IN VITRO STUDY ON INJECTABLE ALGINATE-STRONTIUM HYDROGEL FOR BONE TISSUE ENGINEERING_Chinese Journal of Reparative and Reconstructive Surgery

Authors：

TU Yiji ¹ , WU Tianlong ¹ , YE Aifang ¹ , XU Junhuai ¹ , GUO Fei ¹ , CHENG Xigao ²

1Medical School, Nanchang University, Nanchang Jiangxi, 330000, P.R.China;;
2No.1 Department of Orthopedics, the Second Affiliated Hospital of Nanchang University. Corresponding author: CHENG Xigao, E-mail: cxgyaf1004@sina.com;

Corresponding author：

Keywords：

Bone tissue engineering; Alginate-strontium hydrogel; Alginate-calcium hydrogel; Scaffold material

DOI：

10.7507/1002-1892.20130329

Video：

Export PDF Favorites Scan Get Citation

Abstract Full text Figures/Tables Video References Cited by

Objective To investigate the application potential of alginate-strontium (Sr) hydrogel as an injectable scaffold material in bone tissue engineering. Methods The alginate-Sr/-calcium (Ca) hydrogel beads were fabricated by adding 2.0wt% alginate sodium to 0.2 mol/L SrCl2/CaCl2 solution dropwise. Microstructure, modulus of compression, swelling rate, and degradability of alginate-Sr/-Ca hydrogels were tested. Bone marrow mesenchymal stem cells (BMSCs) were isolated from femoral bones of rabbits by flushing of marrow cavity. BMSCs at passage 5 were seeded onto the alginate-Sr hydrogel (experimental group) and alginate-Ca hydrogel (control group), and the viability and proliferation of BMSCs in 2 alginate hydrogels were assessed. The osteogenic differentiation of cells embeded in 2 alginate hydrogels was evaluated by alkaline phosphate (ALP) activity, osteoblast specific gene [Osterix (OSX), collagen type I, and Runx2] expression level and calcium deposition by fluorescent quantitative RT-PCR and alizarin red staining, Von Kossa staining. The BMSCs which were embeded in alginate-Ca hydrogel and cultured with common growth medium were harvested as blank control group. Results The micromorphology of alginate-Sr hydrogel was similar to that of the alginate-Ca hydrogel, with homogeneous pore structure; the modulus of compression of alginate-Sr hydrogel and alginate-Ca hydrogel was (186.53 ± 8.37) and (152.14 ± 7.45) kPa respectively, showing significant difference (t=6.853, P=0.002); there was no significant difference (t=0.737, P=0.502) in swelling rate between alginate-Sr hydrogel (14.32% ± 1.53%) and alginate-Ca hydrogel (15.25% ± 1.64%). The degradabilities of 2 alginate hydrogels were good; the degradation rate of alginate-Sr hydrogel was significantly lower than that of alginate-Ca hydrogel on the 20th, 25th, and 30th days (P lt; 0.05). At 1-4 days, the morphology of cells on 2 alginate hydrogels was spherical and then the shape was spindle or stellate. When three-dimensional cultured for 21 days, the DNA content of BMSCs in experimental group [(4.38 ± 0.24) g] was significantly higher than that in control group [(3.25 ± 0.21) g ] (t=8.108, P=0.001). On the 12th day after osteogenic differentiation, the ALP activity in experimental group was (15.28 ± 1.26) U/L, which was significantly higher than that in control group [(12.07 ± 1.12) U/L] (P lt; 0.05). Likewise, the mRNA expressions of OSX, collagen type I, and Runx2 in experimental group were significantly higher than those in control group (P lt; 0.05). On the 21th day after osteogenic differentiation, alizarin red staining and Von Kossa staining showed calcium deposition in 2 groups; the calcium nodules and phosphate deposition in experimental group were significantly higher than those in control group (P lt; 0.05). Conclusion Alginate-Sr hydrogel has good physicochemical properties and can promote the proliferation and osteogenic differentiation of BMSCs, so it is an excellent injectable scaffold material for bone tissue engineering.

