In Vitro Characterization of Hydroxyapatite-Based Coatings Doped with Mg or Zn Electrochemically Deposited on Nanostructured Titanium.

Biomaterials are an important and integrated part of modern medicine, and their development and improvement are essential. The fundamental requirement of a biomaterial is found to be in its interaction with the surrounding environment, with which it must coexist. The aim of this study was to assess the biological characteristics of hydroxyapatite (HAp)-based coatings doped with Mg and Zn ions obtained by the pulsed galvanostatic electrochemical method on the surface of pure titanium (cp-Ti) functionalized with titanium dioxide nanotubes (NTs TiO2) obtained by anodic oxidation. The obtained results highlighted that the addition of Zn or Mg into the HAp structure enhances the in vitro response of the cp-Ti surface functionalized with NT TiO2. The contact angle and surface free energy showed that all the developed surfaces have a hydrophilic character in comparison with the cp-Ti surface. The HAp-based coatings doped with Zn registered superior values than the ones with Mg, in terms of biomineralization, electrochemical behavior, and cell interaction. Overall, it can be said that the addition of Mg or Zn can enhance the in vitro behavior of the HAp-based coatings in accordance with clinical requirements. Antibacterial tests showed that the proposed HAp-Mg coatings had no efficiency against Escherichia coli, while the HAp-Zn coatings registered the highest antibacterial efficiency.

Generation of an Immortalized Human Adipose-Derived Mesenchymal Stromal Cell Line Suitable for Wound Healing Therapy.

Adipose-derived mesenchymal stromal cells (ADSC) are a promising source for cellular therapy of chronic wounds. However, the limited life span during in vitro expansion impedes their extensive use in clinical applications and basic research. We hypothesize that by introduction of an ectopic expression of telomerase into ADSC, the cells' lifespans could be significantly extended. To test this hypothesis, we aimed at engineering an immortalized human ADSC line using a lentiviral transduction with human telomerase (hTERT). ADSC were transduced with a third-generation lentiviral system and a hTERT codifying plasmid (pLV-hTERT-IRES-hygro). A population characterized by increased hTERT expression, extensive proliferative potential and remarkable (potent) multilineage differentiation capacity was selected. The properties for wound healing of this immortalized ADSC line were assessed after 17 passages. Their secretome induced the proliferation and migration of keratinocytes, dermal fibroblasts, and endothelial cells similarly to untransduced ADSC. Moreover, they sustained the complete re-epithelialization of a full thickness wound performed on a skin organotypic model. In summary, the engineered immortalized ADSC maintain the beneficial properties of parent cells and could represent a valuable and suitable tool for wound healing in particular, and for skin regenerative therapy in general.

Mesenchymal stromal cell-derived factors promote the colonization of collagen 3D scaffolds with human skin cells.

The development of stem cell technology in combination with advances in biomaterials has opened new ways of producing engineered tissue substitutes. In this study, we investigated whether the therapeutic potential of an acellular porous scaffold made of type I collagen can be improved by the addition of a powerful trophic agent in the form of mesenchymal stromal cells conditioned medium (MSC-CM) in order to be used as an acellular scaffold for skin wound healing treatment. Our experiments showed that MSC-CM sustained the adherence of keratinocytes and fibroblasts as well as the proliferation of keratinocytes. Moreover, MSC-CM had chemoattractant properties for keratinocytes and endothelial cells, attributable to the content of trophic and pro-angiogenic factors. Also, for the dermal fibroblasts cultured on collagen scaffold in the presence of MSC-CM versus serum control, the ratio between collagen III and I mRNAs increased by 2-fold. Furthermore, the gene expression for α-smooth muscle actin, tissue inhibitor of metalloproteinase-1 and 2 and matrix metalloproteinase-14 was significantly increased by approximately 2-fold. In conclusion, factors existing in MSC-CM improve the colonization of collagen 3D scaffolds, by sustaining the adherence and proliferation of keratinocytes and by inducing a pro-healing phenotype in fibroblasts.

Characterization of electron beam deposited Nb2O5 coatings for biomedical applications.

Niobium oxide coatings deposited on Ti6Al4V substrates by electron beam deposition and annealed in air at 600 °C and 800 °C were evaluated for their suitability towards dental, maxillofacial or orthopaedic implant applications. A detailed physico-chemical properties investigation was carried out in order to determine their elemental and phase composition, surface morphology and roughness, mechanical properties, wettability, and corrosion resistance in simulated body fluid solution (pH = 7.4) at room temperature. The biocompatibility of the bare Ti6Al4V substrate and coated surfaces was evaluated by testing the cellular adhesion and viability/proliferation of human osteosarcoma cells (MG-63) after 72 h of incubation. The coatings annealed at 800 °C exhibit more phase pure nanocrystalline Nb2O5 surfaces with enhanced wettability, reduced porosity and enhanced corrosion resistance properties making them good candidate for dental, maxillofacial or orthopaedic implant applications.

Heterogeneity of human fibroblasts isolated from hypertrophic scar.

