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The on-demand replacement of multifunctional hydrogel wound dressings helps to avoid bacterial colonization, and the on-demand painless peeling of tissue adhesive hydrogels on the wound site remains a major challenge to be solved. In this work, we design and develop a series of multifunctional dynamic Schiff base network hydrogels composed of cystamine-modified hyaluronic acid, benzaldehyde-functionalized poly(ethylene glycol)-co-poly(glycerol sebacate), and polydopamine@polypyrrole nanocomposite (PDA@PPy) with mild on-demand removability to enhance drug-resistant bacteria-infected wound healing. These hydrogels exhibited ideal injectable and self-healing properties, excellent tissue adhesion, in vivo hemostasis, good antioxidation, and conductivity. PDA@PPy inspired by melanin endows hydrogels with excellent antioxidant capacity, UV-blocking ability, and photothermal anti-infection ability. Based on the dynamic oxidation-reduction response of disulfide bonds inspired by the dissociation of the tertiary spatial structure transformation of poly-polypeptide chains, these hydrogels can achieve rapid painless on-demand removal under mild conditions by adding dithiothreitol. These multifunctional hydrogels significantly promoted collagen deposition and angiogenesis in the MRSA-infected full-thickness skin repair experiment. All the results showed that these multifunctional hydrogels with painless on-demand removal property showed great potential in clinical treatment of infected wounds.

Lipid nanoparticles (LNPs) are the leading technology for RNA delivery, given the success of the Pfizer/BioNTech and Moderna COVID-19 mRNA (mRNA) vaccines, and small interfering RNA (siRNA) therapies (patisiran). However, optimization of LNP process parameters and compositions for larger RNA payloads such as self-amplifying RNA (saRNA), which can have complex secondary structures, have not been carried out. Furthermore, the interactions between process parameters, critical quality attributes (CQAs), and function, such as protein expression and cellular activation, are not well understood. Here, we used two iterations of design of experiments (DoE) (definitive screening design and Box-Behnken design) to optimize saRNA formulations using the leading, FDA-approved ionizable lipids (MC3, ALC-0315, and SM-102). We observed that PEG is required to preserve the CQAs and that saRNA is more challenging to encapsulate and preserve than mRNA. We identified three formulations to minimize cellular activation, maximize cellular activation, or meet a CQA profile while maximizing protein expression. The significant parameters and design of the response surface modeling and multiple response optimization may be useful for designing formulations for a range of applications, such as vaccines or protein replacement therapies, for larger RNA cargoes.

This study has attempted to systematically investigate the influence of nanoclay and graphene oxide (GO) on thermal, mechanical, hydrophobic, and, most importantly, biological properties of poly(glycerol sebacate)/gelatin (PGS/gel) nanocomposites. The PGS/gel copolymer nanocomposites were successfully synthesized via in situ polymerization, approved by rudimentary characterization methods. The nanofillers were appropriately dispersed within the elastomeric matrix according to morphological studies. Also, the fillers posed as a hydrophobic entity that slightly decreased the hydrophilic properties of PGS/gel. This could be sensed clearly in hybrid composite due to the robust network of GO and clay. Water contact angle values for gelatin-contained nanocomposites were reported in the range of 38.42° to 66.7°, indicating the hydrophilic nature of the prepared samples. Thermal and mechanical studies of nanocomposites displayed rather contradicting results as the former improved while a slight decrease in the latter was noticed compared to the pristine specimens. In dry conditions, their storage modulus was in the range of 0.94-6.4 MPa, making them suitable for mimicking some soft tissues. The swelling ratio for nanocomposites containing nanoparticles was associated with an ascending trend so that GO improved the swelling rate by up to 45%. Biological analyses, such as Ames and in vitro cell viability tests, exhibited promising outcomes. As for the mutagenesis effect, the PGS and hybrid samples showed negative results. The presence of functional groups on the nanofillers' surface positively influenced the cells' metabolic activity as well as its attachment to the matrix. After 7 days, the cell proliferation rate resulted in an 82% improvement for the GO-containing nanocomposite, significantly higher than its neat counterpart (65%). This study has shown the feasibility of the prepared bio-elastomer nanocomposites for diverse tissue engineering applications.

