Constructing injectable tissue-engineered cartilage using cytodex-3 microcarrier and alginate hydrogel

doi:10.3969/j.issn.2095-4344.2016.34.014

Abstract

Abstract:

BACKGROUND: Alginate hydrogel and microcarrier both can be used as injectable scaffolds, but their shortcomings such as poor mechanical property and poor plasticity remain unresolved.
OBJIECTIVE: To explore the feasibility of constructing an injectable tissue-engineered cartilage with cytodex-3 microcarrier/alginate hydrogel composite.
METHODS: Injectable cytodex-3 microcarrier/alginate hydrogel composite scaffold and injectable alginate hydrogel scaffold were established, and the mechanical properties of the two scaffolds were detected. Chondrocytes-seeded cytodex-3 microcarrier was obtained after incubated in the bioreactor, and then composited with alginate hydrogel as experimental group; chondrocytes were co-cultured with alginate hydrogel as control group. Subsequently, cell viability and ability of DNA and glycosaminoglycan synthesis were detected.
RESULTS AND CONCLUSION: The Young’s modulus of the experimental group was significantly higher than that of the control group (P < 0.05). And in the control group, chondrocytes were in a round shape and evenly distributed in the alginate hydrogel; in the experimental group, chondrocytes adhered on the scaffold surface and evenly distributed in the scaffold. After 1 day of culture, both viable and numerous dead chondrocytes could be found in both two scaffolds; and after 14-day culture, there were no dead chondrocytes in both two scaffolds, abundant proliferating chondrocytes maintained a high cell viability, and the number of chondrocytes in the experimental group was significantly higer than that of the control group. What’s more, the contents of DNA and glycosaminoglycans were in a rise with time in both two groups, which were significantly higher in the experimental group than the control group (P < 0.05). These results suggest that the cytodex-3 microcarrier/alginate hydrogel composite is a promising injectable scaffold in cartilage tissue engineering.

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: Chondrocytes, Stents, Tissue Engineering

CLC Number:

R318

Yin He-yong, Sun Zhen, Li Pan, Yu Xiao-ming, Xu Yi-chi, Sun Xun, Xiao Bo, Wang Yu, Wang Ai-yuan, Guo Quan-yi, Xu Wen-jing, Lu Shi-bi, Peng Jiang. Constructing injectable tissue-engineered cartilage using cytodex-3 microcarrier and alginate hydrogel[J]. Chinese Journal of Tissue Engineering Research, 2016, 20(34): 5104-5109.

