PGS Scaffolds Research Articles

Comparative study of iodine-doped and undoped pyrrole grafting with plasma on poly (glycerol sebacate) scaffolds and its human dental pulp stem cells compatibility.

This study addresses the morphological and chemical characterization of PGS scaffolds after (6, 12, 18, 24, and 30 min) residence in undoped pyrrole plasma (PGS-PPy) and the evaluation of cell viability with human dental pulp stem cells (hDPSCs). The results were compared with a previous study that used iodine-doped pyrrole (PGS-PPy/I). Analyses through SEM and AFM revealed alterations in the topography and quantity of deposited PPy particles. FTIR spectra of PGS-PPy scaffolds confirmed the presence of characteristic absorption peaks of PPy, with higher intensities observed in the nitrile and -C≡C- groups compared to PGS-PPy/I scaffolds, while raman spectra indicated a lower presence of polaron N+ groups. On the other hand, PGS scaffolds modified with PPy exhibited lower cytotoxicity compared to PGS-PPy/I scaffolds, as evidenced by the Live/Dead assay. Furthermore, the PGS-PPy scaffolds at 6 and 12 min, and particularly the PGS-PPy/I scaffold at 6 min, showed the best results in terms of cell viability by the fifth day of culture. The findings of this study suggest that undoped pyrrole plasma modification for short durations could also be a viable option to enhance the interaction with hDPSCs, especially when the treatment times range between 6 min and 12 min.

Gelatin-modified 3D printed PGS elastic hierarchical porous scaffold for cartilage regeneration.

Regenerative cartilage replacements are increasingly required in clinical settings for various defect repairs, including bronchial cartilage deficiency, articular cartilage injury, and microtia reconstruction. Poly (glycerol sebacate) (PGS) is a widely used bioelastomer that has been developed for various regenerative medicine applications because of its excellent elasticity, biodegradability, and biocompatibility. However, because of inadequate active groups, strong hydrophobicity, and limited ink extrusion accuracy, 3D printed PGS scaffolds may cause insufficient bioactivity, inefficient cell inoculation, and inconsistent cellular composition, which seriously hinders its further cartilage regenerative application. Here, we combined 3D printed PGS frameworks with an encapsulated gelatin hydrogel to fabricate a PGS@Gel composite scaffold. PGS@Gel scaffolds have a controllable porous microstructure, with suitable pore sizes and enhanced hydrophilia, which could significantly promote the cells' penetration and adhesion for efficient chondrocyte inoculation. Furthermore, the outstanding elasticity and fatigue durability of the PGS framework enabled the regenerated cartilage built by the PGS@Gel scaffolds to resist the dynamic in vivo environment and maintain its original morphology. Importantly, PGS@Gel scaffolds increased the rate of cartilage regeneration concurrent with scaffold degradation. The scaffold was gradually degraded and integrated to form uniform, dense, and mature regenerated cartilage tissue with little scaffold residue.

3D Printed Chondrogenic Functionalized PGS Bioactive Scaffold for Cartilage Regeneration.

Tissue engineering is emerging as a promising approach for cartilage regeneration and repair. Endowing scaffolds with cartilaginous bioactivity to obtain bionic microenvironment and regulating the matching of scaffold degradation and regeneration play a crucial role in cartilage regeneration. Poly(glycerol sebacate) (PGS) is a representative thermosetting bioelastomer known for its elasticity, biodegradability, and biocompatibility and is widely used in tissue engineering. However, the modification and drug loading of the PGS scaffold is still a key challenge due to its high temperature curing conditions and limited reactive groups, which seriously hinders its further functional application. Here, a simple versatile new strategy of super swelling-absorption and cross-linked networks locking is presented to successfully create the 3Dprinted PGS-CS/Gel scaffold for the first time based on FDA-approved PGS, gelatin (Gel) and chondroitin sulfate (CS). The PGS-CS/Gel scaffold exhibits the desirable synergistic properties of well-organized hierarchical structures, excellent elasticity, improved hydrophilicity, and cartilaginous bioactivity, which can promote the adhesion, proliferation, and migration of chondrocytes. Importantly, the rate of cartilage regeneration can be well-matched with degradation of PGS-CS/Gel scaffold, and achieve uniform and mature cartilage tissue without scaffold residual. The bioactive scaffold can successfully repair cartilage in a rabbit trochlear groove defect model indicating a promising prospect of clinical transformation.

