Polystyrene Tissue Culture Plates Research Articles

Virucidal Efficacy of Gaseous Ozone Against Type 1 Herpes Simplex Virus (HSV-1)

ABSTRACT Viruses represent a major threat to human health and are capable of spreading either via direct exposure or contamination of inanimate surfaces. Certain viruses have been shown to remain viable on surfaces of dental implants and devices, fabrics and on plastics for hours, days and weeks representing a source of continuous viral transmission which must be mitigated. In this study, we compared the effect of varying concentrations of gaseous ozone (40, 20, 10, 5, and 1 ppm) on Herpes Simplex Virus (HSV-1) coated on the surfaces of clear polystyrene tissue culture (TC) plates and stainless-steel disc at varying time intervals (1.5, 3, 6, and 8 h) at >90% relative humidity and at 21–25 ˙C (±2˙C). Test samples were placed at a height of 1 and 1.5 m from ozone source. Viral viability after ozone exposure was determined using Vero cells and the viral titer was quantified using the Spearman–Karber’s method. Overall, inactivation of HSV-1 was dependent on ozone concentration and the duration of exposure. At 20 and 40 ppm, ozone rapidly depleted virus viability after 6 h. At 5 and 10 ppm a time-dependent reduction in viral infectivity was observed. At 10 ppm, viral titer decreased from approximately 5.2 pfu/mL at T = 0 to 3.5 and 2.1 pfu/mL after 1.5- and 6-h exposure on plastic surfaces. Similarly, viral titer decreased from approximately 5.6 pfu/mL at T = 0 to 3.0 to 2.0 pfu/mL at same time frame on steel surfaces. This study builds on earlier research in the field and demonstrates the ability of gaseous ozone to inactivate HSV-1 on two surfaces commonly found in food, medical and dental settings. This study recommends that further work is completed to identify optimum ozone, humidity, and temperature combinations to inactivate such viruses on surfaces found within these environments.

Responses of Rat Mesenchymal Stromal Cells to Nanocellulose with Different Functional Groups.

Cellulose nanofibrils (CNFs) are multiscale hydrophilic biocompatible polysaccharide materials derived from wood and plants. TEMPO-mediated oxidation of CNFs (TO-CNF) turns some of the primary hydroxyl groups to carboxylate and aldehyde groups. Unlike carboxylic functional groups, there is little or no information about the biological role of the aldehyde groups on the surface of wood-based CNFs. In this work, we replaced the aldehyde groups in the TO-CNF samples with carboxyl groups by another oxidation treatment (TO-O-CNF) or with primary alcohols with terminal hydroxyl groups by a reduction reaction (TO-R-CNF). Rat mesenchymal stem/stromal cells (MSCs) derived from bone marrow were seeded on polystyrene tissue culture plates (TCP) coated with CNFs with and without aldehyde groups. TCP and TCP coated with bacterial nanocellulose (BNC) were used as control groups. Protein adsorption measurements demonstrated that more proteins were adsorbed from cell culture media on all CNF surfaces compared to BNC. Live/dead and lactate dehydrogenase assays confirmed that all nanocellulose biomaterials supported excellent cell viability. Interestingly, TO-R-CNF samples, which have no aldehyde groups, showed better cell spreading than BNC and comparable results to TCP. Unlike TO-O-CNF surfaces, which have no aldehyde groups either, TO-R-CNF stimulated cells, in osteogenic medium, to have higher alkaline phosphatase activity and to form more biomineralization than TCP and TO-CNF groups. These findings indicate that the presence of aldehyde groups (280 ± 14 μmol/g) on the surface of TEMPO-oxidized CNFs might have little or no effect on attachment, proliferation, and osteogenic differentiation of MSCs.

