Preparation and HL-7702 cell functionality of titania/chitosan composite scaffolds

Titania/chitosan composite scaffolds were prepared through a freeze-drying technique. The composite scaffolds were highly porous with the average pore size of 120–300 μm, and the titania (TiO₂) powders were uniformly dispersed on the surface of the pore walls. The compressive strength of the composite scaffolds was significantly improved compared to that of pure chitosan scaffolds. Composite scaffold with 0.3 of TiO₂/chitosan weight ratio showed the maximum compressive strength of 159.7 ± 21 kPa. Hepatic immortal cell line HL-7702 was used as seeding cells on the scaffolds, and after different culture periods, cell attachment and function was analyzed. HL-7702 cells attached on the pore walls of the scaffolds with the spheroid shape after 1 day of culture, but more cell aggregations formed within the TiO₂/chitosan composite scaffolds as compared to pure chitosan scaffolds. Liver-specific functions, albumin secretion and urea synthesis were detected using a spectrometric method. The results showed that albumin secretion and urea synthesis rate of HL-7702 cells slightly decreased with the culture time, and there was no significant difference between composite scaffolds and pure chitosan scaffolds. In conclusion, the TiO₂/chitosan composite scaffolds possessed an improved mechanical strength compared to pure chitosan scaffolds and supported the attachment and functional expression of hepatocyte, implying their potential application in liver tissue engineering.     

Robotic dispensing of composite scaffolds and in vitro responses of bone marrow stromal cells

The development of bioactive scaffolds with a designed pore configuration is of particular importance in bone tissue engineering. In this study, bone scaffolds with a controlled pore structure and a bioactive composition were produced using a robotic dispensing technique. A poly(ε-caprolactone) (PCL) and hydroxyapatite (HA) composite solution (PCL/HA = 1) was constructed into a 3-dimensional (3D) porous scaffold by fiber deposition and layer-by-layer assembly using a computer-aided robocasting machine. The in vitro tissue cell compatibility was examined using rat bone marrow stromal cells (rBMSCs). The adhesion and growth of cells onto the robotic dispensed scaffolds were observed to be limited by applying the conventional cell seeding technique. However, the initially adhered cells were viable on the scaffold surface. The alkaline phosphatase activity of the cells was significantly higher on the HA–PCL than on the PCL and control culture dish, suggesting that the robotic dispensed HA–PCL scaffold should stimulate the osteogenic differentiation of rBMSCs. Moreover, the expression of a series of bone-associated genes, including alkaline phosphatase and collagen type I, was highly up-regulated on the HA–PCL scaffold as compared to that on the pure PCL scaffold. Overall, the robotic dispensed HA–PCL is considered to find potential use as a bioactive 3D scaffold for bone tissue engineering. Seok-Jung Hong and Ishik Jeong contributed equally.     

Radiation synthesis of gelatin/CM-chitosan/β-tricalcium phosphate composite scaffold for bone tissue engineering

A series of biodegradable composite scaffolds was fabricated from an aqueous solution of gelatin, carboxymethyl chitosan (CM-chitosan) and β-tricalcium phosphate (β-TCP) by radiation-induced crosslinking at ambient temperature. Ultrasonic treatment on the polymer solutions significantly influenced the distribution of β-TCP particles. An ultrasonic time of 20 min, followed by 30 kGy irradiation induced a crosslinked scaffold with homogeneous distribution of β-TCP particles, interconnected porous structure, sound swelling capacity and mechanical strength. Fourier Transform Infrared Spectroscopy and X-ray Diffraction analysis indicated that β-TCP successfully incorporated with the network of gelatin and CM-chitosan. In vivo implantation of the scaffold into the mandible of beagle dog revealed that the scaffolds had excellent biocompatibility and the presence of β-TCP can accelerate bone regeneration. The comprehensive results of this study paved way for the application of gelatin/CM-chitosan/β-TCP composite scaffolds as candidate of bone tissue engineering material.     

