeneration. Materials Science and Engineering: C, 97: 602-612.
  45.  Suganya Bharathi B., Stalin T. 2019. Cerium oxide and peppermint oil loaded polyethylene oxide/graphene oxide electrospun nanofibrous mats as antibacterial wound dressings. Materials Today Communications, 21: 100664.
  46.  Tang L., Wei W., Wang X., Qian J., Li J., He A., Yang L., Jiang X., Li X., Wei J. 2018. LAPONITE® nanorods regulating degradability, acidic-alkaline microenvironment, apatite mineralization and MC3T3-E1 cells responses to poly (butylene succinate) based bio-nanocomposite scaffolds. RSC Advances, 8(20):10794-10805.
  47.  Tao L., Zhonglong L., Ming X., Zezheng Y., Zhiyuan L., Xiaojun Z., Jinwu W. 2017. In vitro and in vivo studies of a gelatin/carboxymethyl hitosan/LAPONITE composite scaffold for bone tissue engineering. RSC Advances, 7(85): 54100-54110.
  48.  Tomás H., Alves C.S., Rodrigues J. 2018. Laponite®: A key nanoplatform for biomedical applications?. Nanomedicine: Nanotechnology, Biology and Medicine, 14(7): 2407-2420.
  49.  Wang C., Wang S., Li K., Ju Y., Li J., Zhang Y., Li J., Liu X., Shi X., Zhao Q. 2014. Preparation of laponite bioceramics for potential bone tissue engineering applications. PloS One, 9(6): e99585.
  50.  Wang G., Maciel D., Wu Y., Rodrigues J., Shi X., Yuan Y., Liu C., Tomás H., Li Y. 2014. Amphiphilic polymer-mediated formation of laponite-based nanohybrids with robust stability and pH sensitivity for anticancer drug delivery. ACS Applied Materials and interfaces, 6(19): 16687-16695.
  51.  Wang S., Castro R., An X., Song C., Luo Y., Shen M., Tomá, H., Zhu M., Shi X. 2012. Electrospun laponite-doped poly (lactic-co-glycolic acid) nanofibers for osteogenic differentiation of human mesenchymal stem cells. Journal of Materials Chemistry, 22(44): 23357-23367.
  52.  Wang S., Zheng F., Huang Y., Fang Y., Shen M., Zhu M., Shi X. 2012. Encapsulation of amoxicillin within laponite-doped poly (lactic-co-glycolic acid) nanofibers: preparation, characterization, and antibacterial activity. ACS Applied Materials and Interfaces, 4(11): 6393-6401.
  53.  Wang S.D., Zhang S.Z., Liu H., Zhang Y.Z. 2014. Controlled release of antibiotics encapsulated in the electrospinning polylactide nanofibrous scaffold and their antibacterial and biocompatible properties. Materials Research Express, 1(2): 025406.
  54.  Wang Y., Cui W., Chou J., Wen S., Sun Y., Zhang H. 2018. Electrospun nanosilicates-based organic/inorganic nanofibers for potential bone tissue engineering. Colloids and Surfaces B: Biointerfaces, 172: 90-97.
  55.  Xiong Z.Q., Li X.D., Fu F., Li Y.N. 2019. Performance evaluation of laponite as a mud-making material for drilling fluids. Petroleum Science, 16(4): 890-900.
  56.  Xu W., Shen R., Yan Y., Gao J. 2017. Preparation and characterization of electrospun alginate/PLA nanofibers as tissue engineering material by emulsion eletrospinning. Journal of the Mechanical Behavior of Biomedical Materials, 65: 428-438.
  57.  Xu Y., Zou L., Lu H., Kang T. 2017. Effect of different solvent systems on PHBV/PEO electrospun fibers. RSC Advances, 7(7): 4000-4010.
  58.  Yao Q., Cosme J.G., Xu T., Miszuk J.M., Picciani P.H., Fong H., Sun H. 2017. Three dimensional electrospun PCL/PLA blend nanofibrous scaffolds with significantly improved stem cells osteogenic differentiation and cranial bone formation. Biomaterials, 115: 115-127.
  59.  Zhai X., Hou C., Pan H., Lu W.W., Liu W., Ruan C. 2018. Nanoclay incorporated polyethylene-glycol nanocomposite hydrogels for stimulating in vitro and in vivo osteogenesis. Journal of Biomedical Nanotechnology, 14(4): 662-674.
  60.  Zhou G.X., Yuan M.W., Jiang L., Yuan M.L., Li H.L. 2013. The Preparation and Property Research on Laponite-Poly (L-Lactide) Composite Film. In Advanced Materials Research, 750: 1919-1923.