Development and characterization of oriented scaffolds derived from cartilage extracellular matrix

doi:10.7518/hxkq.2017.01.007

Abstract

Abstract:

Objective This study aimed to prepare oriented scaffolds derived from a cartilage extracellular matrix (CECM) and to investigate their physicochemical property and compatibility with adipose-derived stem cells (ADSCs). Methods A fresh porcine articular cartilage was cut into pieces. Cartilage nanofibers with diameters of 50-500 nm were collected through homogenization and centrifugation. These nanofibers were then decellularized by using Triton X-100 to produce 6% CECM. The oriented scaffolds derived from the nanoscale CECM were fabricated through unidirectional solidification and lyophili-zation. Afterward, these scaffolds were crosslinked. The physical and chemical performances and cell compatibility of CECM-oriented scaffolds were evaluated. Results The cross-sections of the scaffolds contained homogeneous reticular porous struc-tures with nanofibers on the walls of the pores, and the longitudinal sections revealed vertical tubular structures. Hematoxylin-eosin staining revealed that the scaffolds were red without blue. Toluidine blue, safranin O, and Sirius red staining showed positive results. The porosity, water absorption rate, and vertical compressive elastic modulus of the scaffolds were 95.455%±0.910%, 95.889%±1.071%, and (40.208±5.097) kPa, respectively. Conclusion The components of the oriented scaffolds derived from CECM are similar to those of native cartilage with favorable biocompatibility. The porous structures and sizes of the scaffolds are suitable for the adhesion, prolife-ration, and infiltration of ADSCs. The oriented scaffolds derived from CECM are relatively optimal for cartilage tissue engineering.

Key words: cartilage, extracellular matrix, oriented scaffolds, tissue engineering, unidirectional solidification, lyophilization

CLC Number:

Kun Li, Yanhong Zhao, Chen Xu, Lianyong Wang, Qiang Yang, Hongfa Li, Binhong Teng. Development and characterization of oriented scaffolds derived from cartilage extracellular matrix[J]. West China Journal of Stomatology, 2017, 35(1): 51-56.

