Fabrication and development of artificial osteochondral constructs based on cancellous bone/hydrogel hybrid scaffold.

Using tissue engineering techniques, an artificial osteochondral construct was successfully fabricated to treat large osteochondral defects. In this study, porcine cancellous bones and chitosan/gelatin hydrogel scaffolds were used as substitutes to mimic bone and cartilage, respectively. The porosity and distribution of pore size in porcine bone was measured and the degradation ratio and swelling ratio for chitosan/gelatin hydrogel scaffolds was also determined in vitro. Surface morphology was analyzed with the scanning electron microscope (SEM). The physicochemical properties and the composition were tested by using an infrared instrument. A double layer composite scaffold was constructed via seeding adipose-derived stem cells (ADSCs) induced to chondrocytes and osteoblasts, followed by inoculation in cancellous bones and hydrogel scaffolds. Cell proliferation was assessed through Dead/Live staining and cellular activity was analyzed with IpWin5 software. Cell growth, adhesion and formation of extracellular matrix in composite scaffolds blank cancellous bones or hydrogel scaffolds were also analyzed. SEM analysis revealed a super porous internal structure of cancellous bone scaffolds and pore size was measured at an average of 410 ± 59 µm while porosity was recorded at 70.6 ± 1.7 %. In the hydrogel scaffold, the average pore size was measured at 117 ± 21 µm and the porosity and swelling rate were recorded at 83.4 ± 0.8 % and 362.0 ± 2.4 %, respectively. Furthermore, the remaining hydrogel weighed 80.76 ± 1.6 % of the original dry weight after hydration in PBS for 6 weeks. In summary, the cancellous bone and hydrogel composite scaffold is a promising biomaterial which shows an essential physical performance and strength with excellent osteochondral tissue interaction in situ. ADSCs are a suitable cell source for osteochondral composite reconstruction. Moreover, the bi-layered scaffold significantly enhanced cell proliferation compared to the cells seeded on either single scaffold. Therefore, a bi-layered composite scaffold is an appropriate candidate for fabrication of osteochondral tissue.

A 3D bioprinted decellularized extracellular matrix/gelatin/quaternized chitosan scaffold assembling with poly(ionic liquid)s for skin tissue engineering.

Currently, a suitable bioink for 3D bioprinting and capable of mimicking the microenvironment of native skin and preventing bacterial infection remains a major challenge in skin tissue engineering. In this study, we prepared a tissue-specific extracellular matrix-based bioink, and dECM/Gel/QCS (dGQ) 3D scaffold assembling with poly(ionic liquid)s (PILs) (dGQP) was obtained by an extrusion 3D bioprinting technology and dynamic hydrogen bonding method. The morphologies, mechanical properties, porosity, hydrophilicity, biodegradation, hemostatic effect, antibacterial ability, and biocompatibility of the hybrid scaffolds were characterized and evaluated. Results showed that the rapid release (2 h) of PILs on the dGQP scaffold can quickly kill gram-negative (E. coli) and gram-positive (S. aureus) bacteria with almost 100 % antibacterial activity and maintained a stable sterile environment for a long time (7 d), which was superior to the dGQ scaffold. The hemostasis and hemolysis test showed that the dGQP scaffold had a good hemostatic effect and excellent hemocompatibility. In vitro cytocompatibility studies showed that although the cell growth on dGQP scaffold was slow in the early stage, the cells proliferated rapidly since day 4 and had high ECM secretion at day 7. Overall, this advanced dGQP scaffold has a considerable potential to be applied in skin tissue engineering.

Low-Temperature Hydrothermal Synthesis of Novel 3D Hybrid Nanostructures on Titanium Surface with Mechano-bactericidal Performance.

Titanium is extensively employed in modern medicines as orthopedic and dental implants, but implant failures frequently occur because of bacterial infections. Herein, three types of 3D nanostructured titanium surfaces with nanowire clusters (NWC), nanowire/sheet clusters (NW/SC) and nanosheet clusters (NSC), were fabricated using the low-temperature hydrothermal synthesis under normal pressure, and assessed for the sterilization against two common human pathogens. The results show that the NWC and NSC surfaces merely display good bactericidal activity against Escherichia coli, whereas the NW/SC surface represents optimal bactericidal efficiency against both Escherichia coli (98.6 ± 1.23%) and Staphylococcus aureus (69.82 ± 2.79%). That is attributed to the hybrid geometric nanostructure of NW/SC, i.e., the pyramidal structures of ∼23 nm in tip diameter formed with tall clustered wires, and the sharper sheets of ∼8 nm in thickness in-between these nanopyramids. This nanostructure displays a unique mechano-bactericidal performance via the synergistic effect of capturing the bacteria cells and penetrating the cell membrane. This study proves that the low-temperature hydrothermal synthesized hybrid mechano-bactericidal titanium surfaces provide a promising solution for the construction of bactericidal biomedical implants.

