Basic research

Evaluation of JDBM porous scaffold coated with DCPD in promoting angiogenesis and repairing bone defects

  • Qing WANG ,
  • Wei WANG ,
  • Da-jun JIANG ,
  • Wei-tao JIA
Expand
  • Department of Orthopedic Surgery, Shanghai Sixth People′s Hospital, Shanghai Jiao Tong University, Shanghai 200233, China

Online published: 2021-06-29

Supported by

National Natural Science Foundation of China(81572105);Shanghai Municipal Education Commission—Gaofeng Clinical Medicine Grant Support(20172026);Funding Project for Talent Development in Shanghai(2017035)

Abstract

Objective

·To evaluate the biological effects of JDBM (Mg-Nd-Zn-Zr) scaffold coated with DCPD (CaHPO4·2H2O) on angiogenesis and repairing bone defects in vivo and in vitro.

Methods

·The JDBM-DCPD and JDBM-MgF2 scaffolds were constructed by using template replication method and chemical deposition method, and the characteristics of the scaffolds were observed by micro-CT and scan electron microscope. The bone marrow mesenchymal stem cells (BMSCs) were seeded on the scaffolds and the biocompatibility of scaffolds was evaluated by CCK-8 experiment and cell adhesion experiment. Transwell cell migration experiment and tube formation experiment were used to detect the effects of scaffold extracts on the migration and tube-forming ability of endothelial cell line Ea.hy926 cells, and immunofluorescence was used to further observe the secretion of vascular endothelial growth factor (VEGF). Alkaline phosphatase staining and alizarin red staining were used to detect the effect of the extracts on the osteogenic ability of BMSCs. Furthermore, the model of critical bone defect of femoral condyle was constructed in SD rats. The JDBM-DCPD and JDBM-MgF2 scaffolds were implanted into the defects, respectively, and the effects of scaffolds on the osteogenesis and vascularization were assessed by Microfil vascular perfusion, micro-CT scanning, and tissue section staining after 8 weeks of operation.

Results

·The main spherical pore size of JDBM-DCPD scaffold was 400?450 μm and the calcium and phosphorus particles were evenly distributed on the pore wall with the size of 15?25 μm. BMSCs adhered and grew well on the surface of JDBM-DCPD scaffold. Compared with the control group and the JDBM-MgF2 scaffold extract, the JDBM-DCPD scaffold extract could significantly promote the migration, tube formation and VEGF expression of Ea.hy926 cells, and significantly enhance the early and late osteogenic differentiation of BMSCs in vitro. After 8 weeks of implantation, the JDBM-DCPD scaffold dramatically facilitated the regeneration of new bone and new vessels in the defect area compared with the JDBM-MgF2 scaffold.

Conclusion

·The JDBM-DCPD scaffold exhibits excellent vascularization effects both in vivo and in vitro, especially early vascularization effect after implantation and bone regeneration promotion in vivo.

Cite this article

Qing WANG , Wei WANG , Da-jun JIANG , Wei-tao JIA . Evaluation of JDBM porous scaffold coated with DCPD in promoting angiogenesis and repairing bone defects[J]. Journal of Shanghai Jiao Tong University (Medical Science), 2021 , 41(6) : 732 -740 . DOI: 10.3969/j.issn.1674-8115.2021.06.005