Novel porous scaffolds of poly(lactic acid) produced by phase-separation using room temperature ionic liquid and the assessments of biocompatibility

Here we prepared three-dimensional (3D) porous-structured biodegradable polymer scaffolds for tissue regeneration using room temperature ionic liquids (RTILs) as a novel porogen, and addressed their biological properties, including in vitro cell growth and differentiation and in vivo tissue compatibility. RTIL based on 1-butyl-3-methylimidazolium ([bmim]) bearing hydrophilic anion Cl was introduced within the polymer structure to provide a pore network. A mixture of poly(lactic acid) (PLA) with RTIL dissolved in an organic solvent formed a bi-continuous network during the drying process. Selective dissolution of the RTIL phase was facilitated in ethanol, which resulted in a porous network of the polymer phase with complete removal of the RTIL. The RTILs-assisted porous scaffolds showed a typical open-channeled network with pore sizes over 100 μm and porosities of about 86–94%. For the biocompatibility assessments of the scaffolds, mesenchymal stem cells (MSCs) derived from rat bone marrow were seeded onto the PLA scaffold, and the cell proliferation and osteoblastic differentiation behaviors were examined. Results showed a typical on-going increase in the cell population with a level comparable to that observed on the tissue culture plastic control, indicating good cell compatibility. When cultured in an osteogenic medium, the alkaline phosphatase (ALP) activity of the cells on the PLA scaffolds was stimulated to increase with time from 7 to 14 days, in a similar manner to that on the control. Moreover, the expression of genes related to osteoblasts, including collagen type I, osteocalcin and bone sialoprotein, was stimulated on the 3D PLA scaffold during culture for up to 14 days, with levels higher than those on the control, suggesting the developed scaffold provided a 3D matrix condition for osteogenesis. An in vivo pilot study conducted subcutaneously in rat for 4 weeks revealed good tissue compatibility of the scaffold, with the ingrowth of cells and formation of collageneous tissue around and deep within the pores of the scaffold and no significant inflammatory reaction. Taken together, this novel method of using RTILs as a pore generator is considered to be useful in the development of biocompatible porous polymer scaffolds for tissue regeneration.     

A novel bioactive three-dimensional β-tricalcium phosphate/chitosan scaffold for periodontal tissue engineering

The development of suitable bioactive three-dimensional scaffold for the promotion of cellular proliferation and differentiation is critical in periodontal tissue engineering. In this study,porous β-tricalcium phosphate/chitosan composite scaffolds were prepared through a freeze-drying method. These scaffolds were evaluated by analysis of microscopic structure, porosity, and cytocompatibility. The gene expression of bone sialoprotein (BSP) and cementum attachment protein (CAP) was detected with RT-PCR after human periodontal ligament cells (HPLCs) were seeded in these scaffolds. Then cell–scaffold complexes were implanted subcutaneously into athymic mice. The protein expression of alkaline phosphatase (ALP) and osteopontin (OPN) was detected in vivo. Results indicated that composite scaffolds displayed a homogeneous three-dimensional microstructure; suitable pore size (120 μm) and high porosity (91.07%). The composite scaffold showed higher proliferation rate than the pure chitosan scaffold, and up-regulated the gene expression of BSP and CAP. In vivo, HPLCs in the composite scaffold not only proliferated but also recruited vascular tissue ingrowth. The protein expression of ALP and OPN was up-regulated in the composite scaffold. Therefore, it was suggested that the composite scaffold could promote the differentiation of HPLCs towards osteoblast and cementoblast phenotypes.     

