Use of chitosan and β-tricalcium phosphate, alone and in combination, for bone healing in rabbits

A S Azevedo, M J C Sá, M V L Fook, P I Nóbrega Neto, O B Sousa, S S Azevedo, M W Teixeira, F S Costa, A L Araújo, A S Azevedo, M J C Sá, M V L Fook, P I Nóbrega Neto, O B Sousa, S S Azevedo, M W Teixeira, F S Costa, A L Araújo

Abstract

The aim of this research was to evaluate the process of bone regeneration in rabbits, using chitosan and beta-tricalcium phosphate (β-TCP) independently and in combination. A total of 12 New Zealand rabbits of both sexes, with average weight of 3.0 ± 0.57 kg were used. Animals were randomly divided into two experimental time points, with six animals euthanized 45 days after surgery and six euthanized 90 days after surgery. We performed two osteotomies in each tibia. The left tibia was used for the chitosan (QUI) and control groups, and the right tibia was used for the β-TCP alone and in combination with chitosan (QUI+TCP) groups. Tomographic evaluation showed no statistically significant difference among groups; however radiopacity was higher in the treated groups. Comparative descriptive histological evaluation found that treatment groups stimulated a more pronounced tissue repair reaction and accelerated bone repair. Morphometric analysis showed that treatment groups presented statistically higher bone formation compared with the control group.

Figures

Fig. 1
Fig. 1
Mean and standard deviation of radiodensity in Hounsfield units (HU) at 45 and 90 days. Osteotomized area received the treatments; C control, QUI chitosan only, QUI+TCP chitosan in combination with tricalcium phosphate (TCP), TCP TCP only
Fig. 2
Fig. 2
Descriptive histological evaluation at day 45. a (Control group); red circle indicates osteotomized site filled by newly formed immature bone. b (QUI group); red circle shows implantation of chitosan surrounded by intense cellular reaction, dark arrows highlight blood vessels. c (TCP group); red circle indicates osteotomized site filled by newly formed bone tissue. d (TCP+QUI group); red circle indicates the implant site with intense cellular reaction around the implant and black circle highlights osteotomized area filled by newly formed bone tissue. Obj. ×4 (Color figure online)
Fig. 3
Fig. 3
Descriptive histological evaluation at day 90. a (Control group); red circle indicates osteotomized site filled by newly formed immature bone with little cellular reaction. b (QUI group); red circle shows implantation of chitosan surrounded by intense cellular reaction and dark circle shows newly formed bone tissue. c (TCP group); red circle indicates osteotomized site filled by newly formed bone during the organization process. d (TCP+QUI group); red circle indicates the osteotomy site filled by newly formed bone, in the organization process, and black arrows show cellular reaction. Obj. ×4 (Color figure online)
Fig. 4
Fig. 4
Mean and standard deviation of newly formed bone tissue (mm2) at the treatment interface. Treatments; C control, QUI chitosan only, QUI+TCP chitosan combined with tricalcium phosphate, TCP TCP only)

