scholarly journals 3D-Printed Poly(ε-Caprolactone)/Hydroxyapatite Scaffolds Modified with Alkaline Hydrolysis Enhance Osteogenesis In Vitro

Polymers ◽  
2021 ◽  
Vol 13 (2) ◽  
pp. 257
Author(s):  
Sangbae Park ◽  
Jae Eun Kim ◽  
Jinsub Han ◽  
Seung Jeong ◽  
Jae Woon Lim ◽  
...  

The 3D-printed bioactive ceramic incorporated Poly(ε-caprolactone) (PCL) scaffolds show great promise as synthetic bone graft substitutes. However, 3D-printed scaffolds still lack adequate surface properties for cells to be attached to them. In this study, we modified the surface characteristics of 3D-printed poly(ε-caprolactone)/hydroxyapatite scaffolds using O2 plasma and sodium hydroxide. The surface property of the alkaline hydrolyzed and O2 plasma-treated PCL/HA scaffolds were evaluated using field-emission scanning microscopy (FE-SEM), Alizarin Red S (ARS) staining, and water contact angle analysis, respectively. The in vitro behavior of the scaffolds was investigated using human dental pulp-derived stem cells (hDPSCs). Cell proliferation of hDPSCs on the scaffolds was evaluated via immunocytochemistry (ICC) and water-soluble tetrazolium salt (WST-1) assay. Osteogenic differentiation of hDPSCs on the scaffolds was further investigated using ARS staining and Western blot analysis. The result of this study shows that alkaline treatment is beneficial for exposing hydroxyapatite particles embedded in the scaffolds compared to O2 plasma treatment, which promotes cell proliferation and differentiation of hDPSCs.

Materials ◽  
2019 ◽  
Vol 12 (20) ◽  
pp. 3419 ◽  
Author(s):  
Boldbayar Ganbold ◽  
Seong-Joo Heo ◽  
Jai-Young Koak ◽  
Seong-Kyun Kim ◽  
Jaejin Cho

Recently, the selective laser melting (SLM) method of manufacturing three dimensional (3D) dental prosthetics by applying a laser to metal powder has been widely used in the field of dentistry. This study investigated human adipose derived stem cell (hADSC) behavior on a 3D printed cobalt-chrome (Co-Cr) alloy and its surface characteristics and compared them those of a nickel-chrome (Ni-Cr) alloy. Alloys were divided into four groups according to the material and manufacturing methods. Co-Cr disks were manufactured with three different methods: a conventional casting method, a metal milling method, and an SLM method. Ni-Cr disks were manufactured with a conventional casting method. The surface roughness and compositions of the disks were assessed. hADSCs were then cultured on the disks. Cell morphologies on the disks were analyzed by a field emission scanning electron microscope (FE-SEM). Cell proliferation was assessed with a bromodeoxyuridine (BrdU) assay kit. Cell viability was evaluated with a water-soluble tetrazolium salt (WST) assay kit. There were no differences in surface roughness between all groups. The cells were well attached to the disks, and morphologies of the cells were similar. The cell proliferation and viability of the Ni-Cr disks were significantly lower than the other groups. However, the Co-Cr disks showed no differences in their different fabricating methods. In conclusion, the biocompatibility of 3D printed Co-Cr alloys showed comparable results compared to that of the conventional casting method, and these alloys were more biocompatible than Ni-Cr alloys.


2019 ◽  
Vol 2019 ◽  
pp. 1-8
Author(s):  
Wei Xiong ◽  
Yue-kun Shen ◽  
Peng Dong ◽  
Ying Xiao ◽  
Xiong-qing Huang ◽  
...  

Sustained release of anesthesia has shown great promise in the treatment of chronic pain in patients. In this research, we used neutralized ropivacaine as an anesthesia and poly(ε-caprolactone) (PCL) with different architectures to systematically study how these architectures affect the release of ropivacaine. After optimizing the parameters of the preparation of microspheres, ropivacaine-loaded 1-PCL microspheres and 4-PCL microspheres were obtained. Fourier Transform infrared spectra (FT-IR) and X-ray diffraction spectra (XRD) confirmed that ropivacaine was encapsulated within the microsphere rather than inserted on the surface of the microsphere. Ropivacaine was found to be buried deeper in the 1-PCL microsphere than in the 4-PCL microsphere. In vitro release assay revealed that small crystalline grains interfered with ropivacaine release in 4-PCL microspheres during the initial release period, but then two kinds of microspheres showed a similar ropivacaine release rate. We basically proved that the architecture of PCL has a negligible effect on ropivacaine release. Cell proliferation test revealed that the release of products from the microspheres resulted in insignificant toxicity towards mammalian cells.


