Electrospun polycaprolactone/collagen type<b>Ⅰ</b>nanofibers oriented patch for rotator cuff repairing_Chinese Journal of Reparative and Reconstructive Surgery

Authors：

GAO Chao ^1,2 , LI Chaoming ³ , XU Yulong ³ , WANG Zhenyong ^1,2 , LI Haojiang ² , LUO Xujiang ² , PENG Liqing ² , ZHANG Bin ² , SHEN Shi ² , LIU Shuyun ² , SUI Xiang ² ,  GUO Quanyi ² ,  YANG Jianhua ⁴

1. Department of Orthopedics, the First Affiliated Hospital of Jiamusi University, Jiamusi Heilongjiang, 154007, P.R.China;
2. Institute of Orthopedics, Chinese PLA General Hospital, Beijing Key Lab of Regenerative Medicine in Orthopedics, Key Lab of Musculoskeletal Trauma & War Injuries of Chinese PLA, Beijing, 100853, P.R.China;
3. College of Mechanical and Electric Engineering, Beijing University of Chemical Technology, Beijing, 100029, P.R.China;
4. Department of Orthopedics, Longgang District People’s Hospital of Shenzhen, Shenzhen Guangdong, 518172, P.R.China;

Corresponding?author：

GUO Quanyi, Email: doctorguo_301@163.com; YANG Jianhua, Email: jianhua01@163.com

Keywords：

Electrospinning; rotator cuff patch; tendon tissue engineering; polycaprolactone; collagen type Ⅰ

DOI：

10.7507/1002-1892.201811034

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Objective Electrospinning technique was used to manufacture polycaprolactone (PCL)/collagen typeⅠ nanofibers orientated patches and to study their physical and chemical characterization, discussing their feasibility as synthetic patches for rotator cuff repairing.Methods PCL patches were prepared by electrospinning with 10% PCL electrospinning solution (control group) and PCL/collagen typeⅠorientated nanofibers patches were prepared by electrospinning with PCL electrospinning solution with 25% collagen type Ⅰ(experimental group). The morphology and microstructure of the two patches were observed by gross and scanning electron microscopy, and the diameter and porosity of the fibers were measured; the mechanical properties of the patches were tested by uniaxial tensile test; the composition of the patches was analyzed by Fourier transform infrared spectroscopy; and the contact angle of the patch surface was measured. Two kinds of patch extracts were co-cultured with the third generation of rabbit tendon stem cells. Cell counting kit 8 (CCK-8) was used to detect the toxicity and cell proliferation of the materials. Normal cultured cells were used as blank control group. Rabbit tendon stem cells were co-cultured with the two patches and stained with dead/living cells after 3 days of in vitro culture, and laser confocal scanning microscopy was used to observe the cell adhesion and activity on the patch.Results Gross and scanning electron microscopy showed that the two patch fibers were arranged in orientation. The diameter of patch fibers in the experimental group was significantly smaller than that in the control group (t=26.907, P=0.000), while the porosity in the experimental group was significantly larger than that in the control group (t=2.506, P=0.032). The tensile strength and Young’s modulus of the patch in the experimental group were significantly higher than those in the control group (t=3.705, P=0.029; t=4.064, P=0.034). Infrared spectrum analysis showed that PCL and collagen type Ⅰ were successfully mixed in the experimental group. The surface contact angle of the patch in the experimental group was (73.88±4.97)°, which was hydrophilic, while that in the control group was (128.46±5.10) °, which was hydrophobic. There was a significant difference in the surface contact angle between the two groups (t=21.705, P=0.002). CCK-8 test showed that with the prolongation of culture time, the cell absorbance (A) value increased gradually in each group, and there was no significant difference between the experimental group and the control group at each time point (P>0.05). Laser confocal scanning microscopy showed that rabbit tendon stem cells could adhere and grow on the surface of both patches after 3 days of culture. The number of cells adhered to the surface of the patches in the experimental group was more than that in the control group, and the activity was better.Conclusion PCL/ collagen type Ⅰ nanofibers orientated patch prepared by electrospinning technology has excellent physical and chemical properties, cell adhesion, and no cytotoxicity. It can be used as an ideal scaffold material in tendon tissue engineering for rotator cuff repair in the future.

