The main function of mesenchymal stem cells in the body is to facilitate the restoration and regeneration of damaged tissues. They are known for the ability to differentiate into tissue originating from the mesoderm, which among others includes connective tissues. Due to this feature are MSCs being intensively examined. Different directions of differentiation can be induced by treatment of specific polypeptides, so called growth factors. In the field of tissue engineering are growth factors used to induce and accelerate the healing processes. They may be incorporated into the nanofiber carrier which is inserted into the site of injury. Cells in this area would thus be stimulated by surrounding 3D microenvironment. At the same time the scaffold provides a supply of growth factors which are able to affect metabolism, motility and differentiation of present cells. In order to induce osteogenic differentiation of human MSCs the following bioactive substances were used: TGF-β, bFGF, HGF, IGF-1, VEGF and the BMP-2 and the organic acid taurine. During 21 days lasting experiments, were these molecules added to the medium in various combinations and in the case of taurine also at two different concentrations. Cells were cultured on plastic. The best effect on cellular metabolism of MSCs, evaluated by MTS...

Tissue engineering is a promising approach that combines the interactions of biomaterials, cells, and growth factors to stimulate tissue growth and regeneration. As such, selecting a suitable biomaterial is vital to the success of the procedure. Ideally, the material should show similarity to the extracellular matrix in the structure and relative stiffness, and biofunctionality beside others to provide a comfortable environment for the cells. Additionally, the biomaterial properties should allow for the effective diffusion of relevant growth factors and nutrients throughout the material to enable cell growth. Because peptides are composed of amino acids found naturally within the human body, they are considered non-toxic and biocompatible. Ultrashort peptides are peptides with three to seven amino acids that can be self-assembled into helical fibers forming scaffolds of supramolecular structures. These peptide hydrogels formed a highly porous network of nanofibers which can quickly solidify into nanofibrous hydrogels that resemble the extracellular matrix (ECM) and provide a 3D environment for cells with suitable mechanical properties. Furthermore, we can easily tune the stiffness of these peptide hydrogels by just increasing peptide concentration, thus providing a wide range of peptide hydrogels with different stiffness for 3D cell culture applications. Herein we describe the use of ultrashort peptide hydrogels for the maintenance and the differentiation of human mesenchymal stem cells into the osteogenic lineage. Furthermore, we develop a three dimensional (3D) biomimicry acute myeloid leukemia (AML) disease model using biomaterial from a tetramer ultrashort self-assembling peptide. In addition, we evaluate the potential application of peptide hydrogels as a hemostatic agent. The results presented in this study suggest that our biomimetic ultrashort tetrapeptide hydrogels are an excellent candidate for tissue engineering and biomedical applications.