Figures/Tables 4

References

[1] Johnstone B,Alini M,Cucchiarini M,et al.Tissue engineering for articular cartilage repair the state of the art. Eur Cells Mater.2013;25:248-267.
[2] Matsunaga D,Akizuki S,Takizawa T,et al.Repair of articular cartilage and clinical outcome after osteotomy with microfracture or abrasion arthroplasty for medial gonarthrosis.Knee.2007;14(6):465-471.
[3] 刘奕,谢林.组织工程技术修复损伤关节软骨的研究与应用[J].中国组织工程研究,2013,17(41):7310-7316.
[4] Eyrich D,Brandl F,Appel B,et al.Long-term stable fibrin gels for cartilage engineering.Biomaterials. 2007; 28(1):55-65.
[5] Balakrishnan B,Jayakrishnan A.Self-cross-linking biopolymers as injectable in situ forming biodegradable scaffolds.Biomaterials.2005;26:3941-3951.
[6] 徐虎,胡蕴玉,刘松波,等.可注射性藻酸钙凝胶载体组织工程性软骨的实验研究[J].中华骨科杂志, 2000,20(9): 19-22.
[7] 李卫平,韩运,黄建荣,等.可注射性材料藻酸钠体外培养幼猪关节软骨细胞及形成组织工程化软骨的能力[J].中国组织工程研究与临床康复,2007,11(14):2705-2707.
[8] 周建林,方洪松,彭昊,等.自体、异体骨软骨移植及组织工程材料修复的关节软骨损伤[J].中国组织工程研究, 2015, 19(34):5530-5535.
[9] 朱琳,刘海霞,公衍道,等.利用细胞装配系统直接形成组织工程三维复合支架[J].中国组织工程研究与临床康复, 2007,11(48):9650-9654.
[10] 康红军,卢世璧,许文静,等.人脂肪干细胞结合微载体在生物反应器中向软骨细胞分化[J].中国矫形外科杂志, 2007, 15(10):773-775,803.
[11] 宁斌,田周斌,贾堂宏.微载体培养技术在骨与软骨组织工程研究中的应用[J].中国组织工程研究, 2012,16(8): 1459-1462.
[12] Shao X,Goh JC,Hutmacher DW,et al.Repair of large articular osteochondral defects using hybrid scaffolds and bone marrow-derived mesenchymal stem cells in a rabbit model. Tissue Eng. 2006;12: 1539-1551.
[13] Yang Q,Peng J,Guo Q,et al.A cartilage ECM-derived 3-D porous acellular matrix scaffold for in vivo cartilage tissue engineering with PKH26-labeled chondrogenic bone marrow-derived mesenchymal stem cells. Biomaterials.2008;29:2378-2387.
[14] Hong Y,Gao C,Xie Y,et al.Collagen-coated polylactide microspheres as chondrocyte microcarriers. Biomaterials.2005;26:6305-6313.
[15] Bouwmeester PS,Kuijer R,Homminga GN,et al.A retrospective analysis of two independent prospective cartilagerepair studies: autogenous perichondrial grafting versus subchondraldrilling 10 years post-surgery. J Orthop Res.2002;20(2):267-273.
[16] Harris JD,Siston RA,Pan X,et al. Flanigan, Autologous chondrocyte implantation: a systematic review.J Bone Joint Surg Am.2010;92(12):2220-2233.
[17] Chung HJ,Go DH,Bae JW,et al.Synthesis and characterization of Pluronic grafted chitosan copolymer as a novel injectable biomaterial.Curr Appl Phys. 2005; 5:485-488.
[18] Choi YS,Park S,Suh H.Adipose tissue engineering usingmesenchymal stem cells attached to injectable PLGA spheres.Biomaterials.2005;26:5855-5863.
[19] Matricali GA,Dereymaeker GPE,Luyten FP.Donor site morbidity after articular cartilage repair procedures : a review.Acta Orthop Belg.2010;76:669-674.
[20] Schrobback K, Klein TJ,Crawford R,et al.Effects of oxygen and culture system on in vitro propagation and redifferentiation of osteoarthritic human articular chondrocytes.Cell Tissue Res.2012;347:649-663.
[21] Cummings LJ,Waters SK, Tissue growth in a rotating bioreactor. Part II: ?uid ?ow and nutrient transport problems. Math Med Biol.2007;24:169-208.
[22] Li YY,Cheng HW,Cheung KMet al.Mesenchymal stem cell-collagen microspheres for articular cartilage repair: cell density and differentiation status.Acta Biomater. 2014;10:1919-1929.
[23] Malda J,van Blitterswijk CA,Grojec M,et al.Expansion of bovine chondrocytes on microcarriers enhances redifferentiation.Tissue Eng. 2003;9:939-948.
[24] Schrobback K,Klein TJ,Schuetz M,et el.Adult human articular chon-drocytes in a microcarrier-based culture system: expansion and redifferentiation.J Orthop Res 2011;29:539-546.
[25] Mathieu M,Vigier S,Labour MN,et al.Induction of mesenchymal stem cell differentiation and cartilage formation by cross-linker-free collagen microspheres. Eur Cell Mater.2014;28:82-96.
[26] Hong Y,Gao C,Xie Y,et al.Collagen-coated polylactide microspheres as chondrocyte microcarriers. Biomaterials.2005;26:6305-6313.