Preliminary Application Research of 3D Bioprinting in Craniofacial Reconstruction.

In recent years, 3-dimensional (3D) printing has been widely used in regenerative medicine research and other fields because of its ability to customize macroscopic morphology and precisely control microstructure. Polymer scaffolds are 1 of the commonly used 3D bioprinting materials for defect repair and have recently been a research focus. Our article explored the bone-formation accelerating effect of 3D-printed porous scaffold Poly(glycerol sebacate) [PGS] in the critical bone defect of an enhancing rabbit mandibular model. Also, we overview and summarize the classification of 3D bioprinting materials and prospects for their various application scenarios in craniofacial reconstruction surgery. A PGS elastomer scaffold was prepared by polymerizing equimolar amounts of sebacic acid and glycerol using a biological 3D printer. Six male New Zealand white rabbits were prepared (3 for the control group and 3 for the PGS group), each weighing 3kg. Osteotomy was performed at the anterior edge of the ascending ramus of the mandible with a bone saw to open the 8mm defect. Defects of the control group were empty, and defects of the PGS group were put into 8mm-wide PGS elastomer scaffolds. The rabbits were euthanized 6 weeks after the operation, and the postoperative mandibles were collected. Information (presence or absence of pus from infection, nonunion, degree of macroscopic bone healing) was recorded, and the skeletal tissue was fixed in a paraformaldehyde solution. The mandible on the enhanced side was significantly longer than that on the opposite side, and the contralateral incisor was hyperplasia. The mandibles of rabbits in each group healed well, and there was no obvious local infection and purulence. The gross specimen appearance showed that both ends of the defect were connected. When comparing the reconstructed mandibles of the two groups, it is apparent that the width and thickness of the new bone in the PGS group were significantly better than that in the control group. This article verifies the effect of 3D polypore PGS scaffolds in animal craniomaxillofacial bone defects and introduces various application scenarios of 3D printing materials in craniomaxillofacial reconstruction surgery. There are quite good application prospects for 3D bioprinting in animal experiments and even clinical treatment of craniofacial defects.

Stress-Induced Intensification of Deoxyshikonin Production in Rindera graeca Hairy Root Cultures with Ester-Based Scaffolds.

In vitro plant cell and tissue culture systems allow for controlling a wide range of culture environmental factors selectively influencing biomass growth and the yield of secondary metabolites. Among the most efficient methods, complex supplementation of the culture medium with elicitors, precursors, and other functional substances may significantly enhance valuable metabolite productivity through a stress induction mechanism. In the search for novel techniques in plant experimental biotechnology, the goal of the study was to evaluate stress-inducing properties of novel biodegradable ester-based scaffolds made of poly(glycerol sebacate) (PGS) and poly(lactic acid) (PLA) influencing on the growth and deoxyshikonin productivity of Rindera graeca hairy roots immobilized on the experimental constructs. Rindera graeca hairy roots were maintained under the dark condition for 28 days in three independent systems, i.e., (i) non-immobilized biomass (a reference system), (ii) biomass immobilized on PGS scaffolds, and (iii) biomass immobilized on PLA scaffolds. The stress-inducing properties of the applied polymerized esters selectively impacted R. graeca hairy roots. The PGS scaffolds caused the production of deoxyshikonin, which does not occur in other culture systems, and PLA promoted biomass proliferation by doubling its increase compared to the reference system.