Differential Susceptibility of Mixed Polymicrobial Biofilms Involving Ocular Coccoid Bacteria (Staphylococcus aureus and S. epidermidis) and a Filamentous Fungus (Fusarium solani) on Ex Vivo Human Corneas

Biofilms confer several advantages to the organisms associated with them, such as increased resistances to antibacterial and antifungal compounds compared to free living cells. Compared to monomicrobial biofilms involving a single microorganism, biofilms composed of microorganisms affiliated to bacterial and fungal kingdoms are predominant in nature. Despite the predominance of polymicrobial biofilms, and more so mixed polymicrobial biofilms, they are rarely studied. The objective of the current study is to evaluate the potential of ocular bacteria and a filamentous fungus to form monomicrobial and mixed polymicrobial biofilms on synthetic and natural substrates and to monitor their response to antibiotics. In this sense, we demonstrated that the ocular pathogens Staphylococcus aureus, S. epidermidis, and Fusarium solani form monomicrobial and mixed polymicrobial biofilms both on tissue culture polystyrene plates and on ex vivo human corneas from cadavers using confocal microscopy and scanning electron microscopy. Additionally, the mixed polymicrobial biofilms involving the above ocular bacteria and a filamentous fungus were less susceptible to different antibacterials and antifungals in relation to the corresponding control planktonic cells. Further, the MICs to the screened antibacterials and antifungals in polymicrobial biofilms involving a bacterium or a fungus was either increased, decreased, or unchanged compared to the corresponding individual bacterial or fungal biofilm. The results would be useful to the ophthalmologist to plan effective treatment regimens for the eye since these are common pathogens of the eye causing keratitis, endophthalmitis, conjunctivitis, etc.

The Impact of Polyethylene Glycol-Modified Chitosan Scaffolds on the Proliferation and Differentiation of Osteoblasts.

The objective of this study was to investigate the influence of polyethylene glycol (PEG) incorporated chitosan scaffolds on osteoblasts proliferation and differentiation. The chitosan polymer was initially modified by the predetermined concentration of the photoreactive azido group for UV-crosslinking and with RGD peptides (N-acetyl-GRGDSPGYG-amide). The PEG was mixed at different ratios (0, 10, and 20 wt%) with modified chitosan in 96-well tissue culture polystyrene plates to prepare CHI-100, CHI-90, and CHI-80 scaffolds. PEG-containing scaffolds exhibited bigger pore size and higher water content compared to unmodified chitosan scaffolds. After 10 days of incubation, the cell number of CHI-90 (1.1 × 106 cells/scaffold) surpasses that of CHI-100 (9.2 × 105 cells/scaffold) and the cell number of CHI-80 (7.6 × 105 cells/scaffold) were significantly lower. The ALP activity of CHI-90 was the highest on the fifth day indicating the favored osteoblasts' early-stage differentiation. Moreover, after 14 days of osteogenic culture, calcium deposition in the CHI-90 scaffolds (2.7 μmol Ca/scaffold) was significantly higher than the control (2.2 μmol Ca/scaffold) whereas on CHI-80 was 1.9 μmol/scaffold. The results demonstrate that PEG-incorporated chitosan scaffolds favored osteoblasts proliferation and differentiation; however, mixing relatively excess PEG (≥20% wt.) had a negative impact on osteoblasts proliferation and differentiation.

Challenges and Opportunities Modeling the Dynamic Tumor Matrisome.

We need novel strategies to target the complexity of cancer and, particularly, of metastatic disease. As an example of this complexity, certain tissues are particularly hospitable environments for metastases, whereas others do not contain fertile microenvironments to support cancer cell growth. Continuing evidence that the extracellular matrix (ECM) of tissues is one of a host of factors necessary to support cancer cell growth at both primary and secondary tissue sites is emerging. Research on cancer metastasis has largely been focused on the molecular adaptations of tumor cells in various cytokine and growth factor environments on 2-dimensional tissue culture polystyrene plates. Intravital imaging, conversely, has transformed our ability to watch, in real time, tumor cell invasion, intravasation, extravasation, and growth. Because the interstitial ECM that supports all cells in the tumor microenvironment changes over time scales outside the possible window of typical intravital imaging, bioengineers are continuously developing both simple and sophisticated in vitro controlled environments to study tumor (and other) cell interactions with this matrix. In this perspective, we focus on the cellular unit responsible for upholding the pathologic homeostasis of tumor-bearing organs, cancer-associated fibroblasts (CAFs), and their self-generated ECM. The latter, together with tumoral and other cell secreted factors, constitute the "tumor matrisome". We share the challenges and opportunities for modeling this dynamic CAF/ECM unit, the tools and techniques available, and how the tumor matrisome is remodeled (e.g., via ECM proteases). We posit that increasing information on tumor matrisome dynamics may lead the field to alternative strategies for personalized medicine outside genomics.