A biological functional hybrid scaffold based on decellularized extracellular matrix/gelatin/chitosan with high biocompatibility and antibacterial activity for skin tissue engineering.

Nowadays, decellularized extracellular matrix (dECM) has received widespread attention due to its diversity in providing the unique structural and functional components to support cell growth, and finding material with good biocompatibility and anti-infection capability for skin tissue engineering is still a challenge. In this study, a novel dECM/Gel/CS scaffold with appropriate mechanical strength, good antibacterial activity and high biocompatibility was prepared using a one-pot method. The results showed that the immune components such as cells and DNA (about 98.1%) were successfully removed from the porcine skin tissue. The dECM/Gel/CS scaffolds exhibited an interconnected pore structure and had a high porosity (>90%) to promote cell growth. Moreover, the appropriate elastic modulus (≥482.17 kPa) and degradability (≥80.04% for 15 days) of the scaffolds offered stout "houses" for cell proliferation and suitable degradation rate to match the new tissue formation in skin tissue engineering. Furthermore, the addition of chitosan endowed the scaffold with good antibacterial activity, water and protein absorption capacity to avoid wound infection, and maintain the moisture and nutrition balance. In vitro cytocompatibility studies showed that the presence of dECM effectively enhanced the cell proliferation. Overall, the advanced dECM/Gel/CS scaffold has considerable potential to be applied in skin tissue engineering.

Preparation, fabrication and biocompatibility of novel injectable temperature-sensitive chitosan/glycerophosphate/collagen hydrogels.

This paper introduces a novel type of injectable temperature-sensitive chitosan/glycerophosphate/collagen (C/GP/Co) hydrogel that possesses great biocompatibility for the culture of adipose tissue-derived stem cells. The C/GP/Co hydrogel is prepared by mixing 2.2% (v/v) chitosan with 50% (w/w) ß-glycerophosphate at different proportions and afterwards adding 2 mg/ml of collagen. The gelation time of the prepared solution at 37°C was found to be of around 12 min. The inner structure of the hydrogel presented a porous spongy structure, as observed by scanning electron microscopy. Moreover, the osmolality of the medium in contact with the hydrogel was in the range of 310-330 mmol kg(-1). These analyses have shown that the C/GP/Co hydrogels are structurally feasible for cell culture, while their biocompatibility was further examined. Human adipose tissue-derived stem cells (ADSCs) were seeded into the developed C/GP and C/GP/Co hydrogels (The ratios of C/GP and C/GP/Co were 5:1 and 5:1:6, respectively), and the cellular growth was periodically observed under an inverted microscope. The proliferation of ADSCs was detected using cck-8 kits, while cell apoptosis was determined by a Live/Dead Viability/Cytotoxicity kit. After 7 days of culture, cells within the C/GP/Co hydrogels displayed a typical adherent cell morphology and good proliferation with very high cellular viability. It was thus demonstrated that the novel C/GP/Co hydrogel herein described possess excellent cellular compatibility, representing a new alternative as a scaffold for tissue engineering, with the added advantage of being a gel at the body's temperature that turns liquid at room temperature.

Electrohydrodynamic jet 3D printing of PCL/PVP composite scaffold for cell culture.

Controlled printing of biodegradable and bioresorbable polymers at desired 3D scaffold is of great importance for cell growth and tissue regeneration. In this work, a novel electrohydrodynamic jet 3D printing technology with the resultant effect of electrohydrodynamic force and thermal convection was developed, and its feasibility to fabricate controllable filament composite scaffolds was verified. This method introduces an effective thermal field under the needle to simultaneously enhance the ink viscosity, jetting morphology controllability and printing structure solidify. The fabrication mechanisms of thermal convection on jetting morphology and printed structures feature were investigated through theoretical analysis and experimental characterization. Under optimized conditions, a stable and finer jet was formed; then with the use of this jet, various 3D structures were directly printed at a high aspect ratio ~30. Furthermore, the PCL/PVP composite scaffolds with the controllable filament diameter (~10 µm) which is closed to living cells were printed. Cell culture experiments showed that the printed scaffolds had excellent cell biocompatibility and facilitated cellular proliferation in vitro. It is a great potential that the developed electrohydrodynamic jet 3D printing technology might provide a novel approach to directly print composite synthetic biopolymers into flexibly scale structures for tissue engineering applications.

