In vitro osteogenic potential of human bone marrow stromal cells cultivated in porous scaffolds from mineralized collagen

Abstract

Porous 3D structures from mineralized collagen were fabricated applying a procedure in which collagen fibril reassembly and precipitation of nanocrystalline hydroxyapatite (HA) occur simultaneously. The resulting matrices were evaluated in vitro with respect to their suitability as scaffolds for bone tissue engineering. We found a high capacity of the material to bind serum proteins as well as to absorb Ca2+ ions, which could be advantageous to promote cell attachment, growth, and differentiation. Human bone marrow stromal cells (hBMSCs) were seeded onto the 3D scaffolds and cultivated for 4 weeks in the presence and absence of osteogenic supplements. We studied viability, proliferation, and osteogenic differentiation in terms of total lactate dehydrogenase (LDH) activity, DNA content, and alkaline phosphatase (ALP) activity. Furthermore, the expression for bone-related genes (ALP, bone sialo protein II (BSP II), and osteocalcin) was analyzed. In our investigation we found a 2.5-fold to 5-fold raise in DNA content and an increase of ALP activity for osteogenic induced hBMSC on collagen HA scaffolds. The expression of ALP and BSP II in these cells was also stimulated in the course of cultivation; however, we did not detect an upregulation of osteocalcin gene expression. These data suggest, that porous collagen HA scaffolds are suitable for the expansion and osteogenic differentiation of hBMSC and are therefore promising candidates for application as bone grafts.

Heparin modification of a biomimetic bone matrix modulates osteogenic and angiogenic cell response in vitro

“…The utilisation of the regenerative potential of stem cells in combination with an engineered extracellular matrix (scaffold) and stimulating biological factors is intended to enhance bone formation and hence meet currently unsolved clinical problems (Black et al, 2015). One of the aspects addressed by research is the development of suitable scaffold matrices constituting the physical microenvironment for cells.…”

“…The natural extracellular matrix (ECM) does not only act as structure for cell anchorage but can also actively provide signalling cues to direct cellular processes, act as reservoir for growth factors and modulate their activity (Rosso et al, 2004;Chan and Leong, 2008). Mimicking the ECM is a common strategy of TE; regarding bone tissue, scaffolds composed of calcium phosphates and collagen type I as well as composites thereof have been proven capable of assisting bone regeneration (Szpalski et al, 2012;Black et al, 2015;Pina et al, 2015;Yunus Basha et al, 2015).…”

“…3D scaffolds with an interconnecting porosity suitable for cell colonisation can be fabricated by freeze-drying of the mineralised collagen suspension and chemical crosslinking (Gelinsky et al, 2008). In vitro studies revealed that this nanocomposite is able to support proliferation and osteogenic differentiation of human bone marrow-derived stromal cells (hBMSC) (Bernhardt et al, 2009). Resorption of the material and new bone formation were observed in vivo (Yokoyama et al, 2005;Scholz et al, 2010).…”

Cell-Biomaterial Interactions Reproducing a Niche

“…In this context, cells are considered as a key element to achieve the regeneration of the target tissue, since very few biomaterials are osteoinductive by themselves (Goshima, Goldberg et al 1991;Ohgushi, Dohi et al 1993;Boo, Yamada et al 2002;Cancedda, Bianchi et al 2003;Endres, Hutmacher et al 2003;Livingston, Gordon et al 2003;Derubeis and Cancedda 2004;Warren, Nacamuli et al 2004;Arinzeh, Tran et al 2005;Kimelman, Pelled et al 2006;Bernhardt, Lode et al 2009;Matsushima, Kotobuki et al 2009). The most intriguing concept in modern biomaterials is thus obtaining materials able to mimic a specific eventually pre-existing microenvironment and, therefore, inducing stem/progenitor cells to differentiate in a predetermined manner and to regenerate by themselves the bone tissue according to physiological pathways.…”

“…In this context, the bioengineering challenge become ambitious, since the complex cellbiomaterial interaction moves on multiple spatial and temporal scales. The microenvironmental cues, such as chemical environmental variables, are able to stimulate specific cellular responses at the molecular level already at early time points (Goshima, Goldberg et al 1991;Ohgushi, Dohi et al 1993;Fabbri, Celotti et al 1995;Kon, Muraglia et al 2000;Erbe, Marx et al 2001;Endres, Hutmacher et al 2003;Kasten, Luginbuhl et al 2003;Livingston, Gordon et al 2003;Niemeyer, Krause et al 2003;Arinzeh, Tran et al 2005;Kotobuki, Ioku et al 2005;Kondo, Ogose et al 2006;Fan, Ikoma et al 2007;Mygind, Stiehler et al 2007;Gigante, Manzotti et al 2008;Ng, Tan et al 2008;Bernhardt, Lode et al 2009;Saldana, Sanchez-Salcedo et al 2009). For example, ceramic scaffolds (i.e.…”