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Migration of intervertebral disc cells into dense collagen scaffolds intended for functional replacement.

Bron JL, Mulder HW, Vonk LA, Doulabi BZ, Oudhoff MJ, Smit TH - J Mater Sci Mater Med (2012)

Bottom Line: The aim of current study was to assess whether NP and surrounding annulus fibrosus (AF) cells are capable of migrating into dense collagen scaffolds.We seeded freshly harvested caprine NP and AF cells onto scaffolds consisting of 1.5 and 3.0% type I collagen matrices, prepared by plastic compression, to assess cell invasion.The migration distance appeared both time and density dependent and was higher for NP (25%) compared to AF (10%) cells after 4 weeks.

View Article: PubMed Central - PubMed

Affiliation: Department of Orthopaedic Surgery, VU University Medical Center, Amsterdam, The Netherlands. jl.bron@vumc.nl

ABSTRACT
Invasion of cells from surrounding tissues is a crucial step for regeneration when using a-cellular scaffolds as a replacement of the nucleus pulposus (NP). The aim of current study was to assess whether NP and surrounding annulus fibrosus (AF) cells are capable of migrating into dense collagen scaffolds. We seeded freshly harvested caprine NP and AF cells onto scaffolds consisting of 1.5 and 3.0% type I collagen matrices, prepared by plastic compression, to assess cell invasion. The migration distance appeared both time and density dependent and was higher for NP (25%) compared to AF (10%) cells after 4 weeks. Migration distance was not enhanced by Hst-2, a peptide derived from saliva known to enhance fibroblast migration, and this was confirmed in a scratch assay. In conclusion, we revealed invasion of cells into dense collagen scaffolds and therewith encouraging first steps towards the use of a-cellular scaffolds for NP replacement.

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A confirmation scratch assay with human squamous carcinoma HO-1-N-1 cells reveals a significant enhanced cell migration after the addition of Hst-2
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Fig10: A confirmation scratch assay with human squamous carcinoma HO-1-N-1 cells reveals a significant enhanced cell migration after the addition of Hst-2

Mentions: The scratches had a mean width of 666 μm (+/− 77). No statistically significant differences between AF and NP cells were observed in any of the assays (Fig. 8). The addition of Hst-2 to the medium had no significant effect on scratch closure compared to the negative control (D-Hst-2) or serum free medium. The addition of 10% serum to the medium resulted in complete closure of the scratch in both, assays with NP and AF cells (7). Staining of the F-actin filaments, to visualize the morphology of the cells, revealed different cell shapes for AF and NP cells (Fig. 9). In addition, differences were found between cells inside and outside the scratch. Non-migrated NP cells outside the scratch had a round, chondrocytic, morphology (Fig. 9a), while NP cells that had migrated into the scratch were characterized by long dendritic processes (Fig. 9b) Cells of the AF outside the scratch had a more flattened, fibroblast like, appearance (Fig. 9c), whereas cells inside the scratch were more elongated and aligned with each other (Fig. 9d). To confirm Hst-2 activity, a scratch assay was performed with human oral mucosa derived cells (HO-1-N-1 cell line), known to be sensitive for Hst-2 stimulation [24]. This assay showed a significant increased scratch closure after addition of Hst-2 (Fig. 10).Fig. 8


Migration of intervertebral disc cells into dense collagen scaffolds intended for functional replacement.

Bron JL, Mulder HW, Vonk LA, Doulabi BZ, Oudhoff MJ, Smit TH - J Mater Sci Mater Med (2012)

A confirmation scratch assay with human squamous carcinoma HO-1-N-1 cells reveals a significant enhanced cell migration after the addition of Hst-2
© Copyright Policy
Related In: Results  -  Collection

Show All Figures
getmorefigures.php?uid=PMC3299969&req=5

Fig10: A confirmation scratch assay with human squamous carcinoma HO-1-N-1 cells reveals a significant enhanced cell migration after the addition of Hst-2
Mentions: The scratches had a mean width of 666 μm (+/− 77). No statistically significant differences between AF and NP cells were observed in any of the assays (Fig. 8). The addition of Hst-2 to the medium had no significant effect on scratch closure compared to the negative control (D-Hst-2) or serum free medium. The addition of 10% serum to the medium resulted in complete closure of the scratch in both, assays with NP and AF cells (7). Staining of the F-actin filaments, to visualize the morphology of the cells, revealed different cell shapes for AF and NP cells (Fig. 9). In addition, differences were found between cells inside and outside the scratch. Non-migrated NP cells outside the scratch had a round, chondrocytic, morphology (Fig. 9a), while NP cells that had migrated into the scratch were characterized by long dendritic processes (Fig. 9b) Cells of the AF outside the scratch had a more flattened, fibroblast like, appearance (Fig. 9c), whereas cells inside the scratch were more elongated and aligned with each other (Fig. 9d). To confirm Hst-2 activity, a scratch assay was performed with human oral mucosa derived cells (HO-1-N-1 cell line), known to be sensitive for Hst-2 stimulation [24]. This assay showed a significant increased scratch closure after addition of Hst-2 (Fig. 10).Fig. 8

Bottom Line: The aim of current study was to assess whether NP and surrounding annulus fibrosus (AF) cells are capable of migrating into dense collagen scaffolds.We seeded freshly harvested caprine NP and AF cells onto scaffolds consisting of 1.5 and 3.0% type I collagen matrices, prepared by plastic compression, to assess cell invasion.The migration distance appeared both time and density dependent and was higher for NP (25%) compared to AF (10%) cells after 4 weeks.

View Article: PubMed Central - PubMed

Affiliation: Department of Orthopaedic Surgery, VU University Medical Center, Amsterdam, The Netherlands. jl.bron@vumc.nl

ABSTRACT
Invasion of cells from surrounding tissues is a crucial step for regeneration when using a-cellular scaffolds as a replacement of the nucleus pulposus (NP). The aim of current study was to assess whether NP and surrounding annulus fibrosus (AF) cells are capable of migrating into dense collagen scaffolds. We seeded freshly harvested caprine NP and AF cells onto scaffolds consisting of 1.5 and 3.0% type I collagen matrices, prepared by plastic compression, to assess cell invasion. The migration distance appeared both time and density dependent and was higher for NP (25%) compared to AF (10%) cells after 4 weeks. Migration distance was not enhanced by Hst-2, a peptide derived from saliva known to enhance fibroblast migration, and this was confirmed in a scratch assay. In conclusion, we revealed invasion of cells into dense collagen scaffolds and therewith encouraging first steps towards the use of a-cellular scaffolds for NP replacement.

Show MeSH
Related in: MedlinePlus