Preparation and osteogenic properties of poly (<i>L</i>-lactic acid)/lecithin porous scaffolds with open pore structure _Chinese Journal of Reparative and Reconstructive Surgery

Authors：

CHEN Si ^1,2,3 ,  DU Chang ^1,2,3

1. Department of Biomedical Engineering, School of Materials Science and Engineering, South China University of Technology, Guangzhou Guangdong, 510641, P.R.China;
2. National Engineering Research Center for Tissue Restoration and Reconstruction, South China University of Technology, Guangzhou Guangdong, 510006, P.R.China;
3. Key Laboratory of Biomedical Materials and Engineering of the Ministry of Education, South China University of Technology, Guangzhou Guangdong, 510006, P.R.China;

Corresponding?author：

DU Chang, Email: duchang@scut.edu.cn

Keywords：

Thermally induced phase separation; poly (L-lactic acid); lecithin; porous scaffold; bone tissue engineering

DOI：

10.7507/1002-1892.201804127

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ObjectiveTo investigate the preparation and osteogenic properties of poly (L-lactic acid)(PLLA)/lecithin porous scaffolds with open pore structure.MethodsPLLA/lecithin porous scaffolds with different lecithin contents (0, 5%, 10%, 20%, 30%, 40%, 50%) were prepared by thermally induced phase separation (groups A, B, C, D, E, F, and G, respectively). Scanning electron microscopy (SEM) was used to observe the surface morphology of the scaffolds. Wide-angle X-ray diffraction (XRD) and differential scanning calorimetry (DSC) were used to detect the crystallinity of the scaffolds. The water uptake ability of the scaffolds was measured. The cell growth and viability of bone marrow mesenchymal stem cells (BMSCs) of mouse on each scaffold was assessed by cell counting kit 8 (CCK-8) method. The osteogenic differentiation ability of BMSCs on each scaffold was evaluated by alkaline phosphatase (ALP) activity. Finally, a critical-size rat calvarial bone defect model was used to evaluate the osteogenesis of the scaffolds in vivo. Micro-CT was used to reconstruct the three-dimensional model of the defect area, and the bone volume and bone mineral density were quantitatively analyzed.ResultsSEM results showed that the lecithin could slightly reduce the pore size; when lecithin content was 50%, platelet-like structure could be observed on the scaffolds. Wide angle XRD and DSC showed that the crystallinity of scaffolds gradually decreased with the increase of lecithin content. The water uptake ability test showed that the hydrophilicity of scaffolds increased with the increase of lecithin content. CCK-8 assay showed that cell activity gradually increased with the increase of culture time. After 7 days of culture, the absorbance (A) value of groups C, D, E, and F were significantly higher than that of groups A, B, and G (P<0.05), but no significant difference was found among groups C, D, E, and F (P>0.05). After 14 days of osteogenic induction, with the increase of lecithin content, there was a significant difference in ALP activity of each group. The ALP activity in groups D, E, F, and G were significantly higher than that in groups A, B, and C (P<0.05).In vivo, the results of Micro-CT examination and bone volume and bone mineral density showed that the scaffolds with 30% lecithin had the best repairing effect.ConclusionPrepared by thermally induced phase separation, the cytocompatibility, osteogenic differentiation, and bone repair ability of the PLLA/lecithin porous scaffold is obviously better than that of pure PLLA scaffold. PLLA/lecithin porous scaffold with suitable lecithin content is a promising scaffold material for bone tissue engineering.

Citation： CHEN Si, DU Chang. Preparation and osteogenic properties of poly (L-lactic acid)/lecithin porous scaffolds with open pore structure . Chinese Journal of Reparative and Reconstructive Surgery, 2018, 32(9): 1123-1130. doi: 10.7507/1002-1892.201804127 Copy

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24.	Shao JD, Chen C, Wang YJ, et al. Early stage structural evolution of PLLA porous scaffolds in thermally induced phase separation process and the corresponding biodegradability and biological property. Polym Degrad Stabil, 2012, 97(6): 955-963.
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27.	Dalby MJ, Childs S, Riehle MO, et al. Fibroblast reaction to island topography: changes in cytoskeleton and morphology with time. Biomaterials, 2003, 24(6): 927-935.