Evaluation of the cytotoxic and genotoxic/antigenotoxic effects of resveratrol in human limbal explant cultures.

Resveratrol (RSV) is a natural polyphenol phytoalexin compound and has long been considered to possess antioxidant and anti-inflammatory effects. In order to exploit the protective potential of RSV in anterior segment diseases, we investigated the possible cytotoxic, genotoxic/antigenotoxic effects of human limbal explant cultures to RSV and MMC or H2O2 alone and in combination. A total of 18 limbal explant tissues obtained from three corneal donors were placed on the 12 well tissue culture polystyrene plates and cultured for 14days. Cell growth from limbal explants was observed by inverted phase contrast microscopy. The cytotoxic effects of RSV was studied by neutral red uptake assay. For the assessment of the genotoxic and antigenotoxic effects, basic alkaline technique of comet assay was performed. It was found that the concentrations of RSV up to 100μM did not significantly affect the viability of outgrowth cells of limbal explant during 24h exposure. When compared to negative control, all concentrations of RSV alone caused an increase in DNA strand breakage. Interestingly, 10μg/mL MMC alone caused similar tail intensity and tail moment values with RSV alone. On the other hand, RSV treatment in all doses seemed to decrease the DNA damage induced by either H2O2 or MMC. RSV is an attractive natural compound for the purpose of oxidative stress reduction in ocular surface and can be utilized as a supplement to promote ocular surface regeneration in vivo.

High resolution DLP stereolithography to fabricate biocompatible hydroxyapatite structures that support osteogenesis.

Lithography based additive manufacturing techniques, specifically digital light processing (DLP), are considered innovative manufacturing techniques for orthopaedic implants because of their potential for construction of complex geometries using polymers, metals, and ceramics. Hydroxyapatite (HA) coupons, printed using DLP, were evaluated for biological performance in supporting viability, proliferation, and osteogenic differentiation of the human cell line U2OS and human mesenchymal stem cells (MSCs) up to 35 days in culture to determine feasibility for future use in development of complex scaffold geometries. Contact angle, profilometry, and scanning electron microscopy (SEM) measurements showed the HA coupons to be hydrophilic, porous, and having micro size surface roughness, all within favourable cell culture ranges. The study found no impact of leachable and extractables form the DLP printing process. Cells seeded on coupons exhibited morphologies comparable to conventional tissue culture polystyrene plates. Cell proliferation rates, as determined by direct cell count and the RealTime-GloTM MT Cell Viability Assay, were similar on HA coupons and standard tissue culture polystyrene plates). Osteogenic differentiation of human MSCs on HA coupons was confirmed using alkaline phosphatase, Alizarin Red S and von Kossa staining. The morphology of MSCs cultured in osteogenic medium for 14 to 35 days was similar on HA coupons and tissue culture polystyrene plates, with osteogenic (geometric, cuboidal morphology with dark nodules) and adipogenic (lipid vesicles and deposits) features. We conclude that the DLP process and LithaBone HA400 slurry are biocompatible and are suitable for osteogenic applications. Coupons served as an effective evaluation design in the characterization and visualization of cell responses on DLP printed HA material. Results support the feasibility of future technical development for 3D printing of sophisticated scaffold designs, which can be constructed to meet the mechanical, chemical, and porosity requirements of an artificial bone scaffold.