This study examined poly(glycerol-1,8-octanediol-sebacate) (PGOS) copolymers with low-level substitution of O (1,8-octanediol) for G (glycerol) units (G/O ratios 0.5:0.5, 0.66:0.33, 0.75:0.25, 0.8:0.2, and 0.91:0.09) prepared in bulk by immobilized

Nanocomposites containing clay nanoparticles often present favorable properties such as good mechanical and thermal properties. They frequently have been studied for tissue engineering (TE) and regenerative medicine applications. On the other hand, poly(glycerol sebacate) (PGS), a revolutionary bioelastomer, has exhibited substantial potential as a promising candidate for biomedical application. Here, we present a facile approach to synthesizing stiff, elastomeric nanocomposites from sodium-montmorillonite nano-clay (MMT) in the commercial name of Cloisite Nasup+/supand poly(glycerol sebacate urethane) (PGSU). The strong physical interaction between the intercalated Cloisite Nasup+/supplatelets and PGSU chains resulted in desirable property combinations for TE application to follow. The addition of 5% MMT nano-clay resulted in an over two-fold increase in the tensile modulus, increased the onset thermal decomposition temperature of PGSU matrix by 18°C, and noticeably improved storage modulus of the prepared scaffolds, compared with pure PGSU. As well, Cloisite Nasup+/supenhanced the hydrophilicity and water uptake ability of the samples and accelerated the in-vitro biodegradation rate. Finally, in-vitro cell viability assay using L929 mouse fibroblast cells indicated that incorporating Cloisite Nasup+/supnanoparticles into the PGSU network could improve the cell attachment and proliferation, rendering the synthesized bioelastomers potentially suitable for TE and regenerative medicine applications.

Prevascularization of tissue equivalents is critical for fulfilling the need for sufficient vascular organization for nutrient and gas transport. Hence, endothelial cell culture on biomaterials is of great importance for researchers. Numerous alternate strategies have been suggested in this sense, with cell-based methods being the most commonly employed. In this study, poly (glycerol sebacate) (PGS) elastomers with varying crosslinking ratios were synthesized and their surfaces were patterned with channels by using laser ablation technique. In order to determine an ideal material for cell culture studies, the elastomers were subsequently mechanically, chemically, and biologically characterized. Following that, human umbilical vein endothelial cells (HUVECs) were seeded into the channels established on the PGS membranes and cultured under various culture conditions to establish the optimal culture parameters. Lastly, the endothelial cell responses to the synthesized PGS elastomers were evaluated. Remarkable cell proliferation and impressive cellular organizations were noticed on the constructs created as part of the investigation. On the concrete output of this research, arrangements in various geometries can be created by laser ablation method and the effects of various molecules, drugs or agents on endothelial cells can be evaluated. The platforms produced can be employed as an intermediate biomaterial layer containing endothelial cells for vascularization of tissue-engineered structures, particularly in layer-by-layer tissue engineering approaches.

The targeted delivery of messenger RNA (mRNA) to desired organs remains a great challenge for in vivo applications of mRNA technology. For mRNA vaccines, the targeted delivery to the lymph node (LN) is predicted to reduce side effects and increase the immune response. In this study, we explored an endogenously LN-targeting lipid nanoparticle (LNP) without the modification of any active targeting ligands for developing an mRNA cancer vaccine. The LNP named 113-O12B showed increased and specific expression in the LN compared with LNP formulated with ALC-0315, a synthetic lipid used in the COVID-19 vaccine Comirnaty. The targeted delivery of mRNA to the LN increased the CD8

This study demonstrated that immobilized

In this study, a novel biopolymer based on poly(glycerol sebacic)-urethane (PGS-U) and its nanocomposites containing Cloisite@30B were synthesized by facile approach in which the crosslinking was created by aliphatic hexamethylene diisocyanate (HDI) at room temperature and 80 °C. Moreover, metronidazole and tetracycline drugs were selected as target drugs and loaded into PGSU based nanocomposites. A uniform and continuous microstructure with smooth surface is observed in the case of pristine PGS-U sample. The continuity of microstructure is observed in the case of all bionanocomposites. XRD result confirmed an intercalated morphology for PGSU containing 5 wt% of clay nanoparticles with a d-spacing 3.4 nm. The increment of nanoclay content up to 5%, the ultimate tensile stress and elastic modulus were obtained nearly 0.32 and 0.83 MPa, which the latter was more than eight-fold than that of pristine PGS-U. A sustained release for both dugs was observed by 200 h. The slowest and controlled drug release rate was determined in the case of PGSU containing 5 wt% clay and cured at 80 °C. A non-Fickian diffusion can be concluded in the case of tetracycline release via PGS-U/nanoclay bionanocomposites, while a Fickian process was detected in the case of metronidazole release by PGS-U/nanoclay bionanocomposites. As a result, the designed scaffold showed high flexibility, which makes it an appropriate option for utilization in the treatment of periodontal disease.

