Core‐shell nanofibers: Integrating the bioactivity of gelatin and the mechanical property of polyvinyl alcohol

Coaxial electrospinning is used to fabricate nanofibers with gelatin in the shell and polyvinyl alcohol (PVA) in the core in order to derive mechanical strength from PVA and bioactivity from gelatin. At a 1:1 PVA/gelatin mass ratio, the core‐shell nanofiber scaffolds display a Young's modulus of 168.6 ± 36.5 MPa and a tensile strength of 5.42 ± 1.95 MPa, which are significantly higher than those of the scaffolds composed solely of gelatin or PVA. The Young's modulus and tensile strength of the core‐shell nanofibers are further improved by reducing the PVA/gelatin mass ratio from 1:1 to 1:3. The mechanical analysis of the core‐shell nanofibers suggests that the presence of the gelatin shell may improve the molecular alignment of the PVA core, transforming the semi‐crystalline, plastic PVA into a more crystallized, elastic PVA, and enhancing the mechanical properties of the core. Lastly, the PVA/gelatin core‐shell nanofibers possess cellular viability, proliferation, and adhesion similar to these of the gelatin nanofibers, and show significantly higher proliferation and adhesion than the PVA nanofibers. Taken together, the coaxial electrospinning of nanofibers with a core‐shell structure permits integration of the bioactivity of gelatin and the mechanical strength of PVA in single fibers. © 2013 Wiley Periodicals, Inc. Biopolymers 101: 336–346, 2014.     

Co-electrospun blends of PLGA, gelatin, and elastin as potential nonthrombogenic scaffolds for vascular tissue engineering

In search for novel biomimetic scaffolds for application in vascular tissue engineering, we evaluated a series of fibrous scaffolds prepared by coelectrospinning tertiary blends of poly(lactide-co-glycolide) (PLGA), gelatin, and elastin (PGE). By systematically varying the ratios of PLGA and gelatin, we could fine-tune fiber size and swelling upon hydration as well as the mechanical properties of the scaffolds. Of all PGE blends tested, PGE321 (PLGA, gelatin, elastin v/v/v ratios of 3:2:1) produced the smallest fiber size (317 ± 46 nm, 446 ± 69 nm once hydrated) and exhibited the highest Young's modulus (770 ± 131 kPa) and tensile strength (130 ± 7 kPa). All PGE scaffolds supported the attachment and metabolization of human endothelial cells (ECs) and bovine aortic smooth muscle cells (SMCs) with some variances in EC morphology and cytoskeletal spreading observed at 48 h postseeding, whereas no morphologic differences were observed at confluence (day 8). The rate of metabolization of ECs, but not of SMCs, was lower than that on tissue culture plastic and depended on the specific PGE composition. Importantly, PGE scaffolds were capable of guiding the organotypic distribution of ECs and SMCs on and within the scaffolds, respectively. Moreover, the EC monolayer generated on the PGE scaffold surface was nonthrombogenic and functional, as assessed by the basal and cytokine-inducible levels of mRNA expression and amidolytic activity of tissue factor, a key player in the extrinsic clotting cascade. Taken together, our data indicate the potential application of PGE scaffolds in vascular tissue engineering.     

Fabrication of gelatin nanofibrous scaffolds using ethanol/phosphate buffer saline as a benign solvent

Electrospinning of natural polymer nanofibers useful for biomedical applications often requires the use of cytotoxic organic solvents. In this study, gelatin nanofibers are electrospun from phosphate buffer saline/ethanol binary mixtures as a benign solvent at ambient temperature. The influences of ionic strength, ethanol concentration, and gelatin concentration on the electrospinnability of gelatin solutions and the fiber microarchitectures are analyzed. The electrospun scaffolds retain their morphologies during vapor‐phase crosslinking with glutaraldehyde in ethanol and the subsequent removal of salts contained in the nanofibers via water rinsing. When fully hydrated, the mechanically preconditioned scaffolds display a Young's modulus of 25.5 ± 5.3 kPa, tensile strength of 55.5 ± 13.9 kPa, deformability of 160 ± 15%, and resilience of 89.9 ± 1.8%. When cultured on the gelatin scaffolds, 3T3 fibroblasts displayed spindle‐like morphology, similar to the cell's normal morphology in a 3D extracellular matrix. © 2012 Wiley Periodicals, Inc. Biopolymers 97:1026–1036, 2012.     