ABSTRAKT:Isolering och karaktärisering av mesenkymala stamceller från periodontalligamentet hos friskatänderSYFTE: Att isolera och odla celler från periodontalligamentet samt karaktärisera dem sommesenkymala stamceller.MATERIAL OCH METOD: Friska premolarer gjordes tillgängliga vid ortodontiskaextraktioner. Den mellersta 1/3 av periodontalligamentet skrapades varpå en enzymatiskmetod användes för isolering av individuella celler. Resulterande celler odlades understandardiserade metoder. Karaktärisering av celler skedde genom flödescymetri med 2 olikapaneler av cellyta markörer; en för etablerat positiva uttryck och en för kända negativauttryck hos mesenkymala stamceller. Möjlighet av celler att differentieras in vitro tilladipocyter och osteocyter testades genom tillförsel av specifika substanser till odlingsmediet.RESULTAT: Celler från 11 av 13 tänder isolerades och odlades framgångsrikt adherenta tillodlingsytan i upp till 8 generationer. Celluttryck av de positiva markörerna CD73, CD90 samtCD44 bekräftades genom flödescymetri. Inget uttryck observerades för den negativa panelenCD45, CD34, CD11b, CD19 eller HLA class II. Uttrycket av CD105 kunde inte fastställas pgaofullständigt data. Försök till differentiering av celler till adipocyter och osteocyter visade påfenotypiska förändringar efter 21 dagar.SLUTSATS: Den här studien har bidragit till framgångsrik isolering och delvis karaktäriseringav mesenkymala stamceller från periodontalligamentet hos friska tänder. En icke-invasivmetod av detta slag, resulterande i tillgång till denna cellpopulation utgör ett lovande verktygför framtida studier med goda möjligheter till ytterligare kunskap applicerbart till kliniskasituationer inom tandvården. / ABSTRACT:Isolation and Characterization of Mesenchymal Stem Cells from the Periodontal Ligament ofHealthy TeethAIM: To isolate and culture viable cells from the periodontal ligament and confirming theiridentity as mesenchymal stem cells.METHODS AND MATERIALS: Healthy premolars were collected at the time oforthodontic extractions. The middle 1/3 of the periodontal ligament was scraped andsubsequent cell isolation was performed using an enzymatic method; yielding single cellisolates. Cells were cultured and maintained under standard culture conditions. Cellcharacterization was performed by flow cytometry using two sets of cell surface markers; oneknown to be present and one known to be absent in mesenchymal stem cells. Ability of thecells for in vitro differentiation into adipogenic and osteogenic lineages was tested usingspecifically formulated media supplements.RESULTS: Cells were successfully isolated from 11 of 13 teeth and were maintained asadherent cultures for up to 8 generations. Cellular expression of positive markers; CD73, CD90and CD44 were confirmed by flow cytometry. For the negative marker panel, expression ofCD45, CD34, CD11b, CD19 and HLA class II were not detectable. The expression of CD105was inconclusive. As determined by phenotypic changes, cells appeared to have undergoneadipogenic and osteocytic differentiation at 21 days.CONCLUSION: This study has resulted in successful isolation and partial characterization ofmesenchymal stem cells from the periodontal ligament of healthy teeth. Non-invasive accessto these cells, provides an excellent tool for future studies, potentially leading to beneficialknowledge transferable to the dental clinical situation.

Submitted by Sandro Camargo (sandro.camargo@unipampa.edu.br) on 2015-05-07T22:56:27Z No. of bitstreams: 1 126110041.pdf: 1662239 bytes, checksum: 4c1751ff15692d571713c45fca07289c (MD5) / Made available in DSpace on 2015-05-07T22:56:27Z (GMT). No. of bitstreams: 1 126110041.pdf: 1662239 bytes, checksum: 4c1751ff15692d571713c45fca07289c (MD5) Previous issue date: 2014-03-24 / Células tronco são células indiferenciadas e multipotentes, capazes de se diferenciar nos mais diversos tipos celulares. São definidas por duas características principais, a autorrenovação e o potencial de diferenciação. Devido ao seu potencial de diferenciação, facilidade de isolamento e expansão em cultura, as células tronco mesenquimais (Mesenchymal Stem Cell – MSCs) são largamente utilizadas em testes para o tratamento de inúmeras doenças, como doenças cardíacas, mieloma e osteoartrite. O corticosteroide dexametasona é amplamente utilizado como indutor de diferenciação óssea em células