Effects of concentration variation on the physical properties of alginate-based substrates and cell behavior in culture.

Nowadays alginate capsules exhibit good biocompatibility and high permeability for nutrients and metabolic wastes making them appealing biomaterial for therapeutic cell encapsulation. Further study of the characteristics of alginate beads which are highly dependent on various environmental conditions to create an optimum microenvironment for cells is also critical. Thus, in this study, the effect of concentration variation on the physical properties of alginate-based beads and entrapped-cells behavior was analyzed. Results showed that the increase of Ca ions concentration brought about the decrease of the average diameter, prolongation of dissolution time, reduction of permeability and swelling, and a rise of crosslinking extent and shrinkage of capsules; while raising sodium alginate concentration had an opposite effect on the diameter and shrinkage. Moreover, the addition of gelatin enhanced the penetration and swelling and slowed down the shrinkage of capsules. And MC3T3-E1 cells enclosed in the particles in which the concentration of calcium chloride, sodium alginate and gelatin was 2.5%, 2.0% and 0.5% (w/v %) had preferable abilities of proliferation and higher expression of alkaline phosphatase. Overall, the ability to tailor this system to support in vitro growth of MC3T3-E1 cells might have significance for the future use of other cell types in regenerative medicine.

Multiple comparisons of three different sources of biomaterials in the application of tumor tissue engineering in vitro and in vivo.

Potential anti-cancer drugs are frequently of low efficacy in clinics due to the lack of predictive models or the insufficient employment of existing preclinical test systems. Three-dimensional (3D) in vitro engineered tumor models can better predict the efficacy of novel drugs by reproducing the in vivo tumor microenvironment. In this study, three sources of scaffolds (decellularized lung scaffold, chitosan/gelatin scaffold, and poly-L-lactic acid scaffold) incorporated with breast cancer cells (MCF-7, 4T1) were bioengineered as a platform to study in vitro solid tumor development. The good biocompatibility of three scaffolds favored cell growth and proliferation. Cells in 3D scaffolds were less sensitive to chemotherapy and exhibited characteristics of higher malignancy compared to their 2D counterparts. The expression of breast cancer biomarkers in MCF-7 cells markedly up-regulated in 3D scaffolds in comparison with those in 2D cultures. Cells grown in 3D scaffolds were found to be more tumorigenic and angiogenic in BABL/c mice xenografts than cells grown from monolayers. The results demonstrate that 3D engineered tumor model can better mimic in vivo tumor and can serve as a more appropriate platform for the study and screening of novel cancer therapeutics.

Characterization of human adipose tissue-derived stem cells in vitro culture and in vivo differentiation in a temperature-sensitive chitosan/ß- glycerophosphate/collagen hybrid hydrogel.

In this study, the interaction of human adipose tissue-derived stem cells (ADSCs) with chitosan/ß-glycerophosphate/collagen (C/GP/Co) hybrid hydrogel was test, followed by investigating the capability of engineered adipose tissue formation using this ADSCs seeded hydrogel. The ADSCs were harvested and mixed with a C/GP/Co hydrogel followed by a gelation at 37°C and an in vitro culture. The results showed that the ADSCs within C/GP/Co hydrogels achieved a 30% of expansion over 7days in culture medium and encapsulated cell in C/GP/Co hydrogel demonstrated a characteristic morphology with high viability over 5days. C/GP/Co hydrogel were subcutaneous injected into SD-rats to assess the biocompatibility. The induced ADSCs-C/GP/Co hydrogel and non-induced ADSCs-C/GP/Co hydrogel were subcutaneously injected into nude mice for detecting potential of adipogenic differentiation. It has shown that C/GP/Co hydrogel were well tolerated in SD rats where they had persisted over 4weeks post implantation. Histology analysis indicated that induced ADSCs-C/GP/Co hydrogel has a greater number of adipocytes and vascularized adipose tissues compared with non-induced ADSCs-C/GP/Co hydrogel.

Development of kartogenin-conjugated chitosan-hyaluronic acid hydrogel for nucleus pulposus regeneration.