Citation： GAO Chao, LI Chaoming, XU Yulong, WANG Zhenyong, LI Haojiang, LUO Xujiang, PENG Liqing, ZHANG Bin, SHEN Shi, LIU Shuyun, SUI Xiang, GUO Quanyi, YANG Jianhua. Electrospun polycaprolactone/collagen typeⅠnanofibers oriented patch for rotator cuff repairing. Chinese Journal of Reparative and Reconstructive Surgery, 2019, 33(5): 628-633. doi: 10.7507/1002-1892.201811034 Copy

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4.	Szentivanyi AL, Zernetsch H, Menzel H, et al. A review of developments in electrospinning technology: new opportunities for the design of artificial tissue structures. Int J Artif Organs, 2011, 34(10): 986-997.
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6.	Mun CH, Jung Y, Kim SH, et al. Effects of pulsatile bioreactor culture on vascular smooth muscle cells seeded on electrospun poly (lactide-co-epsilon-caprolactone) scaffold. Artif Organs, 2013, 37(12): E168-178.
7.	Teo WE, He W, Ramakrishna S. Electrospun scaffold tailored for tissue-specific extracellular matrix. Biotechnology J, 2006, 1(9): 918-929.
8.	Lim SH, Mao HQ. Electrospun scaffolds for stem cell engineering. Adv Drug Deliv Rev, 2009, 61(12): 1084-1096.
9.	Seyedjafari E, Soleimani M, Ghaemi N, et al. Nanohydroxyapatite-coated electrospun poly (l-lactide) nanofibers enhance osteogenic differentiation of stem cells and induce ectopic bone formation. Biomacromolecules, 2010, 11(11): 3118-3125.
10.	Kumbar SG, Nukavarapu SP, James R, et al. Electrospun poly (lactic acid-co-glycolic acid) scaffolds for skin tissue engineering. Biomaterials, 2008, 29(30): 4100-4107.
11.	Hajiali H, Shahgasempour S, Naimi-Jamal MR, et al. Electrospun PGA/gelatin nanofibrous scaffolds and their potential application in vascular tissue engineering. Int J Nanomedicine, 2011, 6: 2133-2141.
12.	Karimi A, Karbasi S, Razavi S, et al. Poly (hydroxybutyrate)/chitosan aligned electrospun scaffold as a novel substrate for nerve tissue engineering. Adv Biomed Res, 2018, 7: 44.
13.	Yoon H, Kim G. A three-dimensional polycaprolactone scaffold combined with a drug delivery system consisting of electrospun nanofibers. J Pharm Sci, 2011, 100(2): 424-430.
14.	Ji W, Sun Y, Yang F, et al. Bioactive electrospun scaffolds delivering growth factors and genes for tissue engineering applications. Pharm Res, 2011, 28(6): 1259-1272.
15.	代廣春, 李滎娟, 徐宏亮, 等. 深低溫冷凍對大鼠髕腱組織內肌腱干細胞生物學特性影響的研究. 中國修復重建外科雜志, 2017, 31(7): 845-852.
16.	Sankar S, Sharma CS, Rath SN, et al. Electrospun fibers for recruitment and differentiation of stem cells in regenerative medicine. Biotechnol J, 2017, 12(12). doi: 10.1002/biot.201700263.
17.	Orr SB, Chainani A, Hippensteel KJ, et al. Aligned multilayered electrospun scaffolds for rotator cuff tendon tissue engineering. Acta Biomater, 2015, 24: 117-126.
18.	Lomas AJ, Ryan CN, Sorushanova A, et al. The past, present and future in scaffold-based tendon treatments. Adv Drug Deliv Rev, 2015, 84: 257-277.
19.	Zhang Q, Lv S, Lu J, et al. Characterization of polycaprolactone/collagen fibrous scaffolds by electrospinning and their bioactivity. Int J Biol Macromol, 2015, 76: 94-101.
20.	Qiu Y, Lei J, Koob TJ, et al. Cyclic tension promotes fibroblastic differentiation of human MSCs cultured on collagen-fibre scaffolds. J Tissue Eng Regen Med, 2016, 10(12): 989-999.
21.	沈師, 陳明學, 高爽, 等. 3D打印制備聚己內酯/Ⅰ型膠原組織工程半月板支架及其理化特性的研究. 中國修復重建外科雜志, 2018, 32(9): 1205-1210.
22.	Ghosal K, Manakhov A, Zají?ková L, et al. Structural and surface compatibility study of modified electrospun poly (ε-caprolactone) (PCL) composites for skin tissue engineering. AAPS PharmSciTech, 2017, 18(1): 72-81.
23.	Kopf BS, Ruch S, Berner S, et al. The role of nanostructures and hydrophilicity in osseointegration: In-vitro protein-adsorption and blood-interaction studies. J Biomed Mater Res A, 2015, 103(8): 2661-2672.

