Experimental study on repairing rabbit skull defect with bone morphogenetic protein 2 peptide/functionalized carbon nanotube composite_Chinese Journal of Reparative and Reconstructive Surgery

Authors：

DI Yuntao ¹ , WANG Cunyang ² ^△ ,  ZHU Huixue ¹ , YU Suxiang ³ , REN Yixing ⁴ ,  LI Xiaoming ²

1. Department of Neurosurgery, the Fourth Central Hospital of Baoding City, Baoding Hebei, 072350, P.R.China;
2. Key Laboratory for Biomechanics and Mechanobiology of Ministry of Education, Beijing Advanced Innovation Center for Biomedical Engineering, School of Biological Science and Medical Engineering, Beihang University, Beijing, 100083, P.R.China;
3. Department of Pathology, the Fourth Central Hospital of Baoding City, Baoding Hebei, 072350, P.R.China;
4. Department of Orthopedics, the Fourth Central Hospital of Baoding City, Baoding Hebei, 072350, P.R.China;

Corresponding?author：

ZHU Huixue, Email: Zhuhuixue@126.com; LI Xiaoming, Email: x.m.li@hotmail.com

Keywords：

Bone morphogenetic protein 2; peptide; multi-walled carbon nanotubes; skull defect; three-dimensional reconstruction; bone repair; rabbit

DOI：

10.7507/1002-1892.202009014

Video：

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Objective To observe and compare the effects of peptides on the repair of rabbit skull defects through two different binding modes of non-covalent and covalent, and the combination of carboxyl (-COOH) and amino (-NH₂) groups with materials.Methods Twenty-one 3-month-old male ordinary New Zealand white rabbits were numbered 1 to 42 on the left and right parietal bones. They were divided into 5 groups using a random number table, the control group (group A, 6 sides) and the material group 1, 2, 3, 4 (respectively group B, C, D, E, 9 sides in each group). All animals were prepared with 12-mm-diameter skull defect models, and bone morphogenetic protein 2 (BMP-2) non-covalently bound multiwalled carbon nanotubes (MWCNT)-COOH+poly (L-lactide) (PLLA), BMP-2 non-covalently bound MWCNT-NH₂+PLLA, BMP-2 covalently bound MWCNT-COOH+PLLA, and BMP-2 covalently bound MWCNT-NH₂+PLLA were implanted into the defects of groups B, C, D, and E, respectively. At 4, 8, and 12 weeks after operation, the samples were taken for CT scanning and three-dimensional reconstruction, the ratio of bone tissue regeneration volume to total volume and bone mineral density were measured, and the histological observation of HE staining and Masson trichrome staining were performed to quantitatively analyze the volume ratio of new bone tissue.Results CT scanning and three-dimensional reconstruction showed that with the extension of time, the defects in groups A-E were filled gradually, and the defect in group E was completely filled at 12 weeks after operation. HE staining and Masson trichrome staining showed that the volume of new bone tissue in each group gradually increased with time, and regenerated mature bone tissue appeared in groups D and E at 12 weeks after operation. Quantitative analysis showed that at 4, 8, and 12 weeks after operation, the ratio of bone tissue regeneration volume to total volume, bone mineral density, and the volume ratio of new bone tissue increased gradually over time; and at each time point, the above indexes increased gradually from group A to group E, and the differences between groups were significant (P<0.05).Conclusion Through covalent binding and using -NH₂ to bound peptides with materials, the best bone repair effect can be achieved.