Silk fibroin/chitosan scaffold: preparation,characterization, and culture with HepG2 cell

Tissue engineering requires the development of three-dimensional water-stable scaffolds. In this study, silk fibroin/chitosan (SFCS) scaffold was successfully prepared by freeze-drying method. The scaffold is water-stable, only swelling to a limited extent depending on its composition. Fourier Transform Infrared (FTIR) spectra and X-Ray diffraction curves confirmed the different structure of SFCS scaffolds from both chitosan and silk fibroin. The homogeneous porous structure, together with nano-scale compatibility of the two naturally derived polymers, gives rise to the controllable mechanical properties of SFCS scaffolds. By varying the composition, both the compressive modulus and compressive strength of SFCS scaffolds can be controlled. The porosity of SFCS scaffolds is above 95% when the total concentration of silk fibroin and chitosan is below 6 wt%. The pore sizes of the SFCS scaffolds range from 100 μm to 150 μm, which can be regulated by changing the total concentration. MTT assay showed that SFCS scaffolds can promote the proliferation of HepG2 cells (human hepatoma cell line) significantly. All these results make SFCS scaffold a suitable candidate for tissue engineering.     

Fabrication and biocompatibility of nano non-stoichiometric apatite and poly(ε-caprolactone) composite scaffold by using prototyping controlled process

Nano biocomposite scaffolds of non-stoichiometric apatite (ns-AP) and poly(ε-caprolactone) (PCL) were prepared by a prototyping controlled process (PCP). The results show that the composite scaffolds with 40 wt% ns-AP contained open and well interconnected pores with a size of 400–500 μm, and exhibited a maximum porosity of 76%. The ns-AP particles were not completely embedded in PCL matrix while exposed on the composite surface, which might be useful for cell attachment and growth. Proliferation of MG₆₃ cells was significantly better on the composite scaffolds with porosity of 76% than that those with porosity of 53%, indicating that the scaffolds with high porosity facilitated cell growth, and could promote cell proliferation. The composite scaffolds were implanted into rabbit thighbone defects to investigate the in vivo biocompatibility and osteogenesis. Radiological and histological examination confirmed that the new bony tissue had grown easily into the entire composite scaffold. The results suggest that the well-interconnected pores in the scaffolds might encourage cell proliferation, and migration to stimulate cell functions, thus enhancing bone formation in the scaffolds. This study shows that bioactive and biocompatible ns-AP/PCL composite scaffolds have potential applications in bone tissue engineering.     

In vitro evaluation of borate-based bioactive glass scaffolds prepared by a polymer foam replication method

Borate-based bioactive glass scaffolds with a microstructure similar to that of human trabecular bone were prepared using a polymer foam replication method, and evaluated in vitro for potential bone repair applications. The scaffolds (porosity = 72 ± 3%; pore size = 250–500 μm) had a compressive strength of 6.4 ± 1.0 MPa. The bioactivity of the scaffolds was confirmed by the formation of a hydroxyapatite (HA) layer on the surface of the glass within 7 days in 0.02 M K₂HPO₄ solution at 37 °C. The biocompatibility of the scaffolds was assessed from the response of cells to extracts of the dissolution products of the scaffolds, using assays of MTT hydrolysis, cell viability, and alkaline phosphatase activity. For boron concentrations below a threshold value (0.65 mM), extracts of the glass dissolution products supported the proliferation of bone marrow stromal cells, as well as the proliferation and function of murine MLO-A5 cells, an osteogenic cell line. Scanning electron microscopy showed attachment and continuous increase in the density of MLO-A5 cells cultured on the surface of the glass scaffolds. The results indicate that borate-based bioactive glass could be a potential scaffold material for bone tissue engineering provided that the boron released from the glass could be controlled below a threshold value.     

