Presentation of a novel model of chitosan- polyethylene oxide-nanohydroxyapatite nanofibers together with bone marrow stromal cells to repair and improve minor bone defects

Document Type: Original Article


1 Neuroscience Research Center, Baqiyatallah University of Medical Science, Tehran, Iran

2 Microbial Biotechnology, Faculty of Science, Payame Noor University, Unit of Tehran Center, Tehran, Iran

3 Department of Parasitology and Mycology, School of Medicine, Baqiyatallah University of Medical Science, Tehran, Iran

4 Department of Anatomy, School of Medicine, Baqiyatallah University of Medical Science, Tehran, Iran


Objective(s):Various methods for repairing bone defects are presented. Cell therapy is one of these methods. Bone marrow stromal cells (BMSCs) seem to be suitable for this purpose. On the other hand, lots of biomaterials are used to improve and repair the defect in the body, so in this study we tried to produce a similar structure to the bone by the chitosan and hydroxyapatite.
Materials and Methods: In this study, the solution of chitosan-nanohydroxyapatite-polyethylene oxide (PEO) Nanofibers was produced by electrospinning method, and then the BMSCs were cultured on this solution. A piece of chitosan-nanohydroxyapatite Nanofibers with BMSCs was placed in a hole with the diameter of 1 mm at the distal epiphysis of the rat femur. Then the biomechanical and radiographic studies were performed.
Results: Biomechanical testing results showed that bone strength was significantly higher in the Nanofiber/BMSCs group in comparison with control group. Also the bone strength in nanofiber/BMSCs group was significant, but in nanofiber group was nearly significant. Radiographic studies also showed that the average amount of callus formation (radio opacity) in nanofiber and control group was not significantly different. The callus formation in nanofiber/BMSCs group was increased compared to the control group, and it was not significant in the nanofiber group.
Conclusion: Since chitosan-nanohydroxyapatite nanofibers with BMSCs increases the rate of bone repair, the obtained cell-nanoscaffold shell can be used in tissue engineering and cell therapy, especially for bone defects.


1. Jorgensen CH. Mesenchymal stem cells in arthritis: role of bone marrow microenvironment. Arthritis Res Ther 2010; 12:135.

2. Valtieri M, Sorrentino A. The mesenchymal stromal cell contribution to homeostasis. J Cell Physiol 2008; 217:296-300.

3. Bergfeld SA, DeClerck YA. Bone marrow-derived mesenchymal stem cells and the tumor micro-environment. Cancer Metastasis Rev 2010; 29:249–261.

4. Hideaki K, Hideki A, Arinobu T. Bone marrow strom cells (bone marrow-derived multipotent mesenchymal stromal cells) for bone tissue engineering: Basic science to clinical translation. Int J Biochem Cell Biol 2011; 43:286-289.

5. Salasznyk RM, Williams WA, Boskey A, Batorsky A, Plopper GE. Adhesion to vitronectin and collagen promotes osteogenic differentiation of human mesenchymal stem cells. J Biomed Biotechnol 2004; 1:24-34.

6. Araki T, Nagarajan R, Milbrandt J. Identification of genes induced in peripheral nerve after injury. J Biol Chem 2001; 276:34131-34141.

7. Anita M,  Ranieri C, Rodolfo Q. Clonal mesenchymal progenitors from human bone marrow differentiate in vitro according to a hierarchical model. J Cell Sci 2000; 113: 1161-1166.

8. Tohill M, Mantovani C, Wiberg M, Terenghi G. Rat bone marrow mesenchymal stem cells express glial markers and stimulate nerve regeneration. Neurosci Lett 2004; 362:200-203.

9. HomayoniaH, Hosseini Ravandia SA, Valizadehb M. Electrospinning of chitosan nanofibers: processing optimization. Carbohydr Polym 2009; 77:656–661.

10. Khora E, Yong Lim L. Implantable applications of chitin and Chitosan. Biomaterials 2003; 24:2339-2349.

11. Kumar MNVR. A review of chitin and Chitosan applications. React Funct Polym 2000; 46:1-27.

12. Yogi K, Michibayashi N, Kurikawa N, Nakashima Y, Mizoguchi T, Harada A, et al. Effectiveness of fructose-modified Chitosan as a scaffold for hepatocyte attachment. Biol Pharm Bull 1997; 20:1290–1294.