Injectable constructs for in vivo gelation have many advantages in the regeneration of degenerated nucleus pulposus. In this study, an injectable hydrogel consisting of chitosan (CS) and hyaluronic acid (HA) crosslinked with glycerol phosphate (GP) at different proportions (CS : GP : HA, 6 : 3 : 1, 5 : 3 : 2, 4 : 3 : 3, 3 : 3 : 4, 2 : 3 : 5, 1 : 3 : 6, V : V : V) was developed and employed as a delivery system for kartogenin (KGN), a biocompound that can activate chondrocytes. In vitro gelation time, morphologies, swelling, weight loss, compressive modulus and cumulative release of KGN in hydrogels were studied. For biocompatibility assessments, human adipose-derived stem cells (ADSCs) were encapsulated in these hydrogels. The effects of KGN on stem cell proliferation and differentiation into nucleus pulposus-like cells were examined. The hydrogels with higher concentrations of HA showed a slightly shorter gelation time, higher water uptake, faster weight loss and faster KGN release compared to the hydrogels with lower concentrations of HA. As the KGN-conjugated hydrogel prepared with the proportions 5 : 3 : 2 displayed good mechanical properties, it was chosen as the optimal gel to promote cell proliferation and differentiation. No significant difference was seen in the expression levels of nucleus pulposus markers induced by KGN or TGF-ß. Additionally, inclusion of KGN and TGF-ß together did not produce a synergistic effect in inducing nucleus pulposus properties. In conclusion, we have developed a KGN-conjugated CS/HA hydrogel (5 : 3 : 2) with sustained release of KGN in hydrogel that can promote ADSC proliferation and nucleus pulposus differentiation. This kind of hydrogel may be a simple and effective candidate for the repair of degenerative NP tissue after minimally invasive surgery.

Numerical Simulation of Mass Transfer and Three-Dimensional Fabrication of Tissue-Engineered Cartilages Based on Chitosan/Gelatin Hybrid Hydrogel Scaffold in a Rotating Bioreactor.

Cartilage tissue engineering is believed to provide effective cartilage repair post-injuries or diseases. Biomedical materials play a key role in achieving successful culture and fabrication of cartilage. The physical properties of a chitosan/gelatin hybrid hydrogel scaffold make it an ideal cartilage biomimetic material. In this study, a chitosan/gelatin hybrid hydrogel was chosen to fabricate a tissue-engineered cartilage in vitro by inoculating human adipose-derived stem cells (ADSCs) at both dynamic and traditional static culture conditions. A bioreactor that provides a dynamic culture condition has received greater applications in tissue engineering due to its optimal mass transfer efficiency and its ability to simulate an equivalent physical environment compared to human body. In this study, prior to cell-scaffold fabrication experiment, mathematical simulations were confirmed with a mass transfer of glucose and TGF-ß2 both in rotating wall vessel bioreactor (RWVB) and static culture conditions in early stage of culture via computational fluid dynamic (CFD) method. To further investigate the feasibility of the mass transfer efficiency of the bioreactor, this RWVB was adopted to fabricate three-dimensional cell-hydrogel cartilage constructs in a dynamic environment. The results showed that the mass transfer efficiency of RWVB was faster in achieving a final equilibrium compared to culture in static culture conditions. ADSCs culturing in RWVB expanded three times more compared to that in static condition over 10 days. Induced cell cultivation in a dynamic RWVB showed extensive expression of extracellular matrix, while the cell distribution was found much more uniformly distributing with full infiltration of extracellular matrix inside the porous scaffold. The increased mass transfer efficiency of glucose and TGF-ß2 from RWVB promoted cellular proliferation and chondrogenic differentiation of ADSCs inside chitosan/gelatin hybrid hydrogel scaffolds. The improved mass transfer also accelerated a dynamic fabrication of cell-hydrogel constructs, providing an alternative method in tissue engineering cartilage.

Culture of neural stem cells in calcium alginate beads.

