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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: 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.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.

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

Statistical, histomorphometric evaluation of the von-Kossa stained samples. The area of mineralized matrix is referenced to the respective reference sample for each condition. (For PEI-25k-Mal-B p = 0.163, for PEI-5k-Mal-B p = 0.511)
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Fig12: Statistical, histomorphometric evaluation of the von-Kossa stained samples. The area of mineralized matrix is referenced to the respective reference sample for each condition. (For PEI-25k-Mal-B p = 0.163, for PEI-5k-Mal-B p = 0.511)

Mentions: Despite the long-term effects of cationic core–shell glycoarchitectures on cell viability and proliferation, the differentiation of rdMSC to osteoblasts is not influenced by the nanoparticles (Fig. 5). In opposite to (slightly) lower cell number in the presence of nanoparticles (Fig. 5a), the ALP content of the cells (Fig. 5b) and the production of mineralized matrix (Figs. 6, 12) in the absence and presence of nanoparticles do not differ statistically significant. Further studies if (oligo-)maltose-modified-PEI nanoparticles with higher molecular weights influence osteogenesis in a positive way, as it might be suggested by the histomorphometric evaluation of the von-Kossa stained samples, might have a chance of success if we first succeed in further improvement of biocompatibility. Certainly none of the used (oligo-)maltose-modified-PEI nanoparticles in our case influences the capability of osteogenic differentiation in a negative way. This behavior has been reported for other nanoparticle-systems, for example, using low concentrations of Ag nanoparticles [52]. Again, only the further study by light microscopy will give a clear key feature of both core–shell glycoarchitectures on differentiated rdMSC (Fig. 10). Only in the case of PEI-25k-Mal-B cell morphology is clearly influenced showing the presence of cell debris. This enables us to conclude that the cell function after the differentiation of rdMSC to osteoblasts, checked by the ALP content, is not the deciding experiment (Fig. 5b) to do final conclusion on the long-term biological action of both core–shell glycoarchitectectures. Only the combination of light microscopic study (Fig. 10) and determination of DC-protein measurement (Fig. 5a) and ALP determination (Fig. 5b) gives us a deeper view on the biological action of both nanoparticles where the smaller PEI-5k-Mal-B is the most promising core–shell glycoarchitecture in these experiment series. With this in mind, one may postulate that the core–shell glycoarchitecture PEI-5k-Mal-B with the smaller PEI-5k core can be used as drug delivery system for treating bone disease in locally applied therapy.Fig. 12


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)

Statistical, histomorphometric evaluation of the von-Kossa stained samples. The area of mineralized matrix is referenced to the respective reference sample for each condition. (For PEI-25k-Mal-B p = 0.163, for PEI-5k-Mal-B p = 0.511)
© Copyright Policy - OpenAccess
Related In: Results  -  Collection

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

Fig12: Statistical, histomorphometric evaluation of the von-Kossa stained samples. The area of mineralized matrix is referenced to the respective reference sample for each condition. (For PEI-25k-Mal-B p = 0.163, for PEI-5k-Mal-B p = 0.511)
Mentions: Despite the long-term effects of cationic core–shell glycoarchitectures on cell viability and proliferation, the differentiation of rdMSC to osteoblasts is not influenced by the nanoparticles (Fig. 5). In opposite to (slightly) lower cell number in the presence of nanoparticles (Fig. 5a), the ALP content of the cells (Fig. 5b) and the production of mineralized matrix (Figs. 6, 12) in the absence and presence of nanoparticles do not differ statistically significant. Further studies if (oligo-)maltose-modified-PEI nanoparticles with higher molecular weights influence osteogenesis in a positive way, as it might be suggested by the histomorphometric evaluation of the von-Kossa stained samples, might have a chance of success if we first succeed in further improvement of biocompatibility. Certainly none of the used (oligo-)maltose-modified-PEI nanoparticles in our case influences the capability of osteogenic differentiation in a negative way. This behavior has been reported for other nanoparticle-systems, for example, using low concentrations of Ag nanoparticles [52]. Again, only the further study by light microscopy will give a clear key feature of both core–shell glycoarchitectures on differentiated rdMSC (Fig. 10). Only in the case of PEI-25k-Mal-B cell morphology is clearly influenced showing the presence of cell debris. This enables us to conclude that the cell function after the differentiation of rdMSC to osteoblasts, checked by the ALP content, is not the deciding experiment (Fig. 5b) to do final conclusion on the long-term biological action of both core–shell glycoarchitectectures. Only the combination of light microscopic study (Fig. 10) and determination of DC-protein measurement (Fig. 5a) and ALP determination (Fig. 5b) gives us a deeper view on the biological action of both nanoparticles where the smaller PEI-5k-Mal-B is the most promising core–shell glycoarchitecture in these experiment series. With this in mind, one may postulate that the core–shell glycoarchitecture PEI-5k-Mal-B with the smaller PEI-5k core can be used as drug delivery system for treating bone disease in locally applied therapy.Fig. 12

Bottom Line: 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.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.

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