Materials ◽  
2020 ◽  
Vol 13 (15) ◽  
pp. 3305
Author(s):  
Pablo Kraemer Fernandez ◽  
Alexey Unkovskiy ◽  
Viola Benkendorff ◽  
Andrea Klink ◽  
Sebastian Spintzyk

(1) Background: To date, no information on the polishability of milled and 3D-printed complete denture bases has been provided, which is relevant in terms of plaque accumulation. (2) Methods: three groups (n = 30) were manufactured using the cold-polymerization polymethilmethacrilate, milling (SM) and 3D printing (AM). 10 specimens of each group were left untreated (reference). 10 more specimens were pre-polished (intermediate polishing) and 10 final specimens were highgloss polished. An additional 20 specimens were 3D printed and coated with the liquid resin (coated), 10 of which were additionally polished (coated + polished). For each group Ra and Rz values, gloss value and REM images were obtained. (3). The “highgloss-polished” specimens showed statistically lower Ra and Rz values in the SM, followed by AM and conventional groups. In the AM group statistically lower surfaces roughness was revealed for highgloss-polished, “coated + polished”, and “coated” specimens, respectively. (4) Conclusions: The milled specimens demonstrated superiors surface characteristics than 3D printed and conventionally produced after polishing. The polished specimens demonstrated superior surface characteristics over coated specimens. However, the surface roughness by both polished and coated specimens was within the clinically relevant threshold of 0.2 µm.


2018 ◽  
Vol 5 (8) ◽  
pp. 172033 ◽  
Author(s):  
Zhenfei Huang ◽  
Zhihong Wu ◽  
Bupeng Ma ◽  
Lingjia Yu ◽  
Yu He ◽  
...  

Titanium (Ti) is an ideal bone substitute due to its superior bio-compatibility and remarkable corrosion resistance. However, in order to improve the osteoconduction and osteoinduction capacities in clinical applications, different kinds of surface modifications are typically applied to Ti alloys. In this study, we fabricated a tightly attached polydopamine-assisted Fe 3 O 4 nanoparticle coating on Ti with magnetic properties, aiming to improve the osteogenesis of the Ti substrates. The PDA-assisted Fe 3 O 4 nanoparticle coatings were characterized by scanning electron microscopy, energy dispersive spectroscopy, atomic force microscopy and water contact angle measurements. The cell attachment and proliferation rate of the human bone mesenchymal stem cells (hBMSCs) on the Ti surface significantly improved with the Fe 3 O 4 /PDA coating when compared with the pure Ti without a coating. Furthermore, the results of in vitro alkaline phosphatase (ALP) activity at 7 and 14 days and alizarin red S staining at 14 days showed that the Fe 3 O 4 /PDA coating on Ti promoted the osteogenic differentiation of hBMSCs. Moreover, hBMSCs co-cultured with the Fe 3 O 4 /PDA-coated Ti for approximately 14 days also exhibited a significantly higher mRNA expression level of ALP, osteocalcin and runt-related transcription factor-2 (RUNX2). Our in vitro results revealed that the present PDA-assisted Fe 3 O 4 nanoparticle surface coating is an innovative method for Ti surface modification and shows great potential for clinical applications.


Author(s):  
Índia Olinta de Azevedo Queiroz ◽  
Thiago Machado ◽  
Camila Carneiro Alves ◽  
Victor Gustavo Balera Brito ◽  
Bruno Carvalho de Vasconcelos ◽  
...  