Quantitative grafting of peptide onto the nontoxic biodegradable waterborne polyurethanes to fabricate peptide modified scaffold for soft tissue engineering

Gly-Arg-Gly-Asp-Ser-Pro (GRGDSP) peptide has frequently been used in the biomedical materials to enhance adhesion and proliferation of cells. In this work, we modified the nontoxic biodegradable waterborne polyurethanes (WBPU) with GRGDSP peptide and fabricated 3-D porous scaffold with the modified WBPU to investigate the effect of the immobilized GRGDSP peptide on human umbilical vein endothelial cells (HUVECs) adhesion and proliferation. A facile and reliable approach was first developed to quantitative grafting of GRGDSP onto the WBPU molecular backbone using ethylene glycol diglycidyl ether (EX810) as a connector. Then 3-D porous WBPU scaffolds with various GRGDSP content were fabricated by freeze-drying the emulsion. In both of the HUVECs adhesion and proliferation tests, enhanced cell performance was observed on the GRGDSP grafted scaffolds compared with the unmodified scaffolds and the tissue culture plate (TCP). The adhesion rate and proliferation rate increased with the increase of GRGDSP content in the scaffold and reached a maximum with peptide concentration of 0.85 μmol/g based on the weight of the polyurethanes. These results illustrate the necessity of the effective control of the GRGDSP content in the modified WBPU and support the potential utility of these 3-D porous modified WBPU scaffolds in the soft tissue engineering to guide cell adhesion, proliferation and tissue regeneration.     

The fundamental parameters of chitosan in polymer scaffolds affecting osteoblasts (MC3T3-E1)

The aim of this study was to investigate the degree of deacetylation (DD) and molecular weight (MW) of chitosan within chitosan–collagen scaffolds on mouse osteoblasts (MC3T3-E1). The chitosan–collagen scaffolds were fabricated by freeze-drying technique. The studies on cell attachment and proliferation, alkaline phosphatase (ALP) activity, cell morphology, and mineralized nodule formation by osteoblasts on scaffolds were investigated. No statistically significant difference was found on cell attachment, but the chitosan–collagen scaffolds with low-DD chitosan had a statistically significantly (P < 0.05) higher proliferative effect and ALP activity than those scaffolds with high-DD chitosan, regardless of molecular weight. Scanning electron images demonstrated that MC3T3-E1 cells grew well on all test scaffolds; on the contrary, mineralized nodule formation was not found. In conclusion, the DD of chitosan is a crucial factor for MC3T3-E1 cells and it should be considered in further applications for bone tissue engineering.     

Microstructure and characteristic properties of gelatin/chitosan scaffold prepared by a combined freeze-drying/leaching method

A combined freeze-drying and particulate leaching method for scaffold synthesis showed an improvement in the horizontal microstructure of the gelatin/chitosan scaffolds. Type and concentration of the cross-linking agent, freezing temperature, concentration of the polymeric solution and gelatin/chitosan weight ratio were the variables affecting the scaffold properties. Assessment of the tensile properties of the scaffolds revealed that for a scaffold with 50% chitosan, glutaraldehyde, as a cross-linking agent, created much tighter polymeric network compared to N,N-(3-dimethylaminopropyl)-N′-ethyl carbodiimide (EDC). However, in the case of gelatin scaffolds, EDC was identified as the stronger cross-linker. Compressive behavior of the scaffolds satisfied formulations obtained from the theoretical modeling of the low-density, elastomeric foams. The investigation of the scaffold degradation indicated that the increase in the mechanical strength of the scaffolds would not always reduce their degradation rate.     

Low temperature fabrication of high strength porous calcium phosphate and the evaluation of the osteoconductivity

Porous NaO₂–MgO–CaO–P₂O₅ bioglass doped beta-tri-calcium phosphate (β-TCP) bioceramic possessing high mechanical properties and well pore structure with high porosity and high pore connectivity has been prepared through dipping method with the porous polyurethane as the pore forming template. The sintering mechanism and the mechanical properties of the bioglass doped β-TCP scaffold have been investigated by the X-ray diffraction (XRD) analysis, Scanning electron microscope (SEM) and thermal differential analysis (DTA). The scaffold’s in vivo osteoconductivity has been evaluated by implantation of scaffolds into the femurs of New Zealand rabbits. The results show that the porous structure can achieve the densification process at a low temperature about 950°C by a solid solution sintering mechanism and hence dense macropore scaffold with a compressive strength of 4.32 MPa when the porosity is 75% has been obtained. The in vivo test shows that the Na₂O–MgO–CaO–P₂O₅ bioglass doped porous β-TCP bioceramic has a relatively fast bone formation after implantation; after 1 month implantation new deposited bone tissue has been detected on the strut of the porous scaffold and degraded particles also has been found on the surface of the new formed bone. After 6 months implantation the porous scaffold has been thoroughly covered with new formed bone. Results show that the Na₂O–MgO–CaO–P₂O₅ bioglass doped porous β-TCP bioceramic is potential bone tissue engineering scaffold for orthopedic use.     