Hybrid chitosan–ß‐glycerol phosphate–gelatin nano‐/micro fibrous scaffolds with suitable mechanical and biological properties for tissue engineering

Scaffold‐based tissue engineering is considered as a promising approach in the regenerative medicine. Graft instability of collagen, by causing poor mechanical properties and rapid degradation, and their hard handling remains major challenges to be addressed. In this research, a composite structured nano‐/microfibrous scaffold, made from a mixture of chitosan–ß‐glycerol phosphate–gelatin (chitosan–GP–gelatin) using a standard electrospinning set‐up was developed. Gelatin–acid acetic and chitosan ß‐glycerol phosphate–HCL solutions were prepared at ratios of 30/70, 50/50, 70/30 (w/w) and their mechanical and biological properties were engineered. Furthermore, the pore structure of the fabricated nanofibrous scaffolds was investigated and predicted using a theoretical model. Higher gelatin concentrations in the polymer blend resulted in significant increase in mean pore size and its distribution. Interaction between the scaffold and the contained cells was also monitored and compared in the test and control groups. Scaffolds with higher chitosan concentrations showed higher rate of cell attachment with better proliferation property, compared with gelatin‐only scaffolds. The fabricated scaffolds, unlike many other natural polymers, also exhibit non‐toxic and biodegradable properties in the grafted tissues. In conclusion, the data clearly showed that the fabricated biomaterial is a biologically compatible scaffold with potential to serve as a proper platform for retaining the cultured cells for further application in cell‐based tissue engineering, especially in wound healing practices. These results suggested the potential of using mesoporous composite chitosan–GP–gelatin fibrous scaffolds for engineering three‐dimensional tissues with different inherent cell characteristics. © 2015 Wiley Periodicals, Inc. Biopolymers 105: 163–175, 2016.     

Nanofibrous nonmulberry silk/PVA scaffold for osteoinduction and osseointegration

Poly‐vinyl alcohol and nonmulberry tasar silk fibroin of Antheraea mylitta are blended to fabricate nanofibrous scaffolds for bone regeneration. Nanofibrous matrices are prepared by electrospinning the equal volume ratio blends of silk fibroin (2 and 4 wt%) with poly‐vinyl alcohol solution (10 wt%) and designated as 2SF/PVA and 4SF/PVA, respectively with average nanofiber diameters of 177 ± 13 nm (2SF/PVA) and 193 ± 17 nm (4SF/PVA). Fourier transform infrared spectroscopy confirms retention of the secondary structure of fibroin in blends indicating the structural stability of neo‐matrix. Both thermal stability and contact angle of the blends decrease with increasing fibroin percentage. Conversely, fibroin imparts mechanical stability to the blends; greater tensile strength is observed with increasing fibroin concentration. Blended scaffolds are biodegradable and support well the neo‐bone matrix synthesis by human osteoblast like cells. The findings indicate the potentiality of nanofibrous scaffolds of nonmulberry fibroin as bone scaffolding material. © 2014 Wiley Periodicals, Inc. Biopolymers 103: 271–284, 2015.     

Three-dimensional electrospun gelatin scaffold coseeded with embryonic stem cells and sertoli cells: A promising substrate for in vitro coculture system

In this study, we present an electrospun gelatin (EG) scaffold to mimic the extracellular matrix of the testis. The EG scaffold was synthesized by electrospinning and crosslinked with glutaraldehyde vapor to decrease its water solubility and degradation rate. The scanning electron microscope micrographs showed the homogenous morphology of randomly aligned gelatin fibers. The average diameter of gelatin fibers before and after crosslinking was approximately 180 and 220 nm, respectively. Modulus, tensile strength, and elongation at break values were as 161.8 ± 24.4 MPa, 4.21 ± 0.54 MPa, and 7.06 ± 2.12 MPa, respectively. The crosslinked EG showed 75.2% ± 4.5% weight loss after 14 days with no changes in the pH value of degradation solution. Cytobiocompatibility of the EG for sertoli cells and embryonic stem cells (ESCs) was determined in vitro. Sertoli cells were isolated from mouse testis and characterized by immunostaining and flow cytometry. The effects of EG on proliferation and attachment of both sertoli cells and ESCs were examined. The EG scaffolds exhibited no cytotoxicity for sertoli and ESCs. Both sertoli and ESCs were well attached and grown on EG. Coculture of sertoli and ESCs on EG showed better ESCs adhesion compared with ESCs alone. Our findings indicate the potential of EG as a substrate for proliferation, adhesion, and coculture of sertoli and ESCs and may be considered as a promising engineered microenvironment for in vitro coculture system with the aim of guiding stem cells differentiation toward sperm-producing cells.     