1. Lee EJ, Kasper FK, Mikos AG. Biomaterials for tissue engineering. Ann Biomed Eng, 2014, 42(2): 323-337.
2. Polo-Corrales L, Latorre-Esteves M, Ramirez-Vick JE. Scaffold design for bone regeneration. J Nanosci Nanotechnol, 2014, 14(1): 15-56.
3. Han DK, Hubbell JA. Synthesis of polymer network scaffolds from L-lactide and poly (ethylene glycol) and their interaction with cells. Macromolecules, 1997, 30(20): 6077-6083.
4. Cook AD, Pajvani UB, Hrkach JS, et al. Colorimetric analysis of surface reactive amino groups on poly (lactic acid-co-lysine): poly (lactic acid) blends. Biomaterials, 1997, 18(21): 1417-1424.
5. Wang T, Ji X, Jin L, et al. Fabrication and characterization of heparin-grafted poly-L-lactic acid-chitosan core-shell nanofibers scaffold for vascular gasket. ACS Appl Mater Interfaces, 2013, 5(9): 3757-3763.
6. Ngiam M, Liao S, Patil AJ, et al. Fabrication of mineralized polymeric nanofibrous composites for bone graft materials. Tissue Eng Part A, 2009, 15(3): 535-546.
7. Yang J, Shi G, Bei J, et al. Fabrication and surface modification of macroporous poly (L-lactic acid) and poly (L-lactic-co-glycolic acid) (70/30) cell scaffolds for human skin fibroblast cell culture. J Biomed Mater Res, 2002, 62(3): 438-446.
8. 徐忠華, 吳清玉, 崔福齋. 左旋聚乳酸/卵磷脂共混物膜的血液相容性研究. 中國生物醫學工程學報, 2009, 28(4): 628-631.
9. McKee MG, Layman JM, Cashion MP, et al. Phospholipid nonwoven electrospun membranes. Science, 2006, 311(5759): 353-355.
10. 施雪濤. 骨修復藥物控釋微球支架的多級構建及干細胞介導分化研究. 廣州: 華南理工大學, 2010.
11. Xu ZH, Wu QY. Effect of lecithin content blend with poly (L-lactic acid) on viability and proliferation of mesenchymal stem cells. Mater Sci Eng C, 2009, 29(5): 1593-1598.
12. Park SA, Park KE, Kim W. Preparation of sodium alginate/poly (ethylene oxide) blend nanofibers with lecithin. Macromol Res, 2010, 18(9): 891-896.
13. Zhang M, Wang K, Wang Z, et al. Small-diameter tissue engineered vascular graft made of electrospun PCL/lecithin blend. J Mater Sci Mater Med, 2012, 23(11): 2639-2648.
14. 劉道志, 奚廷斐. 微創介入醫療器械與材料產業的現狀和發展趨勢. 中國醫療器械信息, 2006, 12(12): 1-14.
15. Zhu N, Cui FZ, Hu K, et al. Biomedical modification of poly(L-lactide) by blending with lecithin. J Biomed Mater Res A, 2007, 82(2): 455-461.
16. Wang Y, Cui FZ, Jiao YP, et al. Modification of bone graft by blending with lecithin to improve hydrophilicity and biocompatibility. Biomed Mater, 2008, 3(1): 015012.
17. Miyata T, Masuko T. Crystallization behaviour of poly (L-lactide). Polymer, 1998, 39(22): 5515-5521.
18. Sosnowski S. Poly (L-lactide) microspheres with controlled crystallinity. Polymer, 2001, 42(2): 637-643.
19. Shao J D, Chen S, Du C. Citric acid modification of PLLA nano-fibrous scaffolds to enhance cellular adhesion, proliferation and osteogenic differentiation. Journal Of Materials Chemistry B, 2015, 3(26): 5291-5299.
20. Pan PJ, Zhu B, Kai WH, et al. Polymorphic transition in disordered poly(L-lactide) crystals induced by annealing at elevated temperatures. Macromolecules, 2008, 41(12): 4296-4304.
21. Tsuruga E, Takita H, Itoh H, et al. Pore size of porous hydroxyapatite as the cell-substratum controls BMP-induced osteogenesis. J Biochem, 1997, 121(2): 317-324.
22. Burg KJ, Porter S, Kellam JF. Biomaterial developments for bone tissue engineering. Biomaterials, 2000, 21(23): 2347-2359.
23. He LM, Zhang YQ, Zeng X, et al. Ramakrishna, Fabrication and characterization of poly (L-lactic acid) 3D nanofibrous scaffolds with controlled architecture by liquid-liquid phase separation from a ternary polymer-solvent system. Polymer, 2009, 50(16): 4128-4138.
24. Shao JD, Chen C, Wang YJ, et al. Early stage structural evolution of PLLA porous scaffolds in thermally induced phase separation process and the corresponding biodegradability and biological property. Polym Degrad Stabil, 2012, 97(6): 955-963.
25. Woo KM, Chen VJ, Ma PX. Nano-fibrous scaffolding architecture selectively enhances protein adsorption contributing to cell attachment. J Biomed Mater Res A, 2003, 67(2): 531-537.
26. Groth T, Altankov G, Klosz K. Adhesion of human peripheral blood lymphocytes is dependent on surface wettability and protein preadsorption. Biomaterials, 1994, 15(6): 423-428.
27. Dalby MJ, Childs S, Riehle MO, et al. Fibroblast reaction to island topography: changes in cytoskeleton and morphology with time. Biomaterials, 2003, 24(6): 927-935.

Chinese Journal of Reparative and Reconstructive Surgery

Preparation and osteogenic properties of poly (L-lactic acid)/lecithin porous scaffolds with open pore structure

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