Calcium phosphate fibers coated with collagen: In vivo evaluation of the effects on bone repair

Abstract

The aim of this study was to assess the characteristics of the CaP/Col composites, in powder and fiber form, via scanning electron microscopy (SEM), pH and calcium release evaluation after immersion in SBF and to evaluate the performance of these materials on the bone repair process in a tibial bone defect model. For this, four different formulations (CaP powder – CaPp, CaP powder with collagen – CaPp/Col, CaP fibers – CaPf and CaP fibers with collagen – CaPf/Col) were developed. SEM images indicated that both material forms were successfully coated with collagen and that CaPp and CaPf presented HCA precursor crystals on their surface. Although presenting different forms, FTIR analysis indicated that CaPp and CaPf maintained the characteristic peaks for this class of material. Additionally, the calcium assay study demonstrated a higher Ca uptake for CaPp compared to CaPf for up to 5 days. Furthermore, pH measurements revealed that the collagen coating prevented the acidification of the medium, leading to higher pH values for CaPp/Col and CaPf/Col. The histological analysis showed that CaPf/Col demonstrated a higher amount of newly formed bone in the region of the defect and a reduced presence of material. In summary, the results indicated that the fibrous CaP enriched with the organic part (collagen) glassy scaffold presented good degradability and bone-forming properties and also supported Runx2 and RANKL expression. These results show that the present CaP/Col fibrous composite may be used as a bone graft for inducing bone repair.

Keywords

Composite calcium phosphate collagen fibers bone repair

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References

Hutmacher

D.W.

Schantz

J.T.

Lam

C.X.

Tan

K.C.

and Lim

T.C.

, State of the art and future directions of scaffold-based bone engineering from a biomaterials perspective, Journal of Tissue Engineering and Regenerative Medicine 1(4) (2007), 245–260. doi:10.1002/term.24.

Amini

A.R.

Laurencin

C.T.

and Nukavarapu

S.P.

, Bone tissue engineering: Recent advances and challenges, Critical Reviews in Biomedical Engineering 40(5) (2012), 363–408. doi:10.1615/CritRevBiomedEng.v40.i5.10.

Mesquita

Gomes Pde

Sampaio

Juodzbalys

Afonso

and Fernandes

M.H.

, Surface properties and osteoblastic cytocompatibility of two blasted and acid-etched titanium implant systems with distinct microtopography, Journal of Oral & Maxillofacial Research 3(1) (2012), e4.

Pek

Y.S.

Gao

Arshad

M.S.

Leck

K.J.

and Ying

J.Y.

, Porous collagen-apatite nanocomposite foams as bone regeneration scaffolds, Biomaterials 29(32) (2008), 4300–4305. doi:10.1016/j.biomaterials.2008.07.030.

Hench

L.L.

and David

, Interactions between bioactive glass and collagen: A review and new perspectives, Journal of The Autralian Ceramic Society 49(2) (2013), 1–40.

Renno

A.C.

van de Watering

F.C.

Nejadnik

M.R.

Crovace

M.C.

Zanotto

E.D.

Wolke

J.G.

et al., Incorporation of bioactive glass in calcium phosphate cement: An evaluation, Acta Biomaterialia 9(3) (2013), 5728–5739. doi:10.1016/j.actbio.2012.11.009.

Renno

A.C.M.

Nejadnik

M.R.

van de Watering

F.C.J.

Crovace

M.C.

Zanotto

E.D.

Hoefnagels

J.P.M.

et al., Incorporation of bioactive glass in calcium phosphate cement: Material characterization and in vitro degradation, Journal of Biomedical Materials Research Part A 101A(8) (2013), 2365–2373. doi:10.1002/jbm.a.34531.

Granito

R.N.

Ribeiro

D.A.

Renno

A.C.

Ravagnani

Bossini

P.S.

Peitl-Filho

et al., Effects of biosilicate and bioglass 45S5 on tibial bone consolidation on rats: A biomechanical and a histological study, Journal of Materials Science Materials in Medicine 20(12) (2009), 2521–2526. doi:10.1007/s10856-009-3824-z.

Valerio

Pereira

M.M.

Goes

A.M.

and Leite

M.F.

, The effect of ionic products from bioactive glass dissolution on osteoblast proliferation and collagen production, Biomaterials 25(15) (2004), 2941–2948. doi:10.1016/j.biomaterials.2003.09.086.