Polyurethane/Gelatin Nanofibrils Neural Guidance Conduit Containing Platelet-Rich Plasma and Melatonin for Transplantation of Schwann Cells

The current study aimed to enhance the efficacy of peripheral nerve regeneration using a biodegradable porous neural guidance conduit as a carrier to transplant allogeneic Schwann cells (SCs). The conduit was prepared from polyurethane (PU) and gelatin nanofibrils (GNFs) using thermally induced phase separation technique and filled with melatonin (MLT) and platelet-rich plasma (PRP). The prepared conduit had the porosity of 87.17 ± 1.89%, the contact angle of 78.17 ± 5.30° and the ultimate tensile strength and Young’s modulus of 5.40 ± 0.98 MPa and 3.13 ± 0.65 GPa, respectively. The conduit lost about 14% of its weight after 60 days in distilled water. The produced conduit enhanced the proliferation of SCs demonstrated by a tetrazolium salt-based assay. For functional analysis, the conduit was seeded with 1.50 × 10⁴ SCs (PU/GNFs/PRP/MLT/SCs) and implanted into a 10-mm sciatic nerve defect of Wistar rat. Three control groups were used: (1) PU/GNFs/SCs, (2) PU/GNFs/PRP/SCs, and (3) Autograft. The results of sciatic functional index, hot plate latency, compound muscle action potential amplitude and latency, weight-loss percentage of wet gastrocnemius muscle and histopathological examination using hematoxylin–eosin and Luxol fast blue staining, demonstrated that using the PU/GNFs/PRP/MLT conduit to transplant SCs to the sciatic nerve defect resulted in a higher regenerative outcome than the PU/GNFs and PU/GNFs/PRP conduits.     

Fabrication of a biomimetic spinal cord tissue construct with heterogenous mechanical properties using intrascaffold cell assembly

In tissue engineering studies, scaffolds play a very important role in offering both physical and chemical cues for cell growth and tissue regeneration. However, in some cases, tissue regeneration requires scaffolds with high mechanical properties (e.g., bone and cartilage), while cells need a soft mechanical microenvironment. In this study, to mimic the heterogenous mechanical properties of a spinal cord tissue, a biomimetic rat tissue construct is fabricated. A collagen-coated poly(lactic-co-glycolic acid) scaffold is manufactured using thermally induced phase separation casting. Primary rat neural cells (P01 Wistar rat cortex) with soft hydrogels are later printed within the scaffold using an image-guided intrascaffold cell assembly technique. The scaffolds have unidirectional microporous structure with parallel axial macrochannels (260 ± 4 µm in diameter). Scaffolds showed mechanical properties similar to rat spine (ultimate tensile strength: 0.085 MPa, Young's modulus [stretch]: 0.31 MPa). The bioink composed of gelatin/alginate/fibrinogen is precisely printed into the macrochannels and showed mechanical properties suitable for neural cells (Young's modulus [compressive]: 3.814 kPa). Scaffold interface, cell viability, and immunostaining analyses show uniform distribution of stable, healthy, and elongated neural cells and neurites over 14 culture days in vitro. The results demonstrated that this method can serve as a valuable tool to aid manufacturing of tissue constructs requiring heterogenous mechanical properties for complex cell and/or biomolecule assembly.     

