Characterization of a PLLA-Collagen I Blend Nanofiber Scaffold with Respect to Growth and Osteogenic Differentiation of Human Mesenchymal Stem Cells

Schofer, Markus D.; Boudriot, Ulrich; Leifeld, Irini; Sütterlin, Romina I.; Rudisile, Markus; Wendorff, Joachim H.; Greiner, Andreas; Paletta, Jürgen R. J.; Fuchs-Winkelmann, Susanne

doi:10.1100/tsw.2009.13

Cited by 43 publications

(53 citation statements)

References 36 publications

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“…As described for hMSC [22,47] we found no differences in cell densities of TDF's. This finding was accompanied by a normalisation of CCND expression.…”

Section: Discussionsupporting

confidence: 78%

Influence of nanofibers on growth and gene expression of human tendon derived fibroblast

Theisen

Fuchs-Winkelmann

Knappstein

et al. 2010

BioMedical Engineering OnLine

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BackgroundRotator cuff tears are a common and frequent lesion especially in older patients. The mechanisms of tendon repair are not fully understood. Common therapy options for tendon repair include mini-open or arthroscopic surgery. The use of growth factors in experimental studies is mentioned in the literature. Nanofiber scaffolds, which provide several criteria for the healing process, might be a suitable therapy option for operative treatment. The aim of this study was to explore the effects of nanofiber scaffolds on human tendon derived fibroblasts (TDF's), as well as the gene expression and matrix deposition of these fibroblasts.MethodsNanofibers composed of PLLA and PLLA/Col-I were seeded with human tendon derived fibroblasts and cultivated over a period of 22 days under growth-inductive conditions, and analyzed during the course of culture, with respect to gene expression of different extra cellular matrix components such as collagens, bigylcan and decorin. Furthermore, we measured cell densities and proliferation by using fluorescene microscopy.ResultsPLLA nanofibers possessed a growth inhibitory effect on TDF's. Furthermore, no meaningful influence on the gene expression of collagen I, collagen III and decorin could be observed, while the expression of collagen X increased during the course of cultivation. On the other hand, PLLA/Col-I blend nanofibers had no negative influence on the growth of TDF's. Furthermore, blending PLLA nanofibers with collagen had a positive effect on the gene expression of collagen I, III, X and decorin. Here, gene expression indicated that focal adherence kinases might be involved.ConclusionThis study indicates that the use of nanofibers influence expression of genes associated with the extra cellular matrix formation. The composition of the nanofibers plays a critical role. While PLLA/Col-I blend nanofibers enhance the collagen I and III formation, their expression on PLLA nanofibers was more comparable to controls. However, irrespective of the chemical composition of the fibres, the collagen deposition was altered, an effect which might be associated with a decreased expression of biglycanes.

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“…As described for hMSC [22,47] we found no differences in cell densities of TDF's. This finding was accompanied by a normalisation of CCND expression.…”

Section: Discussionsupporting

confidence: 78%

Influence of nanofibers on growth and gene expression of human tendon derived fibroblast

Theisen

Fuchs-Winkelmann

Knappstein

et al. 2010

BioMedical Engineering OnLine

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“…Recently, non-woven PLLA-and PLLA-based nanofibers have been produced by electrospinning. [13][14][15][16] The high surface area of these nanofibers permit enhanced cell adhesion and proliferation and thus making them a suitable for biomedical application. Other applications of PLLA are in agriculture for mulch films, coatings for paper and cardboard, and fibers for clothing, carpets, sheets and towels, and wall coverings.…”

Section: Introductionmentioning

confidence: 99%

Structural Properties and Enzymatic Degradation Behavior of PLLA and Stereocomplexed PLA Nanofibers

Fundador

Takemura

Iwata

2010

Macro Materials & Eng

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PLLA and stereocomplexed polylactide (sc‐PLA) nanofibers were formed by electrospinning solutions of the polymers in HFIP. A highly semi‐crystalline sc‐PLA nanofiber having only sc crystallites was confirmed by WAXD analysis. The diameters of the nanofibers of both polymers decreased slightly when they were annealed at 60 °C, which was near Tg. Enzyme degradation of both as‐spun PLLA and sc‐PLA nanofibers by proteinase K from Tritirachium album was carried out. The rate of degradation of the nanofibers can be controlled by varying annealing conditions, hence the extent of crystallinity.magnified image