10.

Stevens

Yang

Mohandas

Stucker

and Nguyen

K.T.

, A review of materials, fabrication methods, and strategies used to enhance bone regeneration in engineered bone tissues, Journal of Biomedical Materials Research Part B, Applied Biomaterials 85(2) (2008), 573–582. doi:10.1002/jbm.b.30962.

11.

Lee

K.Y.

Park

Kim

H.M.

Lim

Y.J.

Chun

H.J.

Kim

et al., Ceramic bioactivity: Progresses, challenges and perspectives, Biomedical Materials 1(2) (2006), R31–R37. doi:10.1088/1748-6041/1/2/R01.

12.

Habraken

W.J.

Wolke

J.G.

Mikos

A.G.

and Jansen

J.A.

, Injectable PLGA microsphere/calcium phosphate cements: Physical properties and degradation characteristics, Journal of Biomaterials Science Polymer Edition 17(9) (2006), 1057–1074. doi:10.1163/156856206778366004.

13.

Bodde

E.W.

Habraken

W.J.

Mikos

A.G.

Spauwen

P.H.

and Jansen

J.A.

, Effect of polymer molecular weight on the bone biological activity of biodegradable polymer/calcium phosphate cement composites, Tissue Engineering Part A 15(10) (2009), 3183–3191. doi:10.1089/ten.tea.2008.0694.

14.

Maté Sánchez de Val

J.E.

Calvo Guirado

J.L.

Ramírez Fernández

M.P.

Delgado Ruiz

R.A.

Mazón

and De Aza

P.N.

, In vivo behavior of hydroxyapatite/β-TCP/collagen scaffold in animal model. Histological, histomorphometrical, radiological, and SEM analysis at 15, 30, and 60 days, Clinical Oral Implants Research (2015), to appear. doi:10.1111/clr.12656.

15.

Marelli

Ghezzi

C.E.

Mohn

Stark

W.J.

Barralet

J.E.

Boccaccini

A.R.

et al., Accelerated mineralization of dense collagen-nano bioactive glass hybrid gels increases scaffold stiffness and regulates osteoblastic function, Biomaterials 32(34) (2011), 8915–8926. doi:10.1016/j.biomaterials.2011.08.016.

16.

Mastrogiacomo

Scaglione

Martinetti

Dolcini

Beltrame

Cancedda

et al., Role of scaffold internal structure on in vivo bone formation in macroporous calcium phosphate bioceramics, Biomaterials 27(17) (2006), 3230–3237. doi:10.1016/j.biomaterials.2006.01.031.

17.

Spector

, Biomaterials-based tissue engineering and regenerative medicine solutions to musculoskeletal problems, Swiss Medical Weekly 136(19–20) (2006), 293–301.

18.

Zhu

Chang

Zhang

and Xiao

, Bioactive inorganic-materials/alginate composite microspheres with controllable drug-delivery ability, Journal of Biomedical Materials Research Part B, Applied Biomaterials 94(1) (2010), 32–43.

19.

Boccaccini

A.R.

and Maquet

, Bioresorbable and bioactive polymer/bioglass^® composites with tailored pore structure for tissue engineering applications, Composites Science and Technology 63(16) (2003), 2417–2429. doi:10.1016/S0266-3538(03)00275-6.

20.

Sachot

Castano

Mateos-Timoneda

M.A.

Engel

and Planell

J.A.

, Hierarchically engineered fibrous scaffolds for bone regeneration, Journal of the Royal Society, Interface/The Royal Society 10(88) (2013), 20130684. doi:10.1098/rsif.2013.0684.

21.

Pascu

E.I.

Stokes

and McGuinness

G.B.

, Electrospun composites of PHBV, silk fibroin and nano-hydroxyapatite for bone tissue engineering, Materials Science & Engineering C, Materials for Biological Applications 33(8) (2013), 4905–4916. doi:10.1016/j.msec.2013.08.012.

22.

Souza

M.T.

Zanotto

E.D.

and Peitl-Filho

, Inventor Vitreous composition, bioactive glassy fibers and tissues. Patent Application No. BR 10 2013 020961 9. Universidade Federal de São Carlos, Brazil, 2013.

23.

Brangule

and Gross

K.A.

