Effects of hydroxyapatite/collagen composite on osteogenic differentiation of rat bone marrow derived mesenchymal stem cells

Abstract

Combination of cells and materials opens a new option for tissue repair and regeneration. The aim of this study is to investigate the effects of hydroxyapatite/collagen composite on the osteogenic differentiation of rat bone marrow derived mesenchymal stem cells. The hydroxyapatite/collagen composites are synthesized by bioinspired mineralization and characterized by Fourier transform infrared spectrometry, X-ray diffraction, scanning electron microscope, thermogravimetric analysis and laser particle size analyzer. Different concentrations of hydroxyapatite/collagen are co-cultured with the rat bone marrow derived mesenchymal stem cells in passages 2–4 on a culture plate. Osteogenic differentiation is evaluated using reverse transcription polymerase chain reaction, alkaline phosphatase spectrophotometry as well as western blotting. The results demonstrate that the developed hydroxyapatite/collagen composite has microstructure and composition that are similar to the natural bone matrix. Hydroxyapatite/collagen treatment can induce osteogenic differentiation of rat bone marrow derived mesenchymal stem cells, as confirmed by the expression of osteoblast-related markers at both messenger RNA and protein levels. The concentration of 75 µg/ml may be the optimal inducer. In addition, combining biochemical reagents and hydroxyapatite/collagen has a synergistic interaction on the osteogenic differentiation of rat bone marrow derived mesenchymal stem cells. This provides a new avenue for mechanistic studies of stem cell differentiation and a novel approach to obtain more committed differentiated cells.

Keywords

Hydroxyapatite collagen mesenchymal stem cells differentiation osteoblasts

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References

Wang

. The in vivo bone formation by mesenchymal stem cells in zein scaffolds. Biomaterials 2009; 30: 4369–4376.

Park

Huang

Kurpinski

. Mechanobiology of mesenchymal stem cells and their use in cardiovascular repair. Front Biosci 2007; 12: 5098–5116.

Phillips

Petrie

Creighton

. Human mesenchymal stem cell differentiation on self-assembled monolayers presenting different surface chemistries. Acta Biomater 2010; 6: 12–20.

Pittenger

Mackay

Beck

. Multilineage potential of adult human mesenchymal stem cells. Science 1999; 284: 143–147.

Csaki

Matis

Mobasheri

. Co-culture of canine mesenchymal stem cells with primary bone-derived osteoblasts promotes osteogenic differentiation. Histochem Cell Biol 2009; 131: 251–266.

Heino

Hentunen

Väänänen

. Conditioned medium from osteocytes stimulates the proliferation of bone marrow mesenchymal stem cells and their differentiation into osteoblasts. Exp Cell Res 2004; 294: 458–468.

Locklin

Williamson

Beresford

. In vitro effects of growth factors and dexamethasone on rat marrow stromal cells. Clin Orthop Relat Res 1995; 313: 27–35.

Maegawa

Kawamura

Hirose

. Enhancement of osteoblastic differentiation of mesenchymal stromal cells cultured by selective combination of bone morphogenetic protein-2 (BMP-2) and fibroblast growth factor-2 (FGF-2). J Tissue Eng Regen Med 2007; 1: 306–313.

David

Martin

Lafage-Proust

. Mechanical loading down-regulates peroxisome proliferator-activated receptor gamma in bone marrow stromal cells and favors osteoblastogenesis at the expense of adipogenesis. Endocrinology 2007; 148: 2553–2562.

10.

Schwartz

Simon

Duran

. Pulsed electromagnetic fields enhance BMP-2 dependent osteoblastic differentiation of human mesenchymal stem cells. J Orthop Res 2008; 26: 1250–1255.

11.

Chai

Leong

. Biomaterials approach to expand and direct differentiation of stem cells. Mol Ther 2007; 15: 467–480.

12.

Engler

Sen

Sweeney

. Matrix elasticity directs stem cell lineage specification. Cell 2006; 126: 677–689.

13.

Huang

Zhou

Zheng

. Micro-/nano-sized hydroxyapatite directs differentiation of rat bone marrow derived mesenchymal stem cells towards an osteoblast lineage. Nanoscale 2012; 4: 2484–2490.

14.

Olszta

Cheng

Jee

. Bone structure and formation: a new perspective. Mat Sci Eng R: Rep 2007; 58: 77–116.

15.

Estevez

Escamilla

Vargas

. Synthesis and characterization of porous hybrid biomaterials with improved mechanical properties. J Compos Mater 2011; 46: 1831–1838.