Citation： DI Yuntao, WANG Cunyang, ZHU Huixue, YU Suxiang, REN Yixing, LI Xiaoming. Experimental study on repairing rabbit skull defect with bone morphogenetic protein 2 peptide/functionalized carbon nanotube composite. Chinese Journal of Reparative and Reconstructive Surgery, 2021, 35(3): 286-294. doi: 10.7507/1002-1892.202009014 Copy

1.	Gerhard EM, Wang W, Li C, et al. Design strategies and applications of nacre-based biomaterials. Acta Biomater, 2017, 54: 21-34.
2.	Jazayeri HE, Tahriri M, Razavi M, et al. A current overview of materials and strategies for potential use in maxillofacial tissue regeneration. Mater Sci Eng C Mater Biol Appl, 2017, 70(Pt 1): 913-929.
3.	Li S, Xu Y, Yu J, et al. Enhanced osteogenic activity of poly (ester urea) scaffolds using facile post-3D printing peptide functionalization strategies. Biomaterials, 2017, 141: 176-187.
4.	He B, Zhao J, Ou Y, et al. Biofunctionalized peptide nanofiber-based composite scaffolds for bone regeneration. Mater Sci Eng C Mater Biol Appl, 2018, 90: 728-738.
5.	Wang Z, Chen L, Wang Y, et al. Improved cell adhesion and osteogenesis of op-HA/PLGA composite by poly (dopamine)-assisted immobilization of collagen mimetic peptide and osteogenic growth peptide. ACS Appl Mater Interfaces, 2016, 8(40): 26559-26569.
6.	Li Y, Luo Z, Xu X, et al. Aspirin enhances the osteogenic and anti-inflammatory effects of human mesenchymal stem cells on osteogenic BFP-1 peptide-decorated substrates. J Mater Chem B, 2017, 5(34): 7153-7163.
7.	Eckhart KE, Holt BD, Laurencin MG, et al. Covalent conjugation of bioactive peptides to graphene oxide for biomedical applications. Biomater Sci, 2019, 7(9): 3876-3885.
8.	Liao J, Wu S, Li K, et al. Peptide-modified bone repair materials: Factors influencing osteogenic activity. J Biomed Mater Res A, 2019, 107(7): 1491-1512.
9.	Mohammadi M, Alibolandi M, Abnous K, et al. Fabrication of hybrid scaffold based on hydroxyapatite-biodegradable nanofibers incorporated with liposomal formulation of BMP-2 peptide for bone tissue engineering. Nanomedicine, 2018, 14(7): 1987-1997.
10.	Wang CY, Cao GX, Zhao TX, et al. Terminal group modification of carbon nanotubes determines covalently bound osteogenic peptide performance. ACS Biomater Sci Eng, 2020, 6(2): 865-878.
11.	Wang S, Zhao Z, Yang Y, et al. A high-strength mineralized collagen bone scaffold for large-sized cranial bone defect repair in sheep. Regen Biomater, 2018, 5(5): 283-292.
12.	Hokugo A, Sorice S, Parhami F, et al. A novel oxysterol promotes bone regeneration in rabbit cranial bone defects. J Tissue Eng Regen Med, 2016, 10(7): 591-599.
13.	Costa F, Carvalho IF, Montelaro RC, et al. Covalent immobilization of antimicrobial peptides (AMPs) onto biomaterial surfaces. Acta Biomater, 2011, 7(4): 1431-1440.
14.	Wang J, Fang T, Li M, et al. Intracellular delivery of peptide drugs using viral nanoparticles of bacteriophage P22: covalent loading and cleavable release. J Mater Chem B, 2018, 6(22): 3716-3726.
15.	King WJ, Krebsbach PH. Growth factor delivery: how surface interactions modulate release in vitro and in vivo. Adv Drug Deliv Rev, 2012, 64(12): 1239-1256.
16.	Shuai C, Liu T, Gao C, et al. Mechanical reinforcement of diopside bone scaffolds with carbon nanotubes. Int J Mol Sci, 2014, 15(10): 19319-19329.