Designer self-assembling peptide scaffold stimulates pre-osteoblast attachment, spreading and proliferation

A new peptide scaffold was made by mixing pure RADA16 (Ac-RADARADARADARADA-CONH2) and designer peptide RGDA16 (Ac-RADARGDARADARGDA-CONH2) solutions, and investigate any effect on attachment, spreading and proliferation of pre-osteoblast (MC3T3-E1). The peptides, RADA16 and RGDA16, were custom-synthesized. They were solubilized in deionized water at a concentration of 10 mg/ml (1% w/v), the RGDA16 peptide solution was mixed 1:1 with RADA16 solution and a new peptide solution RGDAmix was produced. The RGDAmix and RADA16 solution were directly loaded in 96-well plates and cover slips, and two different peptide scaffolds were formed with the addition of maintenance medium (α-MEM) in several minutes. About 1.0 × 10⁴ MC3T3-E1 cells were seeded on each hydrogel scaffold, and then the cell morphological changes were observed using a fluorescence microscope at 1 h, 3 h and 24 h timepoint, respectively. Cell attachment was evaluated 1 h, 3 h and 24 h after cell seeding and cell proliferation was determined 4d, 7d and 14d after cell seeding. The RGDAmix scaffold significantly promoted the initial cell attachment compared with the RADA16 scaffold. MC3T3-E1 cells adhered and spread well on both scaffolds, however, cells spread better on the RGDAmix scaffold than on the RADA16 scaffold. Cell proliferation was greatly stimulated when cultured on RGDAmix scaffold. The RGD sequence contained peptide scaffold RGDAmix significantly enhances MC3T3-E1 cells attachment, spreading and proliferation.     

Genipin-crosslinked gelatin scaffolds for articular cartilage tissue engineering with a novel crosslinking method

A novel crosslinking method with directly crosslinking the gelatin gel, being cut to a disc of chosen size beforehand, for the fabrication of porous gelatin scaffold was proposed. This novel method of gel-crosslinking was compared with the traditional methods of mixing-crosslinking and scaffold-crosslinking. The structure of the scaffold fabricated by the gel-crosslinking method shows uniformly distributed and interconnected pores which can be much smaller than those made by the other two methods. All three methods have the last step as freeze-drying; nevertheless, freeze-drying once more will increase the uniformity of the structure and the interconnecting pores. Crosslinking of gelatin was carried out at room temperature with glutaraldehyde (GTA) or genipin (GP). In vitro cell culture of Wistar rat's joint chondrocytes demonstrates that the GTA-crosslinked scaffold is much worse than the GP-crosslinked one; a tissue containing collagen and glycosaminoglycan was produced in the GP-crosslinked scaffold in just 9 days after cell seeding, and a tissue with a cell distribution resembling that of the native cartilage was developed after 30 day cell culture. It was concluded that the novel method is feasible for application in articular cartilage tissue engineering.     