Shape memory poly (glycerol sebacate)‐based electrospun fiber scaffolds for tissue engineering applications: A review

AbstractElectrospun fiber scaffolds has made a huge progress in tissue engineering applications owing to their continuous morphology with variable chemical, physical, and biological properties. In this review paper, we reviewed the electrospinning technique, its processing parameters and characterizations involved. A focus on shape memory poly (glycerol sebacate) (PGS)‐based electrospun fiber scaffolds for tissue engineering field was explored. The stimuli responsive PGS scaffold has gained much attention due to its ability to self‐expanding, self‐deploying, or self‐fitting. Herein, the electrospinning of shape memory PGS‐based scaffolds, particularly their processing parameters, properties, and characterizations have been reviewed. Besides, other issues concerning the technology constraints, challenges, and future directions are also discussed.

Use of cyclic strain bioreactor for the upregulation of key tenocyte gene expression on Poly(glycerol-sebacate) (PGS) sheets.

The inadequate donor source and the difficulty of using natural grafts in tendon repair and regeneration has led researchers to develop biodegradable and biocompatible synthetic based tissue equivalents. Poly(glycerol sebacate) (PGS) is a surface-erodible bioelastomer and has been increasingly investigated in a variety of biomedical applications. In this study, PGS elastomeric sheets were prepared by using a facile microwave method and used as elastomeric platform for the first time under mechanical stimulation to induct the tenocyte gene expression. It is revealed that elastomeric PGS sheets promote progenitor tendon cell structure by increasing proliferation and gene expression with regard to tendon extracellular matrix components. Human tenocytes were seeded onto poly(glycerol-sebacate) sheets and were cultured two days prior to transfer to dynamic culture in a bioreactor system. Cell culture studies were carried out for 12 days under 0%, 3% and 6% strain at 0.33 Hz. The PGS-cell constructs were examined by using Scanning Electron Microscopy (SEM), cell viability via live/dead staining using confocal microscopy, and GAG/DNA analysis. In addition, gene expression was examined using real-time polymerase chain reaction (RT-PCR). Tenocytes cultured upon PGS scaffolds under 6% cyclic strain exhibited tendon-like gene expression profile compared to 3% and 0% strain groups. The results of this study show that PGS is a suitable material in promoting tendon tissue formation under dynamic conditions.

Mechanical performance of elastomeric PGS scaffolds under dynamic conditions

In developing novel scaffolds, addressing mechanical properties is essential especially when future applications involve cyclic mechanical loading. Therefore, it is important to understand the behaviour of its physical properties with the evolution of its weight loss. Poly(glycerol sebacate) (PGS) is a promising material for tissue and biomedical engineering applications due to its biocompatibility, biodegradability and mechanical properties. To understand the impact of the hydrolytic degradation on the density, cross-linking degree and porosity; scaffolds with an average porosity of 93 ± 2% were synthetized by salt leaching technique and submitted to hydrolytic degradation. The scaffold showed a Young modulus of 17.3 ± 3.4 kPa, with a negligible energy loss during the mechanical solicitation. Moreover, a weight loss of 28 ± 2% followed by an increase in the swelling ratio of the scaffold was observed after 8 weeks of hydrolytic degradation. When submitted to cyclic mechanical loading-unloading, the PGS scaffolds present an outstanding fatigue behaviour under dry and wet conditions, with a remarkable resilience to the cyclic mechanical solicitation, and even after 1000 mechanical cycles, the construct was able to recover to its initial geometry. Overall, the PGS scaffolds demonstrate promising mechanical properties for biomedical applications, especially under dynamic conditions.

The effect of salt fusion processing variables on structural, physicochemical and biological properties of poly(glycerol sebacate) scaffolds

Poly(glycerol sebacate), PGS, is a biodegradable elastomer recently proposed in the form of scaffolds for cardiac, vascular, cartilage or neural applications. In the present work, several processing variables for the fabrication of PGS scaffolds by the salt fusion method were systematically studied, namely the pre-polymer/porogen ratio, the salt particles average size, use of tetrahydrofuran to dissolve the pre-polymer for its injection in the porogen template, and the curing pressure. The effect of these variables on their structural, mechanical and biological properties was assessed to select those leading to optimal ones in terms of their potential performance in tissue engineering applications.

