This study is devoted to fabricate a novel hydroxyapatite(HAp)/gelatin scaffold coated with nano-HAp in nano-rod configuration to evaluate its biocompatibility potential. The nano-HAp particles are needle and rod-like with widths ranging between 30 to 60 nm and lengths from 100 to 300 nm, respectively. Because of their higher surface area and higher reactivity, the nano-rod particles were distributed in gelatin much better than spherical and mixed shapes particles. The compressive modulus of the nano-HAp/gelatin scaffolds coated with nano-HAp was comparable with the compressive modulus of a human cancellous bone. The potential performance of the fabricated scaffolds as seeding media was assayed using mesenchymal stem cells (MSCs). MTT (3-(4,5-dimethylthiazol-2-yl)-1,5-diphenyl tetrazulium bromide) assays were performed on days 4 and 7 and the number of the cells per scaffold was determined. On the basis of this assay, all the studied scaffolds exhibited an appropriate environment in which the loaded cells appeared to be proliferated during the cultivation periods. In all fabricated composite scaffolds, marrow-derived MSCs appeared to occupy the scaffolds internal spaces and attach on their surfaces. According to the cell culture experiments, the incorporation of rod-like nano-HAp and coating of scaffolds with nano-HAp particles enabled the prepared scaffolds to possess desirable biocompatibility, high bioactivity, and sufficient mechanical strength in comparison with noncoated HAp samples. This research suggests that the newly developed scaffold has a potential as a suitable scaffold for bone tissue engineering.
The field of tissue engineering is an emerging discipline which applies the basic principles of life sciences and engineering to repair and restore living tissues and organs. The purpose of this study was to investigate the effect of cold and non-thermal plasma surface modification of poly (ϵ-caprolactone) (PCL) scaffolds on fibroblast cell behavior. Nano-fiber PCL was fabricated through electrospinning technique, and some fibers were then treated by cold and non-thermal plasma. The cell-biomaterial interactions were studied by culturing the fibroblast cells on nano-fiber PCL. Scaffold biocompatibility test was assessed using an inverted microscope. The growth and proliferation of fibroblast cells on nano-fiber PCL were analyzed by MTT viability assay. Cellular attachment on the nano-fiber and their morphology were evaluated using scanning electron microscope. The result of cell culture showed that nano-fiber could support the cellular growth and proliferation by developing three-dimensional topography. The present study demonstrated that the nano-fiber surface modification with cold plasma sharply enhanced the fibroblast cell attachment. Thus, cold plasma surface modification greatly raised the bioactivity of scaffolds.
In situ forming drug delivery system is prepared by phase inversion technique using poly (D,L-lacticco-glycolide) and leuprolide acetate dissolved in N-methyl-2-pyrrolidone. The effects of ethyl heptanoate and glycerol additives are important determinant as rate modifying agents on the drug release kinetics in biodegradable in situ forming porous systems of poly(D,L-lactide-co-glycolide) (PLGA) in N-methyl-2-pyrrolidone (NMP). The release performance and porous structure morphology are investigated by scanning electron microscopy and UV-visible spectroscopy techniques to study the effect of additives. The experimental results exhibit the crucial role of ethyl heptanoate and glycerol at different loadings (1, 3, and 5% w/w) on release profile of leuprolide acetate loaded on poly(D,L-lactide-co-glycolide)hydroxylated (PLGA-H). Both additives at different concentrations reduce the burst effect, while increasing duration of drug release. Ethyl heptanoate, however, shows stronger effect than glycerol. The results of morphological studies show that ethyl heptanoate reduces the porosity of the polymer surface and interconnected tear-like structures of the bulk disappear while the sponge-like structures are observed. In this system glycerol reduces the surface porosity intensively, while the interconnected tears change into channel-like structures. Therefore, morphological results confirm the effect of additives on leuprolide release profile.
Collagen I as a major organic component of bone matrix may be important for establishment and maintenance of mesenchymal stem cells (MSCs) in osteogenic 3D culture. To explore this subject, murine marrow-derived MSCs were seeded onto hybrid scaffolds of alginate/gelatin/beta-tricalcium phosphate in a medium either with or without collagen I gel. The cultures were then provided with osteogenic medium and incubated for three weeks during which loading efficiency, cell proliferation and the culture mineralization were quantified and statistically compared. According to the findings, in culture with collagen, although about 60% of the cells left the scaffolds, the remaining cells, however, proliferated extensively with a population doubling number (PDN) equivalent to 2.46 +/- 0.31 and organized as cell aggregations that were heavily mineralized (calcium concentration = 1.017 +/- 0.141 mM per scaffold), whereas in the culture without collagen, about 75% of the cells left the scaffolds, less cell proliferation occurred (PDN = 1.48 +/- 0.29) and no cell aggregation was observed. The calcium concentration in this culture was 0.185 +/- 0.029 mM per scaffold. All these differences were statistically significant (p < 0.001). Taken together, these data suggested that using the collagen I in seeding medium could help mMSCs loading into the scaffold, enhance their subsequent proliferation, and increase calcium deposition in 3D culture system.
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