, Importance of FTIR spectra deconvolution for the analysis of amorphous calcium phosphates, Materials Science and Engineering 77 (2015), 012027.

24.

Beniash

Metzler

R.A.

Lam

R.S.

and Gilbert

P.U.

, Transient amorphous calcium phosphate in forming enamel, Journal of Structural Biology 166(2) (2009), 133–143. doi:10.1016/j.jsb.2009.02.001.

25.

Silva

A.C.

Aparecida

A.H.

and Braga

F.J.C.

, Dispersed hydroxyapatite Bioglass 45S5 composites: Comparative evaluation of the use of bovine bone and synthetic hydroxyapatite, Materials Science Forum 727 (2012), 1147–1152. doi:10.4028/www.scientific.net/MSF.727-728.1147.

26.

Friedrich

and Fritz

, Apparatus and method for production of fibers from glass, slag, and the like meltable materials. Google Patents; 1941.

27.

Braga

Silva

A.C.

Allegrini

Jr. and Ottoni

(eds), Calcium phosphate graft substitute: When the impact of innovation is in the form rather than content, in: 26th Annual Conference of the European Society of Biomaterials, Liverpool, 2014.

28.

Navarro

J.M.F.

, El Vidrio. Madrid: Consejo Superior de Invest. Científicas – Fundación Centro Nacional del Vidrio, 2003.

29.

Kokubo

and Takadama

, How useful is SBF in predicting in vivo bone bioactivity?, Biomaterials 27(15) (2006), 2907–2915. doi:10.1016/j.biomaterials.2006.01.017.

30.

Mooren

R.E.

Hendriks

E.J.

van den Beucken

J.J.

Merkx

M.A.

Meijer

G.J.

Jansen

J.A.

et al., The effect of platelet-rich plasma in vitro on primary cells: Rat osteoblast-like cells and human endothelial cells, Tissue Engineering Part A 16(10) (2010), 3159–3172. doi:10.1089/ten.tea.2009.0832.

31.

Oliveira

Ribeiro

D.A.

Pipi

E.F.

Driusso

Parizotto

N.A.

and Renno

A.C.

, Low level laser therapy does not modulate the outcomes of a highly bioactive glass-ceramic (Biosilicate) on bone consolidation in rats, Journal of Materials Science Materials in Medicine 21(4) (2010), 1379–1384. doi:10.1007/s10856-009-3945-4.

32.

Granito

R.N.

Renno

A.C.

Ravagnani

Bossini

P.S.

Mochiuti

Jorgetti

et al., In vivo biological performance of a novel highly bioactive glass-ceramic (Biosilicate^®): A biomechanical and histomorphometric study in rat tibial defects, Journal of Biomedical Materials Research Part B, Applied Biomaterials 97(1) (2011), 139–147. doi:10.1002/jbm.b.31795.

33.

Hench

L.L.

and Polak

J.M.

, Third-generation biomedical materials, Science 295(5557) (2002), 1014–1017.

34.

Gauthier

Muller

von Stechow

Lamy

Weiss

Bouler

J.M.

et al., In vivo bone regeneration with injectable calcium phosphate biomaterial: A three-dimensional micro-computed tomographic, biomechanical and SEM study, Biomaterials 26(27) (2005), 5444–5453. doi:10.1016/j.biomaterials.2005.01.072.

35.

Link

D.P.

van den Dolder

van den Beucken

J.J.

Cuijpers

V.M.

Wolke

J.G.

Mikos

A.G.

et al., Evaluation of the biocompatibility of calcium phosphate cement/PLGA microparticle composites, Journal of Biomedical Materials Research Part A 87(3) (2008), 760–769. doi:10.1002/jbm.a.31831.

36.

Wise

D.L.

Trantolo

D.J.

Altobelli

D.E.

Yaszemski

M.J.

Gresser

J.D.

and Schwartz

E.R.

, Encyclopedic Handbook of Biomaterials and Bioengineering. Part B: Applications, Marcel Dekker, Inc., New York, 1995.

37.

Urquia Edreira

E.R.

Wolke

J.G.

Jansen

J.A.

and van den Beucken

J.J.

, Influence of ceramic disk material, surface hemispheres, and SBF volume on in vitro mineralization, Journal of Biomedical Materials Research Part A 103(8) (2015), 2740–2746. doi:10.1002/jbm.a.35406.

