In vitro osteogenic differentiation of HOS cells on two types of collagen gels

Takitoh, Takako; Kato, Yoichi; Nakasu, Asako; Tadokoro, Mika; Bessho, Masahiko; Hirose, Motohiro; Ohgushi, Hajime; Mori, Hideki; Hara, Masayuki

doi:10.1016/j.jbiosc.2010.04.009

Cited by 13 publications

(13 citation statements)

References 13 publications

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“…28 They compared conventional collagen gel (neutral gel) and gammacrosslinked collagen gel without collagen fibrils (acidic gel). Compared with their experiments, we used conventional culture dishes and non-crosslinked tilapia collagen membrane with and without fibril formation.…”

Section: Discussionmentioning

confidence: 99%

Rapid oriented fibril formation of fish scale collagen facilitates early osteoblastic differentiation of human mesenchymal stem cells

Matsumoto

Uemura

et al. 2014

J Biomedical Materials Res

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We studied the effect of fibril formation of fish scale collagen on the osteoblastic differentiation of human mesenchymal stem cells (hMSCs). We found that hMSCs adhered easily to tilapia scale collagen, which remarkably accelerated the early stage of osteoblastic differentiation in hMSCs during in vitro cell culture. Osteoblastic markers such as ALP activity, osteopontin, and bone morphogenetic protein 2 were markedly upregulated when the hMSCs were cultured on a tilapia collagen surface, especially in the early osteoblastic differentiation stage. We hypothesized that this phenomenon occurs due to specific fibril formation of tilapia collagen. Thus, we examined the time course of collagen fibril formation using high-speed atomic force microscopy. Moreover, to elucidate the effect of the orientation of fibril formation on the differentiation of hMSCs, we measured ALP activity of hMSCs cultured on two types of tilapia scale collagen membranes with different degrees of fibril formation. The ALP activity in hMSCs cultured on a fibrous collagen membrane was significantly higher than on a non-fibrous collagen membrane even before adding osteoblastic differentiation medium. These results showed that the degree of the fibril formation of tilapia collagen was essential for the osteoblastic differentiation of hMSCs.

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Section: Discussionmentioning

confidence: 99%

Rapid oriented fibril formation of fish scale collagen facilitates early osteoblastic differentiation of human mesenchymal stem cells

Matsumoto

Uemura

et al. 2014

J Biomedical Materials Res

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“…At earlier time points, at 2, 4 or 6 h after seeding, the levels of osteoblast gene expression were similar in cells growing in 2D and 3D collagen cultures (data not shown). In a number of previous studies, 3,8,14,15 collagen gels were formed first and osteoblasts were seeded on top of the gels, allowing for the analysis of cell migration into the gels. Interestingly, while osteoblasts migrated into the gels and formed a 3D cell network, fibroblasts seeded on top of the gels migrated through them and formed a monolayer on the plastic underneath.…”

Section: Enhanced Osteoblastogenesis In 3d Gelsmentioning

confidence: 99%

“…4 Although a number of studies have investigated osteoblasts cultured in 3D collagen gels, they mostly used microscopy for visual analysis of the osteoblast phenotype and alkaline phosphatase activity as a measure of osteoblast function. 1,3,[7][8][9] The aim of the current study was to directly compare osteoblasts growing in parallel in 3D collagen gels and in 2D on plastic and to comprehensively characterize the effects of these growth environments on the expression of osteoblast marker genes, on cell differentiation and on responses to factors that affect the rate of cell proliferation.…”