tronco e está envolvido no aumento da atividade da enzima fosfatase alcalina (ALP). Há quatro genes que codificam a ALP: ALPL, ALPP, ALPPL2 e ALPI. A ALPL é um dos principais marcadores de diferenciação óssea promovida pelo tratamento com dexametasona, esse gene possui três diferentes variantes que são geradas por splicing alternativo. Porém, até hoje não se tem conhecimento de qual dessas variantes atua na deposição de matriz óssea. O objetivo do trabalho foi analisar, por PCR quantitativo (qPCR), a atividade das variantes de ALPL durante a diferenciação de MSCs tratadas com dexametasona. Os primers das três variantes do gene ALPL foram desenhados pelo nosso grupo de pesquisa. Para o experimento, aproximadamente 3x10 4 células por mL foram semeadas em placa de cultura. Para o tratamento utilizou-se dexametasona 10 -7 M, tendo como período de tratamento 0, 3 e 7 dias. Os resultados da qPCR mostraram que os primers desenhados possuem uma eficiência superior a 90% e R 2 = 1, sendo os primers da variante 2 mais eficiente que os primers da variante 1, e os primers de ambas as variantes mais eficientes que os primers de ALP atualmente utilizados. Os primers da ALPL/v3 não apresentaram amplificação. Nossa análise estatística indica que a diferença de expressão entre as células tratadas e os controles se mostrou relevante, apontando para um grande aumento de ALPL/v1 nas células tratadas. A expressão de ALPL/v2, apesar de significativa entre os controles e tratados, é baixa quando comparada à ALPL/v1. Os resultados obtidos neste trabalho indicam que a dexametasona, além de aumentar a atividade do promotor de ALPL, também dá preferência ao splicing alternativo que origina o mRNA da variante 1 do gene. / Stem cells are undifferentiated and pluripotent cells that are able to differentiate into several cell types. Stem cells are distinguished from other cell types by two important characteristics: self-renewal and differentiation potential. Due to their differentiation potential and easiness for isolate and growth in vitro, mesenchymal stem cells (MSCs) are widely used in trials for the treatment of numerous diseases such as heart disease, myeloma and osteoarthritis. The corticosteroid dexamethasone is broadly used to induce bone differentiation in stem cells and is involved in the increased activity of alkaline phosphatase (ALPL). There are four genes coding ALP: ALPL, ALPP, ALPPL2 and ALPI. The ALPL is one of the main markers for bone differentiation stimulated by dexamethasone. This gene has three different variants which are generated by alternative splicing. However, to date it is not known which of these variants act in the bone matrix deposition. In this context, the aim of this work was to analyze the activity of ALPL variants during the differentiation of MSCs treated with dexamethasone. Primers for the three ALPL variants were designed by our group. For the experiment, approximately 3x10 4 cells per ml were seeded in culture plate. For treatment, we used 10 -7 M dexamethasone for 0, 3 and 7 days. The qPCR results showed that the primers designed had an amplification efficiency higher than 90% and R 2 = 1; variant 2 (V2) primers was more efficient than the variant 1 (V1) primers and primers of both variants are more efficient than the currently used ALP primers. The primers of ALPL/v3 showed no amplification. Our statistical data showed that the differences in expression between treated and control cells were significant, indicating an expressive increase of ALPL/v1 in treated cells. The expression of ALPL/v2, although significant between controls and treated cells, was low when compared to ALPL/v1. The results of this study indicate that dexamethasone, besides increasing the activity of the ALPL promoter, gives preference to alternative splicing that originates the variant1 mRNA.

