Evaluate the growth and adhesion of osteoblast cells on nanocomposite scaffold of hydroxyapatite/titania coated with poly hydroxybutyrate

Authors

1 Department of Tissue Engineering, Faculty of Basic Science and Nuclear Engineering, Islamic Azad University, Najafabad Branch, Najafabad, Iran

2 Department of Biomaterials and Tissue Engineering, School of Advance Technology in Medicine, Isfahan University of Medical Sciences, Isfahan, Iran

3 Department of Anatomical Sciences, School of Medicine, Isfahan University of Medical Sciences, Isfahan, Iran

Abstract

Background: The generation of bioartificial bone tissues may help to overcome the problems related to donor site morbidity and size limitations.
Materials and Methods: In this paper, hydroxyapatite (HA) powder was made out of bovine bone by thermal analysis at 900°C and first, and then, porous HA (50 weight percentage) was produced by polyurethane sponge replication method. In order to improve the scaffold mechanical properties, they have been coated with poly hydroxybutyrate. In terms of phase studies, morphology, and specifying agent groups, the specific characterization devices such as X-ray diffraction and Fourier transform infrared, were employed. To compare the behavior of cellular scaffolds, they were divided into four groups of scaffolds. The osteoblast cells were cultured. To perform phase studies, analysis of Methylthiazole tetrazolium (MTT) and Trypan blue were carried out for the viability and attachment on the surface of the scaffold, and the specification of Scanning electron microscopy was employed for the morphology of the cells.
Results: The results of MTT analysis performed on four groups of scaffolds have shown that Titanium oxide (Tio2 ) had no effect on cell growth alone and HA was the main factor of growth and cell osteoblast adhesion on the scaffold. Moreover, the results showed that the use of coating with poly-3-hydroxybutyrate saved the factors and placed the osteoblasts within the pore. Since the main part of bone consists of HA, the TiO2 accelerates the formation of apatite crystals at the scaffold surface which is the evidence for bone tissue regeneration.
Conclusions: It is likely that the relation between HA and TiO2 leads to an increase in osteoblast adhesion and growth of cells on the scaffold surface.

Keywords

1.
Langer R, Vacanti JP. Tissue engineering. Science 1993;260:920-6.  Back to cited text no. 1
[PUBMED]    
2.
Hollinger JO, Einhorn TA, Doll B, Sfeir C, editors. Bone tissue engineering. CRC press; 2004 Oct 14.  Back to cited text no. 2
    
3.
Buckwalter JA, Glimcher MJ, Cooper RR, Recker R. Bone biology. II: Formation, form, modeling, remodeling, and regulation of cell function. Instr Course Lect 1996;45:387-99.  Back to cited text no. 3
[PUBMED]    
4.
Buckwalter JA, Glimcher MJ, Cooper RR, Recker R. Bone biology. I: Structure, blood supply, cells, matrix, and mineralization. Instr Course Lect 1996;45:371-86.  Back to cited text no. 4
[PUBMED]    
5.
Ackerman LV, Spjut HJ, Abell MR. Bones and Joints (Monographs in Pathology). Baltimore: Williams and Wilkins; 1976.  Back to cited text no. 5
    
6.
Aubin JE. Bone stem cells. J Cell Biochem Suppl 1998;30-31:73-82.  Back to cited text no. 6
[PUBMED]    
7.
Owen M. The origin of bone cells. Int Rev Cytol 1970;28:213-38.  Back to cited text no. 7
[PUBMED]    
8.
Heinegård D, Oldberg A. Structure and biology of cartilage and bone matrix noncollagenous macromolecules. FASEB J 1989;3:2042-51.  Back to cited text no. 8
    
9.
Robey PG, Fedarko NS, Hefferan TE, Bianco P, Vetter UK, Grzesik W, et al. Structure and molecular regulation of bone matrix proteins. J Bone Miner Res 1993;8 Suppl 2:S483-7.  Back to cited text no. 9
[PUBMED]    
10.
Huang S, Ingber DE. The structural and mechanical complexity of cell-growth control. Nat Cell Biol 1999;1:E131-8.  Back to cited text no. 10
[PUBMED]    
11.
Lian JB, Stein GS. Development of the osteoblast phenotype: Molecular mechanisms mediating osteoblast growth and differentiation. Iowa Orthop J 1995;15:118-40.  Back to cited text no. 11
[PUBMED]    
12.
Nomura S, Takano-Yamamoto T. Molecular events caused by mechanical stress in bone. Matrix Biol 2000;19:91-6.  Back to cited text no. 12
[PUBMED]    
13.
Reddi AH. Initiation of fracture repair by bone morphogenetic proteins. Clinical orthopaedics and related research. 1998 Oct 1;355:S66-72.  Back to cited text no. 13
    
