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.

Combined Effects of Mechanical Strain and Hydroxyapatite/Collagen Composite on Osteogenic Differentiation of Rat Bone Marrow Derived Mesenchymal Stem Cells

“…The effects of HA/Col composite on the osteogenic differentiation of rBMSCs have been investigated in our previous study, and the results exhibit that the optimal concentration of HA/Col is 75 g/mL [20]. The cells of mechanical strain group in this study are subjected to a cyclic uniaxial stretch (0.5 Hz, 5% elongation).…”

“…Gene Forward primer Reverse primer ALP 5 -CATGTTCCTGGGAGATGGTA-3 5 -GTGTTGTACGTCTTGGAGAGA-3 cbfa1 5 -GCCGGGAATGATGAGAACTA-3 5 -GGACCGTCCACTGTCACTTT-3 COL1A1 5 -TTACTACCGGGCCGATGA-3 5 -CTGCGGATGTTCTCAATCTG-3 GAPDH 5 -TGTTCCTACCCCCAATGTATCCG-3 5 -TGCTTCACCACCTTCTTGATGTCAT-3 an important influence on the adaptation, remodeling, or reconstruction of tissues and cells [14][15][16][17]. Our group and other researchers have showed that hydroxyapatite/collagen (HA/Col) composite could induce the osteogenic differentiation of rat bone marrow derived MSCs (rBMSCs) [18][19][20]. These studies suggest an interesting biomaterial approach to affect the osteogenic differentiation of MSCs.…”

Amorphous calcium phosphate (ACP) in tissue repair process

“…The apatite is the inorganic phase in the bone matrix, configuring and providing natural bone with its mechanical properties. This bone apatite is predominantly composed of HA, (Huang et al, ; Zhou et al, ) yet its raw material comes from amorphous calcium phosphate (ACP) (Boonrungsiman et al, ; Liu et al, ), considered a more stable precursor of carbonated HA, which is the main mineral phase of mineralized tissues, found in the sites during initial mineral deposition of bone and dental tissues (Beniash, Metzler, Lam, & Gilbert, ; Mahamid, Sharir, Addadi, & Weiner, ).…”

Scaffold combination of chitosan and collagen synthesized from chicken feet induces osteoblast and osteoprotegerin expression in bone healing process of mice

“…Treatments for tissue and bone defects incorporating tissue engineering methods, such as the use of bone graft and stem cells, have been developed as an alternative to conventional defect treatments. 4 In recent decades, treatments involving the use of grafts have represented a novel approach to tissue and bone repair. Tissue engineering methods primarily intended for hard tissue repair not only rely on natural osteointegrative processes, but also on a material promoting osteointegration which is the bone graft.…”