Effect of Cell-Derived Matrices on Growth and Differentiation of Human Wharton’s Jelly-Derived Mesenchymal Stem Cells

Cell-derived matrices (CDMs) are scaffolds constructed by decellularization of cellular matrices from different tissues and organs. Since CDMs mimic the extracellular matrices (ECMs) of native tissues, it plays an essential role in the preparation of bioscaffolds. CDM scaffolds from mesenchymal stem cells (MSCs) have been reported to support cell adhesion and proliferation of its own cells. Therefore, in this study we aimed to test if growth of human Wharton’s jelly-derived MSCs may be enhanced when cultured on their own CDMs. To do this, MSCs were induced to generate ECM using ascorbic acid. Thus, obtained matrices were decellularized and characterized quantitatively for changes in their biochemical components (total protein, collagen, glycosaminoglycans) and qualitatively for fibronectin, laminin, and collagen (I & IV) by immunostaining. Our results show the retention of essential ECM components in the decellularized WJ-MSC-derived matrix (WJ-CDM). The influence of WJ-CDM on proliferation and differentiation of WJ-MSCs were evaluated by comparing their growth on collagen and fibronectin-only coated plates. A non-coated tissue culture polystyrene plate (TCPS) served as control. Our cell proliferation results show that no significant changes were observed in the proliferation of MSCs when cultured on WJ-CDM as compared to the bio-coated and non-coated cultures. However, gene expression analysis of the differentiation process showed that osteogenic and adipogenic differentiation potential of the WJ-MSCs was significantly increased upon culturing them on WJ-CDM. In conclusion, the present study reveals that the WJ-MSCs cultured on WJ-CDM may augment osteogenic and adipogenic differentiation.

3D Culture Modeling of Metastatic Breast Cancer Cells in Additive Manufactured Scaffolds.

Cancer biology research is increasingly moving toward innovative in vitro 3D culture models, as conventional and current 2D cell cultures fail to resemble in vivo cancer biology. In the current study, porous 3D scaffolds, designed with two different porosities along with 2D tissue culture polystyrene (TCP) plates were used with a model breast cancer human cell line. The 3D engineered system was evaluated for the optimal seeding method (dynamic versus static), adhesion, and proliferation rate of MDA-MB-231 breast cancer cells. The expression profiles of proliferation-, stemness-, and dormancy-associated cancer markers, namely, ki67, lamin A/C, SOX2, Oct3/4, stanniocalcin 1 (STC1), and stanniocalcin 2 (STC2), were evaluated in the 3D cultured cells and compared to the respective profiles of the cells cultured in the conventional 2D TCP. Our data suggested that static seeding was the optimal seeding method with porosity-dependent efficiency. Moreover, cells cultured in 3D scaffolds displayed a more dormant phenotype in comparison to 2D, which was manifested by the lower proliferation rate, reduced ki67 expression, increased lamin A/C expression, and overexpression of STCs. The possible relationship between the cell affinity to different extracellular matrix (ECM) proteins and the RANK expression levels was also addressed after deriving collagen type I (COL-I) and fibronectin (FN) MDA-MB-231 filial cell lines with enhanced capacity to attach to the respective ECM proteins. The new derivatives exhibited a more mesenchymal like phenotype and higher RANK levels in relation to the parental cells, suggesting a relationship between ECM cell affinity and RANK expression. Therefore, the present 3D cell culture model shows that cancer cells on printed scaffolds can work as better representatives in cancer biology and drug screening related studies.

Effects of nanofiber scaffolds coated with nanoparticulate and micro-particulate FDBA on the morphology, adhesion and proliferation of human mesenchymal stem cells.