In vitro evaluation of biodegradable nHAP‐Chitosan‐Gelatin‐based scaffold for tissue engineering application

The present study focuses on fabrication and characterisation of porous composite scaffold containing hydroxyapatite (HAP), chitosan, and gelatin with an average pore size of 250–1010 nm for improving wound repair and regeneration by Electrospinning method. From the results of X ‐Ray Diffraction (XRD) study, the peaks correspond to crystallographic structure of HAP powder. The presence of functional group bonds of HAP powder, Chitosan and scaffold was studied using Fourier Transform Infrared Spectroscopy (FTIR). The surface morphology of the scaffold was observed using Scanning Electron Microscope (SEM). The Bioactivity of the Nano composite scaffolds was studied using simulated body fluid solution at 37 ± 1°C. The biodegradability test was studied using Tris‐Buffer solution for the prepared nanocomposites [nano Chitosan, nano Chitosan gelatin, Nano based Hydroxyapatite Chitosan gelatin]. The cell migration and potential biocompatibility of nHAP‐chitosan‐gelatin scaffold was assessed via wound scratch assay and were compared to povedeen as control. Cytocompatibility evaluation for Vero Cells using wound scratch assay showed that the fabricated porous nanocomposite scaffold possess higher cell proliferation and growth than that of povedeen. Thus, the study showed that the developed nanocomposite scaffolds are potential candidates for regenerating damaged cell tissue in wound healing process.Inspec keywords: nanofabrication, tissue engineering, electrospinning, wounds, cellular biophysics, scanning electron microscopy, surface morphology, X‐ray diffraction, biomedical materials, nanomedicine, porosity, biodegradable materials, nanoporous materials, calcium compounds, gelatin, nanocomposites, Fourier transform infrared spectra, nanoparticles, precipitation (physical chemistry)Other keywords: average pore size, wound repair, crystallographic structure, HAP powder, functional group bonds, simulated body fluid solution, biodegradability test, Tris‐Buffer solution, cell migration, wound scratch assay, tissue engineering, electrospinning method, X‐ray diffraction, Fourier transform infrared spectroscopy, scanning electron microscopy, biocompatibility, cytocompatibility, porous nanocomposite scaffold, cell tissue, nHAP‐chitosan‐gelatin scaffold composites, wet chemical precipitation method, surface morphology, nanohydroxyapatite‐nanochitosan‐gelatin scaffold composites, cell proliferation, wound healing, (Ca₁₀ (PO₄)₆ (OH)₂)     

Osteogenic effect of hyaluronic acid sodium salt in the pores of a hydroxyapatite scaffold

According to previous reports, a large volume of bone marrow cells (1 × 10⁷ cells/ml) is required for bone regeneration in the pores of a scaffold in vivo. We theorized that immersion of a porous hydroxyapatite (HA) scaffold in hyaluronic acid solution would facilitate bone formation in the scaffold at 1 × 10⁶ cells/ml density of bone marrow cells. The cells were respectively seeded into pores of the cylindrical HA scaffolds with a hollow center after immersion in hyaluronic acid solution or in culture medium. The scaffolds were implanted in the dorsal subcutis of rats for 4 weeks. Thereafter, serially sectioned paraffin specimens were made and observed histologically. Bone formation was observed in many pores of HA scaffold by immersion in hyaluronic acid solution. However, there were no or less pores with new bone formation in the scaffold by immersion in culture medium. The cells were cultured with and without hyaluronic acid in vitro. There was no significant difference in bone formation in vitro with and without hyaluronic acid. The results of this study suggest that hyaluronic acid binds to the cells on the wall of three-dimensional structure and effectively promotes new bone formation.     

Preparation, characterization and in vitro biological study of biomimetic three-dimensional gelatin–montmorillonite/cellulose scaffold for tissue engineering

