Comparison of liposomal formulations incorporating BMP-2 peptide to induce bone tissue engineering

Document Type : Research Paper


1 Department of Pharmaceutics, School of Pharmacy, Mashhad University of Medical Sciences, Mashhad, Iran

2 Pharmaceutical Research Center, Pharmaceutical Technology Institute, Mashhad University of Medical Sciences, Mashhad, Iran

3 Department of Pharmaceutical Biotechnology, School of Pharmacy, Mashhad University of Medical Sciences, Mashhad, Iran

4 Department of Pharmaceutical Nanotechnology, School of Pharmacy, Mashhad University of Medical Sciences, Mashhad, Iran

5 Nanotechnology Research Center, Pharmaceutical Technology Institute, Mashhad University of Medical Sciences, Mashhad, Iran


Objective(s): Fabricating a biomimetic scaffold platform combined with controlled release of bioactive agents is a practical approach for bone tissue engineering. Controlled delivery of peptides and growth factors which play a significant role in osteogenesis is an important issue reducing the associated adverse effects and leading to cost-effectiveness.
Materials and Methods: We developed two liposomal formulations of bone morphogenetic protein-2 (BMP-2) peptide designated as F1 and F2 with controlled release properties. Due to high negative zeta potential of F1 formulation, the surface of the liposomes was decorated with positively charged BMP-2 peptide while the peptide was encapsulated in F2 formulation. Then, we evaluated the hypothesis that whether the electrostatically loaded peptide could act as a ligand and improve the cellular uptake and osteogenic differentiation of mesenchymal stem cells.
Results: Both formulations were less than 100 nm in size. The release study revealed that both formulations showed a sustained release pattern for 21 days. However, the cumulative releases were 60% and 40% in F1 and F2 formulations, respectively. Flow cytometry analysis indicated that cell internalization of F1 liposomes was more than the other formulation. In the next step, F1 and F2 formulations were attached covalently to our previously developed nanofibrous electrospun scaffold and biocompatibility and osteogenic differentiation of each formulation were studied. The results indicated that the proliferation of the cells seeded on F1 liposcaffold was significantly more than F2 liposcaffold at days 1 and 3. Furthermore, F1 liposcaffold showed superior osteogenic differentiation through measurement of alkaline phosphatase activity which could be due to the higher release pattern of F1 liposomes and their improved cellular uptake.
Conclusion: Our findings revealed that controlled release BMP-2 decorated liposomal formulations immobilized on nanofibrous electrospun scaffold platform could be a promising candidate for bone regeneration therapeutics and merits further investigation.