Background:Freeze dried bone allograft nanoparticles on a nanofiber membrane may serve as an ideal scaffold for bone regeneration. This study aimed to assess the biological behavior of human MSCs in terms of proliferation and adhesion to nanoparticulate and microparticulate FDBA scaffolds on PLLA nanofiber membrane. Methods:In this experimental study, PLLA nanofiber scaffolds were synthesized by the electrospinning method. The FDBA nanoparticles were synthesized mechanically. The FDBA nanoparticles and microparticles were loaded on the surface of PLLA nanofiber membrane. A total of 64 scaffold samples in four groups of n-FDBA/PLLA, FDBA/PLLA, PLLA and control were placed in 24-well polystyrene tissue culture plates; 16 wells were allocated to each group. Data were analyzed using one-way ANOVA and Bonferroni test. Results:The proliferation rate of MSCs was significantly higher in the nanoparticulate group compared to the microparticulate group at five days (p = 0.034). Assessment of cell morphology at 24 hours revealed spindle-shaped cells with a higher number of appendages in the nanoparticulate group compared to other groups. Conclusion:MSCs on n-FDBA/PLLA scaffold were morphologically more active and flatter with a higher number of cellular appendages, as compared to FDBA/PLLA. It seems that the nanoparticulate scaffold is superior to the microparticulate scaffold in terms of proliferation, attachment, and morphology of MSCs in vitro.

An in vitro assessment of the responses of human dermal fibroblast seeded on 3D printed thermoplastic polyurethane scaffold

Tissue engineering is a multidisciplinary field is an interdisciplinary field for the design of biological substitutes that can improve, restore, and maintain tissue functions. Thermoplastic polyurethanes (TPUs) are linear polymers which are widely used for tissue engineering due to its flexibility in processing methods, biocompatibility and excellent mechanical properties. They are suitable materials for use in three-dimensional (3D) printing. Dermal fibroblasts are mesenchymal cells which play crucial roles in physiological tissue repair. The present study aimed to investigate the viability, proliferation, adhesion, and type IV collagen expression of human dermal fibroblasts (HDFs) seeded on 3D printed TPU scaffolds in vitro. HDFs were seeded on 3D TPU scaffolds or tissue culture polystyrene plates as control and cultured for 1, 3, 7, and 14 days. 3D TPU scaffolds were prepared using a custom made fused deposition modelling printer. The viability and proliferation of cells was analyzed by WST-1 assay on days 1 and 3. The cell adhesion was evaluated by scanning electron microscopy (SEM) on days 1 and 3. The cell morphology was examined by hematoxylin and eosin (H&E) staining. Expression of type IV collagen was analyzed by immunohistochemical (IHC) staining. The viability of HDFs on 3D TPU scaffolds was lower than their control groups on days 1 and 3, slightly higher on day 3. SEM images showed HDF attachment to the 3D TPU scaffold surface with spindle-shaped morphology. H&E staining demonstrated that HDFs on 3D TPU scaffolds showed smaller morphologies on days 7 and 14 compared to days 1 and 3. Type IV collagen staining was more intense in HDFs on 3D TPU scaffolds on day 1, 3, and 7 compared to day 14. In conclusion, our study shows the biocompatibility and the potential applications of 3D printed TPU scaffolds for skin tissue engineering using fibroblasts.

Biomimetic Cell-Laden MeHA Hydrogels for the Regeneration of Cartilage Tissue.

Methacrylated hyaluronic acid (MeHA) and chondroitin sulfate (CS)-biofunctionalized MeHA (CS-MeHA), were crosslinked in the presence of a matrix metalloproteinase 7 (MMP7)-sensitive peptide. The synthesized hydrogels were embedded with either human mesenchymal stem cells (hMSCs) or chondrocytes, at low concentrations, and subsequently cultured in a stem cell medium (SCM) or chondrogenic induction medium (CiM). The pivotal role of the synthesized hydrogels in promoting the expression of cartilage-related genes and the formation of neocartilage tissue despite the low concentration of encapsulated cells was assessed. It was found that hMSC-laden MeHA hydrogels cultured in an expansion medium exhibited a significant increase in the expression of chondrogenic markers compared to hMSCs cultured on a tissue culture polystyrene plate (TCPS). This favorable outcome was further enhanced for hMSC-laden CS-MeHA hydrogels, indicating the positive effect of the glycosaminoglycan binding peptide on the differentiation of hMSCs towards a chondrogenic phenotype. However, it was shown that an induction medium is necessary to achieve full span chondrogenesis. Finally, the histological analysis of chondrocyte-laden MeHA hydrogels cultured on an ex vivo osteochondral platform revealed the deposition of glycosaminoglycans (GAGs) and the arrangement of chondrocyte clusters in isogenous groups, which is characteristic of hyaline cartilage morphology.