14.
Ducy P. Cbfa1: A molecular switch in osteoblast biology. Dev Dyn 2000;219:461-71.  Back to cited text no. 14
[PUBMED]    
15.
Lieberman JR, Daluiski A, Einhorn TA. The role of growth factors in the repair of bone. Biology and clinical applications. J Bone Joint Surg Am 2002;84-A: 1032-44.  Back to cited text no. 15
[PUBMED]    
16.
Perry CR. Bone repair techniques, bone graft, and bone graft substitutes. Clinical orthopaedics and related research. 1999 Mar 1;360:71-86.  Back to cited text no. 16
    
17.
Gugenheim JJ Jr. The Ilizarov method. Orthopedic and soft tissue applications. Clin Plast Surg 1998;25:567-78.  Back to cited text no. 17
[PUBMED]    
18.
Motoki DS, Mulliken JB. The healing of bone and cartilage. Clin Plast Surg 1990;17:527-44.  Back to cited text no. 18
[PUBMED]    
19.
Polykandriotis E, Stangl R, Hennig HH, Lennerz JK, Frank WM, Loos MD, et al. The composite vastus medialis-patellar complex osseomuscular flap as a salvage procedure after complex trauma of the knee - An anatomical study and clinical application. Br J Plast Surg 2005;58:646-51.  Back to cited text no. 19
[PUBMED]    
20.
Williams SF, Martin DP, Horowitz DM, Peoples OP. PHA applications: Addressing the price performance issue: I. Tissue engineering. Int J Biol Macromol 1999;25:111-21.  Back to cited text no. 20
[PUBMED]    
21.
Kunze C, Freier T, Kramer S, Schmitz KP. Anti-inflammatory prodrugs as plasticizers for biodegradable implant materials based on poly (3-hydroxybutyrate). J Mater Sci Mater Med 2002;13:1051-5.  Back to cited text no. 21
[PUBMED]    
22.
Doi Y, Kitamura S, Abe H. Microbial synthesis and character-ization of poly (3-hydroxybutyrate co-3-hydroxyhexanoate). Macromolecules 1995;28:4822-8.  Back to cited text no. 22
    
23.
Yang X, Zhao K, Chen GQ. Effect of surface treatment on the biocompatibility of microbial polyhydroxyalkanoates. Biomaterials 2002;23:1391-7.  Back to cited text no. 23
[PUBMED]    
24.
Dalby MJ, Di Silvio L, Harper EJ, Bonfield W. Increasing hydroxyapatite incorporation into poly (methylmethacrylate) cement increases osteoblast adhesion and response. Biomaterials 2002;23:569-76.  Back to cited text no. 24
[PUBMED]    
25.
Di Silvio L, Dalby MJ, Bonfield W. Osteoblast behaviour on HA/PE composite surfaces with different HA volumes. Biomaterials 2002;23:101-7.  Back to cited text no. 25
[PUBMED]    
26.
Ramires PA, Romito A, Cosentino F, Milella E. The influence of titania/hydroxyapatite composite coatings on in vitro osteoblasts behaviour. Biomaterials 2001;22:1467-74.  Back to cited text no. 26
[PUBMED]    
27.
Galego N, Rozsa C, Sanchez R, Fung J, Vazquez A, Tomas JS. Characterization and application of poly (beta-hydroxyalkano-ates) family as composite biomaterials. Polym Test 2000;19:485-92.  Back to cited text no. 27
    
28.
Chen LJ, Wang M. Production and evaluation of biodegradable composites based on PHB-PHV copolymer. Biomaterials 2002;23:2631-9.  Back to cited text no. 28
[PUBMED]    
29.
Sankapal BR, Sartale S, Ennaoui A. Chemical and electrochemical synthesis of nanosized TiO 2 anatase for large-area photon conversion. C R Chim 2006;9:702-7.  Back to cited text no. 29
    
30.
Chiba A, Kimura S, Raghukandan K, Morizono Y. Effect of alumina addition on hydroxyapatite biocomposites fabricated by underwater-shock compaction. Materials Science and Engineering: A. 2003 Jun 15;350(1):179-83.  Back to cited text no. 30
    
31.
Kim S, Bang H, Song J, Park S. Effect of fluoride additive on the mechanical properties of hydroxyapatite/alumina composites. Ceram Int 2009;35:1647-50.  Back to cited text no. 31
    
32.
Juang H, Hon M. Fabrication and mechanical properties of hydroxyapatite-alumina composites. Mater Sci Eng 1994;2:77-81.  Back to cited text no. 32
    
33.
Ravarian R, Moztarzadeh F, Solati Hashjin M, Rabiee SM, Khoshakhlagh P, Tahriri M. Synthesis, characterization and bioactivity investigation of bioglass/hydroxyapatite composite. Ceram Int 2010;36:291-7.  Back to cited text no. 33
    