1.Mohammadi M, Alibolandi M, Abnous K, Salmasi Z, Jaafari MR, Ramezani M. Fabrication of hybrid scaffold based on hydroxyapatite-biodegradable nanofibers incorporated with liposomal formulation of BMP-2 peptide for bone tissue engineering. Nanomed-nanotechnol. 2018; 14(7): 1987-1997.
2.Bochicchio B, Barbaro K, De Bonis A, Rau JV, Pepe A. Electrospun poly(d,l-lactide)/gelatin/glass-ceramics tricomponent nanofibrous scaffold for bone tissue engineering. J Biomed Mater Res. 2010; 108(5): 1064-1076.
3.De Witte T-M, Fratila-Apachitei LE, Zadpoor AA, Peppas NA. Bone tissue engineering via growth factor delivery: from scaffolds to complex matrices. Regen Biomater. 2018; 5(4): 197-211.
4.Newman MR, Benoit DSW. Local and targeted drug delivery for bone regeneration. Curr opin biotech. 2016; 40: 125-132.
5. Cui, W Liu, Q Yang, L Wang, K Sun, T Ji, Y Liu, L Yu, W Qu, Y Wang, J Zhao, Z Zhu, J Guo X. Sustained Delivery of BMP-2-Related Peptide from the True Bone Ceramics/Hollow Mesoporous Silica Nanoparticles Scaffold for Bone Tissue Regeneration. Acs biomater-sci eng. 2018; 4(1): 211-221.
6.Bone Regeneration by Controlled Release of Bone Morphogenetic Protein-2: A Rabbit Spinal Fusion Chamber Molecular Study. Tissue Eng Part A. 2019; 25(19-20): 1356-1368.
7. Mohammadi M, Shaegh S A M, Alibolandi M, Ebrahimzadeh M H, Tamayol A, Jaafari M R, Ramezani M. Micro and nanotechnologies for bone regeneration: recent advances and emerging designs. J Control Release. 2018; 274: 35-55.
8.Mont MA, Ragland PS, Biggins B, Friedlaender G, Patel T, Cook S. Use of bone morphogenetic proteins for musculoskeletal applications: an overview. JBJS. 2004; 86 (suppl_2): 41-55.
9.Bouyer M, Guillot R, Lavaud J, Plettinx C, Olivier C, Curry V. Surface delivery of tunable doses of BMP-2 from an adaptable polymeric scaffold induces volumetric bone regeneration. Biomaterials. 2016; 104: 168-181.
10. Lee SS, Hsu EL, Mendoza M, Ghodasra J, Nickoli M S, Ashtekar A, Polavarapu M, Babu J, Riaz R M, Nicolas J D. Gel scaffolds of BMP‐2‐binding peptide amphiphile nanofibers for spinal arthrodesis. Adv Healthc Mater. 2015; 4(1): 131-141.
11.Niu X, Feng Q, Wang M, Guo X, Zheng Q. Porous nano-HA/collagen/PLLA scaffold containing chitosan microspheres for controlled delivery of synthetic peptide derived from BMP-2. J Control Release. 2009; 134(2): 111-117.
12.Moeinzadeh S, Barati D, Sarvestani SK, Karimi T, Jabbari EJTEPA. Experimental and computational investigation of the effect of hydrophobicity on aggregation and osteoinductive potential of BMP-2-derived peptide in a hydrogel matrix. Tissue Eng Part A. 2015; 21(1-2): 134-146.
13. Li J, Hong J, Zheng Q, Guo X, Lan S, Cui F, Pan H, Zou Z, Chen C. Repair of rat cranial bone defects with nHAC/PLLA and BMP-2-related peptide or rhBMP-2. J Orthop Res. 2011; 29(11): 1745-1752.
14.Kirsch T, Sebald W, Dreyer MKJNsb. Crystal structure of the BMP-2–BRIA ectodomain complex. Nat Struct Mol Biol. 2000; 7(6): 492-496.
15. Lee, J Y, Choo, J E, Choi, Y S, Suh, J S, Lee S J, Chung, C P, Park, Y J. Osteoblastic differentiation of human bone marrow stromal cells in self-assembled BMP-2 receptor-binding peptide-amphiphiles. Biomaterials. 2009; 30(21): 3532-3541.
16. Zhou X, Feng W, Qiu K, Chen L, Wang W, Nie W, Mo X, He C. BMP-2 derived peptide and dexamethasone incorporated mesoporous silica nanoparticles for enhanced osteogenic differentiation of bone mesenchymal stem cells. ACS Appl Mater Interfaces. 2015; 7(29): 15777-15789.
17.Jeon O, Song SJ, Yang HS, Bhang S-H, Kang S-W, Sung MA. Long-term delivery enhances in vivo osteogenic efficacy of bone morphogenetic protein-2 compared to short-term delivery. Biochem bioph res co. 2008; 369(2): 774-780.
18.Tan J, Zhang M, Hai Z, Wu C, Lin J, Kuang W. Sustained release of two bioactive factors from supramolecular hydrogel promotes periodontal bone regeneration.
ACS Nano. 2019; 13(5): 5616-5622.
19.Chen L, Shao L, Wang F, Huang Y, Gao FJRa. Enhancement in sustained release of antimicrobial peptide and BMP-2 from degradable three dimensional-printed PLGA scaffold for bone regeneration. RSC Adv. 2019; 9(19): 10494-10507.
20.Jung T, Lee JH, Park S, Kim Y-J, Seo J, Shim H-E. Effect of BMP-2 delivery mode on osteogenic differentiation of stem cells. Stem Cells Int. 2017; 2017: 1-7.