Osteoblasts and fibroblasts attachment to poly(3-hydroxybutyric acid-co-3-hydrovaleric acid) (PHBV) film and electrospun scaffolds.

The cellular response is the most crucial in vitro research. Materials' biocompatibility is determined based on cell proliferation and growth. Moreover, the topography of the scaffold surface is the key to enhance cell attachment and anchoring that importantly control further tissue development. Individual cell types have specific preferences regarding the type of surface and its geometry. In our research, we used poly(3-hydroxybutyric acid-co-3-hydrovaleric acid) PHBV to produce two types of substrate: a 3D structure of electrospun fibers and 2D flat films. The PHBV products were morphologically characterized by scanning electron microscopy (SEM). The cytocompatibility was evaluated with cell viability and proliferation using two different types of cells: human osteoblast-like cells (MG-63) and NIH 3T3 murine fibroblast cells. The behaviour of both cell types was compared on the similar PHBV fiber scaffolds and films using two types of polystyrene (PS) based substrate for the cell culture study: unmodified PS that is not favourable for the attachment of cells and on tissue culture polystyrene (TCPS) plates, which are chemically modify to enhance cells attachment. The results clearly showed high biocompatibility of PHBV as both types of cells showed similar proliferation. These results indicated that PHBV scaffolds are suitable for the development of multifunctional substrates facilitating the growth of different types of tissue regardless of the 3D and 2D designed structures for regeneration purposes.

Guiding Mesenchymal Stem Cells into Myelinating Schwann Cell-Like Phenotypes by Using Electrospun Core-Sheath Nanoyarns.

Nerve guidance conduit (NGC)-infilling substrates have been reported to facilitate the regeneration of injured peripheral nerves (PNs), especially for large nerve gaps. In this study, longitudinally oriented electrospun core-sheath nanoyarns (csNYs), consisting of a polylactic acid microfiber core and an electrospun nanofiber sheath, were fabricated for potential PN tissue engineering applications. Our novel csNY displayed a well-aligned nanofibrous surface topography, resembling the ultrastructure of axons and fascicles of a native PN system, and it also provided a mechanically stable structure. The biological results showed that the csNY significantly enhanced the attachment, growth, and proliferation of human adipose derived mesenchymal stem cells (hADMSC) and also promoted the migration, proliferation, and phenotype maintenance of rabbit Schwann cells (rSCs). Our csNY notably increased the differentiation capability of hADMSC into SC-like cells (hADMSC-SC), in comparison with a 2D tissue culture polystyrene plate. More importantly, when combined with the appropriate induction medium, our csNY promoted hADMSC-SC to express high levels of myelination-associated markers. Overall, this study demonstrates that our csNYs have great potential to serve as not only ideal in vitro culture models for understanding SC-axon interaction and SC myelination but also as promising NGC-infilling substrates for PN regeneration applications.

Role of sugars in formation and maintenance of biofilm in Coagulase negative staphylococcal (CoNS) strains

Two different Coagulase negative staphylococcal environmental isolates BP/SU1(Staphylococcus epidermidis; MTCC accession number:9538) and BP/SU2 (Staphylococcus haemolyticus; MTCC accession number:5731) show a tendency to propagate in the biofilm mode specially on low surface energy mediums. In this study we have co-related the biofilm formation of the two species on polystyrene tissue culture plates with the quantity of utilizable sugar present in the culture medium. The dispersal of the same biofilms with common dispersing agents like NaIO4, DNase, and proteolytic enzymes has also been studied. Results indicate that though polysaccharides undoubtedly enhance the biofilm formation for both the species, their role as biofilm - material adhesive agents are minimal for these two strains.