Pathological wound healing states, such as hypertrophic scarring and keloids, represent a huge clinical and financial burden on healthcare system. The complex biological mechanisms occurring in hypertrophic scarring are still barely understood. To date, there is no satisfactory description of hypertrophic fibroblasts. Therefore, in the present study we focused on the comparatively characterization of the fibroblasts residing in different regions of hypertrophic scars. To achieve this aim, fibroblasts were isolated from normal skin samples (n=4) and hypertrophic scars (n=4). These cell populations were further were used for the evaluation of proliferation and migration capacity, for the gene and protein expression of extracellular matrix protein type I collagen and fibronectin and for the presence of myofibroblasts. Our results demonstrated that perilesional and intralesional fibroblasts isolated from hypertrophic scars could be considered as distinct populations, having different properties. Thus, the intralesional fibroblasts had an increased proliferation capacity and increased gene and protein expression of collagen I and fibronectin. However, the perilesional fibroblasts had augmented mobility as revealed by in vitro scratch test and contained a higher percentage of myofibroblasts [alpha-smooth muscle actin (α-SMA)high cells], in comparison to the intralesional population. In conclusion, our data could provide an explanation regarding the inconsistent efficacy of topic therapies for hypertrophic scars.

In Vitro Biocompatibility of Si Alloyed Multi-Principal Element Carbide Coatings.

In the current study, we have examined the possibility to improve the biocompatibility of the (TiZrNbTaHf)C through replacement of either Ti or Ta by Si. The coatings were deposited on Si and 316L stainless steel substrates by magnetron sputtering in an Ar+CH4 mixed atmosphere and were examined for elemental composition, chemical bonds, surface topography, surface electrical charge and biocompatible characteristics. The net surface charge was evaluated at nano and macroscopic scale by measuring the electrical potential and work function, respectively. The biocompatible tests comprised determination of cell viability and cell attachment to the coated surface. The deposited coatings had C/(metal+Si) ratios close to unity, while a mixture of metallic carbide, free-carbon and oxidized species formed on the film surface. The coatings' surfaces were smooth and no influence of surface roughness on electrical charge or biocompatibility was found. The biocompatible characteristics correlated well with the electrical potential/work function, suggesting a significant role of surface charge in improving biocompatibility, particularly cell attachment to coating's surface. Replacement of either Ti or Ta by Si in the (TiZrNbTaHf)C coating led to an enhanced surface electrical charge, as well as to superior biocompatible properties, with best results for the (TiZrNbSiHf)C coating.

Electrospun TiO2 nanofibers decorated Ti substrate for biomedical application.

Various TiO2 nanofibers on Ti surface have been fabricated via electrospinning and calcination. Due to different elaboration conditions the electrospun fibers have different surface feature morphologies, characterized by scanning electronic microscopy, surface roughness, and contact angle measurements. The results have indicated that the average sample diameters are between 32 and 44 nm, roughness between 61 and 416 nm, and all samples are hydrophilic. As biological evaluation, cell culture with MG63 cell line originally derived from a human osteosarcoma was performed and correlation between nanofibers elaboration, properties and cell response was established. The cell adherence and growth are more evident on Ti samples with more aligned fibers, higher roughness and strong hydrophilic character and such fibers have been elaborated with a high speed rotating cylinder collector, confirming the idea that nanostructure elaboration conditions guide the cells' growth.

The two step nanotube formation on TiZr as scaffolds for cell growth.

Various TiO2 nanotubes on Ti50Zr alloy have been fabricated via a two step anodization method in glycol with 15vol.% H2O and 0.2M NH4F under anodization controlled voltages of 15, 30 and 45V. A new sonication treatment in deionized water with three steps and total sonication time as 1min was performed after the first anodization step in order to remove the oxide layer grown during 2h. The second step of anodization was for 1h and took place at the same conditions. The role of removed layer as a nano-prepatterned surface was evidenced in the formation of highly ordered nanotubular structures and morphological features were analyzed by SEM, AFM and surface wettability. The voltage-controlled anodization leads to various nanoarhitectures, with diameters in between 20 and 80nm. As biological assay, cell culture tests with MG63 cell line originally derived from a human osteosarcoma were performed. A correlation between nanostructure morphological properties as a result of voltage-controlled anodization and cell response was established.

Osteoblast ontogeny and implications for bone pathology: an overview.

Osteoblasts are specialized mesenchyme-derived cells accountable for bone synthesis, remodelling and healing. Differentiation of osteoblasts from mesenchymal stem cells (MSC) towards osteocytes is a multi-step process strictly controlled by various genes, transcription factors and signalling proteins. The aim of this review is to provide an update on the nature of bone-forming osteoblastic cells, highlighting recent data on MSC-osteoblast-osteocyte transformation from a molecular perspective and to discuss osteoblast malfunctions in various bone diseases. We present here the consecutive stages occurring in the differentiation of osteoblasts from MSC, the transcription factors involved and the role of miRNAs in the process. Recent data concerning the pathogenic mechanisms underlying the loss of bone mass and architecture caused by malfunctions in the synthetic activity and metabolism of osteoblasts in osteoporosis, osteogenesis imperfecta, osteoarthritis and rheumatoid arthritis are discussed. The newly acquired knowledge of the ontogeny of osteoblasts will assist in unravelling the abnormalities taking place during their differentiation and will facilitate the prevention and/or treatment of bone diseases by therapy directed against altered molecules and mechanisms.