La myélofibrose primitive (MFP) est un néoplasme myéloprolifératif chromosome Philadelphie négatif rare, mais de pronostic sévère. Elle se caractérise par une prolifération clonale et une mobilisation des cellules souches et progéniteurs hématopoïétiques (CSH/PH) de la moelle osseuse vers la rate et le foie. Cette anomalie de l’hématopoïèse est associée à une pathologie du stroma (myélofibrose, ostéosclérose et néoangiogenèse). L’existence d’anomalies moléculaires de la CSH/PH telles que les mutations de Jak2, Mpl, TET2 ou CALR ne permet pas à elle seule d’expliquer la physiopathologie de la maladie. Les résultats obtenus dans le laboratoire suggèrent que le microenvironnement médullaire au sein des niches hématopoïétiques et en particulier les cellules stromales mésenchymateuses (CSM), participe vraisemblablement à cette dérégulation de l’hématopoïèse, favorisant le développement du clone pathologique. Cependant, aucune preuve tangible d’une altération des CSM médullaires n’a été jusqu’à présent apportée.Dans ce travail, nous avons isolé les CSM de la moelle de patients atteints de MFP et réalisé une caractérisation « complète » de ces cellules : prolifération, phénotype, soutien de l’hématopoïèse, sécrétome, transcriptome, miRNome et capacités de différenciation. Nos résultats ont permis de dégager un faisceau d’arguments en faveur d’une dérégulation de leur différenciation ostéoblastique (DOB). (i) Les cytokines BMP2, RANTES, PDGF, TGF-β1, VEGF et Il-6 sont significativement produites en plus grande quantité par ces cellules. (ii) L’étude du transcriptome a révélé une expression significativement différente d’un ensemble de gènes impliqués dans la DOB tels que RUNX2, DLX5, TWIST1 et NOGGIN. (iii) De nombreux micro-ARN, dont certains sont connus pour être impliqués dans la DOB comme miR-210 ou dans le nichage des cellules souches hématopoïétiques comme miR-34a, sont dérégulés à l’état basal et au cours de cette DOB. (iv) Enfin, l’étude de leurs capacités de différenciation ostéoblastique in vitro et in vivo chez la souris immunodéprimée est en faveur d’une augmentation de ces capacités. Nous avons étudié l’impact du TGF- β1 dans cette DOB. Nous avons mis en évidence que les CSM de malades présentent un état basal d'activation de la voie de signalisation pSmad significativement augmenté, confirmant l’expression endogène de TGF-β1. En utilisant des inhibiteurs spécifiques du récepteur de type I au TGF- β, nous avons montré l’implication de cette cytokine dans les altérations de la DOB. En conclusion, notre travail montre pour la première fois que les CSM des malades de MFP sont anormales et ce indépendamment de la stimulation par le clone hématopoïétique pathologique, suggérant la présence d'anomalies constitutives ou acquises. Ces anomalies impliquent deux acteurs majeurs de la pathologie : le TGF-β1 et l'ostéogenèse. / Primary myelofibrosis (PMF) is a Philadelphia-negative myeloproliferative neoplasm, rare but associated with a poor prognosis. Its features are a clonal proliferation and an egress of hematopoietic stem cells (HSC) from bone marrow to spleen. These abnormalities of hematopoiesis are in relation with a pathological stroma (myelofibrosis, osteosclerosis and neoangiogenesis). Molecular abnormalities present in HSC partially explain the physiopathology of the disease. Results from our lab suggest that the bone marrow micro-environnement, especially mesenchymal stromal cells (MSC), are involved in the deregulation of hematopoiesis, promoting the clonal cells. However, there is no strong evidence of bone marrow MSC alterations reported for now.In our study, we isolated MSC from bone marrow of patients suffering from PMF and performed a broad characterization: proliferation, phenotype, hematopoiesis supporting capacities, secretome, transcriptome and miRNome analysis. Our results highlight arguments in favor of a deregulation of their osteogenic capacities. (i) Cytokines NMP2, RANTES, PDGF, TGF-β1, VEGF and Il-6 were significantly overproduced by MSCs. (ii) Transcriptome analysis revealed a specific signature involving genes participating in osteogenic differentiation such as RUNX2, DLX5, TWIST1 and NOGGIN. (iii) Many micro-RNAs, some know to be involved in osteogenic differentiation regulation, as mir-34a, are deregulated in MSCs and in MSC-derived osteoblasts. (iv) Finally, study of their osteogenic potency in vitro and in vivo in nude mice showed an increasing of their osteogenic potency. We studied the impact of TGF-β1 in this process and showed that PMF MSCs showed a basal expression of Smad pathway significantly increased as compared to control. Using specific inhibitor of TGF-β1 receptor, we demonstrated the implication of this cytokine in the osteogenic impairment.To summarize, our work shows for the first time that MSCs from PMF patients are abnormal, independently from stimulation by clonal cells, suggesting intrinsic abnormalities. These abnormalities involve two main factor of the disease: TGF-β1 and osteogenesis.