Section: Introductionmentioning

confidence: 99%

Enhanced osteoblastogenesis in three-dimensional collagen gels

et al. 2014

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Growth and differentiation of osteoblasts are often studied in cell cultures. In vivo, however, osteoblasts are embedded within a complex three-dimensional (3D) microenvironment, which bears little relation to standard culture flasks. Our study characterizes osteoblast-like cells cultured in 3D collagen gels and compares them with cells in two-dimensional (2D) cultures. Primary rat osteoblasts and MC3T3-E1 cells were seeded within type I collagen gels, and differentiation was determined by mineral staining and gene expression analysis. Cells growing in 3D gels showed positive mineral staining and induction of osteoblast marker genes earlier than cells growing in 2D. A number of genes, including osteocalcin, bone sialoprotein, alkaline phosphatase and dentin matrix protein 1, were already highly upregulated in 3D cultures 24 h after seeding. The early expression of osteoblast genes was dependent on the 3D structure and was not induced in cells growing on collagen-coated dishes in 2D. Comparison of thymidine incorporation between cells in 3D and 2D cultures treated with agents that induce proliferation-transforming growth factor b, platelet-derived growth factor and lactoferrin-showed a much greater response in 3D gels. Cells in 3D cultures were also much more sensitive to inhibition of proliferation by the protein kinase inhibitor imatinib mesylate. The 3D collagen gels better represent the physiological bone environment and offer a number of technical advantages for the study of osteoblasts in vitro. These studies have additional practical implications as 3D collagen gels are considered as a scaffold material in regenerative medicine for the repair of bone defects.

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“…dense collagen scaffolds at 40 mg/mL, was compared to that of the same cells seeded on loose collagen matrices at 3 mg/mL, classically used in previous experiments [12], [13]. Macroscopically, the 40 mg/mL collagen matrices are opaque and rigid.…”

Section: Resultsmentioning

confidence: 99%

“…However they remained far from in vivo conditions. In addition, cell culture scaffolds used in cell biology are sponges [11], hydrogels [12], [13], cements [14], or demineralized bone matrix [15], [16].…”

Section: Introductionmentioning

confidence: 99%

Collagen Osteoid-Like Model Allows Kinetic Gene Expression Studies of Non-Collagenous Proteins in Relation with Mineral Development to Understand Bone Biomineralization

et al. 2013

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Among persisting questions on bone calcification, a major one is the link between protein expression and mineral deposition. A cell culture system is here proposed opening new integrative studies on biomineralization, improving our knowledge on the role played by non-collagenous proteins in bone. This experimental in vitro model consisted in human primary osteoblasts cultured for 60 days at the surface of a 3D collagen scaffold mimicking an osteoid matrix. Various techniques were used to analyze the results at the cellular and molecular level (adhesion and viability tests, histology and electron microscopy, RT- and qPCR) and to characterize the mineral phase (histological staining, EDX, ATG, SAED and RMN). On long term cultures human bone cells seeded on the osteoid-like matrix displayed a clear osteoblast phenotype as revealed by the osteoblast-like morphology, expression of specific protein such as alkaline phosphatase and expression of eight genes classically considered as osteoblast markers, including BGLAP, COL1A1, and BMP2. Von Kossa and alizarine red allowed us to identify divalent calcium ions at the surface of the matrix, EDX revealed the correct Ca/P ratio, and SAED showed the apatite crystal diffraction pattern. In addition RMN led to the conclusion that contaminant phases were absent and that the hydration state of the mineral was similar to fresh bone. A temporal correlation was established between quantified gene expression of DMP1 and IBSP, and the presence of hydroxyapatite, confirming the contribution of these proteins to the mineralization process. In parallel a difference was observed in the expression pattern of SPP1 and BGLAP, which questioned their attributed role in the literature. The present model opens new experimental possibilities to study spatio-temporal relations between bone cells, dense collagen scaffolds, NCPs and hydroxyapatite mineral deposition. It also emphasizes the importance of high collagen density environment in bone cell physiology.

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In vitro osteogenic differentiation of HOS cells on two types of collagen gels

Cited by 13 publications

References 13 publications

Rapid oriented fibril formation of fish scale collagen facilitates early osteoblastic differentiation of human mesenchymal stem cells

Rapid oriented fibril formation of fish scale collagen facilitates early osteoblastic differentiation of human mesenchymal stem cells

Enhanced osteoblastogenesis in three-dimensional collagen gels

Collagen Osteoid-Like Model Allows Kinetic Gene Expression Studies of Non-Collagenous Proteins in Relation with Mineral Development to Understand Bone Biomineralization

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