AbstractThis study aimed to investigate the antimicrobial and biological properties of Ambroxol associated with glycerin (GLI), propylene glycol (PG), and polyethylene glycol (PEG) as a possible vehicle for an experimental tricalcium silicate sealer, with the intention of developing a new biomaterial. Mouse undifferentiated dental pulp cells (OD-21) were cultured, and the effects of different association on cell proliferation and inflammatory cytokine production were investigated. Antimicrobial adhesion of Enterococcus faecalis to setting sealers at 2 h was evaluated. Polyethylene tubes containing experimental sealers and empty tubes were implanted into dorsal connective tissues of 12 male 3- to 4-months-old Wistar rats (250–280 g). After 7 and 30 days, the tubes were removed and processed for histological and immunohistochemical analyses. ANOVA followed by Bonferroni correction and ANOVA followed by Tukey test was used for parametric data and Kruskal–Wallis followed by Dunn for nonparametric (p < 0.05). Cell proliferation was dose-dependent, since all association were cytotoxic at higher concentrations; however, Ambroxol–PEG showed significantly higher cytotoxicity than other association (p < 0.05). In addition, irrespective of the association, no cytokine production was observed in vitro. Ambroxol–GLI reduced bacterial viability, whereas Ambroxol–PEG increased (p < 0.05). Histological examination showed no significant difference in the inflammatory response (p > 0.05) and mineralization ability in all association. Additionally, IL-1β and TNF-α were upregulated on Ambroxol–PEG in relation to Control at 07 days (p < 0.05). Ambroxol–GLI was the best vehicle for experimental tricalcium silicate sealer, as it promoted an increase in antimicrobial activity without altering the inflammatory response or mineralization ability.


2021 ◽  
Vol 11 (23) ◽  
pp. 11273
Author(s):  
Kazuyuki Yusa ◽  
Shigeo Ishikawa ◽  
Tomoharu Hemmi ◽  
Hiroshi Takano ◽  
Masayuki Fukuda ◽  
...  

This study investigates the effects of zoledronic acid (ZA) and compressive force on osteoblast functions, to elucidate the pathogenesis of medication-related osteonecrosis of the jaw (MRONJ). MC3T3-E1 cells were exposed to ZA (1, 10 and 100 µM) to evaluate the effects of ZA on cell proliferation. Furthermore, to investigate the influence of ZA with or without compressive force on osteoblast differentiation, real-time polymerase chain reaction and Alizarin Red S staining were performed. ZA concentrations > 10 μM were highly cytotoxic to MC3T3-E1 cells. Combining 1-μM ZA with compressive force influenced expression levels of osteoblast-related genes and matrix mineralization. The inhibitory effects of ZA on cell proliferation and the combination of ZA and compressive force on osteoblast differentiation may contribute to the pathogenesis of MRONJ.


2020 ◽  
Vol 2020 ◽  
pp. 1-10
Author(s):  
Lina M. Escobar ◽  
Zita Bendahan ◽  
Andrea Bayona ◽  
Jaime E. Castellanos ◽  
María-Clara González

Introduction. The aim of the present study was to determine the effects of vitamins D and E on the proliferation, morphology, and differentiation of human dental pulp stem cells (hDPSCs). Methods. In this in vitro experimental study, hDPSCs were isolated, characterized, and treated with vitamins D and E, individually and in combination, utilizing different doses and treatment periods. Changes in morphology and cell proliferation were evaluated using light microscopy and the resazurin assay, respectively. Osteoblast differentiation was evaluated with alizarin red S staining and expression of RUNX2, Osterix, and Osteocalcin genes using real-time RT-PCR. Results. Compared with untreated cells, the number of cells significantly reduced following treatment with vitamin D (49%), vitamin E (35%), and vitamins D + E (61%) after 144 h. Compared with cell cultures treated with individual vitamins, cells treated with vitamins D + E demonstrated decreased cell confluence, with more extensive and flatter cytoplasm that initiated the formation of a significantly large number of calcified nodules after 7 days of treatment. After 14 days, treatment with vitamins D, E, and D + E increased the transcription of RUNX2, Osterix, and Osteocalcin genes. Conclusions. Vitamins D and E induced osteoblastic differentiation of hDPSCs, as evidenced by the decrease in cell proliferation, morphological changes, and the formation of calcified nodules, increasing the expression of differentiation genes. Concurrent treatment with vitamins D + E induces a synergistic effect in differentiation toward an osteoblastic lineage.


Cells ◽  
2020 ◽  
Vol 9 (3) ◽  
pp. 775 ◽  
Author(s):  
Yingying Cui ◽  
Mengfan Zhang ◽  
Changsen Leng ◽  
Tjasso Blokzijl ◽  
Bernadien H. Jansen ◽  
...  

