Limits...
Effects of dendritic core-shell glycoarchitectures on primary mesenchymal stem cells and osteoblasts obtained from different human donors.

Lautenschläger S, Striegler C, Dakischew O, Schütz I, Szalay G, Schnettler R, Heiß C, Appelhans D, Lips KS - J Nanobiotechnology (2015)

Bottom Line: As well dose as incubation-time dependent effects and interactions have been researched for concentrations between 1 μg/ml to 1 mg/ml and periods of 24 h up to 28 days.In all experiments PEI-5k-Mal-B exhibits a superior biocompatibility compared to PEI-25k-Mal-B.Additionally, for all experiments, results are strongly influenced by a large donor-to-donor variability of the four different rdMSC samples.

View Article: PubMed Central - PubMed

Affiliation: Laboratory for Experimental Trauma Surgery, Justus-Liebig-University Giessen, Schubertstr. 81, 35392, Giessen, Germany. Stefan.Lautenschlaeger@med.uni-giessen.de.

ABSTRACT
The biological impact of novel nano-scaled drug delivery vehicles in highly topical therapies of bone diseases have to be investigated in vitro before starting in vivo trials. Highly desired features for these materials are a good cellular uptake, large transport capacity for drugs and a good bio-compatibility. Essentially the latter has to be addressed as first point on the agenda. We present a study on the biological interaction of maltose-modified poly(ethyleneimine) (PEI-Mal) on primary human mesenchymal stem cell, harvested from reaming debris (rdMSC) and osteoblasts obtained from four different male donors. PEI-Mal-nanoparticles with two different molecular weights of the PEI core (5000 g/mol for PEI-5k-Mal-B and 25,000 g/mol for PEI-25k-Mal-B) have been administered to both cell lines. As well dose as incubation-time dependent effects and interactions have been researched for concentrations between 1 μg/ml to 1 mg/ml and periods of 24 h up to 28 days. Studies conducted by different methods of microscopy as light microscopy, fluorescence microscopy, transmission-electron-microscopy and quantitative assays (LDH and DC-protein) indicate as well a good cellular uptake of the nanoparticles as a particle- and concentration-dependent impact on the cellular macro- and micro-structure of the rdMSC samples. In all experiments PEI-5k-Mal-B exhibits a superior biocompatibility compared to PEI-25k-Mal-B. At higher concentrations PEI-25k-Mal-B is toxic and induces a directly observable mitochondrial damage. The alkaline phosphatase assay (ALP), has been conducted to check on the possible influence of nanoparticles on the differentiation capabilities of rdMSC to osteoblasts. In addition the production of mineralized matrix has been shown by von-Kossa stained samples. No influence of the nanoparticles on the ALP per cell has been detected. Additionally, for all experiments, results are strongly influenced by a large donor-to-donor variability of the four different rdMSC samples. To summarize, while featuring a good cellular uptake, PEI-5k-Mal-B induces only minimal adverse effects and features clearly superior biocompatibility compared to the larger PEI-25k-Mal-B.

No MeSH data available.


Related in: MedlinePlus

CLSM images of cell cultures treated with rhodamin B marked nanoparticles (green). The nucleus has been DAPI dyed (blue), the actin filaments were stained with phalloidin coupled TRITS (red). a Displays untreated reference, only the nucleus and the actin-filaments can be observed. b Rhodamin B labeled Rh-PEI-5k-Mal-B nanoparticles, surrounding the nucleus. c The rhodamin-marked nanoparticles Rh-PEI-25k-Mal-B feature a perinuclear distribution, in this sample the nucleus has not been stained
© Copyright Policy - OpenAccess
Related In: Results  -  Collection

License 1 - License 2
getmorefigures.php?uid=PMC4597403&req=5

Fig7: CLSM images of cell cultures treated with rhodamin B marked nanoparticles (green). The nucleus has been DAPI dyed (blue), the actin filaments were stained with phalloidin coupled TRITS (red). a Displays untreated reference, only the nucleus and the actin-filaments can be observed. b Rhodamin B labeled Rh-PEI-5k-Mal-B nanoparticles, surrounding the nucleus. c The rhodamin-marked nanoparticles Rh-PEI-25k-Mal-B feature a perinuclear distribution, in this sample the nucleus has not been stained

Mentions: To proof the cellular uptake of PEI-5k-Mal-B and PEI-25k-Mal-B by CLSM, both were labeled by Rhodamine B (Rh-PEI-5k-Mal-B or Rh-PEI-25k-Mal-B, green) and administrated to rdMSC for 24 h with concentrations of 0.1 mg/ml. The nucleus and the actin filaments were stained with either DAPI (blue) or Phalloidin coupled TRITS (red). The reference (Fig. 7a) shows no signs of nanoparticles. As shown in Fig. 7b, sample has been treated with Rh-PEI-5k-Mal-B where the nanoparticles feature a peri-nuclear distribution. For samples incubated with Rh-PEI-25k-Mal-B (Fig. 7c), again a peri-nuclear particle distribution can be found, in this case the nucleus has not been stained (Fig. 7c).Fig. 7


Effects of dendritic core-shell glycoarchitectures on primary mesenchymal stem cells and osteoblasts obtained from different human donors.