Citation： TU Yiji,WU Tianlong,YE Aifang,XU Junhuai,GUO Fei,CHENG Xigao. IN VITRO STUDY ON INJECTABLE ALGINATE-STRONTIUM HYDROGEL FOR BONE TISSUE ENGINEERING. Chinese Journal of Reparative and Reconstructive Surgery, 2013, 27(12): 1499-1505. doi: 10.7507/1002-1892.20130329 Copy

1.	Scott TG, Blackburn G, Ashley M, et al. Advances in bionanomaterials for bone tissue engineering. J Nanosci Nanotechnol, 2013, 13(1): 1-22.
2.	Jakob F, Ebert R, Ignatius A, et al. Bone tissue engineering in osteoporosis. Maturitas, 2013, 75(2): 118-124.
3.	Honda H, Tamai N, Naka N, et al. Bone tissue engineering with bone marrow-derived stromal cells integrated with concentrated growth factor in Rattus norvegicus calvaria defect model. J Artif Organs, 2013. [Epub ahead of print].
4.	参考文献.
5.	Haidar ZS, Hamdy RC, Tabrizian M. Delivery of recombinant bone morphogenetic proteins for bone regeneration and repair. Part B: Delivery systems for BMPs in orthopaedic and craniofacial tissue engineering. Biotechnol Lett, 2009, 31(12): 1825-1835.
6.	Drury JL, Boontheekul T, Mooney DJ. Cellular cross-linking of peptide modified hydrogels. J Biomech Eng, 2005, 127(2): 220-228.
7.	Ohsumi H, Hirata H, Nagakura T, et al. Enhancement of perineurial repair and inhibition of nerve adhesion by viscous injectable pure alginate sol. Plast Reconstr Surg, 2005, 116(3): 823-830.
8.	Singh B, Sharma DK, Gupta A. The controlled and sustained release of a fungicide from starch and alginate beads. J Environ Sci Health B, 2009, 44(2): 113-122.
9.	Poldervaart MT, Wang H, van der Stok J, et al. Sustained release of BMP-2 in bioprinted alginate for osteogenicity in mice and rats. PLoS One, 2013, 8(8): e72610.
10.	Reyes R, Delgado A, Sánchez E, et al. Repair of an osteochondral defect by sustained delivery of BMP-2 or TGFβ1 from a bilayered alginate-PLGA scaffold. J Tissue Eng Regen Med, 2012. [Epub ahead of print].
11.	Park Y, Sugimoto M, Watrin A, et al. BMP-2 induces the expression of chondrocyte-specific genes in bovine synovium-derived progenitor cells cultured in three-dimensional alginate hydrogel. Osteoarthritis Cartilage, 2005, 13(6): 527-536.
12.	Pors Nielsen S. The biological role of strontium. Bone, 2004, 35(3): 583-588.
13.	Augst AD, Kong HJ, Mooney DJ. Alginate hydrogels as biomaterials. Macromol Biosci, 2006, 6(8): 623-633.
14.	Cowan CM, Soo C, Ting K, et al. Evolving concepts in bone tissue engineering. Curr Top Dev Biol, 2005, 66: 239-285.
15.	Paige KT, Cima LG, Yaremchuk MJ, et al. De novo cartilage generation using calcium alginate-chondrocyte constructs. Plast Reconstr Surg, 1996, 97(1): 168-180.
16.	Xia Y, Mei F, Duan Y, et al. Bone tissue engineering using bone marrow stromal cells and an injectable sodium alginate/gelatin scaffold. J Biomed Mater Res A, 2012, 100(4): 1044-1050.
17.	Smidsrød O, Skjåk-Braek G. Alginate as immobilization matrix for cells. Trends Biotechnol, 1990, 8(3): 71-78.
18.	Wee S, Gombotz WR. Protein release from alginate matrices. Adv Drug Deliv Rev, 1998, 31(3): 267-285.
19.	Peng S, Zhou G, Luk KD, et al. Strontium promotes osteogenic differentiation of mesenchymal stem cells through the Ras/MAPK signaling pathway. Cell Physiol Biochem, 2009, 23(1-3): 165-174.
20.	Capuccini C, Torricelli P, Sima F, et al. Strontium-substituted hydroxyapatite coatings synthesized by pulsed-laser deposition: in vitro osteoblast and osteoclast response. Acta Biomater, 2008, 4(6): 1885-1893.
21.	Barzegar-Jalali M, Hanaee J, Omidi Y, et al. Preparation and evaluation of sustained release calcium alginate beads and matrix tablets of acetazolamide. Drug Res (Stuttg), 2013, 63(2): 60-64.
22.	Ashton RS, Banarjee A, Punyani S, et al. Scaffolds based on degradable alginate hydrogels and poly (lactide-co-glycolide) microspheres for stem cell culture. Biomaterials, 2007, 28(36): 5518-5525.
23.	Mitalipova MM, Rao RR, Hoyer DM, et al. Preserving the genetic integrity of human embryonic stem cells. Nat Biotechnol, 2005, 23(1): 19-20.