Complete recombinant silk-elastinlike protein-based tissue scaffold

Due to their improved biocompatibility and specificity over synthetic materials, protein-based biomaterials, either derived from natural sources or genetically engineered, have been widely fabricated into nanofibrous scaffolds for tissue engineering applications. However, their inferior mechanical properties often require the reinforcement of protein-based tissue scaffolds using synthetic polymers. In this study, we report the electrospinning of a completely recombinant silk-elastinlike protein-based tissue scaffold with excellent mechanical properties and biocompatibility. In particular, SELP-47K containing tandemly repeated polypeptide sequences derived from native silk and elastin was electrospun into nanofibrous scaffolds, and stabilized via chemical vapor treatment and mechanical preconditioning. When fully hydrated in 1× PBS at 37 °C, mechanically preconditioned SELP-47K scaffolds displayed elastic moduli of 3.4-13.2 MPa, ultimate tensile strengths of 5.7-13.5 MPa, deformabilities of 100-130% strain, and resilience of 80.6-86.9%, closely matching or exceeding those of protein-synthetic blend polymeric scaffolds. Additionally, SELP-47K nanofibrous scaffolds promoted cell attachment and growth, demonstrating their in vitro biocompatibility.     

On‐demand three‐dimensional freeform fabrication of multi‐layered hydrogel scaffold with fluidic channels

One of the challenges in tissue engineering is to provide adequate supplies of oxygen and nutrients to cells within the engineered tissue construct. Soft‐lithographic techniques have allowed the generation of hydrogel scaffolds containing a network of fluidic channels, but at the cost of complicated and often time‐consuming manufacturing steps. We report a three‐dimensional (3D) direct printing technique to construct hydrogel scaffolds containing fluidic channels. Cells can also be printed on to and embedded in the scaffold with this technique. Collagen hydrogel precursor was printed and subsequently crosslinked via nebulized sodium bicarbonate solution. A heated gelatin solution, which served as a sacrificial element for the fluidic channels, was printed between the collagen layers. The process was repeated layer‐by‐layer to form a 3D hydrogel block. The printed hydrogel block was heated to 37°C, which allowed the gelatin to be selectively liquefied and drained, generating a hollow channel within the collagen scaffold. The dermal fibroblasts grown in a scaffold containing fluidic channels showed significantly elevated cell viability compared to the ones without any channels. The on‐demand capability to print fluidic channel structures and cells in a 3D hydrogel scaffold offers flexibility in generating perfusable 3D artificial tissue composites. Biotechnol. Bioeng. 2010;105: 1178–1186. © 2009 Wiley Periodicals, Inc.     

Electrospun tecophilic/gelatin nanofibers with potential for small diameter blood vessel tissue engineering

Tissue engineering techniques particularly using electrospun scaffolds have been intensively used in recent years for the development of small diameter vascular grafts. However, the development of a completely successful scaffold that fulfills multiple requirements to guarantee complete vascular regeneration remains challenging. In this study, a hydrophilic and compliant polyurethane namely Tecophilic (TP) blended with gelatin (gel) at a weight ratio of 70:30 (TP(70)/gel(30)) was electrospun to fabricate a tubular composite scaffold with biomechanical properties closely simulating those of native blood vessels. Hydrophilic properties of the composite scaffold induced non‐thrombogenicity while the incorporation of gelatin molecules within the scaffold greatly improved the capacity of the scaffold to serve as an adhesive substrate for vascular smooth muscle cells (SMCs), in comparison to pure TP. Preservation of the contractile phenotype of SMCs seeded on electrospun TP(70)/gel(30) was yet another promising feature of this scaffold. The nanostructured TP(70)/gel(30) demonstrated potential feasibility toward functioning as a vascular graft. © 2014 Wiley Periodicals, Inc. Biopolymers 101: 1165–1180, 2014.     

Cytocompatibility of electrospun nanofiber tubular scaffolds for small diameter tissue engineering blood vessels

A tubular scaffold was fabricated by using electrospun polymer solution blends of pNSR32 (recombinant spider silk protein), PCL (polycaprolactone) and Gt (gelatin). The physicochemical properties and cytocompatibility of these scaffolds were investigated. Afterwards, the pNSR32/PCL/Gt tubular scaffold (inner diameter = 3 mm) showed high porosity of 86.2 ± 2.9%, pore size of 2423 ± 979 nm and average fibre diameter of 166 ± 85 nm. Water uptake and contact angle of the scaffolds reached 112.0 ± 4.4% and 45.7 ± 13.7°, respectively. SDRAECs (Sprague Dawley Rat Aortic Endothelial Cells) grew and proliferated well and phenotype could be maintained on the composite scaffolds after they had been cultured on the composite scaffolds for 7 days. Compared with pure PCL scaffolds a greater density of viable cells was seen on the composites, especially the pNSR32/PCL/Gt scaffolds.     

