Study on the preparation of polycaprolactone/type<b>Ⅰ</b>collagen tissue engineered meniscus scaffold by three-dimensional printing and its physiochemical properties _Chinese Journal of Reparative and Reconstructive Surgery

Authors：

SHEN Shi ^1,2 , CHEN Mingxue ² , GAO Shuang ² , GUO Weimin ² , WANG Zhenyong ² , LI Haojiang ² , LI Xu ² , ZHANG Bin ^1,2 , XIAN Hai ^1,2 , ZHANG Xueliang ² , LIU Shuyun ² , HAO Libo ² ,  ZHUO Naiqiang ¹ ,  GUO Quanyi ²

1. Department of Orthopedics and Joint Surgery, the Affiliated Hospital of Southwest Medical University, Luzhou Sichuan, 646000, P.R.China;
2. Institute of Orthopedics, General Hospital of Chinese PLA, Beijing Key Laboratory of Regenerative Medicine in Orthopedics, Key Laboratory of Musculoskeletal Trauma & War Injuries, PLA, Beijing, 100853, P.R.China;

Corresponding author：

ZHUO Naiqiang, Email: znq0101@163.com; GUO Quanyi, Email: doctorguo_301@163.com

Keywords：

Meniscus tissue engineering; low temperature deposition technique; Three-dimensional printing technology; scaffold material; physicochemical property

DOI：

10.7507/1002-1892.201803074

Video：

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ObjectiveTo manufacture a polycaprolactone (PCL)/type Ⅰ collagen (COL Ⅰ) tissue engineered meniscus scaffold (hereinafter referred to as PCL/COL Ⅰ meniscus scaffold) by three-dimensional (3D) printing with low temperature deposition technique and to study its physicochemical properties.MethodsFirst, the 15% PCL/4% COLⅠ composite solution and 15% PCL simple solution were prepared. Then, 15% PCL/4% COL Ⅰmeniscus scaffold and 15% PCL meniscal scaffold were prepared by using 3D printing with low temperature deposition techniques. The morphology and microstructure of the scaffolds were observed by gross observation and scanning electron microscope. The compression modulus and tensile modulus of the scaffolds were measured by biomechanical test. The components of the scaffolds were analyzed by Fourier transform infrared spectroscopy (FTIR). The contact angle of the scaffold surface was measured. The meniscus cells of rabbits were cultured with the two scaffold extracts and scaffolds, respectively. After cultured, the cell proliferations were detected by cell counting kit 8 (CCK-8), and the normal cultured cells were used as controls. Cell adhesion and growth of scaffold-cell complex were observed by scanning electron microscope.ResultsAccording to the gross and scanning electron microscope observations, two scaffolds had orientated 3D microstructures and pores, but the surface of the PCL/COLⅠ meniscus scaffold was rougher than the PCL meniscus scaffold. Biomechanical analysis showed that the tensile modulus and compression modulus of the PCL/COL Ⅰ meniscus scaffold were not significantly different from those of the PCL meniscus scaffold (P>0.05). FTIR analysis results showed that COL Ⅰ and PCL were successful mixed in PCL/ COL Ⅰ meniscus scaffolds. The contact angle of PCL/COLⅠ meniscus scaffold [(83.19±7.49)°] was significantly lower than that of PCL meniscus scaffold [(111.13±5.70)°] (t=6.638, P=0.000). The results of the CCK-8 assay indicated that with time, the number of cells cultured in two scaffold extracts showed an increasing trend, and there was no significant difference when compared with the control group (P>0.05). Scanning electron microscope observation showed that the cells attached on the PCL/ COL Ⅰ meniscus scaffold more than that on the PCL scaffold.ConclusionPCL/COLⅠmeniscus scaffolds are prepared by 3D printing with low temperature deposition technique, which has excellent physicochemical properties without cytotoxicity. PCL/COLⅠmeniscus scaffold is expected to be used as the material for meniscus tissue engineering.