Endometrium Injury: PGS Scaffolds Promote the In Vivo Survival and Directional Differentiation of Bone Marrow Mesenchymal Stem Cells Restoring the Morphology and Function of Wounded Rat Uterus (Adv. Healthcare Mater. 5/2019)

Endometrium Injury: PGS Scaffolds Promote the In Vivo Survival and Directional Differentiation of Bone Marrow Mesenchymal Stem Cells Restoring the Morphology and Function of Wounded Rat Uterus (Adv. Healthcare Mater. 5/2019)

PGS Scaffolds Promote the In Vivo Survival and Directional Differentiation of Bone Marrow Mesenchymal Stem Cells Restoring the Morphology and Function of Wounded Rat Uterus.

Intrauterine adhesion (IUA) causing infertility and recurrent miscarriage of reproductive female mammals usually results from endometrium injury. Nevertheless, there is no efficient therapeutic method to avoid IUA. Bone marrow derived mesenchymal stem cells (BMSCs) are an important cell source for tissue regeneration. This study designs and explores the ability of BMSC-loaded elastic poly(glycerol sebacate) (PGS) scaffold to prevent IUA and compares the effect of PGS with poly(lactic-co-glycolic acid) (PLGA) and collagen scaffolds in resumption of damaged rat uteruses. The 3D architecture provided by PGS scaffolds favors the attachment and growth of rat BMSCs. In vivo bioluminescence imaging shows that compared with direct BMSC intrauterine injection, PLGA, and collagen scaffolds, the PGS scaffold significantly prolongs the retention time of BMSCs in a wounded rat uterus model. More importantly, BMSCs can directly differentiate into endometrial stromal cells after transplantation of PGS/BMSCs constructs, but not PLGA/BMSCs and collagen/BMSCs. It is found that the level of transforming growth factor β1 (TGF-β1), basic fibroblast growth factor (bFGF), vascular endothelial growth factor, and insulin-like growth factors in the injured endometrium adjacent to PGS/BMSCs constructs is higher than those of rats receiving PLGA/BMSCs, collagen/BMSCs, or BMSCs intrauterine transplantation. Besides, transplantation of PGS/BMSCs leads to better morphology recovery of the damaged uterus than PLGA/BMSCs and collagen/BMSCs. The receptive fertility of PGS/BMSCs is 72.2 ± 6.4%, similar to the one of collagen/BMSCs, but significantly higher than 42.3 ± 3.9% in PLGA/BMSCs. Taken together, PGS/BMSCs may be a promising candidate for preventing IUA.

Microwave-assisted facile fabrication of porous poly (glycerol sebacate) scaffolds

The biodegradable elastomeric polyester poly(glycerol sebacate) (PGS) was developed for soft-tissue engineering. It has been used in various research applications such as wound healing, cartilage tissue engineering, and vascular grafting due to its biocompatibility and elastomeric properties. However conventional PGS manufacture is generally limited by the laborious reaction conditions needed for curing which requires elevated reaction temperatures, high vacuum and multi-day reaction times. In this study, we developed a microwave irradiation methodology to fabricate PGS scaffolds under milder conditions with curing times that are 8 times faster than conventional methods. In particular, we determined microwave reaction temperatures and times for maximum crosslinking of PGS elastomers, demonstrating that PGS is fully crosslinked using gradual heating up to 160 °C for 3 h. Porosity and mechanical properties of these microwave-cured PGS elastomers were shown to be similar to PGS elastomers fabricated by the conventional polycondensation method (150 °C under 30 Torr for 24 h). To move one step closer to clinical application, we also examined the biocompatibility of microwave-cured PGS using in vitro cell viability assays with primary baboon arterial smooth muscle cells (SMCs). These combined results show microwave curing of PGS is a viable alternative to conventional curing.