Fabrication and evaluation of poly(epsilon-caprolactone)/silk fibroin blend nanofibrous scaffold

In this study we investigated the blend electrospinning of poly(?‐caprolactone) (PCL) and silk fibroin (SF) to improve the biodegradability and biocompatibility of PCL‐based nanofibrous scaffolds. Optimal conditions to fabricate PCL/SF (50/50) blend nanofiber were established for electrospinning using formic acid as a cosolvent and three‐dimensional (3D) PCL/SF blend nanofibrous scaffolds were prepared by a modified electrospinning process using methanol coagulation bath. The physical properties of 2D PCL/SF blend nanofiber mats and 3D highly porous blend nanofibrous scaffolds were measured and compared. To evaluate cytocompatibility of the 3D blend scaffolds as compared to 3D PCL nanofibrous scaffold, normal human dermal fibroblasts were cultured. It is concluded that biodegradability and cytocompatibility could be improved for the 3D highly porous PCL/SF (50/50) blend nanofibrous scaffold prepared by blending PCL with SF in electrospinning. In addition to the blending of PCL and SF, the 3D structure and high porosity of electrospun nanofiber assemblies may also be important factors for enhancing the performance of scaffolds. © 2011 Wiley Periodicals, Inc. Biopolymers 97: 265–275, 2012.     

Electrospun poly (ɛ-caprolactone)/silk fibroin core-sheath nanofibers and their potential applications in tissue engineering and drug release

One of the key tenets of tissue engineering is to develop scaffold materials with favorable biodegradability, surface properties, outstanding mechanical strength and controlled drug release property. In this study, we generated core-sheath nanofibers composed of poly (?-caprolactone) (PCL) and silk fibroin (SF) blends via emulsion electrospinning. Nanofibrous scaffolds were characterized by combined techniques of scanning electron microscopy (SEM), transmission electron microscopy (TEM), Fourier-transform infrared spectroscopy (FTIR), X-ray diffraction (XRD), differential scanning calorimetry (DSC), contact angle and tensile measurements. An in vitro FITC release study was conducted to evaluate sustained release potential of the core-sheath structured nanofibers. We found that the conformation of SF contained in PCL/SF composite nanofibers was transformed from random coil to β-sheet when treated with methanol, leading to improved crystallinity and tensile strength of nanofibrous scaffolds. The hydrophobicity and diameter of nanofibers decreased when we increased the content of SF in PCL/SF composite nanofibers. Furthermore, we evaluated the potential of fabricated PCL/SF composite nanofibers as scaffold in vitro. The results confirmed that fabricated PCL/SF scaffolds improved cell attachment and proliferation. Our results demonstrated the feasibility to generate core-sheath nanofibers composed of PCL and SF using a single-nozzle technique. The produced nanofibrous scaffolds with sustained drug release have potential application in tissue engineering.     

Preparation and characterization of cockle shell aragonite nanocomposite porous 3D scaffolds for bone repair

The demands for applicable tissue-engineered scaffolds that can be used to repair load-bearing segmental bone defects (SBDs) is vital and in increasing demand. In this study, seven different combinations of 3 dimensional (3D) novel nanocomposite porous structured scaffolds were fabricated to rebuild SBDs using an extraordinary blend of cockle shells (CaCo₃) nanoparticles (CCN), gelatin, dextran and dextrin to structure an ideal bone scaffold with adequate degradation rate using the Freeze Drying Method (FDM) and labeled as 5211, 5400, 6211, 6300, 7101, 7200 and 8100. The micron sized cockle shells powder obtained (75 µm) was made into nanoparticles using mechano-chemical, top-down method of nanoparticles synthesis with the presence of the surfactant BS-12 (dodecyl dimethyl bataine). The phase purity and crystallographic structures, the chemical functionality and the thermal characterization of the scaffolds’ powder were recognized using X-Ray Diffractometer (XRD), Fourier transform infrared (FTIR) spectrophotometer and Differential Scanning Calorimetry (DSC) respectively. Characterizations of the scaffolds were assessed by Scanning Electron Microscopy (SEM), Degradation Manner, Water Absorption Test, Swelling Test, Mechanical Test and Porosity Test. Top-down method produced cockle shell nanoparticles having averagely range 37.8±3–55.2±9 nm in size, which were determined using Transmission Electron Microscope (TEM). A mainly aragonite form of calcium carbonate was identified in both XRD and FTIR for all scaffolds, while the melting (Tm) and transition (Tg) temperatures were identified using DSC with the range of Tm 62.4–75.5 °C and of Tg 230.6–232.5 °C. The newly prepared scaffolds were with the following characteristics: (i) good biocompatibility and biodegradability, (ii) appropriate surface chemistry and (iii) highly porous, with interconnected pore network. Engineering analyses showed that scaffold 5211 possessed 3D interconnected homogenous porous structure with a porosity of about 49%, pore sizes ranging from 8.97 to 337 µm, mechanical strength 20.3 MPa, Young's Modulus 271±63 MPa and enzymatic degradation rate 22.7 within 14 days.     