Citation： SHEN Shi, CHEN Mingxue, GAO Shuang, GUO Weimin, WANG Zhenyong, LI Haojiang, LI Xu, ZHANG Bin, XIAN Hai, ZHANG Xueliang, LIU Shuyun, HAO Libo, ZHUO Naiqiang, GUO Quanyi. Study on the preparation of polycaprolactone/typeⅠcollagen tissue engineered meniscus scaffold by three-dimensional printing and its physiochemical properties . Chinese Journal of Reparative and Reconstructive Surgery, 2018, 32(9): 1205-1210. doi: 10.7507/1002-1892.201803074 Copy

1.	Pereira H, Caridade SG, Frias AM, et al. Biomechanical and cellular segmental characterization of human meniscus: building the basis for Tissue Engineering therapies. Osteoarthritis Cartilage, 2014, 22(9): 1271-1281.
2.	Abrams GD, Frank RM, Gupta AK, et al. Trends in meniscus repair and meniscectomy in the United States, 2005-2011. Am J Sports Med, 2013, 41(10): 2333-2339.
3.	Hasan J, Fisher J, Ingham E. Current strategies in meniscal regeneration. J Biomed Mate Res B Appl Biomater, 2014, 102(3): 619-634.
4.	Howell R, Kumar NS, Patel N, et al. Degenerative meniscus: Pathogenesis, diagnosis, and treatment options. World J Orthop, 2014, 5(5): 597-602.
5.	Mordecai SC, Al-Hadithy N, Ware HE, et al. Treatment of meniscal tears: An evidence based approach. World J Orthop, 2014, 5(3): 233-241.
6.	Pereira H, Frias AM, Oliveira JM, et al. Tissue engineering and regenerative medicine strategies in meniscus lesions. Arthroscopy, 2011, 27(12): 1706-1719.
7.	Guo W, Liu S, Zhu Y, et al. Advances and prospects in tissue-engineered meniscal scaffolds for meniscus regeneration. Stem Cells Int, 2015, 2015: 517520.
8.	Ohno T, Tanisaka K, Hiraoka Y, et al. Effect of type Ⅰ and type Ⅱ collagen sponges as 3D scaffolds for hyaline cartilage-like tissue regeneration on phenotypic control of seeded chondrocytes in vitro. Materials Science and Engineering: C, 2004, 24(3): 407-411.
9.	Nehrer S, Breinan HA, Ramappa A, et al. Canine chondrocytes seeded in type Ⅰ and type Ⅱ collagen implants investigated in vitro. J Biomed Mater Res, 1997, 38(2): 95-104.
10.	Mota C, Puppi D, Chiellini F, et al. Additive manufacturing techniques for the production of tissue engineering constructs. Journal of Tissue Engineering & Regenerative Medicine, 2015, 9(3): 174-190.
11.	陈明学, 郭维民, 沈师, 等. 半月板细胞外基质-海藻酸水凝胶的制备及其对半月板细胞的影响. 中国医药生物技术, 2017, 12(6): 505-512.
12.	Niu W, Guo W, Han S, et al. Cell-based strategies for meniscus tissue engineering. Stem Cells International, 2016, 2016: 4717184.
13.	Sun F, Zhou H, Lee J. Various preparation methods of highly porous hydroxyapatite/polymer nanoscale biocomposites for bone regeneration. Acta Biomaterialia, 2011, 7(11): 3813-3828.
14.	Zhao C, Tan A, Pastorin G, et al. Nanomaterial scaffolds for stem cell proliferation and differentiation in tissue engineering. Biotechnology Advances, 2013, 31(5): 654-668.
15.	Wang YQ, Cai JY. Enhanced cell affinity of poly(l-lactic acid) modified by base hydrolysis: Wettability and surface roughness at nanometer scale. Current Applied Physics, 2007, 7: e108-e111.
16.	Chung TW, Liu DZ, Wang SY, et al. Enhancement of the growth of human endothelial cells by surface roughness at nanometer scale. Biomaterials, 2003, 24(25): 4655-4661.
17.	Fautrel B, Hilliquin P, Rozenberg S, et al. Impact of osteoarthritis: results of a nationwide survey of 10,000 patients consulting for OA. Joint Bone Spine, 2005, 72(3): 235-240.
18.	Ghosal K, Thomas S, Kalarikkal N, et al. Collagen coated electrospun polycaprolactone (PCL) with titanium dioxide (TiO₂) from an environmentally benign solvent: preliminary physico-chemical studies for skin substitute. Journal of Polymer Research, 2014, 21(5): 1-5.
19.	Kirchhof K, Groth T. Surface modification of biomaterials to control adhesion of cells. Clin Hemorheol Microcirc, 2008, 39(1-4): 247-251.