A descoberta das células tronco adultas tornou possível a sua aplicação clínica em protocolos de medicina regenerativa e de engenharia de tecidos para a reparação de órgãos danificados ou pouco funcionais. Neste sentido, são conhecidas muitas fontes diferentes de células tronco adultas, entre elas o tecido epitelial, o tecido muscular e a medula óssea. Recentemente, o tecido adiposo adulto foi reconhecido como uma fonte alternativa, acessível e rica em células tronco mesenquimais (MSCs), também denominadas de células tronco derivadas do tecido adiposo (hADSCs). As hADSCs são as células tronco adultas que apresentam a maior plasticidade, e são facilmente isoladas pela característica de aderência a substratos plásticos. Sob condições específicas de cultivo, as hADSCs podem diferenciar-se em células precursoras osteoblásticas. Dentre as proteínas necessárias para a diferenciação das hADSCs, o papel do complexo minichromosome maintenance (MCM) não é muito bem conhecido. A proteína MCM2 é parte do complexo MCM, fazendo parte do complexo pré-replicativo (pre-RC). Em células proliferativas, o gene MCM2 apresenta alta expressão, sendo que o seu produto gênico está relacionado com os mecanismos de reparação de DNA. Sendo assim, analisamos as mudanças na expressão do gene MCM2 durante a osteodiferenciação das hADSCs em diferentes meios de cultura, contendo BMP-4 ou OM. Os genes relacionados com a osteodiferenciação, como osteocalcina (OC), osteopontina (OP) e fosfatase alcalina (ALP) foram analisados por RT-PCR, por PCR em tempo real e citoquímica. Os dados obtidos pelas técnicas citoquímicas e de biologia molecular indicaram uma diminuição da expressão do gene MCM2 durante a osteodiferenciação, que pode estar relacionada com danos ao DNA ou senescência nas hADSCs. / The discovery of adult stem cells is a promise field in regenerative medicine and tissue engineering. There are many sources of adult stem cells among which of whom are the skin, muscle and bone marrow. Currently, the adipose tissue has been an alternative source of mesenchymal stem cells (MSCs), termed human adipose-derived stem cells (hADSCs). The MSCs are the most plastic adult stem cells known until now and are easily isolated by adherence in plastic substrates. Under specifics conditions of culture hADSCs can be differentiated in osteoblast cell precursors. Among proteins needed for hADSCs differentiation, the roles of minichromosome maintenance (MCM) complex remains poorly understood. The protein complex MCM2-7 is part of DNA pre-replicative complex (pre-RC), being abundant in proliferating cells and involved in DNA repair mechanisms. In this sense, the Mcm2 is essential for the activity of the MCM complex. Thus, we analysed MCM2 gene expression changes during osteoinduction by culturing hADSCs in different media containing BMP-4 or in osteogenic medium. Genes related to osteodifferentiation, like osteocalcin (OC), osteopontin (OP) and alkaline phosphatase (ALP) were analysed by molecular and cytochemistry approaches during osteodifferentiation. Interestingly, we observed a decrease in the MCM2 gene level osteoinduction of hADSCs in osteogenic medium, which could be related to an increase in genome damage and senescence in hADSCs.