17.	Mikael PE, Amini AR, Basu J, et al. Functionalized carbon nanotube reinforced scaffolds for bone regenerative engineering: fabrication, in vitro and in vivo evaluation. Biomed Mater, 2014, 9(3): 035001.
18.	Xu J, Hu X, Jiang S, et al. The application of multi-walled carbon nanotubes in bone tissue repair hybrid scaffolds and the effect on cell growth in vitro. Polymers (Basel), 2019, 11(2): 230.
19.	Luo F, Pan L, Hong G, et al. In vitro and in vivo characterization of multi-walled carbon nanotubes/polycaprolactone composite scaffolds for bone tissue engineering applications. J Biomater Tissue Eng, 2017, 7(9): 787-797.
20.	Curran JM, Chen R, Hunt JA. Controlling the phenotype and function of mesenchymal stem cells in vitro by adhesion to silane-modified clean glass surfaces. Biomaterials, 2005, 26(34): 7057-7067.
21.	Kikuchi A, Taira H, Tsuruta T, et al. Adsorbed serum protein mediated adhesion and growth behavior of bovine aortic endothelial cells on polyamine graft copolymer surfaces. J Biomater Sci Polym Ed, 1996, 8(2): 77-90.
22.	Wilson CJ, Clegg RE, Leavesley DI, et al. Mediation of biomaterial-cell interactions by adsorbed proteins: a review. Tissue Eng, 2005, 11(1-2): 1-18.
23.	Keselowsky BG, Collard DM, Garcia AJ. Surface chemistry modulates fibronectin conformation and directs integrin binding and specificity to control cell adhesion. J Biomed Mater Res Part A, 2003, 66(2): 247-259.
24.	Li X, Liu H, Niu X, et al. The use of carbon nanotubes to induce osteogenic differentiation of human adipose-derived MSCs in vitro and ectopic bone formation in vivo. Biomaterials, 2012, 33(19): 4818-4827.
25.	Pahlevanzadeh F, Bakhsheshi-Rad HR, Ismail AF, et al. Development of PMMA-Mon-CNT bone cement with superior mechanical properties and favorable biological properties for use in bone-defect treatment. Mater Lett, 2019, 240: 9-12.
26.	Pan WY, Xiao X, Li JL, et al. The comparison of biocompatibility and osteoinductivity between multi-walled and single-walled carbon nanotube/PHBV composites. J Mater Sci-Mater Med, 2018, 29(12): 189.
27.	Shrestha BK, Shrestha S, Tiwari AP, et al. Bio-inspired hybrid scaffold of zinc oxide-functionalized multi-wall carbon nanotubes reinforced polyurethane nanofibers for bone tissue engineering. Mater Des, 2017, 133: 69-81.
28.	Rahimi Z, Zinatizadeh AAL, Zinadini S. Preparation of high antibiofouling amino functionalized MWCNTs/PES nanocomposite ultrafiltration membrane for application in membrane bioreactor. J Ind Eng Chem, 2015, 29: 366-374.
29.	Usui Y, Aoki K, Narita N, et al. Carbon nanotubes with high bone-tissue compatibility and bone-formation acceleration effects. Small, 2008, 4(2): 240-246.
30.	Du Z, Wang C, Zhang R, et al. Applications of graphene and its derivatives in bone repair: Advantages for promoting bone formation and providing real-time detection, challenges and future prospects. Int J Nanomedicine, 2020, 15: 7523-7551.
31.	彭雙麟, 姚志浩, 羅道文, 等. 多孔雙相磷酸鈣陶瓷修復骨質疏松癥大鼠顱骨極量缺損的實驗研究. 口腔醫學研究, 2019, 35(4): 377-381.
32.	宮瑋玉, 劉紹清, 董艷梅, 等. 納米生物活性玻璃促進兔顱骨臨界骨缺損修復. 北京大學學報 (醫學版), 2018, 50(1): 42-48.
33.	Moncal KK, Aydin RST, Abulaban M, et al. Collagen-infilled 3D printed scaffolds loaded with miR-148b-transfected bone marrow stem cells improve calvarial bone regeneration in rats. Mater Sci Eng C Mater Biol Appl, 2019, 105: 110128.