References

    1. Alghamdi HS, Bosco R, Van Den Beucken JJJP, Walboomers XF, Jansen JA. Osteogenicity of titanium implants coated with calcium phosphate or collagen type-I in osteoporotic rats. Biomaterials. 2013;34:3747–3757. doi: 10.1016/j.biomaterials.2013.02.033.
    1. Reis ECC, Borges APB, Araújo MVF, Mendes VC, Guan L, Davies JE. Periodontal regeneration using a bilayered PLGA/calcium phosphate construct. Biomaterials. 2011;32:9244–9253. doi: 10.1016/j.biomaterials.2011.08.040.
    1. Guastaldi AC, Aparecida AH. Fosfatos de cálcio de interesse biológico: importância como biomateriais, propriedades e métodos de obtenção de recobrimentos. Quimica Nova. 2010;33:1352–1358. doi: 10.1590/S0100-40422010000600025.
    1. Bohner M. Resorbable biomaterials as bone graft substitutes. Mater Today. 2010;13:24–30. doi: 10.1016/S1369-7021(10)70014-6.
    1. Khor E, Lim LY. Implantable applications of chitin and chitosan. Biomaterials. 2003;24:2339–2349. doi: 10.1016/S0142-9612(03)00026-7.
    1. Gorzelanny C, Pöppelmann B, Pappelbaum K, Moerschbacher BM, Schneider SW. Human macrophage activation triggered by chitotriosidase-mediated chitin and chitosan degradation. Biomaterials. 2010;31:8556–8563. doi: 10.1016/j.biomaterials.2010.07.100.
    1. Ueno H, Nakamura F, Murakami M, Okumura M, Kodosawa T, Fujinaga T. Evaluation effects of chitosan for the extracellular matrix production by fibroblasts and the growth factors production by macrophages. Biomaterials. 2001;22:2125–2130. doi: 10.1016/S0142-9612(00)00401-4.
    1. Peluso G, Petillo O, Ranieri M, Santin M, Ambrosia L, Calabró D, et al. Chitosan-mediated stimulation of macrophage function. Biomaterials. 1994;15:1215–1220. doi: 10.1016/0142-9612(94)90272-0.
    1. Azevedo AS, Sa MJC, Costa Neto PI, Fook MVL, Portela RA, Azevedo SS. Avaliação de diferentes proporções de fosfato de cálcio naregeneração do tecido ósseo de coelhos: estudo clínico-cirúrgico, radiológico e histológico. Braz J Vet Res Anim Sci. 2012;49:12–18.
    1. Ge Z, Baguenard S, Lim LY, Wee A, Khor E. Hydroxyapatite–chitin materials as potential tissue engineered bone substitutes. Biomaterials. 2004;25:1049–1058. doi: 10.1016/S0142-9612(03)00612-4.
    1. Zar JH. Biostatistical analysis. New Jersey: Prentice Hall; 1999.
    1. Adams JE. Quantitative computed tomography. Eur J Radiol. 2009;71:415–424. doi: 10.1016/j.ejrad.2009.04.074.
    1. Park YJ, Lee YM, Park SM, Shenn SY, Chung CP, Lee SJ. Platelet derived growth factor releasing chitosan sponge for periodontal bone regeneration. Biomaterials. 2000;21:153–159. doi: 10.1016/S0142-9612(99)00143-X.
    1. Ehara A, Ogata K, Imazato S, Ebisu S, Nakano T, Umakoshi Y. Effects of a-TCP and TetCP on MC3T3-E1 proliferation, differentiation and mineralization. Biomaterials. 2003;24:831–836. doi: 10.1016/S0142-9612(02)00411-8.
    1. Liu H, Peng H, Wu Y, Zhang C, Cai Y, Xu G, et al. The promotion of bone regeneration by nanofibrous hydroxyapatite/chitosan scaffolds by effects on integrin-BMP/Smad signaling pathway in BMSCs. Biomaterials. 2013;34:4404–4417. doi: 10.1016/j.biomaterials.2013.02.048.
    1. Erisken C, Kalyon DM, Wang H. Functionally graded electrospun polycaprolactone and beta-tricalcium phosphate nanocomposites for tissue engineering applications. Biomaterials. 2008;29:4065–4073. doi: 10.1016/j.biomaterials.2008.06.022.
    1. Lim H-P, Mercado-Pagan AE, Yun K-D, Kang S-S, Choi T-H, Bishop J, Koh J-T, Maloney W, Lee K-M, Yang YP, Park S-W. The effect of rhBMP-2 and PRP delivery by biodegradable β-tricalcium phosphate scaffolds on new bone formation in a non-through rabbit cranial defect model. J Mater Sci Mater Med. 2013;24:1895–1903. doi: 10.1007/s10856-013-4939-9.
    1. Frohbergh ME, Katsman A, Botta GP, Lazarovici P, Schauer CL, Wegst UG, et al. Electrospun hydroxyapatite-containing chitosan nanofibers cross linked with genipin for bone tissue engineering. Biomaterials. 2012;33:9167–9178. doi: 10.1016/j.biomaterials.2012.09.009.
    1. Azevedo AS, Sá MJC, Fook MVL, Nobrega Neto PI, Sousa OB, Azevedo SS. Hydroxiapatita e quitosana isoladas e associadas à medula óssea no reparo do tecido ósseo em coelhos. Ciência Rural. 2013;43:1265–1270. doi: 10.1590/S0103-84782013000700019.

Source: PubMed

Patrocinadores e Colaboradores

Condições médicas

Intervenções de drogas

3
Se inscrever