References

1 Giannoudis PV, Dinopoulos H, Tsiridis E. Bone substitutes: an update[J]. Injury, 2005, 36(): S20-S27.
2 Moore WR, Graves SE, Bain GI. Synthetic bone graft substitutes[J]. ANZ J Surg, 2001, 71(6): 354-361.
3 Samartzis D, Shen FH, Goldberg EJ, et al. Is autograft the gold standard in achieving radiographic fusion in one-level anterior cervical discectomy and fusion with rigid anterior plate fixation?[J]. Spine, 2005, 30(15): 1756-1761.
4 Jin L, Li P, Wang YC, et al. Studies of superb microvascular imaging and contrast-enhanced ultrasonography in the evaluation of vascularization in early bone regeneration[J]. J Ultrasound Med, 2019, 38(11): 2963-2971.
5 Brandi ML, Collin-Osdoby P. Vascular biology and the skeleton[J]. J Bone Miner Res, 2006, 21(2): 183-192.
6 Parfitt AM. The mechanism of coupling: a role for the vasculature[J]. Bone, 2000, 26(4): 319-323.
7 Novosel EC, Kleinhans C, Kluger PJ. Vascularization is the key challenge in tissue engineering[J]. Adv Drug Deliv Rev, 2011, 63(4/5): 300-311.
8 Rouwkema J, Rivron NC, van Blitterswijk CA. Vascularization in tissue engineering[J]. Trends Biotechnol, 2008, 26(8): 434-441.
9 Kanczler JM, Oreffo RO. Osteogenesis and angiogenesis: the potential for engineering bone[J]. Eur Cell Mater, 2008, 15: 100-114.
10 Thevenot P, Nair A, Dey J, et al. Method to analyze three-dimensional cell distribution and infiltration in degradable scaffolds[J]. Tissue Eng Part C Methods, 2008, 14(4): 319-331.
11 Staiger MP, Pietak AM, Huadmai J, et al. Magnesium and its alloys as orthopedic biomaterials: a review[J]. Biomaterials, 2006, 27(9): 1728-1734.
12 Velasco MA, Narváez-Tovar CA, Garzón-Alvarado DA. Design, materials, and mechanobiology of biodegradable scaffolds for bone tissue engineering[J]. Biomed Res Int, 2015, 2015: 729076.
13 Kunjukunju S, Roy A, Ramanathan M, et al. A layer-by-layer approach to natural polymer-derived bioactive coatings on magnesium alloys[J]. Acta Biomater, 2013, 9(10): 8690-8703.
14 Zhang J, Ma X, Lin D, et al. Magnesium modification of a calcium phosphate cement alters bone marrow stromal cell behavior via an integrin-mediated mechanism[J]. Biomaterials, 2015, 53: 251-264.
15 Qin H, Zhao Y, An Z, et al. Enhanced antibacterial properties, biocompatibility, and corrosion resistance of degradable Mg-Nd-Zn-Zr alloy[J]. Biomaterials, 2015, 53: 211-220.
16 Kong X, Wang L, Li G, et al. Mg-based bone implants show promising osteoinductivity and controllable degradation: a long-term study in a goat femoral condyle fracture model[J]. Mater Sci Eng C Mater Biol Appl, 2018, 86: 42-47.
17 Guan X, Xiong M, Zeng F, et al. Enhancement of osteogenesis and biodegradation control by brushite coating on Mg-Nd-Zn-Zr alloy for mandibular bone repair[J]. ACS Appl Mater Interfaces, 2014, 6(23): 21525-21533.
18 Tamimi F, Sheikh Z, Barralet J. Dicalcium phosphate cements: brushite and monetite[J]. Acta Biomater, 2012, 8(2): 474-487.
19 Apelt D, Theiss F, El-Warrak AO, et al. In vivo behavior of three different injectable hydraulic calcium phosphate cements[J]. Biomaterials, 2004, 25(7): 1439-1451.
20 Malhotra A, Habibovic P. Calcium phosphates and angiogenesis: implications and advances for bone regeneration[J]. Trends Biotechnol, 2016, 34(12): 983-992.
21 Wang W, Jia G, Wang Q, et al. The in vitro and in vivo biological effects and osteogenic activity of novel biodegradable porous Mg alloy scaffolds[J]. Mater Des, 2020, 189: 108514.