Surface Nanocoating with Plant‐Derived Pectins Improves Fibroblast Response In Vitro

Pectins, plant‐derived polysaccharides, are novel candidates for biomaterial nanocoatings. Their chemical structure imitates the polysaccharides from the extracellular matrix of mammals, providing bio‐specific cell adhesion. It is possible to control the structure of pectin rhamnogalacturonan‐I (RG‐I) by enzymatic modification. RG‐Is has been proposed for surface nanocoating of orthopedic and dental titanium implants due to its effect on osseointegration with stimulation of bone and connective tissue healing. The aim of the current study is to evaluate in vitro, the impact of plant‐derived pectin RG‐I nanocoating on the cellular response of human fibroblasts. In this study, unmodified RG‐I and enzymatically modified RG‐I with low amount of arabinose and high amount of galactose are examined. All in vitro studies are performed on tissue culture polystyrene plates, whose surfaces are coated with unmodified and modified RG‐I. The results show that the nanocoating with pectin RG‐Is increas proliferation, cell metabolic activity, and gene expression of COL1A1, FN1, FGFR1, BMP7, and MMP2 as compared to control surface, without pectin coatings. The results show that nanocoating with RG‐Is has the capacity to improve fibroblast proliferation, differentiation, and extracellular matrix protein production, and may therefore be considered as a potential candidate for further improvement of bone and connective tissue regeneration.

Pectin coating of titanium and polystyrene surfaces modulates the macrophage inflammatory response

Titanium has been used with success for bone anchoring of dental implants. However, when implant surfaces are exposed to the oral environment, the progression of peri-implantitis triggered by specific oral bacteria has been reported. Bacterial colonization of implants leads to prolonged immune cell activation and bone resorption. A new strategy to improve implant biocompatibility and prevent peri-implantitis is to develop pectin surface nanocoatings. These plant-derived polysaccharides are promising candidates for surface nanocoatings of titanium implants due to their osteogenic and anti-inflammatory properties. Therefore, the aim of the study was to evaluate the in vitro effect of nanocoating with plant-derived rhamnogalacturonan-I (RG-I) on pro- and anti-inflammatory responses of primary human monocyte-derived macrophages (HMDMs) induced by Escherichia coli LPS and Porphyromonas gingivalis bacteria. In the present study, two different types of surface materials, tissue culture polystyrene (TCPS) plates and titanium (Ti) discs, coated with pectic polysaccharides, potato unmodified RG-I (PU) and potato dearabinanated RG-I (PA), have been examined. The inflammatory responses of HMDMs after E. coli LPS/P. gingivalis stimulation were investigated through gene expression measurements of pro- and anti-inflammatory cytokines. The results showed that PU and PA decreased expression of the proinflammatory genes tumour necrosis factor-alpha (TNFA), interleukin-1 beta (IL1B) and interleukin-8 (IL8) in activated HMDMs cultured on TCPS/Ti surfaces. In contrast, the effects on anti-inflammatory interleukin-10 (IL10) gene expression were not significant. The results indicate that RG-Is should be considered as a candidate for organic nanocoatings of titanium implant surfaces in order to limit host proinflammatory responses and improve bone healing. DOI: http://dx.doi.org/10.5281/zenodo.1250541

Aligned nanofiber material supports cell growth and increases osteogenesis in canine adipose-derived mesenchymal stem cells in vitro.

Tissue engineering shows great promise for the treatment of degenerative diseases, including bone repair. Polymer nanofibers provide a three-dimensional (3-D) scaffold for attachment and growth of mesenchymal stem cells. Increasing evidence supports that fiber alignment on scaffolds plays a major role in the viability and differentiation of stem cells. We compared the cell viability of canine adipose tissue-derived mesenchymal stem cells (cADMSCs) cultured in the aligned- (NanoAligned™) and random- (NanoECM™) oriented polycaprolactone (PCL) nanofiber-coated plates to control polystyrene tissue culture plates using a proliferation assay. Ability of the plates to induce differentiation of cADMSCs into osteocytes, adipocytes, and neurons was evaluated based on expression of the osteocyte markers, COL1A1 and osterix; adipocyte markers PPARγ2 and LPL; and neuronal marker nestin using RT-PCR. Proliferation results demonstrated that aligned-oriented PCL nanofiber-coated plates were more suitable substrate for cADMSCs after 7 days in culture compared to random-oriented PCL nanofiber-coated or control plates. Additionally, we demonstrated that both 3-D PCL nanofiber-coated plates were a better scaffold for cADMSCs differentiation into osteocytes compared to control plates. In conclusion, our results confirm that PCL nanofiber is a suitable tissue engineering material for use in regenerative medicine for canine patients in vivo. © 2018 Wiley Periodicals, Inc. J Biomed Mater Res Part A: 106A: 1780-1788, 2018.