Neural stem cells (NSCs) with the capacity of extensive self-renewal and multilineage differentiation have attracted more and more attention in research as NSCs will play an important role in the nerve disease treatment and nerve injury repair. The shortage of NSCs, both their sources and their numbers, however, is the biggest challenge for their clinic application, and hence, in vitro culture and expansion of NSCs is vitally important to realize their potentials. In this work, mouse-derived NSCs were cultured in three-dimensional calcium alginate beads (Ca-Alg-Bs). Gelling conditions, cell density, and cell harvest were determined by the exploration of formation and dissociation parameters for Ca-Alg-Bs. Additionally, the recovered and the subsequent induced cells were identified by immunofluorescence staining of Nestin, beta-tubulin, and GFAP. The results show that the 2-mm diameter Ca-Alg-Bs, prepared with 1.5% sodium alginate solution and 3.5% CaCl2 solution and with gelling for 10 min, is suitable for the NSCs culture. The seeding density of 0.8 x 10(5) cells x mL-1 for the encapsulation of NSCs resulted in the most expansion, and the NSCs almost doubled during the experiment. The average cell recovery rate is over 88.5%, with the Ca-Alg-Bs dissolving in 55 mM sodium citrate solution for 10 min. The recovered cells cultured in the Ca-Alg-Bs still expressed Nestin and had the capacity of multilineage differentiation into neurons and glial cells and, thus, remained to be NSCs. These results demonstrate that NSC expansion within Ca-Alg-Bs is feasible and provides further possibilities for NSC expansion in bioreactors of the scale of clinical relevance.

Fabrication and detection of tissue engineered bone aggregates based on encapsulated human ADSCs within hybrid calcium alginate/bone powder gel-beads in a spinner flask.

Traditional treatment for bone diseases limits their clinical application due to undesirable host immune rejection, limited donator sources and severe pain and suffering for patients. Bone tissue engineering therefore is expected to be a more effective way in treating bone diseases. In the present study, hybrid calcium alginate/bone powder gel-beads with a uniform size distribution, good biocompatibility and osteoinductive capability, were prepared to be used as an in-vitro niche-like matrix. The beads were optimized using 2.5% (w/v) sodium alginate solution, 4.5% (w/v) CaCl2 solution and 5.0mg/mL bone powder using an easy-to-use method. Human ADSCs were cultured and induced into chondrocytes and osteoblasts, respectively. The cells were characterized by histological staining showing the ADSCs were able to maintain their characteristic morphology with multipotent differentiation ability. ADSCs at density of 5 × 10(6)cells/mL were encapsulated into the gel-beads aiming to explore cell expansion under different conditions and the osteogenic induction of ADSCs was verified by specific staining. Results demonstrated that the encapsulated ADSCs expanded 5.6 folds in 10 days under dynamic condition via spinner flask, and were able to differentiate into osteoblasts (OBs) with extensive mineralized nodules forming the bone aggregates over 3 weeks postosteogenic induction. In summary, hybrid gel-beads encapsulating ADSCs are proved to be feasible as a new method to fabricate tissue engineered bone aggregation with potential to treat skeletal injury in the near future.

In vitro culture and harvest of BMMSCs on the surface of a novel thermosensitive glass microcarrier.

Traditional two-dimensional (2D) static culture environment for stem cells followed by enzymatic cell detachment or mechanical treatment is routinely used in research laboratories. However, this method is not ideal as stem cells expand slowly, with cell damage and partial loss of specific stemness. For this reason, a better culture condition is urgently needed to improve stem cell recovery. A novel thermosensitive P(NIPAAm-co-HPM)-g-TMSPM-g-microcarrier was prepared here as a three-dimensional (3D) culture substitute. This novel microcarrier was prepared by grafting NIPAAm and HPM to the surface of glass microcarrier using TMSPM through surface free radical copolymerization. The prepared material was tested in cell culture and via cooling harvest method. We found that NIPAAm was successfully grafted on to the surface of the microcarriers, providing an excellent biocompatible environment for BMMSC adhesion and growth. More importantly, BMMSCs could be fully removed from the thermosensitive glass microcarriers with remained cell viability.

Development and fabrication of a two-layer tissue engineered osteochondral composite using hybrid hydrogel-cancellous bone scaffolds in a spinner flask.