1. Scott TG, Blackburn G, Ashley M, et al. Advances in bionanomaterials for bone tissue engineering. J Nanosci Nanotechnol, 2013, 13(1): 1-22.
2. Jakob F, Ebert R, Ignatius A, et al. Bone tissue engineering in osteoporosis. Maturitas, 2013, 75(2): 118-124.
3. Honda H, Tamai N, Naka N, et al. Bone tissue engineering with bone marrow-derived stromal cells integrated with concentrated growth factor in Rattus norvegicus calvaria defect model. J Artif Organs, 2013. [Epub ahead of print].
4. 参考文献.
5. Haidar ZS, Hamdy RC, Tabrizian M. Delivery of recombinant bone morphogenetic proteins for bone regeneration and repair. Part B: Delivery systems for BMPs in orthopaedic and craniofacial tissue engineering. Biotechnol Lett, 2009, 31(12): 1825-1835.
6. Drury JL, Boontheekul T, Mooney DJ. Cellular cross-linking of peptide modified hydrogels. J Biomech Eng, 2005, 127(2): 220-228.
7. Ohsumi H, Hirata H, Nagakura T, et al. Enhancement of perineurial repair and inhibition of nerve adhesion by viscous injectable pure alginate sol. Plast Reconstr Surg, 2005, 116(3): 823-830.
8. Singh B, Sharma DK, Gupta A. The controlled and sustained release of a fungicide from starch and alginate beads. J Environ Sci Health B, 2009, 44(2): 113-122.
9. Poldervaart MT, Wang H, van der Stok J, et al. Sustained release of BMP-2 in bioprinted alginate for osteogenicity in mice and rats. PLoS One, 2013, 8(8): e72610.
10. Reyes R, Delgado A, Sánchez E, et al. Repair of an osteochondral defect by sustained delivery of BMP-2 or TGFβ1 from a bilayered alginate-PLGA scaffold. J Tissue Eng Regen Med, 2012. [Epub ahead of print].
11. Park Y, Sugimoto M, Watrin A, et al. BMP-2 induces the expression of chondrocyte-specific genes in bovine synovium-derived progenitor cells cultured in three-dimensional alginate hydrogel. Osteoarthritis Cartilage, 2005, 13(6): 527-536.
12. Pors Nielsen S. The biological role of strontium. Bone, 2004, 35(3): 583-588.
13. Augst AD, Kong HJ, Mooney DJ. Alginate hydrogels as biomaterials. Macromol Biosci, 2006, 6(8): 623-633.
14. Cowan CM, Soo C, Ting K, et al. Evolving concepts in bone tissue engineering. Curr Top Dev Biol, 2005, 66: 239-285.
15. Paige KT, Cima LG, Yaremchuk MJ, et al. De novo cartilage generation using calcium alginate-chondrocyte constructs. Plast Reconstr Surg, 1996, 97(1): 168-180.
16. Xia Y, Mei F, Duan Y, et al. Bone tissue engineering using bone marrow stromal cells and an injectable sodium alginate/gelatin scaffold. J Biomed Mater Res A, 2012, 100(4): 1044-1050.
17. Smidsrød O, Skjåk-Braek G. Alginate as immobilization matrix for cells. Trends Biotechnol, 1990, 8(3): 71-78.
18. Wee S, Gombotz WR. Protein release from alginate matrices. Adv Drug Deliv Rev, 1998, 31(3): 267-285.
19. Peng S, Zhou G, Luk KD, et al. Strontium promotes osteogenic differentiation of mesenchymal stem cells through the Ras/MAPK signaling pathway. Cell Physiol Biochem, 2009, 23(1-3): 165-174.
20. Capuccini C, Torricelli P, Sima F, et al. Strontium-substituted hydroxyapatite coatings synthesized by pulsed-laser deposition: in vitro osteoblast and osteoclast response. Acta Biomater, 2008, 4(6): 1885-1893.
21. Barzegar-Jalali M, Hanaee J, Omidi Y, et al. Preparation and evaluation of sustained release calcium alginate beads and matrix tablets of acetazolamide. Drug Res (Stuttg), 2013, 63(2): 60-64.
22. Ashton RS, Banarjee A, Punyani S, et al. Scaffolds based on degradable alginate hydrogels and poly (lactide-co-glycolide) microspheres for stem cell culture. Biomaterials, 2007, 28(36): 5518-5525.
23. Mitalipova MM, Rao RR, Hoyer DM, et al. Preserving the genetic integrity of human embryonic stem cells. Nat Biotechnol, 2005, 23(1): 19-20.

Chinese Journal of Reparative and Reconstructive Surgery

IN VITRO STUDY ON INJECTABLE ALGINATE-STRONTIUM HYDROGEL FOR BONE TISSUE ENGINEERING

Abstract Full text Figures/Tables Video References Cited by

Previous Article

Next Article

Format

Content