1. Gerhard EM, Wang W, Li C, et al. Design strategies and applications of nacre-based biomaterials. Acta Biomater, 2017, 54: 21-34.
2. Jazayeri HE, Tahriri M, Razavi M, et al. A current overview of materials and strategies for potential use in maxillofacial tissue regeneration. Mater Sci Eng C Mater Biol Appl, 2017, 70(Pt 1): 913-929.
3. Li S, Xu Y, Yu J, et al. Enhanced osteogenic activity of poly (ester urea) scaffolds using facile post-3D printing peptide functionalization strategies. Biomaterials, 2017, 141: 176-187.
4. He B, Zhao J, Ou Y, et al. Biofunctionalized peptide nanofiber-based composite scaffolds for bone regeneration. Mater Sci Eng C Mater Biol Appl, 2018, 90: 728-738.
5. Wang Z, Chen L, Wang Y, et al. Improved cell adhesion and osteogenesis of op-HA/PLGA composite by poly (dopamine)-assisted immobilization of collagen mimetic peptide and osteogenic growth peptide. ACS Appl Mater Interfaces, 2016, 8(40): 26559-26569.
6. Li Y, Luo Z, Xu X, et al. Aspirin enhances the osteogenic and anti-inflammatory effects of human mesenchymal stem cells on osteogenic BFP-1 peptide-decorated substrates. J Mater Chem B, 2017, 5(34): 7153-7163.
7. Eckhart KE, Holt BD, Laurencin MG, et al. Covalent conjugation of bioactive peptides to graphene oxide for biomedical applications. Biomater Sci, 2019, 7(9): 3876-3885.
8. Liao J, Wu S, Li K, et al. Peptide-modified bone repair materials: Factors influencing osteogenic activity. J Biomed Mater Res A, 2019, 107(7): 1491-1512.
9. Mohammadi M, Alibolandi M, Abnous K, et al. Fabrication of hybrid scaffold based on hydroxyapatite-biodegradable nanofibers incorporated with liposomal formulation of BMP-2 peptide for bone tissue engineering. Nanomedicine, 2018, 14(7): 1987-1997.
10. Wang CY, Cao GX, Zhao TX, et al. Terminal group modification of carbon nanotubes determines covalently bound osteogenic peptide performance. ACS Biomater Sci Eng, 2020, 6(2): 865-878.
11. Wang S, Zhao Z, Yang Y, et al. A high-strength mineralized collagen bone scaffold for large-sized cranial bone defect repair in sheep. Regen Biomater, 2018, 5(5): 283-292.
12. Hokugo A, Sorice S, Parhami F, et al. A novel oxysterol promotes bone regeneration in rabbit cranial bone defects. J Tissue Eng Regen Med, 2016, 10(7): 591-599.
13. Costa F, Carvalho IF, Montelaro RC, et al. Covalent immobilization of antimicrobial peptides (AMPs) onto biomaterial surfaces. Acta Biomater, 2011, 7(4): 1431-1440.
14. Wang J, Fang T, Li M, et al. Intracellular delivery of peptide drugs using viral nanoparticles of bacteriophage P22: covalent loading and cleavable release. J Mater Chem B, 2018, 6(22): 3716-3726.
15. King WJ, Krebsbach PH. Growth factor delivery: how surface interactions modulate release in vitro and in vivo. Adv Drug Deliv Rev, 2012, 64(12): 1239-1256.
16. Shuai C, Liu T, Gao C, et al. Mechanical reinforcement of diopside bone scaffolds with carbon nanotubes. Int J Mol Sci, 2014, 15(10): 19319-19329.
17. Mikael PE, Amini AR, Basu J, et al. Functionalized carbon nanotube reinforced scaffolds for bone regenerative engineering: fabrication, in vitro and in vivo evaluation. Biomed Mater, 2014, 9(3): 035001.
18. Xu J, Hu X, Jiang S, et al. The application of multi-walled carbon nanotubes in bone tissue repair hybrid scaffolds and the effect on cell growth in vitro. Polymers (Basel), 2019, 11(2): 230.