A descoberta das células tronco adultas tornou possível a sua aplicação clínica em protocolos de medicina regenerativa e de engenharia de tecidos para a reparação de órgãos danificados ou pouco funcionais. Neste sentido, são conhecidas muitas fontes diferentes de células tronco adultas, entre elas o tecido epitelial, o tecido muscular e a medula óssea. Recentemente, o tecido adiposo adulto foi reconhecido como uma fonte alternativa, acessível e rica em células tronco mesenquimais (MSCs), também denominadas de células tronco derivadas do tecido adiposo (hADSCs). As hADSCs são as células tronco adultas que apresentam a maior plasticidade, e são facilmente isoladas pela característica de aderência a substratos plásticos. Sob condições específicas de cultivo, as hADSCs podem diferenciar-se em células precursoras osteoblásticas. Dentre as proteínas necessárias para a diferenciação das hADSCs, o papel do complexo minichromosome maintenance (MCM) não é muito bem conhecido. A proteína MCM2 é parte do complexo MCM, fazendo parte do complexo pré-replicativo (pre-RC). Em células proliferativas, o gene MCM2 apresenta alta expressão, sendo que o seu produto gênico está relacionado com os mecanismos de reparação de DNA. Sendo assim, analisamos as mudanças na expressão do gene MCM2 durante a osteodiferenciação das hADSCs em diferentes meios de cultura, contendo BMP-4 ou OM. Os genes relacionados com a osteodiferenciação, como osteocalcina (OC), osteopontina (OP) e fosfatase alcalina (ALP) foram analisados por RT-PCR, por PCR em tempo real e citoquímica. Os dados obtidos pelas técnicas citoquímicas e de biologia molecular indicaram uma diminuição da expressão do gene MCM2 durante a osteodiferenciação, que pode estar relacionada com danos ao DNA ou senescência nas hADSCs. / The discovery of adult stem cells is a promise field in regenerative medicine and tissue engineering. There are many sources of adult stem cells among which of whom are the skin, muscle and bone marrow. Currently, the adipose tissue has been an alternative source of mesenchymal stem cells (MSCs), termed human adipose-derived stem cells (hADSCs). The MSCs are the most plastic adult stem cells known until now and are easily isolated by adherence in plastic substrates. Under specifics conditions of culture hADSCs can be differentiated in osteoblast cell precursors. Among proteins needed for hADSCs differentiation, the roles of minichromosome maintenance (MCM) complex remains poorly understood. The protein complex MCM2-7 is part of DNA pre-replicative complex (pre-RC), being abundant in proliferating cells and involved in DNA repair mechanisms. In this sense, the Mcm2 is essential for the activity of the MCM complex. Thus, we analysed MCM2 gene expression changes during osteoinduction by culturing hADSCs in different media containing BMP-4 or in osteogenic medium. Genes related to osteodifferentiation, like osteocalcin (OC), osteopontin (OP) and alkaline phosphatase (ALP) were analysed by molecular and cytochemistry approaches during osteodifferentiation. Interestingly, we observed a decrease in the MCM2 gene level osteoinduction of hADSCs in osteogenic medium, which could be related to an increase in genome damage and senescence in hADSCs.

A descoberta das células tronco adultas tornou possível a sua aplicação clínica em protocolos de medicina regenerativa e de engenharia de tecidos para a reparação de órgãos danificados ou pouco funcionais. Neste sentido, são conhecidas muitas fontes diferentes de células tronco adultas, entre elas o tecido epitelial, o tecido muscular e a medula óssea. Recentemente, o tecido adiposo adulto foi reconhecido como uma fonte alternativa, acessível e rica em células tronco mesenquimais (MSCs), também denominadas de células tronco derivadas do tecido adiposo (hADSCs). As hADSCs são as células tronco adultas que apresentam a maior plasticidade, e são facilmente isoladas pela característica de aderência a substratos plásticos. Sob condições específicas de cultivo, as hADSCs podem diferenciar-se em células precursoras osteoblásticas. Dentre as proteínas necessárias para a diferenciação das hADSCs, o papel do complexo minichromosome maintenance (MCM) não é muito bem conhecido. A proteína MCM2 é parte do complexo MCM, fazendo parte do complexo pré-replicativo (pre-RC). Em células proliferativas, o gene MCM2 apresenta alta expressão, sendo que o seu produto gênico está relacionado com os mecanismos de reparação de DNA. Sendo assim, analisamos as mudanças na expressão do gene MCM2 durante