Biological treatment using engineered osteochondral composites has received growing attention for the repair of cartilage defects. Osteochondral composites combined with a dynamic culture provide great potential for improving the quality of constructs and cartilage regeneration as dynamic conditions mimic the in vivo condition where cells were constantly subjected to mechanical and chemical stimulation. In the present study, biophasic composites were produced in vitro consisting of cell-hydrogel (CH) and cell-cancellous bone (CB) constructs, followed by culturing in a dynamic system in a spinner flask. The aim of this study was to investigate cell behaviors (i.e. cell growth, differentiation, distribution and matrix deposition) cultured in different constructs under static and dynamic circumstances. As a result, we found that mechanical stimulation promoted osteogenic and chondrogenic differentiation of cells as indicated by the increased expression of ALP and glycosaminoglycan (GAG) in either bone or cartilage substitute materials. Dynamic culture yielded a preferable extracellular matrix production, particularly in hydrogel scaffolds. In addition, the enhanced mass transfer contributed to the interface formation, cells infiltration and distribution in the osteochondral composites. This study demonstrates that osteochondral composites incorporated with a dynamic culture improved the performance of the constructs, providing the basis for a promising tool and a better strategy for the rapid fabrication of osteochondral substitutes and regeneration of injured cartilage.

Thermo-responsive poly(N-isopropylacrylamide)-grafted hollow fiber membranes for osteoblasts culture and non-invasive harvest.

Hollow fiber membrane (HFM) culture system is one of the most important bioreactors for the large-scale culture and expansion of therapeutic cells. However, enzymatic and mechanical treatments are traditionally applied to harvest the expanded cells from HFMs, which inevitably causes harm to the cells. In this study, thermo-responsive cellulose acetate HFMs for cell culture and non-invasive harvest were prepared for the first time via free radical polymerization in the presence of cerium (IV). ATR-FTIR and elemental analysis results indicated that the poly(N-isopropylacrylamide) (PNIPAAm) was covalently grafted on HFMs successfully. Dynamic contact angle measurements at different temperatures revealed that the magnitude of volume phase transition was decreased with increasing grafted amount of PNIPAAm. And the amount of serum protein adsorbed on HFMs surface also displayed the same pattern. Meanwhile osteoblasts adhered and spread well on the surface of PNIPAAm-grafted HFMs at 37 °C. And Calcein-AM/PI staining, AB assay, ALP activity and OCN protein expression level all showed that PNIPAAm-grafted HFMs had good cell compatibility. After incubation at 20 °C for 120 min, the adhering cells on PNIPAAm-grafted HFMs turned to be round and detached after being gently pipetted. These results suggest that thermo-responsive HFMs are attractive cell culture substrates which enable cell culture, expansion and the recovery without proteolytic enzyme treatment for the application in tissue engineering and regenerative medicine.

Three-dimensional dynamic fabrication of engineered cartilage based on chitosan/gelatin hybrid hydrogel scaffold in a spinner flask with a special designed steel frame.

Cartilage transplantation using in vitro tissue engineered cartilage is considered a promising treatment for articular cartilage defects. In this study, we assessed the advantages of adipose derived stem cells (ADSCs) combined with chitosan/gelatin hybrid hydrogel scaffolds, which acted as a cartilage biomimetic scaffold, to fabricate a tissue engineered cartilage dynamically in vitro and compared this with traditional static culture. Physical properties of the hydrogel scaffolds were evaluated and ADSCs were inoculated into the hydrogel at a density of 1×10(7) cells/mL and cultured in a spinner flask with a special designed steel framework and feed with chondrogenic inductive media for two weeks. The results showed that the average pore size, porosity, swelling rate and elasticity modulus of hybrid scaffolds with good biocompatibility were 118.25±19.51 µm, 82.60±2.34%, 361.28±0.47% and 61.2±0.16 kPa, respectively. ADSCs grew well in chitosan/gelatin hybrid scaffold and successfully differentiated into chondrocytes, showing that the scaffolds were suitable for tissue engineering applications in cartilage regeneration. Induced cells cultivated in a dynamic spinner flask with a special designed steel frame expressed more proteoglycans and the cell distribution was much more uniform with the scaffold being filled mostly with extracellular matrix produced by cells. A spinner flask with framework promoted proliferation and chondrogenic differentiation of ADSCs within chitosan/gelatin hybrid scaffolds and accelerated dynamic fabrication of cell-hydrogel constructs, which could be a selective and good method to construct tissue engineered cartilage in vitro.

Biodegradable radiopaque iodinated poly(ester urethane)s containing poly(ε-caprolactone) blocks: synthesis, characterization, and biocompatibility.

