Bone morphogenetic protein sustained-release microspheres combined with   platelet-rich gel contribute to the proliferation and differentiation of bone marrow stromal stem cells

doi:10.3969/j.issn.2095-4344.2016.47.009

Abstract

Abstract:

BACKGROUND: Various growth factors together participate in the different stages of bone healing, but the current tissue-engineered materials limited on single type and concentration cannot obtain satisfactory outcomes. There is still a lack of study on the novel tissue-engineered material releasing two or more growth factors in order.
OBJECTIVE: To observe the effect of platelet-rich gel (PRG) combined with poly(lactide-glycolide acid) (PLGA) microspheres encapsulating bone morphogenetic protein-2 (BMP-2) on the proliferation and osteoblastic differentiation of bone marrow stromal stem cells.
METHODS: PLGA microspheres loading BMP-2 were prepared by multiple emulsion method, and the encapsulation rate, morphology and volume of microspheres as well as in vitro releasing rate of BMP-2 were measured. The eligible microspheres combined with the activated PRG were co-cultured with human bone marrow stromal stem cells on the PLGA three-dimensional scaffold. Additionally, bone marrow stromal stem cells were divided into six groups including platelet poor gel, PRG, BMP-2/platelet poor gel, BMP-2/PRG, BMP-2 microspheres/platelet poor gel and BMP-2 microspheres/PRG groups, and the preprocessed PLGA scaffold was added into each group. The DNA content, alkaline phosphatase activity and mRNA expression levels of osteopontin and osteocalcin were observed.
RESULTS AND CONCLUSION: The DNA content of bone marrow stromal stem cells was higher in all PRG scaffolds compared with the platelet poor gel scaffolds. The alkaline phosphatase activity and mRNA expression levels of osteopontin and osteocalcin in the BMP-2 microspheres/platelet poor gel and BMP-2 microspheres/PRG groups were markedly higher than those in the other four groups. These results suggest that the BMP-2 sustained-release microspheres combined with PRG are able to enhance the proliferation and differentiation of bone marrow stromal stem cells.

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: Platelet-Rich Plasma, Bone Morphogenetic Proteins, Tissue Engineering

CLC Number:

R318

Lu Xiao-lang, Zheng Yi-jing, Cheng Tao, Ye Chao, Hong Jian-jun. Bone morphogenetic protein sustained-release microspheres combined with platelet-rich gel contribute to the proliferation and differentiation of bone marrow stromal stem cells[J]. Chinese Journal of Tissue Engineering Research, 2016, 20(47): 7057-7063.

