Effects of different organic solvents on slow-release recombinant human bone morphogenetic protein-2 microcapsules

doi:10.3969/j.issn.2095-4344.2015.21.007

Abstract

Abstract:

BACKGROUND: In literatures, the recombinant human bone morphogenetic protein-2 (rhBMP-2) loaded microcapsules can be fabricated by double emulsion solvent evaporation method with different organic solvents, such as methylene chloride, ethyl acetate or their mixture. But so far it is not determined yet which one is better.
OBJECTIVE: To optimize the preparation method of microcapsules encapsulating rhBMP-2 and to compare the effects of different organic solvents on the microcapsules.
METHODS: Polylactic acid-polyethylene glycol-polylactic acid copolymer as microcapsules was used to prepare rhBMP-2 loaded microcapsules with double emulsion solvent evaporation method. Four kinds of organic solvents, methylene chloride (group A), mixture of methylene chloride and ethyl acetate (group B), ethyl acetate (group C) and acetyl acetone (group D) were chosen as oil phases to compare their effects on microcapsule’s morphology, diameter, and encapsulation efficiency. Passage 3 bone marrow mesenchymal stem cells from rats were
co-cultured with prepared microcapsules for 14 days, and then alkaline phosphatase activity was detected.
RESULTS AND CONCLUSION: Compared with the other organic solvents, dichloromethane could cause microcapsules with the smaller and more uniform shape (4-10 microns) and the highest encapsulation efficiency; the microcapsules prepared by mixture of methylene chloride and ethyl acetate had relatively wide size distribution and moderate encapsulation efficiency; the microcapsules prepared by acetylacetone were difficult to form and keep the bioactivity of rhBMP-2. After cultured with rat bone marrow mesenchymal stem cells for 14 days, the alkaline phosphatase activity in groups A, B and C was significantly higher than that in group D and there was no significant difference between group A and group B; the alkaline phosphatase activity in groups A and B was significantly higher than that in group C (P < 0.05). The results show the rhBMP-2-loaded microcapsules prepared by methylene chloride as organic solvent have good shape, high encapsulation efficiency, and good biological activity.

中国组织工程研究杂志出版内容重点：生物材料；骨生物材料; 口腔生物材料; 纳米材料; 缓释材料; 材料相容性；组织工程

Key words: Methylene Chloride, Bone Morphogenetic Proteins, Capsules

CLC Number:

R318

Li Xia-lin, Yi Wei-hong, Jin An-min, Min Shao-xiong. Effects of different organic solvents on slow-release recombinant human bone morphogenetic protein-2 microcapsules [J]. Chinese Journal of Tissue Engineering Research, 2015, 19(21): 3317-3322.

Figures/Tables 3

References

[1] Lei L,Wang S,Wu H,et al.Optimization of release pattern of FGF-2 and BMP-2 for osteogenic differentiation of low-population density hMSCs.J Biomed Mater Res A. 2015; 103(1):252-261.

[2] Lim SM,Eom HN,Jiang HH,et al.Evaluation of PEGylated exendin-4 released from poly (lactic-co-glycolic acid) microspheres for antidiabetic therapy.J Pharm Sci. 2015; 104(1):72-80.

[3] Simon-Yarza T,Formiga FR,Tamayo E,et al.PEGylated-PLGA microparticles containing VEGF for long term drug delivery.Int J Pharm. 2013;440(1):13-18.

[4] Li X, Min S, Zhao X,et al. Optimization of entrapping conditions to improve the release of BMP-2 from PELA carriers by response surface methodology. Biomed Mater. 2014;10:015002.

[5] Vyslouzil J,Dolezel P,Kejdusova M,et al.Long-term controlled release of PLGA microparticles containing antidepressant mirtazapine. Pharm Dev Technol. 2014:1-8. [Epub ahead of print]

[6] Shah RB, Schwendeman SP. A biomimetic approach to active self-microencapsulation of proteins in PLGA.J Control Release. 2014;196:60-70.

[7] Ogbonna JD,Kenechukwu FC,Nwobi CS,et al.Formulation, in vitro and in vivo evaluation of halofantrine-loaded solid lipid microparticles.Pharm Dev Technol. 2014:1-8.[Epub ahead of print]

[8] 林楚伟,周胜华,杜优优.全骨髓贴壁并差速传代分离纯化大鼠骨髓间充质干细胞:与密度梯度离心法的比较[J].中国组织工程与临床康复,2010,14(14):2508-2512.

[9] McKay WF,Peckham SM,Badura JM.A comprehensive clinical review of recombinant human bone morphogenetic protein-2 (INFUSE Bone Graft). Int Orthop. 2007;31(6):729-734.