1. Pereira H, Caridade SG, Frias AM, et al. Biomechanical and cellular segmental characterization of human meniscus: building the basis for Tissue Engineering therapies. Osteoarthritis Cartilage, 2014, 22(9): 1271-1281.
2. Abrams GD, Frank RM, Gupta AK, et al. Trends in meniscus repair and meniscectomy in the United States, 2005-2011. Am J Sports Med, 2013, 41(10): 2333-2339.
3. Hasan J, Fisher J, Ingham E. Current strategies in meniscal regeneration. J Biomed Mate Res B Appl Biomater, 2014, 102(3): 619-634.
4. Howell R, Kumar NS, Patel N, et al. Degenerative meniscus: Pathogenesis, diagnosis, and treatment options. World J Orthop, 2014, 5(5): 597-602.
5. Mordecai SC, Al-Hadithy N, Ware HE, et al. Treatment of meniscal tears: An evidence based approach. World J Orthop, 2014, 5(3): 233-241.
6. Pereira H, Frias AM, Oliveira JM, et al. Tissue engineering and regenerative medicine strategies in meniscus lesions. Arthroscopy, 2011, 27(12): 1706-1719.
7. Guo W, Liu S, Zhu Y, et al. Advances and prospects in tissue-engineered meniscal scaffolds for meniscus regeneration. Stem Cells Int, 2015, 2015: 517520.
8. Ohno T, Tanisaka K, Hiraoka Y, et al. Effect of type Ⅰ and type Ⅱ collagen sponges as 3D scaffolds for hyaline cartilage-like tissue regeneration on phenotypic control of seeded chondrocytes in vitro. Materials Science and Engineering: C, 2004, 24(3): 407-411.
9. Nehrer S, Breinan HA, Ramappa A, et al. Canine chondrocytes seeded in type Ⅰ and type Ⅱ collagen implants investigated in vitro. J Biomed Mater Res, 1997, 38(2): 95-104.
10. Mota C, Puppi D, Chiellini F, et al. Additive manufacturing techniques for the production of tissue engineering constructs. Journal of Tissue Engineering & Regenerative Medicine, 2015, 9(3): 174-190.
11. 陈明学, 郭维民, 沈师, 等. 半月板细胞外基质-海藻酸水凝胶的制备及其对半月板细胞的影响. 中国医药生物技术, 2017, 12(6): 505-512.
12. Niu W, Guo W, Han S, et al. Cell-based strategies for meniscus tissue engineering. Stem Cells International, 2016, 2016: 4717184.
13. Sun F, Zhou H, Lee J. Various preparation methods of highly porous hydroxyapatite/polymer nanoscale biocomposites for bone regeneration. Acta Biomaterialia, 2011, 7(11): 3813-3828.
14. Zhao C, Tan A, Pastorin G, et al. Nanomaterial scaffolds for stem cell proliferation and differentiation in tissue engineering. Biotechnology Advances, 2013, 31(5): 654-668.
15. Wang YQ, Cai JY. Enhanced cell affinity of poly(l-lactic acid) modified by base hydrolysis: Wettability and surface roughness at nanometer scale. Current Applied Physics, 2007, 7: e108-e111.
16. Chung TW, Liu DZ, Wang SY, et al. Enhancement of the growth of human endothelial cells by surface roughness at nanometer scale. Biomaterials, 2003, 24(25): 4655-4661.
17. Fautrel B, Hilliquin P, Rozenberg S, et al. Impact of osteoarthritis: results of a nationwide survey of 10,000 patients consulting for OA. Joint Bone Spine, 2005, 72(3): 235-240.
18. Ghosal K, Thomas S, Kalarikkal N, et al. Collagen coated electrospun polycaprolactone (PCL) with titanium dioxide (TiO₂) from an environmentally benign solvent: preliminary physico-chemical studies for skin substitute. Journal of Polymer Research, 2014, 21(5): 1-5.
19. Kirchhof K, Groth T. Surface modification of biomaterials to control adhesion of cells. Clin Hemorheol Microcirc, 2008, 39(1-4): 247-251.

Chinese Journal of Reparative and Reconstructive Surgery

Study on the preparation of polycaprolactone/typeⅠcollagen tissue engineered meniscus scaffold by three-dimensional printing and its physiochemical properties

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