1. Maffulli N. Rupture of the Achilles tendon. J Bone Joint Surg (Am), 1999, 81(7): 1019-1036.
2. Wang JH. Mechanobiology of tendon. J Biomech, 2006, 39(9): 1563-1582.
3. Zhao C, Tan A, Pastorin G, et al. Nanomaterial scaffolds for stem cell proliferation and differentiation in tissue engineering. Biotechnol Adv, 2013, 31(5): 654-668.
4. Szentivanyi AL, Zernetsch H, Menzel H, et al. A review of developments in electrospinning technology: new opportunities for the design of artificial tissue structures. Int J Artif Organs, 2011, 34(10): 986-997.
5. Shuakat MN, Lin T. Recent developments in electrospinning of nanofiber yarns. J Nanosci Nanotechnol, 2014, 14(2): 1389-1408.
6. Mun CH, Jung Y, Kim SH, et al. Effects of pulsatile bioreactor culture on vascular smooth muscle cells seeded on electrospun poly (lactide-co-epsilon-caprolactone) scaffold. Artif Organs, 2013, 37(12): E168-178.
7. Teo WE, He W, Ramakrishna S. Electrospun scaffold tailored for tissue-specific extracellular matrix. Biotechnology J, 2006, 1(9): 918-929.
8. Lim SH, Mao HQ. Electrospun scaffolds for stem cell engineering. Adv Drug Deliv Rev, 2009, 61(12): 1084-1096.
9. Seyedjafari E, Soleimani M, Ghaemi N, et al. Nanohydroxyapatite-coated electrospun poly (l-lactide) nanofibers enhance osteogenic differentiation of stem cells and induce ectopic bone formation. Biomacromolecules, 2010, 11(11): 3118-3125.
10. Kumbar SG, Nukavarapu SP, James R, et al. Electrospun poly (lactic acid-co-glycolic acid) scaffolds for skin tissue engineering. Biomaterials, 2008, 29(30): 4100-4107.
11. Hajiali H, Shahgasempour S, Naimi-Jamal MR, et al. Electrospun PGA/gelatin nanofibrous scaffolds and their potential application in vascular tissue engineering. Int J Nanomedicine, 2011, 6: 2133-2141.
12. Karimi A, Karbasi S, Razavi S, et al. Poly (hydroxybutyrate)/chitosan aligned electrospun scaffold as a novel substrate for nerve tissue engineering. Adv Biomed Res, 2018, 7: 44.
13. Yoon H, Kim G. A three-dimensional polycaprolactone scaffold combined with a drug delivery system consisting of electrospun nanofibers. J Pharm Sci, 2011, 100(2): 424-430.
14. Ji W, Sun Y, Yang F, et al. Bioactive electrospun scaffolds delivering growth factors and genes for tissue engineering applications. Pharm Res, 2011, 28(6): 1259-1272.
15. 代廣春, 李滎娟, 徐宏亮, 等. 深低溫冷凍對大鼠髕腱組織內肌腱干細胞生物學特性影響的研究. 中國修復重建外科雜志, 2017, 31(7): 845-852.
16. Sankar S, Sharma CS, Rath SN, et al. Electrospun fibers for recruitment and differentiation of stem cells in regenerative medicine. Biotechnol J, 2017, 12(12). doi: 10.1002/biot.201700263.
17. Orr SB, Chainani A, Hippensteel KJ, et al. Aligned multilayered electrospun scaffolds for rotator cuff tendon tissue engineering. Acta Biomater, 2015, 24: 117-126.
18. Lomas AJ, Ryan CN, Sorushanova A, et al. The past, present and future in scaffold-based tendon treatments. Adv Drug Deliv Rev, 2015, 84: 257-277.
19. Zhang Q, Lv S, Lu J, et al. Characterization of polycaprolactone/collagen fibrous scaffolds by electrospinning and their bioactivity. Int J Biol Macromol, 2015, 76: 94-101.
20. Qiu Y, Lei J, Koob TJ, et al. Cyclic tension promotes fibroblastic differentiation of human MSCs cultured on collagen-fibre scaffolds. J Tissue Eng Regen Med, 2016, 10(12): 989-999.
21. 沈師, 陳明學, 高爽, 等. 3D打印制備聚己內酯/Ⅰ型膠原組織工程半月板支架及其理化特性的研究. 中國修復重建外科雜志, 2018, 32(9): 1205-1210.
22. Ghosal K, Manakhov A, Zají?ková L, et al. Structural and surface compatibility study of modified electrospun poly (ε-caprolactone) (PCL) composites for skin tissue engineering. AAPS PharmSciTech, 2017, 18(1): 72-81.
23. Kopf BS, Ruch S, Berner S, et al. The role of nanostructures and hydrophilicity in osseointegration: In-vitro protein-adsorption and blood-interaction studies. J Biomed Mater Res A, 2015, 103(8): 2661-2672.

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