[10] Lebl DR.Bone morphogenetic protein in complex cervical spine surgery: A safe biologic adjunct? World J Orthop.2013; 4(2):53-57.

[11] Lad SP,Bagley JH,Karikari IO,et al. Cancer after spinal fusion: the role of bone morphogenetic protein.Neurosurgery. 2013; 73: 440-449.

[12] Calori GM,Donati D,Di Bella C,et al.Bone morphogenetic proteins and tissue engineering: future directions. Injury.2009; 40 Suppl 3:S67-76.

[13] Kempen DH,Kruyt MC, Lu L,et al.Effect of autologous bone marrow stromal cell seeding and bone morphogenetic protein-2 delivery on ectopic bone formation in a microsphere/ poly(propylene fumarate) composite.Tissue Eng Part A. 2009; 15(3):587-594.

[14] Kim SS,Gwak SJ,Kim BS.Orthotopic bone formation by implantation of apatite-coated poly(lactide-co-glycolide)/ hydroxyapatite composite particulates and bone morphogenetic protein-2.J Biomed Mater Res A. 2008; 87(1):245-253.

[15] Zhao J,Wang S,Bao J,et al.Trehalose maintains bioactivity and promotes sustained release of BMP-2 from lyophilized CDHA scaffolds for enhanced osteogenesis in vitro and in vivo. PloS one.2013;8:e54645.

[16] White LJ, Kirby GT,Cox HC,et al.Accelerating protein release from microparticles for regenerative medicine applications. Mater Sci Eng C Mater Biol Appl. 2013;33(5):2578-2583.

[17] Chen L,Lu X,Li S,et al.Sustained delivery of BMP-2 and platelet-rich plasma-released growth factors contributes to osteogenesis of human adipose-derived stem cells. Orthopedics.2012;35:e1402-1409.

[18] Hernandez A,Reyes R,Sanchez E,et al.In vivo osteogenic response to different ratios of BMP-2 and VEGF released from a biodegradable porous system. J Biomed Mater Res A.2012;100(9):2382-2391.

[19] Molavi O,Mahmud A,Hamdy S,et al.Development of a poly(d,l-lactic-co-glycolic acid) nanoparticle formulation of STAT3 inhibitor JSI-124: implication for cancer immunotherapy. Mol Pharm.2010;7(2):364-374.

[20] Zhang L,Lan X,Xu M,et al.Improvement in angiogenesis and osteogenesis with modified cannulated screws combined with VEGF/PLGA/fibrin glue in femoral neck fractures.J Mater Sci Mater Med. 2014;25(4):1165-1172.

[21] Yan J,Yang S,Sun H,et al.Effects of releasing recombinant human growth and differentiation factor-5 from poly(lactic-co-glycolic acid) microspheres for repair of the rat degenerated intervertebral disc.J Biomater Appl. 2013;29(1): 72-80.

[22] Gainza G,Aguirre JJ,Pedraz JL,et al.rhEGF-loaded PLGA-Alginate microspheres enhance the healing of full-thickness excisional wounds in diabetised Wistar rats. Eur J Pharm Sci.2013;50(3-4):243-252.

[23] Zou GK,Song YL,Zhou W,et al.Effects of local delivery of bFGF from PLGA microspheres on osseointegration around implants in diabetic rats.Oral Surg Oral Med Oral Pathol Oral Radiol. 2012;114(3):284-289.

[24] Jelvehgari M, Montazam SH. Comparison of microencapsulation by emulsion-solvent extraction/ evaporation technique using derivatives cellulose and acrylate-methacrylate copolymer as carriers.Jundishapur J Nat Pharm Prod. 2012;7(4):144-152.

[25] Gupta V,Davis M,Hope-Weeks LJ,et al.PLGA microparticles encapsulating prostaglandin E1-hydroxypropyl-beta- cyclodextrin (PGE1-HPbetaCD) complex for the treatment of pulmonary arterial hypertension (PAH). Pharm Res. 2011; 28(7): 1733-1749.

[26] Sheng Y,Yuan Y,Shan X,et al. Study on mass transfer behavior of hemoglobin-based nanocapsule surface. Sheng wu yi xue gong cheng xue za zhi = Journal of biomedical engineering.Sheng Wu Yi Xue Gong Cheng Xue Za Zhi. 2008;25(4):879-884.

[27] Zhao J,Liu CS,Yuan Y,et al.Preparation of hemoglobin-loaded nano-sized particles with porous structure as oxygen carriers. Biomaterials.2007;28:1414-1422.

[28] Kim HK, Park TG. Microencapsulation of human growth hormone within biodegradable polyester microspheres: protein aggregation stability and incomplete release mechanism. Biotechnol Bioeng.1999;65(6):659-667.