Design and characterization of dexamethasone-loaded poly (glycerol sebacate)-poly caprolactone/gelatin scaffold by coaxial electro spinning for soft tissue engineering

The aim of this research was to fabricate dexamethasone (Dex)-loaded poly (glycerol sebacate) (PGS)-poly (caprolactone) (PCL)/gelatin (Gt) (PGS-PCL/Gt-Dex) fibrous scaffolds in the form of core/shell structure which have potential application in soft tissues. In this regard, after synthesize and characterizations of PGS, PGS-PCL and gelatin fibrous scaffolds were separately developed in order to optimize the electrospinning parameters. In the next step, coaxial electrospun fibrous scaffold of PGS-PCL/Gt fibrous scaffold with PGS-PCL as core and Gt as shell was developed and its mechanical, physical and chemical properties were characterized. Moreover, degradability, hydrophilicity and biocompatibility of PGS-PCL/Gt fibrous scaffold were evaluated. In addition, Dex was encapsulated in PGS-PCL/Gt fibrous scaffold and drug release was assessed for tissue engineering application. Results demonstrated the formation of coaxial fibrous scaffold with average porosity of 79% and average fiber size of 294nm. Moreover, PGS-PCL/Gt fibrous scaffold revealed lower elastic modulus, ultimate tensile and ultimate elongation than those of PGS-PCL scaffold and more close to mechanical properties of natural tissue. Furthermore, lower contact angle of PGS-PCL/Gt than that of PGS-PCL demonstrated improved surface hydrophilicity of scaffold. DEX release was sustained over a period time of 30days from the scaffolds via three steps consisting of an initial burst release, secondary linear phase release pattern with slower rate over 20days followed by an apparent zero-order release phase. MTT observations demonstrated that there was no evidence of toxicity in the samples with and without Dex. Our findings indicated that core/shell PGS-PCL/Gt-Dex fibrous could be used as a carrier for the sustained release of drugs relevant for tissue engineering which makes it appropriate for soft tissue engineering.

Fabrication of a mechanically anisotropic poly(glycerol sebacate) membrane for tissue engineering.

Poly(glycerol sebacate) (PGS) has been used successfully as a scaffolding material for soft tissue engineering. PGS scaffolds, however, are usually mechanically isotropic, which may restrict their use in tissue repairs as many soft tissues in the body have anisotropic mechanical behaviors. Although various methods have been used to fabricate anisotropic scaffolds, it remains challenging to make anisotropic scaffolds from thermoset PGS. Here a new, simple method to fabricate an anisotropic PGS membrane which can then be used to construct thicker three-dimensional anisotropic scaffolds was developed. First, an aligned sacrificial poly(vinyl alcohol) fibrous membrane was prepared by electrospinning. The fibrous membrane was then partially immersed in PGS prepolymer solution, resulting in a composite membrane upon drying. After curing, the sacrificial fibers within the membrane were removed by water, supposedly leaving aligned cylindrical pores in the membrane. Both SEM and AFM illustrated aligned grooves on the surface of the resultant PGS membrane, indicating the successful removal of sacrificial fibers. The PGS membrane was validated to be mechanically anisotropic using uniaxial tensile testing along and perpendicular to the predominant pore direction. The in vitro cytocompatibility of the PGS membrane was confirmed. As a demonstration of its potential application in vascular tissue engineering, a tubular scaffold was constructed by wrapping a stack of two axisymmetric pieces of the anisotropic PGS membranes on a mandrel. The compliance of the scaffold was found to depend on the pitch angle of its double helical structure, imitating the anisotropic mechanical behavior of the arterial media. © 2017 Wiley Periodicals, Inc. J Biomed Mater Res Part B: Appl Biomater, 106B: 760-770, 2018.

Nanoengineered Osteoinductive and Elastomeric Scaffolds for Bone Tissue Engineering.