Effect of adding nano‐titanium dioxide on the microstructure,mechanical properties and in vitro bioactivity of a freeze cast merwinite scaffold

In the present research, merwinite (M) scaffolds with and without nano‐titanium dioxide (titania) were synthesized by water‐based freeze casting method. Two different amounts (7.5 and 10 wt%) of n‐TiO₂ were added to M scaffolds. They were sintered at temperature of 1573.15°K and at cooling rate of 4°K/min. The changes in physical and mechanical properties were investigated. The results showed that although M and M containing 7.5 wt% n‐TiO₂ (MT7.5) scaffolds had approximately the same microstructures in terms of pore size and wall thickness, these factors were different for sample MT10. In overall, the porosity, volume and linear shrinkage were decreased by adding different weight ratios of n‐TiO₂ into the M structure. According to the obtained mechanical results, the optimum mechanical performance was related to the sample MT7.5 (E = 51 MPa and σ = 2 MPa) with respect to the other samples, i.e.: M (E = 47 MPa and σ = 1.8 MPa) and MT10 (E = 32 MPa and σ = 1.4 MPa). The acellular in vitro bioactivity experiment confirmed apatite formation on the surfaces of all samples for various periods of soaking time. Based on cell study, the sample which possessed favorable mechanical behavior (MT7.5) supported attachment and proliferation of osteoblastic cells. These results revealed that the MT7.5 scaffold with improved mechanical and biological properties could have a potential to be used in bone substitute. © 2015 American Institute of Chemical Engineers Biotechnol. Prog., 31:550–556, 2015     

Gelatin blending improves the performance of poly(3-hydroxybutyrate-co-3-hydroxyhexanoate) films for biomedical application

To improve the performance of poly(3-hydroxybutyrate-co-3-hydroxyhexanoate) (PHBHHx), gelatin was blended with PHBHHx at different ratios. With increasing gelatin content, the weight loss of gelatin/PHBHHx blend in simulated body fluid at 37 degrees C was accelerated. After 2 months, there was about 15% weight loss in PHBHHx blending with 30% gelatin. Scanning electron microscopy and X-ray diffraction results showed that gelatin blending increased the surface porosity and decreased the crystallinity, which may be responsible for the acceleration of the weight loss. Second harmonic generation results indicated that 10% gelatin blending had less disruption to PHBHHx spatial structure, resulting in better tensile mechanical properties. At the same time, increased surface porosity and decreased crystallinity caused by gelatin incorporation may be beneficial for cell growth compared with pure PHBHHx. All these indicated that gelatin incorporation may improve the performances of PHBHHx to meet the need of different situations during medical implantation.     