Biodegradable radiopaque iodinated poly(ester-urethane) (I-PU), consisting of poly(ε-caprolactone) (PCL) diol and iodinated bisphenol A (IBPA), has been successfully synthesized via a coupling reaction of PCL-diisocyanate and IBPA with varying compositions. The IBPA with four iodine atoms per molecule was applied as a chain extender to endow the I-PUs with intrinsic X-ray visibility. The chemical structure and molecular weights of I-PUs were characterized by Fourier transform infrared spectroscopy (FT-IR), proton-nuclear magnetic resonance, and gel permeation chromatography (GPC). The effects of IBPA on the physical properties of I-PUs were systematically studied by means of differential scanning calorimetry (DSC), thermogravimetric analysis (TGA), and wide-angle X-ray diffraction (WAXD). The DSC results showed that the crystallization of PCL segments in I-PUs was restrained with increasing amount of IBPA, which was also confirmed by WAXD. In the X-radiography analysis, all the synthesized I-PUs exhibited high radiopacity compared with an aluminum wedge of equivalent thickness. Enzymatic degradation tests showed that the incorporation of IBPA prolonged the degradation of I-PUs and distinct mass loss and degradation happened in the third month. Basic cytocompatibility conducted using rat adipose-derived cells proved that all the I-PUs and their biodegradation products were nontoxic. The radiopaque I-PUs is expected to possess a significant advantage over the traditional polymer counterparts in some related biomedical fields.

Fabrication and evaluation of a sustained-release chitosan-based scaffold embedded with PLGA microspheres.

Nutrient depletion within three-dimensional (3D) scaffolds is one of the major hurdles in the use of this technology to grow cells for applications in tissue engineering. In order to help in addressing it, we herein propose to use the controlled release of encapsulated nutrients within polymer microspheres into chitosan-based 3D scaffolds, wherein the microspheres are embedded. This method has allowed maintaining a stable concentration of nutrients within the scaffolds over the long term. The polymer microspheres were prepared using multiple emulsions (w/o/w), in which bovine serum albumin (BSA) and poly (lactic-co-glycolic) acid (PLGA) were regarded as the protein pattern and the exoperidium material, respectively. These were then mixed with a chitosan solution in order to form the scaffolds by cryo-desiccation. The release of BSA, entrapped within the embedded microspheres, was monitored with time using a BCA kit. The morphology and structure of the PLGA microspheres containing BSA before and after embedding within the scaffold were observed under a scanning electron microscope (SEM). These had a round shape with diameters in the range of 27-55 µm, whereas the chitosan-based scaffolds had a uniform porous structure with the microspheres uniformly dispersed within their 3D structure and without any morphological change. In addition, the porosity, water absorption and degradation rate at 37 °C in an aqueous environment of 1% chitosan-based scaffolds were (92.99±2.51) %, (89.66±0.66) % and (73.77±3.21) %, respectively. The studies of BSA release from the embedded microspheres have shown a sustained and cumulative tendency with little initial burst, with (20.24±0.83) % of the initial amount released after 168 h (an average rate of 0.12%/h). The protein concentration within the chitosan-based scaffolds after 168 h was found to be (11.44±1.81)×10(-2) mg/mL. This novel chitosan-based scaffold embedded with PLGA microspheres has proven to be a promising technique for the development of new and improved tissue engineering scaffolds.

Comparison of mesenchymal stem cells released from poly(N-isopropylacrylamide) copolymer film and by trypsinization.

Temperature-responsive platforms containing poly(N-isopropylacrylamide) (PNIPAAm) have been developed as an effective substitute for enzymatic treatment to recover adherent cells, but it remains unclear whether this alternative harvesting method tends to support stem cells preserving them being primitive. This study mainly investigated the biological properties of mesenchymal stem cells derived from rat bone marrow and human adipose tissue (BM-MSCs and AT-MSCs) after being cultured on PNIPAAm copolymer films and recovered by temperature drop, and compared the cells harvested from glass coverslips with trypsinization as controls. The experimental results demonstrated that after three serial passages, the released MSCs from thermal liftoff showed no significant differences in cell morphology, immunophenotype and osteogenesis for BM-MSCs or adipogenesis for AT-MSCs, but had higher viability, stronger proliferation and higher adipogenic differentiation for BM-MSCs or higher osteogenic differentiation for AT-MSCs compared with the trypsinization group. Besides, more proteins remained around or within the cell membranes upon temperature drop. It is concluded that cell detachment with more extracellular matrix proteins facilitates the maintenance of membrane proteins, and accordingly preserves MSC properties related to viability, proliferation and differentiation to some extent. This indicates that the PNIPAAm copolymer films and their matching cooling treatment can be used as effective alternatives to the existing culture substrates and traditional enzymatic digestion for MSCs.