Silk fibroin/chitosan thin film promotes osteogenic and adipogenic differentiation of rat bone marrow-derived mesenchymal stem cells.

As a biodegradable polymer thin film, silk fibroin/chitosan composite film overcomes the defects of pure silk fibroin and chitosan films, respectively, and shows remarkable biocompatibility, appropriate hydrophilicity and mechanical properties. Silk fibroin/chitosan thin film can be used not only as metal implant coating for bone injury repair, but also as tissue engineering scaffold for skin, cornea, adipose, and other soft tissue injury repair. However, the biocompatibility of silk fibroin/chitosan thin film for mesenchymal stem cells, a kind of important seed cell of tissue engineering and regenerative medicine, is rarely reported. In this study, silk fibroin/chitosan film was prepared by solvent casting method, and the rat bone marrow-derived mesenchymal stem cells were cultured on the silk fibroin/chitosan thin film. Osteogenic and adipogenic differentiation of rat bone marrow-derived mesenchymal stem cells were induced, respectively. The proliferation ability, osteogenic and adipogenic differentiation abilities of rat bone marrow-derived mesenchymal stem cells were systematically compared between silk fibroin/chitosan thin film and polystyrene tissue culture plates. The results showed that silk fibroin/chitosan thin film not only provided a comparable environment for the growth and proliferation of rat bone marrow-derived mesenchymal stem cells but also promoted their osteogenic and adipogenic differentiation. This work provided information of rat bone marrow-derived mesenchymal stem cells behavior on silk fibroin/chitosan thin film and extended the application of silk fibroin/chitosan thin film. Based on the results, we suggested that the silk fibroin/chitosan thin film could be a promising material for tissue engineering of bone, cartilage, adipose, and skin.

Pectin nanocoating reduces proinflammatory fibroblast response to bacteria.

Implant failures are primarily related to bacterial infections and inflammation. Nanocoating of implant devices with organic molecules is a method used for improving their integration into host tissues and limiting inflammation. Bioengineered plant-derived rhamnogalacturonan-Is (RG-Is) from pectins improve tissue regeneration and exhibit anti-inflammatory properties. Therefore, the aim of this study is to evaluate the in vitro effect of RG-I nanocoating on human gingival primary fibroblast (HGF) activity and proinflammatory response following Porphyromonas gingivalis (P. gingivalis) infection. Infected HGFs were incubated on tissue culture polystyrene (TCPS) plates coated with unmodified RG-I isolated from potato pectin (PU) and dearabinanated RG-I (PA). HGF morphology, proliferation, metabolic activity, and expression of genes responsible for extracellular matrix (ECM) turnover and proinflammatory response were examined. Following the P. gingivalis infection, PU and PA significantly promoted HGF proliferation and metabolic activity. Moreover, gene expression levels of IL1B, IL8, TNFA, and MMP2 decreased in the infected cells cultured on PU and PA, whereas the expression of COL1A1, FN1, and FGFR1 was upregulated. The results indicate that RG-Is are promising candidates for nanocoating of an implant surface, can reduce inflammation, and enhance implant integration, particularly in medically compromised patients with chronic inflammatory diseases such as periodontitis and rheumatoid arthritis. © 2017 Wiley Periodicals, Inc. J Biomed Mater Res Part A: 105A: 3475-3481, 2017.