38.

Dominique

G.P.

, Biomechanics and Biomaterials in Orthopedics, Springer-Verlag, London, 2004.

39.

Taylor

A.C.

, Responses of cells to pH changes in the medium, The Journal of Cell Biology 15 (1962), 201–209. doi:10.1083/jcb.15.2.201.

40.

Ruhé

Hedberg

Padron

N.T.

Spauwen

Jansen

and Mikos

, Biocompatibility and degradation of poly(DL-lactic-co-glycolic acid)/calcium phosphate cement composites, J. Biomed. Mater. Res. A 74A(4) (2005), 533–544. doi:10.1002/jbm.a.30341.

41.

and Wang

, Improved injectability and in vitro degradation of a calcium phosphate cement containing poly(lactide-co-glycolide) microspheres, Acta Biomater. 4(6) (2008), 1837–1845. doi:10.1016/j.actbio.2008.05.009.

42.

van de Watering

F.C.J.

van den Beucken

J.J.J.P.

Walboomers

X.F.

and Jansen

J.A.

, Calcium phosphate/poly(d,l-lactic-co-glycolic acid) composite bone substitute materials: Evaluation of temporal degradation and bone ingrowth in a rat critical-sized cranial defect, Clin. Oral. Implants Res. 23(2) (2012), 151–159. doi:10.1111/j.1600-0501.2011.02218.x.

43.

Zhou

Z.Q.

D.P.

Liang

W.G.

Wang

and Zhu

Z.Z.

, Preparation and characterization of a novel injectable strontium-containing calcium phosphate cement with collagen, Chinese Journal of Traumatology (Zhonghua chuang shang za zhi/Chinese Medical Association) 18(1) (2015), 33–38.

44.

Gabbai-Armelin

P.R.

Souza

M.T.

Kido

H.W.

Tim

C.R.

Bossini

P.S.

Magri

A.M.

et al., Effect of a new bioactive fibrous glassy scaffold on bone repair, Journal of Materials Science Materials in Medicine 26(5) (2015), 177. doi:10.1007/s10856-015-5516-1.

45.

Afzal

Polak

and Buttery

, Endothelial nitric oxide synthase in the control of osteoblastic mineralizing activity and bone integrity, The Journal of Pathology 202(4) (2004), 503–510. doi:10.1002/path.1536.

46.

Zhang

Schwarz

E.M.

Young

D.A.

Puzas

J.E.

Rosier

R.N.

and O’Keefe

R.J.

, Cyclooxygenase-2 regulates mesenchymal cell differentiation into the osteoblast lineage and is critically involved in bone repair, The Journal of Clinical Investigation 109(11) (2002), 1405–1415. doi:10.1172/JCI0215681.

47.

Lemaire

Tobin

F.L.

Greller

L.D.

Cho

C.R.

and Suva

L.J.

, Modeling the interactions between osteoblast and osteoclast activities in bone remodeling, Journal of Theoretical Biology 229(3) (2004), 293–309. doi:10.1016/j.jtbi.2004.03.023.

48.

Kearns

A.E.

Khosla

and Kostenuik

P.J.

, Receptor activator of nuclear factor kappaB ligand and osteoprotegerin regulation of bone remodeling in health and disease, Endocrine Reviews 29(2) (2008), 155–192. doi:10.1210/er.2007-0014.

49.

Anandarajah

A.P.

, Role of RANKL in bone diseases, Trends in Endocrinology and Metabolism: TEM 20(2) (2009), 88–94. doi:10.1016/j.tem.2008.10.007.

50.

Pinto

K.N.

Tim

C.R.

Crovace

M.C.

Matsumoto

M.A.

Parizotto

N.A.

Zanotto

E.D.

et al., Effects of biosilicate^® scaffolds and low-level laser therapy on the process of bone healing, Photomedicine and Laser Surgery 31(6) (2013), 252–260. doi:10.1089/pho.2012.3435.

51.

Kondo

Ogose

Tokunaga

Ito

Arai

Kudo

et al., Bone formation and resorption of highly purified beta-tricalcium phosphate in the rat femoral condyle, Biomaterials 26(28) (2005), 5600–5608. doi:10.1016/j.biomaterials.2005.02.026.