Synthesis and fabrication of porous and elastomeric nanocomposite scaffolds from biodegradable poly(glycerol sebacate) (PGS) and osteoinductive nanosilicates is reported. Nanosilicates are mineral-based two-dimensional (2D) nanomaterials with high surface area which reinforced PGS network. The addition of nanosilicates to PGS resulted in mechanically stiff and elastomeric nanocomposites. The degradation rate and mechanical stiffness of nanocomposite network could be modulated by addition of nanosilicates. Nanocomposite scaffolds supported cell adhesion, spreading, and proliferation and promoted osteogenic differentiation of preosteoblasts. The addition of nanosilicates to PGS scaffolds increased alkaline phosphatase (ALP) activity and production of matrix mineralization. In vivo studies demonstrated biocompatibility and biodegradability of nanocomposite scaffolds. Overall, the combination of elasticity and tailorable stiffness, tunable degradation profiles, and the osteoinductive capability of the scaffolds offer a promising approach for bone tissue engineering.

Porous poly(glycerol sebacate) (PGS) elastomer scaffolds for skin tissue engineering

Poly (glycerol sebacate) (PGS) elastomer scaffolds with different porosity for skin tissue engineering were fabricated via particulate leaching. The introduction of pores lowers the hydrophilicity but improves the water uptake capability of PGS. The gel content of PGS increases with the increase of salt mass ratio, but the degree of swelling goes the opposite way due to the existence of the porous structure. The degradation rate of PGS can be tailored and controlled by the porous structure, which is of great value for its applications in tissue engineering. The feasibility of these porous PGS scaffolds for skin tissue engineering was evaluated by seeding mouse dermal fibroblasts (MDFs) onto the scaffold. In vitro cell culture results indicate good attachment, proliferation and deep penetration of MDFs into porous PGS scaffolds, which confirms the excellent biocompatibility of these scaffolds.

Poly(Limonene Thioether) Scaffold for Tissue Engineering.

A photocurable thiol-ene network polymer, poly(limonene thioether) (PLT32o), is synthesized, characterized, fabricated into tissue engineering scaffolds, and demonstrated in vitro and in vivo. Micromolded PLT32o grids exhibit compliant, elastomeric mechanical behavior similar to grids made of poly(glycerol sebacate) (PGS), an established biomaterial. Multilayered PL32o scaffolds with regular, geometrically defined pore architectures support heart cell seeding and culture in a manner similar to multilayered PGS scaffolds. Subcutaneous implantation of multilayered PLT32o scaffolds with cultured heart cells provides long-term 3D structural support and retains the exogenous cells, whereas PGS scaffolds lose both their structural integrity and the exogenous cells over 31 d in vivo. PLT32o membrane implants retain their dry mass, whereas PGS implants lose 70 percent of their dry mass by day 31. Macrophages are initially recruited to PLT32o and PGS membrane implants but are no longer present by day 31. Facile synthesis and processing in combination with the capability to support heart cells in vitro and in vivo suggest that PLT32o can offer advantages for tissue engineering applications where prolonged in vivo maintenance of 3D structural integrity and elastomeric mechanical behavior are required.

In vitro human chondrocyte culture on plasma-treated poly(glycerol sebacate) scaffolds