13. Zhang Y, Zhang MQ. Calcium phosphate/Chitosan composite scaffolds for controlled in vitro antibiotic drug release. J Biomed Mater Res 2002; 62:378–386.

14. Zhang Y, ZhangMQ. Three-dimensional macroporous calcium phosphate bioceramics with nested Chitosan sponges for load-bearing bone implants. J Biomed Mater Res 2002; 61: 1–8.

15. Zhang Y, Zhang MQ. Synthesis and characterization of macroporous Chitosan/calcium phosphate composite scaffolds for tissue engineering. J Biomed Mater Res 2001; 55:304–312.

16. Bhattaraia N, Edmondsona D, Veiseha O, Matsenb FA, Zhang M. Electrospun chitosan-based nanofibers and their cellular compatibility. Biomaterials 2005; 26:6176–6184.

17. Fouda MFA, Nemat A, Gawish A, Baiuomy AR. Does the coating of titanium implants by hydroxyapatite affect the elaboration of free radicals. An Experimental Study. Austr J Basic Appl Sci 2009; 3:1122-1129.

18. Seung Y. The survival and migration pattern of the bone marrow stromal cells after intracevebral transplantation in rats. J Korean Neurosurg 2004; 400-404.

19. Madsen JE, Hukkhanen M. Fracture healing and callus innervations after peripheral nervive resection in rats. Clin Orthopaed Relat Res 1998; 351:230-240.

20. Boskey AL, Wians FH, Hauschka PV. The effect of osteocalcin on in vitro lipid-induced hydroxyapatite formation and seeded hydroxyapatite growth. Calcif Tissue Int 1985; 37:57-62.

21. Yanzhong ZH, Jayarama R, Adel E, Seeram R, Bo SU, Teck L. Electrospun biomimetic nanocomposite nanofibers of hydroxyapatite/Chitosan for bone tissue engineering. Biomaterials 2008; 29:4314-4322.

22. Faheem A, Naseer A, Muzafar A, Soo Jin P, Dae Kwang P and  Hak Y. Synthesis of Polyvinyl alcohol (PVA) nanofibers Incorporating Hydroxyapatite nanoparticles as Future Implant Materials. 2010; 18: 59-66.

23. Han L, Anthony A, Montserrat C, Megan C, James B, Salim C, et al. Composite tissue engineering on
polycaprolactone nanofiber scaffolds. Ann Plast Surg 2009; 62:505-512.

24. Zhao LR, Duan WM, Reyes M, Keene CD, Verfaillie CM, Low WC. Human bone marrow stem cells exhibit neural phenotypes and ameliorate neurological deficits after grafting into the ischemic brain of rats. Exp Neurol 2002; 174:11-20.

25. Bossolasco P, Cova L, Calzarossa C, Rimoldi SG, Borsotti C, Deliliers GL, et al. Neuro-glial differentiation of human bone marrow stem cells in vitro. Exp Neurol 2005; 193:312-325.

26. Lamoury FM, Croitoru-Lamoury J, Brew BJ. Undifferentiated mouse mesenchymal stem cells spontaneously express neural and stem cell markers Oct-4 and Rex-1. Cytotherapy 2006; 210:228-242.

27. Stockmann P, Wilmowsky J, Nkenke V, Felszeghy C, Friedrich E, et al Guided bone regeneration in pig calvarial bone defects using autologous mesenchymal stem/progenitor cells-a comparison of different tissue sources. J Craniomaxillofac Sur 2012; 40:310-320.

28. Cao L, Liu G, Gan Y, Fan Q, Yang F, ZhangX, et al. The use of autologous enriched bone marrow MSCs to enhance osteoporotic bone defect repair in long-term estrogen deficient goats. Biomaterials 2012; 33:5076-5084.

29. Li F, Wang X, Niyibizi C. Bone marrow stromal cells contribute to bone formation following infusion into femoral cavities of a mouse model of osteogenesis imperfecta. Bone 2010; 47:546-555.