a osteodiferenciação das hADSCs em diferentes meios de cultura, contendo BMP-4 ou OM. Os genes relacionados com a osteodiferenciação, como osteocalcina (OC), osteopontina (OP) e fosfatase alcalina (ALP) foram analisados por RT-PCR, por PCR em tempo real e citoquímica. Os dados obtidos pelas técnicas citoquímicas e de biologia molecular indicaram uma diminuição da expressão do gene MCM2 durante a osteodiferenciação, que pode estar relacionada com danos ao DNA ou senescência nas hADSCs. / The discovery of adult stem cells is a promise field in regenerative medicine and tissue engineering. There are many sources of adult stem cells among which of whom are the skin, muscle and bone marrow. Currently, the adipose tissue has been an alternative source of mesenchymal stem cells (MSCs), termed human adipose-derived stem cells (hADSCs). The MSCs are the most plastic adult stem cells known until now and are easily isolated by adherence in plastic substrates. Under specifics conditions of culture hADSCs can be differentiated in osteoblast cell precursors. Among proteins needed for hADSCs differentiation, the roles of minichromosome maintenance (MCM) complex remains poorly understood. The protein complex MCM2-7 is part of DNA pre-replicative complex (pre-RC), being abundant in proliferating cells and involved in DNA repair mechanisms. In this sense, the Mcm2 is essential for the activity of the MCM complex. Thus, we analysed MCM2 gene expression changes during osteoinduction by culturing hADSCs in different media containing BMP-4 or in osteogenic medium. Genes related to osteodifferentiation, like osteocalcin (OC), osteopontin (OP) and alkaline phosphatase (ALP) were analysed by molecular and cytochemistry approaches during osteodifferentiation. Interestingly, we observed a decrease in the MCM2 gene level osteoinduction of hADSCs in osteogenic medium, which could be related to an increase in genome damage and senescence in hADSCs.

Problems with musculoskeletal system, such as of developmental disorders, fractures or damage of the bone by age, inflammatory or tumor diseases, are still increasing in orthopaedics. Sometimes the bone tissue is not capable to completely regenerate to exert its physiological function in the organism. For this reason, using the bone replacements is necessary and common nowadays. Despite of an intensive research and testing of a wide range of the potential biomaterials and their combinations, the usage of metal materials for construction of the bone implants, still remains to be the gold standard. Ti-6Al-4V alloy is one of the commercialy used metal materials, which is known for the high mechanical and chemical resistance and a good biocompatibility. For a good biological response of the patient's organism for the bone implant, is an ability of osteointegration into the surrounding bone tissue, the key. This ability can be influenced in the case of the metals, by their surface structure. As it is known from earlier studies, the surface topography of the material is very important for the adhesion and proliferation of the bone cells, which are able to discriminate, very sensitively, between various stages of the material surface roughness. For this reason we have focused on studying of an influence...

The present study analyzes the capacity of collagen (coll)/sulfated glycosaminoglycan (sGAG)-based surface coatings containing bioactive glass nanoparticles (BGN) in promoting the osteogenic differentiation of human mesenchymal stroma cells (hMSC). Physicochemical characteristics of these coatings and their effects on proliferation and osteogenic differentiation of hMSC were investigated. BGN were stably incorporated into the artificial extracellular matrices (aECM). Oscillatory rheology showed predominantly elastic, gel-like properties of the coatings. The complex viscosity increased depending on the GAG component and was further elevated by adding BGN. BGN-containing aECM showed a release of silicon ions as well as an uptake of calcium ions. hMSC were able to proliferate on coll and coll/sGAG coatings, while cellular growth was delayed on aECM containing BGN. However, a stimulating effect of BGN on ALP activity and calcium deposition was shown. Furthermore, a synergistic effect of sGAG and BGN was found for some donors. Our findings demonstrated the promising potential of aECM and BGN combinations in promoting bone regeneration. Still, future work is required to further optimize the BGN/aECM combination for increasing its combined osteogenic effect.