Porous poly(glycerol sebacate) (PGS) scaffolds were prepared using a salt leaching technique and subsequently surface modified by a low oxygen plasma treatment prior to the use in the in vitro culture of human chondrocytes. Condensation polymerization of glycerol and sebacic acid used at various mole ratios, i.e. 1:1, 1:1.25, and 1:1.5, was initially conducted to prepare PGS prepolymers. Porous elastomeric PGS scaffolds were directly fabricated from the mixtures of each prepolymer and 90% (w/w) NaCl particles and then subjected to the plasma treatment to enhance the surface hydrophilicity of the materials. The properties of both untreated and plasma-treated PGS scaffolds were comparatively evaluated, in terms of surface morphology, surface chemical composition, porosity, and storage modulus using scanning electron microscopy (SEM), X-ray photoelectron spectroscopy, micro-computed tomography, and dynamic mechanical analysis, respectively. The responses of chondrocytes cultured on individual PGS scaffolds were assessed, in terms of cell proliferation and ECM production. The results revealed that average pore sizes and porosity of the scaffolds were increased with an increasing sebacic acid concentration used. The storage moduli of the scaffolds were raised after the plasma treatment, possibly due to the further crosslinking of PGS upon treatment. Moreover, the scaffold prepared with a higher sebacic acid content demonstrated a greater capability of promoting cell infiltration, proliferation, and ECM production, especially when it was plasma-treated; the greatest HA, sGAG, uronic acid, and collagen contents were detected in matrix of this scaffold. The H & E and safranin O staining results also strongly supported this finding. The storage modulus of the scaffold was intensified after incubation with the chondrocytes for 21 days, indicating the accretion and retention of matrix ECM on the cell-cultured scaffold.

Highly elastic and suturable electrospun poly(glycerol sebacate) fibrous scaffolds

Poly(glycerol sebacate) (PGS) is a thermally-crosslinked elastomer suitable for tissue regeneration due to its elasticity, degradability, and pro-regenerative inflammatory response. Pores in PGS scaffolds are typically introduced by porogen leaching, which compromises strength. Methods for producing fibrous PGS scaffolds are very limited. Electrospinning is the most widely used method for laboratory scale production of fibrous scaffolds. Electrospinning PGS by itself is challenging, necessitating a carrier polymer which can affect material properties if not removed. We report a simple electrospinning method to produce distinct PGS fibers while maintaining the desired mechanical and cytocompatibility properties of thermally crosslinked PGS. Fibrous PGS demonstrated 5 times higher tensile strength and increased suture retention compared to porous PGS foams. Additionally, similar modulus and elastic recovery were observed. A final advantage of fibrous PGS sheets is the ability to create multi-laminate constructs due to fiber bonding that occurs during thermal crosslinking. Taken together, these highly elastic fibrous PGS scaffolds will enable new approaches in tissue engineering and regenerative medicine.

Repair of critical-sized bone defects with anti-miR-31-expressing bone marrow stromal stem cells and poly(glycerol sebacate) scaffolds

The repair of critical-sized defects (CSDs) is a significant challenge in bone tissue engineering. Combining the use of progenitor cells with gene therapy represents a promising approach for bone regeneration. MicroRNAs play important roles in most gene regulatory networks, regulate the endogenous expression of multiple growth factors and simultaneously modulate stem cell differentiation. Our previous study showed that knocking down miR-31 promotes the osteogenesis of bone marrow stromal stem cells (BMSCs). To investigate the therapeutic potential of cells engineered to express anti-miR-31 for CSD repair, lentiviral vectors encoding negative control, miR-31 precursor and anti-sense sequences were constructed and transduced into osteo-inductive BMSCs. The expression of osteogenic-specific genes, alkaline phosphatase activity and Alizarin Red S staining were investigated to evaluate the effects of miR-31 on the cell fate of BMSCs over a 3-week period. In addition, miR-31-modified BMSCs seeded on poly(glycerol sebacate) (PGS) scaffolds were used to repair 8 mm critical-sized calvarial defects in rats. The results showed that miR-31 suppression significantly increased the expression of osteogenic-specific genes in vitro at the mRNA and protein levels, and that robust new bone formation with high local bone mineral density was observed in the anti-miR groups in vivo. Moreover, the PGS scaffolds carrying anti-miR-31-expressing BMSCs exhibited good biocompatibility and a high regeneration rate (~60%) within in vivo bone defects. Our results suggest that miR-31 gene delivery affects the potential of BMSCs for osteogenic differentiation and bone regeneration and that PGS is a potential substrate for genetically modified, tissue-engineered bone in the repair of large bone defects.