Figures/Tables 5

References 23

[1]	Getgood A, Bhullar T, Rushton N.Current concepts in arti-cular cartilage repair[J]. Orthop Trauma, 2009, 23(3):189-200.
[2]	Haleem AM, Chu CR.Advances in tissue engineering tech-niques for articular cartilage repair[J]. Oper Tech Orthop, 2010, 20(2):76-89.
[3]	Danisovic L, Varga I, Zamborsky R, et al.The tissue engi-neering of articular cartilage: cells, scaffolds and stimulating factors[J]. Exp Biol Med (Maywood), 2012, 237(1):10-17.
[4]	Tortelli F, Cancedda R.Three-dimensional cultures of osteo-genic and chondrogenic cells: a tissue engineering approach to mimic bone and cartilage in vitro[J]. Eur Cell Mater, 2009, 17:1-14.
[5]	Cheng CW, Solorio LD, Alsberg E.Decellularized tissue and cell-derived extracellular matrices as scaffolds for ortho-paedic tissue engineering[J]. Biotechnol Adv, 2014, 32(2):462-484.
[6]	Luo Z, Jiang L, Xu Y, et al.Mechano growth factor (MGF) and transforming growth factor (TGF)-β3 functionalized silk scaffolds enhance articular hyaline cartilage regeneration in rabbit model[J]. Biomaterials, 2015, 52:463-475.
[7]	Hollister SJ, Murphy WL.Scaffold translation: barriers be-tween concept and clinic[J]. Tissue Eng Part B Rev, 2011, 17(6):459-474.
[8]	Cui L, Wu Y, Cen L, et al.Repair of articular cartilage defect in non-weight bearing areas using adipose derived stem cells loaded polyglycolic acid mesh[J]. Biomaterials, 2009, 30(14):2683-2693.
[9]	Morelli S, Salerno S, Holopainen J, et al.Osteogenic and osteoclastogenic differentiation of co-cultured cells in poly-lactic acid-nanohydroxyapatite fiber scaffolds[J]. J Biotech-nol, 2015, 204:53-62.
[10]	Zhang K, Zhang Y, Yan S, et al.Repair of an articular car-tilage defect using adipose-derived stem cells loaded on a polyelectrolyte complex scaffold based on poly(l-glutamic acid) and chitosan[J]. Acta Biomater, 2013, 9(7):7276-7288.
[11]	Chang NJ, Lin CC, Shie MY, et al.Positive effects of cell-free porous PLGA implants and early loading exercise on hyaline cartilage regeneration in rabbits[J]. Acta Biomater, 2015, 28:128-137.
[12]	鹿亮, 郭全义, 杨启友, 等. 猪来源关节软骨脱细胞支架的生物安全性研究[J]. 中国医药生物技术, 2010, 5(1):19-23.
	Lu L, Guo QY, Yang QY, et al.Study on the biological safety of acellular scaffold derived from porcine articular cartilage[J]. Chin Med Biotechnol, 2010, 5(1):19-23.
[13]	Gilbert TW, Sellaro TL, Badylak SF.Decellularization of tissues and organs[J]. Biomaterials, 2006, 27(19):3675-3683.
[14]	马浩, 王奇, 石海燕, 等. 不同方法制备脱细胞猪主动脉瓣膜支架的组织结构[J]. 中国组织工程研究, 2012, 16(29):5355-5360.
	Ma H, Wang Q, Shi HY, et al.Histological structure of por-cine aortic valve scaffolds decellularized by using different methods[J]. Chin J Tiss Eng Res, 2012, 16(29):5355-5360.
[15]	Brody LT.Knee osteoarthritis: clinical connections to arti-cular cartilage structure and function[J]. Phys Ther Sport, 2015, 16(4):301-316.
[16]	Almeida HV, Liu Y, Cunniffe GM, et al.Controlled release of transforming growth factor-β3 from cartilage-extra-cellular-matrix-derived scaffolds to promote chondrogenesis of human-joint-tissue-derived stem cells[J]. Acta Biomater, 2014, 10(10):4400-4409.
[17]	陈富林, 毛天球, 丁桂聪, 等. 转化生长因子-β诱导骨髓基质细胞团体外形成软骨的观察[J]. 华西口腔医学杂志, 2003, 21(2):92-94.
	Chen FL, Mao TQ, Ding GC, et al.Experimental study on the chondrogenesis potentiality of marrow stromal cell under the induction of transforming growth factor-β[J]. West Chin J Stomatol, 2003, 21(2):92-94.
[18]	Matsiko A, Levingstone TJ, Gleeson JP, et al.Incorpora-tion of TGF-beta 3 within collagen-hyaluronic acid scaffolds improves their chondrogenic potential[J]. Adv Healthc Mater, 2015, 4(8):1175-1179.
[19]	孙良, 李丕宝, 栾保华. 脱细胞基质软骨支架的制备[J]. 中国组织工程研究, 2015, 19(16):2494-2499.
	Sun L, Li PB, Luan BH.Preparation of an acellular cartilage matrix scaffold[J]. Chin J Tiss Eng Res, 2015, 19(16):2494-2499.
[20]	Luo L, Eswaramoorthy R, Mulhall KJ, et al.Decellulariza-tion of porcine articular cartilage explants and their subse-quent repopulation with human chondroprogenitor cells[J]. J Mech Behav Biomed Mater, 2015, 55:21-31.
[21]	Jia S, Liu L, Pan W, et al.Oriented cartilage extracellular matrix-derived scaffold for cartilage tissue engineering[J]. J Biosci Bioeng, 2012, 113(5):647-653.
[22]	姚军, 卢世壁, 彭江, 等. 关节软骨细胞外基质源性软骨组织工程取向支架的制备[J]. 中国组织工程研究与临床康复, 2009, 13(3):432-436.
	Yao J, Lu SB, Peng J, et al.Preparation of articular cartilage extracellular matrix derived oriented scaffold for cartilage tissue engineering[J]. J Clin Rehabil Tiss Eng Res, 2009, 13(3):432-436.
[23]	Chung C, Burdick JA.Engineering cartilage tissue[J]. Adv Drug Deliv Rev, 2008, 60(2):243-262.