34.
Peltola T, Pätsi M, Rahiala H, Kangasniemi I, Yli-Urpo A. Calcium phosphate induction by sol-gel-derived titania coatings on titanium substrates in vitro. J Biomed Mater Res 1998;41:504-10.  Back to cited text no. 34
    
35.
Foroughi MR, Karbasi S, Ebrahimi-Kahrizsangi R. Physical and mechanical properties of a poly-3-hydroxybutyrate-coated nanocrystalline hydroxyapatite scaffold for bone tissue engineering. J Porous Mater 2012;19:667-75.  Back to cited text no. 35
    
36.
Ramay HR, Zhang M. Preparation of porous hydroxyapatite scaffolds by combination of the gel-casting and polymer sponge methods. Biomaterials 2003;24:3293-302.  Back to cited text no. 36
[PUBMED]    
37.
Wang YW, Wu Q, Chen J, Chen GQ. Evaluation of three-dimensional scaffolds made of blends of hydroxyapatite and poly (3-hydroxybutyrate-co-3-hydroxyhexanoate) for bone reconstruction. Biomaterials 2005;26:899-904.  Back to cited text no. 37
[PUBMED]    
38.
Misra SK, Valappil SP, Roy I, Boccaccini AR. Polyhydroxyalkanoate (PHA)/inorganic phase composites for tissue engineering applications. Biomacromolecules 2006;7:2249-58.  Back to cited text no. 38
[PUBMED]    
39.
Evans MD, Steele JG. Polymer surface chemistry and a novel attachment mechanism in corneal epithelial cells. J Biomed Mater Res 1998;40:621-30.  Back to cited text no. 39
[PUBMED]    
40.
Hu SG, Jou CH, Yang MC. Protein adsorption, fibroblast activity and antibacterial properties of poly (3-hydroxybutyric acid-co-3-hydroxyvaleric acid) grafted with chitosan and chitooligosaccharide after immobilized with hyaluronic acid. Biomaterials 2003;24:2685-93.  Back to cited text no. 40
[PUBMED]    
41.
Chen C, Zhou XS, Zhuang YG, Dong LS. Thermal behavior and intermolecular interactions in blends of poly (3-hydroxy-butyrate) and maleated poly(3-hydroxybutyrate) with chito-san. J Appl Polym Sci 2004;10:35-47.  Back to cited text no. 41
    
42.
O′Brien FJ, Harley BA, Yannas IV, Gibson LJ. The effect of pore size on cell adhesion in collagen-GAG scaffolds. Biomaterials 2005;26:433-41.  Back to cited text no. 42
[PUBMED]    
43.
Indolfi L, Baker AB, Edelman ER. The role of scaffold microarchitecture in engineering endothelial cell immunomodulation. Biomaterials 2012;33:7019-27.  Back to cited text no. 43
    
44.
Ma PX, Langer R. Fabrication of biodegradable polymer foams for cell transplantation and tissue engineering. Methods Mol Med 1999;18:47-56.  Back to cited text no. 44
[PUBMED]    
45.
Vaccaro AR. The role of the osteoconductive scaffold in synthetic bone graft. Orthopedics 2002;25 5 Suppl:s571-8.  Back to cited text no. 45
    
46.
Foroughi MR, Karbasi S, Kahrizsangi RE. Physical and mechanical properties of a poly-3-hydroxybutyrate-coated nanocrystalstaline hydroxyapatite scaffold for bone tissue engineering. J Porous Master 2012;19:667-675.  Back to cited text no. 46
    
47.
Montazeri M, Karbasi S, Foroughi MR, Monshi A, Ebrahimi-Kahrizsangi R. Evaluation of mechanical property and bioactivity of nano-bioglass 45S5 scaffold coated with poly-3-hydroxybutyrate. J Mater Sci Mater Med 2015;26:62.  Back to cited text no. 47
[PUBMED]    
48.
Saadat A, Behnamghader A, Karbasi S, Abedi D, Soleimani M, Shafiee A. Comparison of acellular and cellular bioactivity of poly 3-hydroxybutyrate/hydroxyapatite nanocomposite and poly 3-hydroxybutyrate scaffolds. Biotechnology and bioprocess engineering. 2013 Jun 1;18(3):587-93.  Back to cited text no. 48
    
49.
Zhang S, Prabhakaran MP, Qin X, Ramakrishna S. Poly-3-hydroxybutyrate-co-3-hydroxyvalerate containing scaffolds and their integration with osteoblasts as a model for bone tissue engineering. J Biomater Appl 2015;29:1394-406.  Back to cited text no. 49
[PUBMED]