16.

Ngiam

Liao

Patil

. The fabrication of nano-hydroxyapatite on PLGA and PLGA/collagen nanofibrous composite scaffolds and their effects in osteoblastic behavior for bone tissue engineering. Bone 2009; 45: 4–16.

17.

Niu

Feng

Wang

. Porous nano-HA/collagen/PLLA scaffold containing chitosan microspheres for controlled delivery of synthetic peptide derived from BMP-2. J Control Release 2009; 134: 111–117.

18.

Zhao

Huang

Jin

. A novel route for collagen/hydroxyapatite preparation by enzymatic decomposition of urea. J Compos Mater 2010; 44: 2127–2133.

19.

Van den Dolder

Jansen

. The response of osteoblast-like cells towards collagen type I coating immobilized by p-nitrophenylchloroformate to titanium. J Biomed Mater Res A 2007; 83: 712–719.

20.

Karageorgiou

Kaplan

. Porosity of 3D biomaterial scaffolds and osteogenesis. Biomaterials 2005; 26: 5474–5491.

21.

Murugan

Sampath Kumar

Yang

. Hydroxyl carbonateapatite hybrid bone composites using carbohydrate polymer. J Compos Mater 2005; 39: 1159–1167.

22.

Niu

Fan

Liu

. Repair of bone defect in femoral condyle using microencapsulated chitosan, nanohydroxyapatite/collagen and poly(L-lactide)-based microsphere-scaffold delivery system. Artif Organs 2011; 35: E119–E128.

23.

Huang

Jia

Bai

. Effect of fluid shear stress on cardiomyogenic differentiation of rat bone marrow mesenchymal stem cells. Arch Med Res 2010; 41: 497–505.

24.

Huang

Zheng

Gong

. Effect of cyclic strain on cardiomyogenic differentiation of rat bone marrow derived mesenchymal stem cells. PLoS One 2012; 7: e34960–e34960.

25.

Doyle

Bendit

Blout

. Infrared spectroscopy of collagen and collagen-like polypeptides. Biopolymers 1975; 14: 937–957.

26.

Camacho

West

Torzilli

. FTIR microscopic imaging of collagen and proteoglycan in bovine cartilage. Biopolymers 2001; 62: 1–8.

27.

Marinucci

Balloni

Becchetti

. Effects of hydroxyapatite and Biostite on osteogenic induction of hMSC. Ann Biomed Eng 2010; 38: 640–648.

28.

Stevens

Yang

Mohandas

. A review of materials, fabrication methods, and strategies used to enhance bone regeneration in engineered bone tissues. J Biomed Mater Res B Appl Biomater 2008; 85: 573–582.

29.

Prosecká

Rampichová

Vojtová

. Optimized conditions for mesenchymal stem cells to differentiate into osteoblasts on a collagen/hydroxyapatite matrix. J Biomed Mater Res A 2011; 99: 307–315.

30.

Ducheyne

Qiu

. Bioactive ceramics: the effect of surface reactivity on bone formation and bone cell function. Biomaterials 1999; 20: 2287–2303.

31.

Burg

Porter

Kellam

. Biomaterial developments for bone tissue engineering. Biomaterials 2000; 21: 2347–2359.

32.

Zayzafoon

Gathings

McDonald

. Modeled microgravity inhibits osteogenic differentiation of human mesenchymal stem cells and increases adipogenesis. Endocrinology 2004; 145: 2421–2432.

33.

Huang

Jin

Zhang

. In vitro and in vivo evaluation of akermanite bioceramics for bone regeneration. Biomaterials 2009; 30: 5041–5048.

34.

Zou

. Mechanical strain induces osteogenic differentiation: Cbfa1 and Ets-1 expression in stretched rat mesenchymal stem cells. Int J Oral Maxillofac Surg 2008; 37: 453–458.

35.

Schneider

Puellen

Kramann

. The osteogenic differentiation of adult bone marrow and perinatal umbilical mesenchymal stem cells and matrix remodelling in three-dimensional collagen scaffolds. Biomaterials 2010; 31: 467–480.

36.

Motskin

Wright

Muller

. Hydroxyapatite nano and microparticles: correlation of particle properties with cytotoxicity and biostability. Biomaterials 2009; 30: 3307–3317.

37.

Johnson

Frierson

Keller

. Porous ceramics as bone graft substitutes in long bone defects: a biomechanical, histological, and radiographic analysis. J Orthop Res 1996; 14: 351–369.