Intestinal fibrosis is a common complication of inflammatory bowel disease. So far, there is no safe and effective drug for intestinal fibrosis. Pirfenidone is an anti-fibrotic compound available for the treatment of idiopathic pulmonary fibrosis. Here, we explored the anti-proliferative and anti-fibrotic properties of pirfenidone on primary human intestinal fibroblasts (p-hIFs). p-hIFs were cultured in the absence and presence of pirfenidone. Cell proliferation was measured by a real-time cell analyzer (xCELLigence) and BrdU incorporation. Cell motility was monitored by live cell imaging. Cytotoxicity and cell viability were analyzed by Sytox green, Caspase-3 and Water Soluble Tetrazolium Salt-1 (WST-1) assays. Gene expression of fibrosis markers was determined by quantitative reverse transcription PCR (RT-qPCR). The mammalian target of rapamycin (mTOR) signaling was analyzed by Western blotting and type I collagen protein expression additionally by immunofluorescence microscopy. Pirfenidone dose-dependently inhibited p-hIF proliferation and motility, without inducing cell death. Pirfenidone suppressed mRNA levels of genes that contribute to extracellular matrix production, as well as basal and TGF-β1-induced collagen I protein production, which was associated with inhibition of the rapamycin-sensitive mTOR/p70S6K pathway in p-hIFs. Thus, pirfenidone inhibits the proliferation of intestinal fibroblasts and suppresses collagen I production through the TGF-β1/mTOR/p70S6K signaling pathway, which might be a novel and safe anti-fibrotic strategy to treat intestinal fibrosis.


Nanomaterials ◽  
2019 ◽  
Vol 9 (3) ◽  
pp. 367 ◽  
Author(s):  
Chuan Yin ◽  
Sélène Rozet ◽  
Rino Okamoto ◽  
Mikihisa Kondo ◽  
Yasushi Tamada ◽  
...  

In this study, the physical properties and the biocompatibility of electrospun silicone-modified polyurethane (PUSX) nanofibers were discussed and compared with PUSX films. To investigate the effects of different structures on the physical properties, tensile strength, elongation at break, Young’s modulus, water retention, water contact angle (WCA) and thermal conductivity measurements were performed. To prove the in vitro biocompatibility of the materials, cell adhesion, cell proliferation, and cytotoxicity were studied by NIH3T3 mouse embryonic fibroblasts cells following by lactate dehydrogenase (LDH) analysis. As a conclusion, the mechanical properties, water retention, and WCA were proven to be able to be controlled and improved by adjusting the structure of PUSX. A higher hydrophobicity and lower thermal conductivity were found in PUSX nanofibers compared with polyurethane (PU) nanofibers and films. An in vitro biocompatibility evaluation shows that the cell proliferation can be performed on both PUSX nanofibers and films. However, within a short period, cells prefer to attach and entangle on PUSX nanofibers rather than PUSX films. PUSX nanofibers were proven to be a nontoxic alternative for PU nano-membranes or films in the biomedical field, because of the controllable physical properties and the biocompatibility.


Pharmaceutics ◽  
2019 ◽  
Vol 11 (10) ◽  
pp. 500 ◽  
Author(s):  
Denise Beconcini ◽  
Francesca Felice ◽  
Ylenia Zambito ◽  
Angela Fabiano ◽  
Anna Maria Piras ◽  
...  

This study aimed at evaluating the anti-inflammatory effect of natural cherry extract (CE), either free or encapsulated in nanoparticles (NPs) based on chitosan derivatives (Ch-der) or poly(lactic-co-glycolic acid) (PLGA), on human umbilical vein endothelial cells (HUVEC). CE from Prunus avium L. was characterized for total polyphenols, flavonoids, and anthocyanins content. CE and CE-loaded NP cytotoxicity and protective effect on lipopolysaccharide (LPS)-stressed HUVEC were tested by water-soluble tetrazolium salt (WST-1) assay. Pro- and anti-inflammatory cytokines (TNF-α, IL-6, IL-10, and PGE2) released by HUVEC were quantified by enzyme-linked immunosorbent assay (ELISA). All NP types were internalized into HUVEC after 2 h incubation and promoted the anti-inflammatory effect of free CE at the concentration of 2 µg gallic acid equivalents (GAE)/mL. CE-loaded Ch-der NPs showed the highest in vitro uptake and anti-inflammatory activity, blunting the secretion of IL-6, TNF-α, and PGE2 cytokines. Moreover, all NPs reduced the production of nitric oxide and NLRP3 inflammasome, and had a stronger anti-inflammatory effect than the major corticosteroid dexamethasone. In particular, the results demonstrate that natural CE protects endothelial cells from inflammatory stress when encapsulated in NPs based on quaternary ammonium chitosan. The CE beneficial effects were directly related with in vitro internalization of CE-loaded NPs.


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