Figures/Tables 5

References

[1]Yang LJ, Jin Y. Immunohistochemical observations on bone morphogenetic protein in normal and abnormal conditions. Clin Orthop Relat Res. 1990;(257): 249-256.
[2]Kawamura M, Urist MR. Human fibrin is a physiologic delivery system for bone morphogenetic protein. Clin Orthop Relat Res. 1988;(235):302-310.
[3]Yuan T, Zhang C, Zeng B. Treatment of chronic femoral osteomyelitis with platelet-rich plasma (PRP): a case report. Transfus Apher Sci. 2008;38(2):167-173.
[4]Pietramaggiori G, Scherer SS, Mathews JC, et al. Healing modulation induced by freeze-dried platelet-rich plasma and micronized allogenic dermis in a diabetic wound model. Wound Repair Regen. 2008; 16(2):218-225.
[5]Anitua E, Andía I, Sanchez M, et al. Autologous preparations rich in growth factors promote proliferation and induce VEGF and HGF production by human tendon cells in culture. J Orthop Res. 2005;23(2):281-286.
[6]Sun W, Lin H, Xie H, et al. Collagen membranes loaded with collagen-binding human PDGF-BB accelerate wound healing in a rabbit dermal ischemic ulcer model. Growth Factors. 2007;25(5):309-318.
[7]Kim SJ, Kim SY, Kwon CH, et al. Differential effect of FGF and PDGF on cell proliferation and migration in osteoblastic cells. Growth Factors. 2007;25(2):77-86.
[8]Basmanav FB, Kose GT, Hasirci V. Sequential growth factor delivery from complexed microspheres for bone tissue engineering. Biomaterials. 2008;29(31):4195-4204.
[9]Chen FM, Zhang M, Wu ZF. Toward delivery of multiple growth factors in tissue engineering. Biomaterials. 2010;31(24):6279-6308.
[10]Jaklenec A, Hinckfuss A, Bilgen B, et al. Sequential release of bioactive IGF-I and TGF-beta 1 from PLGA microsphere-based scaffolds. Biomaterials. 2008; 29(10):1518-1525.
[11]Holland TA, Bodde EW, Cuijpers VM, et al. Degradable hydrogel scaffolds for in vivo delivery of single and dual growth factors in cartilage repair. Osteoarthritis Cartilage. 2007;15(2):187-197.
[12]Yamada Y, Ueda M, Naiki T, et al. Autogenous injectable bone for regeneration with mesenchymal stem cells and platelet-rich plasma: tissue-engineered bone regeneration. Tissue Eng. 2004;10(5-6):955-964.
[13]Sarkar MR, Augat P, Shefelbine SJ, et al. Bone formation in a long bone defect model using a platelet-rich plasma-loaded collagen scaffold. Biomaterials. 2006;27(9):1817-1823.
[14]Weibrich G, Hansen T, Kleis W, et al. Effect of platelet concentration in platelet-rich plasma on peri-implant bone regeneration. Bone. 2004;34(4):665-671.
[15]Everts PA, Knape JT, Weibrich G, et al. Platelet-rich plasma and platelet gel: a review. J Extra Corpor Technol. 2006;38(2):174-187.
[16]Gruber R, Karreth F, Kandler B, et al. Platelet-released supernatants increase migration and proliferation, and decrease osteogenic differentiation of bone marrow-derived mesenchymal progenitor cells under in vitro conditions. Platelets. 2004;15(1):29-35.
[17]Bouletreau PJ, Warren SM, Spector JA, et al. Factors in the fracture microenvironment induce primary osteoblast angiogenic cytokine production. Plast Reconstr Surg. 2002;110(1):139-148.
[18]Carano RA, Filvaroff EH. Angiogenesis and bone repair. Drug Discov Today. 2003;8(21):980-989.
[19]Schallmoser K, Strunk D. Preparation of pooled human platelet lysate (pHPL) as an efficient supplement for animal serum-free human stem cell cultures. J Vis Exp. 2009;(32):pii: 1523.
[20]Mishra A, Tummala P, King A, et al. Buffered platelet-rich plasma enhances mesenchymal stem cell proliferation and chondrogenic differentiation. Tissue Eng Part C Methods. 2009;15(3):431-435.
[21]Drengk A, Zapf A, Stürmer EK, et al. Influence of platelet-rich plasma on chondrogenic differentiation and proliferation of chondrocytes and mesenchymal stem cells. Cells Tissues Organs. 2009;189(5):317-326.
[22]Calori GM, Tagliabue L, Gala L, et al. Application of rhBMP-7 and platelet-rich plasma in the treatment of long bone non-unions: a prospective randomised clinical study on 120 patients. Injury. 2008;39(12): 1391-1402.
[23]Jung RE, Schmoekel HG, Zwahlen R, et al. Platelet-rich plasma and fibrin as delivery systems for recombinant human bone morphogenetic protein-2. Clin Oral Implants Res. 2005;16(6):676-682.
[24]Tomoyasu A, Higashio K, Kanomata K, et al. Platelet-rich plasma stimulates osteoblastic differentiation in the presence of BMPs. Biochem Biophys Res Commun. 2007;361(1):62-67.
[25]Zhu SJ, Choi BH, Jung JH, et al. A comparative histologic analysis of tissue-engineered bone using platelet-rich plasma and platelet-enriched fibrin glue. Oral Surg Oral Med Oral Pathol Oral Radiol Endod. 2006;102(2):175-179.