Lautenschläger S, Striegler C, Dakischew O, Schütz I, Szalay G, Schnettler R, Heiß C, Appelhans D, Lips KS - J Nanobiotechnology (2015)

CLSM images of cell cultures treated with rhodamin B marked nanoparticles (green). The nucleus has been DAPI dyed (blue), the actin filaments were stained with phalloidin coupled TRITS (red). a Displays untreated reference, only the nucleus and the actin-filaments can be observed. b Rhodamin B labeled Rh-PEI-5k-Mal-B nanoparticles, surrounding the nucleus. c The rhodamin-marked nanoparticles Rh-PEI-25k-Mal-B feature a perinuclear distribution, in this sample the nucleus has not been stained
© Copyright Policy - OpenAccess
Related In: Results  -  Collection

License 1 - License 2
Show All Figures
getmorefigures.php?uid=PMC4597403&req=5

Fig7: CLSM images of cell cultures treated with rhodamin B marked nanoparticles (green). The nucleus has been DAPI dyed (blue), the actin filaments were stained with phalloidin coupled TRITS (red). a Displays untreated reference, only the nucleus and the actin-filaments can be observed. b Rhodamin B labeled Rh-PEI-5k-Mal-B nanoparticles, surrounding the nucleus. c The rhodamin-marked nanoparticles Rh-PEI-25k-Mal-B feature a perinuclear distribution, in this sample the nucleus has not been stained
Mentions: To proof the cellular uptake of PEI-5k-Mal-B and PEI-25k-Mal-B by CLSM, both were labeled by Rhodamine B (Rh-PEI-5k-Mal-B or Rh-PEI-25k-Mal-B, green) and administrated to rdMSC for 24 h with concentrations of 0.1 mg/ml. The nucleus and the actin filaments were stained with either DAPI (blue) or Phalloidin coupled TRITS (red). The reference (Fig. 7a) shows no signs of nanoparticles. As shown in Fig. 7b, sample has been treated with Rh-PEI-5k-Mal-B where the nanoparticles feature a peri-nuclear distribution. For samples incubated with Rh-PEI-25k-Mal-B (Fig. 7c), again a peri-nuclear particle distribution can be found, in this case the nucleus has not been stained (Fig. 7c).Fig. 7

Bottom Line: As well dose as incubation-time dependent effects and interactions have been researched for concentrations between 1 μg/ml to 1 mg/ml and periods of 24 h up to 28 days.In all experiments PEI-5k-Mal-B exhibits a superior biocompatibility compared to PEI-25k-Mal-B.Additionally, for all experiments, results are strongly influenced by a large donor-to-donor variability of the four different rdMSC samples.

View Article: PubMed Central - PubMed

Affiliation: Laboratory for Experimental Trauma Surgery, Justus-Liebig-University Giessen, Schubertstr. 81, 35392, Giessen, Germany. Stefan.Lautenschlaeger@med.uni-giessen.de.

ABSTRACT
The biological impact of novel nano-scaled drug delivery vehicles in highly topical therapies of bone diseases have to be investigated in vitro before starting in vivo trials. Highly desired features for these materials are a good cellular uptake, large transport capacity for drugs and a good bio-compatibility. Essentially the latter has to be addressed as first point on the agenda. We present a study on the biological interaction of maltose-modified poly(ethyleneimine) (PEI-Mal) on primary human mesenchymal stem cell, harvested from reaming debris (rdMSC) and osteoblasts obtained from four different male donors. PEI-Mal-nanoparticles with two different molecular weights of the PEI core (5000 g/mol for PEI-5k-Mal-B and 25,000 g/mol for PEI-25k-Mal-B) have been administered to both cell lines. As well dose as incubation-time dependent effects and interactions have been researched for concentrations between 1 μg/ml to 1 mg/ml and periods of 24 h up to 28 days. Studies conducted by different methods of microscopy as light microscopy, fluorescence microscopy, transmission-electron-microscopy and quantitative assays (LDH and DC-protein) indicate as well a good cellular uptake of the nanoparticles as a particle- and concentration-dependent impact on the cellular macro- and micro-structure of the rdMSC samples. In all experiments PEI-5k-Mal-B exhibits a superior biocompatibility compared to PEI-25k-Mal-B. At higher concentrations PEI-25k-Mal-B is toxic and induces a directly observable mitochondrial damage. The alkaline phosphatase assay (ALP), has been conducted to check on the possible influence of nanoparticles on the differentiation capabilities of rdMSC to osteoblasts. In addition the production of mineralized matrix has been shown by von-Kossa stained samples. No influence of the nanoparticles on the ALP per cell has been detected. Additionally, for all experiments, results are strongly influenced by a large donor-to-donor variability of the four different rdMSC samples. To summarize, while featuring a good cellular uptake, PEI-5k-Mal-B induces only minimal adverse effects and features clearly superior biocompatibility compared to the larger PEI-25k-Mal-B.

No MeSH data available.


Related in: MedlinePlus