22 Pijuan J, Barceló C, Moreno DF, et al. In vitro cell migration, invasion, and adhesion assays: from cell imaging to data analysis[J]. Front Cell Dev Biol, 2019, 7: 107.
23 Hu H, Chen Y, Zou Z, et al. Panax notoginseng saponins prevent bone loss by promoting angiogenesis in an osteoporotic mouse model[J]. Biomed Res Int, 2020, 2020: 8412468.
24 Dai C, Guo H, Lu J, et al. Osteogenic evaluation of calcium/magnesium-doped mesoporous silica scaffold with incorporation of rhBMP-2 by synchrotron radiation-based μCT[J]. Biomaterials, 2011, 32(33): 8506-8517.
25 Griffith LG. Emerging design principles in biomaterials and scaffolds for tissue engineering[J]. Ann N Y Acad Sci, 2002, 961: 83-95.
26 Karageorgiou V, Kaplan D. Porosity of 3D biomaterial scaffolds and osteogenesis[J]. Biomaterials, 2005, 26(27): 5474-5491.
27 Chiesa R, Sandrini E, Santin M, et al. Osteointegration of titanium and its alloys by anodic spark deposition and other electrochemical techniques: a review[J]. J Appl Biomater Biomech, 2003, 1(2): 91-107.
28 Liang C, Wang H, Yang J, et al. Femtosecond laser-induced micropattern and Ca/P deposition on Ti implant surface and its acceleration on early osseointegration[J]. ACS Appl Mater Interfaces, 2013, 5(16): 8179-8186.
29 Ma H, Luo J, Sun Z, et al. 3D printing of biomaterials with mussel-inspired nanostructures for tumor therapy and tissue regeneration[J]. Biomaterials, 2016, 111: 138-148.
30 Xu F, Ding H, Song F, et al. Effects of preparation methods on the bone formation potential of apatite-coated chitosan microspheres[J]. J Biomater Sci Polym Ed, 2014, 25(18): 2080-2093.
31 Yu W, Zhao H, Ding Z, et al. In vitro and in vivo evaluation of MgF2 coated AZ31 magnesium alloy porous scaffolds for bone regeneration[J]. Colloids Surf B Biointerfaces, 2017, 149: 330-340.
32 Lafage-Proust MH, Prisby R, Roche B, et al. Bone vascularization and remodeling[J]. Joint Bone Spine, 2010, 77(6): 521-524.
33 Kusumbe AP, Ramasamy SK, Adams RH. Coupling of angiogenesis and osteogenesis by a specific vessel subtype in bone[J]. Nature, 2014, 507(7492): 323-328.
34 Dhandapani R, Krishnan PD, Zennifer A, et al. Additive manufacturing of biodegradable porous orthopaedic screw[J]. Bioact Mater, 2020, 5(3): 458-467.
35 Xie H, Cui Z, Wang L, et al. PDGF-BB secreted by preosteoclasts induces angiogenesis during coupling with osteogenesis[J]. Nat Med, 2014, 20(11): 1270-1278.
36 Liu Q, Zhou YF, Li ZB. PDGF?BB promotes the differentiation and proliferation of MC3T3?E1 cells through the Src/JAK2 signaling pathway[J]. Mol Med Rep, 2018, 18(4): 3719-3726.
37 Liu W, Guo S, Tang Z, et al. Magnesium promotes bone formation and angiogenesis by enhancing MC3T3-E1 secretion of PDGF-BB[J]. Biochem Biophys Res Commun, 2020, 528(4): 664-670.
38 Saghiri MA, Asatourian A, Orangi J, et al. Functional role of inorganic trace elements in angiogenesis—part Ⅰ: N, Fe, Se, P, Au, and Ca[J]. Crit Rev Oncol Hematol, 2015, 96(1): 129-142.
39 Song GL, Song SZ. A possible biodegradable magnesium implant material[J]. Adv Eng Mater, 2007, 9(4): 298-302.
40 Gu XN, Zheng YF, Chen LJ. Influence of artificial biological fluid composition on the biocorrosion of potential orthopedic Mg-Ca, Az31, Az91 alloys[J]. Biomed Mater, 2009, 4(6): 065011.
41 Fischer J, Prosenc MH, Wolff M, et al. Interference of magnesium corrosion with tetrazolium-based cytotoxicity assays[J]. Acta Biomater, 2010, 6(5): 1813-1823.
Outlines

/