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“…Extraction of RNA, synthesis of cDNA, as well as quantitative polymerase chain reaction (qPCR) analysis were performed as described earlier [17,18,19,20]. Primers (purchased at TIB Biomol, Berlin, Germany), cycle temperatures, and incubation times for VEGF [23], BMP-2, and 18s rRNA [24,25,26] were previously detailed.…”

Section: Gene Expression Analysismentioning

confidence: 99%

“…The preparation of PLLA nanofibers and the incorporation of BMP-2 into the nanofibers, as well as their characterization by scanning electron microscopy (SEM) and analysis of static contact angles of water, has been reported earlier [17,18,19,20].…”

Section: Construction Of Nanofibers and Characterizationmentioning

confidence: 99%

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Lack of Obvious Influence of PLLA Nanofibers on the Gene Expression of BMP-2 and VEGF during Growth and Differentiation of Human Mesenchymal Stem Cells

Schofer

Fuchs-Winkelmann²,

Wack³

et al. 2009

The Scientific World JOURNAL

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Growth factors like bone morphogenetic protein 2 (BMP-2) and vascular endothelial growth factor (VEGF) play an important role in bone remodeling and fracture repair. Therefore, with respect to tissue engineering, an artificial graft should have no negative impact on the expression of these factors. In this context, the aim of this study was to analyze the impact of poly(L-lactic acid) (PLLA) nanofibers on VEGF and BMP-2 gene expression during the time course of human mesenchymal stem cell (hMSC) differentiation towards osteoblasts. PLLA matrices were seeded with hMSCs and cultivated over a period of 22 days under growth and osteoinductive conditions, and analyzed during the course of culture, with respect to gene expression of VEGF and BMP-2. Furthermore, BMP-2-enwoven PLLA nanofibers were used in order to elucidate whether initial down-regulation of growth factor expression could be compensated. Although there was a great interpatient variability with respect to the expression of VEGF and BMP-2, PLLA nanofibers tend to result in a down-regulation in BMP-2 expression during the early phase of cultivation. This effect was diminished in the case of VEGF gene expression. The initial down-regulation was overcome when BMP-2 was directly incorporated into the PLLA nanofibers by electrospinning. Furthermore, the incorporation of BMP-2 into the PLLA nanofibers resulted in an increase in VEGF gene expression. Summarized, the results indicate that the PLLA nanofibers have little effect on growth factor production. An enhancement in gene expression of BMP-2 and VEGF can be achieved by an incorporation of BMP-2 into the PLLA nanofibers.KEYWORDS: nanofibers, tissue engineering, human mesenchymal stem cells, PLLA, BMP, VEGF Schofer et al.: Growth Factor Gene Expession in hMSCs TheScientificWorldJOURNAL (2009) 9, 313-319 314 INTRODUCTIONThe reconstruction of large bony defects after injury or tumor resection often requires the use of graft material. Besides autologous bone grafts, artificial scaffolds based on synthetic biomaterials such as metals, ceramics, polymers, and composites have been developed [1,2]. Among them, scaffolds based on nanofibers are becoming more and more important [3,4]. These nanofibers can be produced by a broad spectrum of polymers, including biocompatible and biodegradable polymers, such as poly(glycolic acid) (PGA), poly(L-lactic acid) (PLLA), poly(ε caprolactone) (PCL), polyurethanes, polyphosphazenes, collagen, gelatin, and chitosan, as well as copolymers from the corresponding monomers in various compositions [5,6]. This results in the production of a broad spectrum of nanofiber-based scaffolds with different mechanical and biophysical properties. These fibers allow the differentiation of human mesenchymal stem cells (hMSCs) towards osteoblasts in principal [7,8,9]. Furthermore, they can be supplemented with growth factors like bone morphogenetic protein (BMP) or vascular endothelial growth factor (VEGF) in order to enhance osteoinductivity or angiogenesis.BMP-2 is a well-studied member of ...

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Characterization of a PLLA-Collagen I Blend Nanofiber Scaffold with Respect to Growth and Osteogenic Differentiation of Human Mesenchymal Stem Cells

Cited by 43 publications

References 36 publications

Influence of nanofibers on growth and gene expression of human tendon derived fibroblast

Influence of nanofibers on growth and gene expression of human tendon derived fibroblast

Structural Properties and Enzymatic Degradation Behavior of PLLA and Stereocomplexed PLA Nanofibers

Lack of Obvious Influence of PLLA Nanofibers on the Gene Expression of BMP-2 and VEGF during Growth and Differentiation of Human Mesenchymal Stem Cells

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