19. Luo F, Pan L, Hong G, et al. In vitro and in vivo characterization of multi-walled carbon nanotubes/polycaprolactone composite scaffolds for bone tissue engineering applications. J Biomater Tissue Eng, 2017, 7(9): 787-797.
20. Curran JM, Chen R, Hunt JA. Controlling the phenotype and function of mesenchymal stem cells in vitro by adhesion to silane-modified clean glass surfaces. Biomaterials, 2005, 26(34): 7057-7067.
21. Kikuchi A, Taira H, Tsuruta T, et al. Adsorbed serum protein mediated adhesion and growth behavior of bovine aortic endothelial cells on polyamine graft copolymer surfaces. J Biomater Sci Polym Ed, 1996, 8(2): 77-90.
22. Wilson CJ, Clegg RE, Leavesley DI, et al. Mediation of biomaterial-cell interactions by adsorbed proteins: a review. Tissue Eng, 2005, 11(1-2): 1-18.
23. Keselowsky BG, Collard DM, Garcia AJ. Surface chemistry modulates fibronectin conformation and directs integrin binding and specificity to control cell adhesion. J Biomed Mater Res Part A, 2003, 66(2): 247-259.
24. Li X, Liu H, Niu X, et al. The use of carbon nanotubes to induce osteogenic differentiation of human adipose-derived MSCs in vitro and ectopic bone formation in vivo. Biomaterials, 2012, 33(19): 4818-4827.
25. Pahlevanzadeh F, Bakhsheshi-Rad HR, Ismail AF, et al. Development of PMMA-Mon-CNT bone cement with superior mechanical properties and favorable biological properties for use in bone-defect treatment. Mater Lett, 2019, 240: 9-12.
26. Pan WY, Xiao X, Li JL, et al. The comparison of biocompatibility and osteoinductivity between multi-walled and single-walled carbon nanotube/PHBV composites. J Mater Sci-Mater Med, 2018, 29(12): 189.
27. Shrestha BK, Shrestha S, Tiwari AP, et al. Bio-inspired hybrid scaffold of zinc oxide-functionalized multi-wall carbon nanotubes reinforced polyurethane nanofibers for bone tissue engineering. Mater Des, 2017, 133: 69-81.
28. Rahimi Z, Zinatizadeh AAL, Zinadini S. Preparation of high antibiofouling amino functionalized MWCNTs/PES nanocomposite ultrafiltration membrane for application in membrane bioreactor. J Ind Eng Chem, 2015, 29: 366-374.
29. Usui Y, Aoki K, Narita N, et al. Carbon nanotubes with high bone-tissue compatibility and bone-formation acceleration effects. Small, 2008, 4(2): 240-246.
30. Du Z, Wang C, Zhang R, et al. Applications of graphene and its derivatives in bone repair: Advantages for promoting bone formation and providing real-time detection, challenges and future prospects. Int J Nanomedicine, 2020, 15: 7523-7551.
31. 彭雙麟, 姚志浩, 羅道文, 等. 多孔雙相磷酸鈣陶瓷修復骨質疏松癥大鼠顱骨極量缺損的實驗研究. 口腔醫學研究, 2019, 35(4): 377-381.
32. 宮瑋玉, 劉紹清, 董艷梅, 等. 納米生物活性玻璃促進兔顱骨臨界骨缺損修復. 北京大學學報 (醫學版), 2018, 50(1): 42-48.
33. Moncal KK, Aydin RST, Abulaban M, et al. Collagen-infilled 3D printed scaffolds loaded with miR-148b-transfected bone marrow stem cells improve calvarial bone regeneration in rats. Mater Sci Eng C Mater Biol Appl, 2019, 105: 110128.

Chinese Journal of Reparative and Reconstructive Surgery

Experimental study on repairing rabbit skull defect with bone morphogenetic protein 2 peptide/functionalized carbon nanotube composite

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