This work focused on studying the effect of blending gelatin (Gel) with Cellulose (Cel), in the presence of montmorillonite (MMT), on the swelling behavior, in vitro degradation and surface morphology. Additionally, the effect of the prepared biocomposites on the characteristics of the human osteosarcoma cells (Saos-2), including proliferation, scaffold/cells interactions, apoptosis and their potential of the cells to induce osteogenesis and differentiation was evaluated. The crosslinked biocomposites with glutaraldehyde (GA) or N,N-methylene-bisacrylamide (MBA) was prepared via an intercalation process and freeze-drying technique. Properties including SEM morphology, X-ray diffraction characterization and in vitro biodegradation were investigated. The successful generation of 3-D biomimetic porous scaffolds incorporating Saos-2 cells indicated their potential for de novo bone formation that exploits cell–matrix interactions. In vitro studies revealed that the scaffolds containing 12 and 6% MMT crosslinked by 5 and 0.5% GA seem to be the two most efficient and effective biodegradable scaffolds, which promoted Saos-2 cells proliferation, migration, expansion, adhesion, penetration, spreading, and differentiation, respectively. MMT improved cytocompatibility between the osteoblasts and the biocomposite. In vitro analysis indicated good biocompatibility of the scaffold and presents the scaffold as a new potential candidate as suitable biohybrid material for tissue engineering.     

A novel method for the fabrication of homogeneous hydroxyapatite/collagen nanocomposite and nanocomposite scaffold with hierarchical porosity

Homogeneous nanocomposites composed of hydroxyapatite (HAp) and collagen were synthesized using a novel in situ precipitation method through dual template-driven. The morphological and componential properties of nanocomposites were investigated. The HAp particulates, in sizes of about 50–100 nm, were distributed homogeneously in the organic collagen hydrogel. Highly magnified TEM observation showed that HAp inorganic particles were composed of fine sub-particles (2–5 nm) without regular crystallographic orientation. Based on these homogeneous nanocomposites, a novel HAp/collagen nanocomposite scaffold with hierarchical porosity was prepared by multilevel freeze-drying technique. Compared to other conventional scaffolds for tissue engineering, this novel in situ method endows synthesized composite scaffolds with unique morphology—ultrafine HAp particles dispersed homogenously in collagen at nano level and the foam scaffold with hierarchical pore structures. The mechanical performance increased obviously compared with neat collagen. These results provided an efficient approach toward new biomimetic tissue scaffold for the biomedical applications with enhanced intensity/bioactivity and controlled resorption rates. This novel method, we expect, will lead to a wide application in many other hydrogel systems and may be useful for fabrication of various homogeneous inorganic/organic nanocomposites.     

Electrospinning of PLGA/gelatin randomly-oriented and aligned nanofibers as potential scaffold in tissue engineering

Electrospinning technique can be used to produce the three-dimensional nanofibrous scaffold similar to natural extracellular matrix, which satisfies particular requirements of tissue engineering scaffold. Randomly-oriented and aligned poly(lactic-co-glycolic acid) (PLGA) and PLGA/gelatin biocomposite scaffolds were successfully produced by electrospinning in the present study. The resulting nanofibrous scaffolds exhibited smooth surface and high porous structure. Blending PLGA with gelatin enhanced the hydrophilicity but decreased the average fiber diameter and the mechanical properties of the scaffolds under the same electrospinning condition. The cell culture results showed that the elongation of the osteoblast on the aligned nanofibrous scaffold was parallel to the fiber arrangement and the cell number was similar to that of randomly-oriented scaffold, indicating that the aligned nanofibrous scaffold provide a beneficial approach for the bone regeneration.     

Biomimetic design and fabrication of porous chitosan–gelatin liver scaffolds with hierarchical channel network

The presence of a hierarchical channel network in tissue engineering scaffold is essential to construct metabolically demanding liver tissue with thick and complex structures. In this research, chitosan–gelatin (C/G) scaffolds with fine three-dimensional channels were fabricated using indirect solid freeform fabrication and freeze-drying techniques. Fabrication processes were studied to create predesigned hierarchical channel network inside C/G scaffolds and achieve desired porous structure. Static in-vitro cell culture test showed that HepG2 cells attached on both micro-pores and micro-channels in C/G scaffolds successfully. HepG2 proliferated at much higher rates on C/G scaffolds with channel network, compared with those without channels. This approach demonstrated a promising way to engineer liver scaffolds with hierarchical channel network, and may lead to the development of thick and complex liver tissue equivalent in the future.