Nanocomposite scaffold seeded with mesenchymal stem cells for bone repair

The mechanical property of bone tissue scaffolds is one of the most important aspects in bone tissue engineering that has remained problematic. In our previous study, we fabricated a three‐dimensional scaffold from nano‐hydroxyapatite/gelatin (nHA/Gel) and investigated its efficiency in promoting bone regeneration both in vitro and in vivo. In the present study, the effect of adding silicon carbide (SiC) on the mechanical and biological behaviors of the nHA/Gel/SiC and bone regeneration in vivo were determined. nHA and SiC were synthesized and characterized by the X‐ray diffraction pattern and transmission electron microscope image. Layer solvent casting, freeze drying, and lamination techniques were applied to prepare these scaffolds. Then, the biocompatibility and cell adhesion behavior of the synthesized nHA/Gel/SiC scaffolds were investigated. For in vivo studies, rats were categorized into three groups: blank defect, blank scaffold, and rat bone marrow mesenchymal stem cells (rBM‐MSCs)/scaffold. After 1, 4, and 12 weeks post‐injury, the rats were sacrificed and the calvaria were harvested. Sections with a thickness of 5 µm thickness were prepared and stained with hematoxylin–eosin and Masson's Trichrome, and immunohistochemistry was performed. Our results showed that SiC effectively increased the mechanical properties of the nHA/Gel/SiC scaffold. No significant differences were observed in biocompatibility, cell adhesion, and cytotoxicity of the nHA/Gel/SiC in comparison with the nHA/Gel nanocomposite. Based on histological and immunohistochemical studies, both osteogenesis and collagenization were significantly higher in the rBM‐MSCs/scaffold group, quantitatively and qualitatively. The present study strongly suggests the potential of SiC as an alternative strategy to improve the mechanical and biological properties of bone tissue engineering scaffolds, and shows that the pre‐seeded nHA/Gel/SiC scaffold with rBM‐MSCs improves osteogenesis in the engineered bone implant.     

3D-printed PLA/Gel hybrid in liver tissue engineering: Effects of architecture on biological functions

The liver is one of the vital organs in the body, and the gold standard of treatment for liver function impairment is liver transplantation, which poses many challenges. The specific three-dimensional (3D) structure of liver, which significantly impacts the growth and function of its cells, has made biofabrication with the 3D printing of scaffolds suitable for this approach. In this study, to investigate the effect of scaffold geometry on the performance of HepG2 cells, poly-lactic acid (PLA) polymer was used as the input of the fused deposition modeling (FDM) 3D-printing machine. Samples with simple square and bioinspired hexagonal cross-sectional designs were printed. One percent and 2% of gelatin coating were applied to the 3D printed PLA to improve the wettability and surface properties of the scaffold. Scanning electron microscopy pictures were used to analyze the structural properties of PLA–Gel hybrid scaffolds, energy dispersive spectroscopy to investigate the presence of gelatin, water contact angle measurement for wettability, and weight loss for degradation. In vitro tests were performed by culturing HepG2 cells on the scaffold to evaluate the cell adhesion, viability, cytotoxicity, and specific liver functions. Then, high-precision scaffolds were printed and the presence of gelatin was detected. Also, the effect of geometry on cell function was confirmed in viability, adhesion, and functional tests. The albumin and urea production of the Hexagonal PLA scaffold was about 1.22 ± 0.02-fold higher than the square design in 3 days. This study will hopefully advance our understanding of liver tissue engineering toward a promising perspective for liver regeneration.     

Microfluidic synthesis of composite cross‐gradient materials for investigating cell–biomaterial interactions

Combinatorial material synthesis is a powerful approach for creating composite material libraries for the high‐throughput screening of cell–material interactions. Although current combinatorial screening platforms have been tremendously successful in identifying target (termed “hit”) materials from composite material libraries, new material synthesis approaches are needed to further optimize the concentrations and blending ratios of the component materials. Here we employed a microfluidic platform to rapidly synthesize composite materials containing cross‐gradients of gelatin and chitosan for investigating cell–biomaterial interactions. The microfluidic synthesis of the cross‐gradient was optimized experimentally and theoretically to produce quantitatively controllable variations in the concentrations and blending ratios of the two components. The anisotropic chemical compositions of the gelatin/chitosan cross‐gradients were characterized by Fourier transform infrared spectrometry and X‐ray photoelectron spectrometry. The three‐dimensional (3D) porous gelatin/chitosan cross‐gradient materials were shown to regulate the cellular morphology and proliferation of smooth muscle cells (SMCs) in a gradient‐dependent manner. We envision that our microfluidic cross‐gradient platform may accelerate the material development processes involved in a wide range of biomedical applications. Biotechnol. Bioeng. 2011; 108:175–185. © 2010 Wiley Periodicals, Inc.