Poly(glycerol sebacate) (PGS) is an aliphatic polyester which attracted significant scientific attention in recent years due to its vast potential in biomedical applications with regard to tissue engineering. It has been presented in the literature in the form of 2D films, porous scaffolds or nonwovens, to name just a few. Moreover, various applications have been proposed as a component of composite materials or polymer blends. Its physicochemical properties can be significantly adjusted by means of synthesis and post-synthetic modifications, including cross-linking or chemical modification, such as copolymerization. Many scientists have discussed PGS as a new-generation polymer for biomedical applications. Its regenerative potential has been confirmed, in particular, in tissue engineering of soft tissues (including nerve, cartilage and cardiac tissues). Therefore, we must anticipate a growing importance of PGS in contemporary biomedical applications. This brief review aims to familiarize the readers with this relatively new polymeric material for tissue engineering applications.

Glycolipids can be synthetized in deep eutectic solvents (DESs) as they possess low water content allowing a reversed lipase activity and thus enables ester formation. Based on this principle, honey can also serve as a media for glycolipid synthesis. Indeed, this supersaturated sugar solution is comparable in terms of physicochemical properties to the sugar-based DESs. Honey-based products being commercially available for therapeutic applications, it appears interesting to enhance its bioactivity. In the current work, we investigate if enriching medical grade honey with in situ enzymatically-synthetized glycolipids can improve the antimicrobial property of the mixture. The tested mixtures are composed of Manuka honey that is enriched with octanoate, decanoate, laurate, and myristate sugar esters, respectively dubbed GOH, GDH, GLH, and GMH. To characterize the bioactivity of those mixtures, first a qualitative screening using an agar well diffusion assay has been performed with methicillin-resistant

Ionizable cationic lipids are essential for efficient in vivo delivery of RNA by lipid nanoparticles (LNPs). DLin-MC3-DMA (MC3), ALC-0315, and SM-102 are the only ionizable cationic lipids currently clinically approved for RNA therapies. ALC-0315 and SM-102 are structurally similar lipids used in SARS-CoV-2 mRNA vaccines, while MC3 is used in a siRNA therapy to knock down transthyretin in hepatocytes. Hepatocytes and hepatic stellate cells (HSCs) are particularly attractive targets for RNA therapy because they synthesize many plasma proteins, including those that influence blood coagulation. While LNPs preferentially accumulate in the liver, evaluating the ability of different ionizable cationic lipids to deliver RNA cargo into distinct cell populations is important for designing RNA-LNP therapies with minimal hepatotoxicity. Here, we directly compared LNPs containing either ALC-0315 or MC3 to knock-down coagulation factor VII (FVII) in hepatocytes and ADAMTS13 in HSCs. At a dose of 1 mg/kg of siRNA in mice, LNPs with ALC-0315 achieved a two-fold and ten-fold greater knockdown of FVII and ADAMTS13, respectively, compared to LNPs with MC3. At a high dose (5 mg/kg), ALC-0315 LNPs increased markers of liver toxicity (ALT and bile acids) while the same dose of MC3 LNPs did not. These results demonstrate that ALC-0315 LNPs achieves potent siRNA-mediated knockdown of target proteins in hepatocytes and HSCs, in mice, though markers of liver toxicity can be observed after a high dose. This study provides an initial comparison that may inform the development of ionizable cationic LNP therapeutics with maximal efficacy and limited toxicity.

This paper describes a synthesis of ALC-0315 by a sequence that more than doubles the overall yield relative to the published one, and that employs much cleaner reactions, thereby facilitating purifications to a considerable extent.

Rhamnolipids (RLs) are biosurfactants with significant tensioactive and emulsifying properties. They are mainly composed by mono-RL and di-RL components. Although there are numerous studies concerning their molecular properties, information is scarce regarding the mechanisms by which each of the two components interacts with cell membranes. Herein, we performed phase-contrast and fluorescence microscopy experiments on plasma membrane models represented by giant-unilamellar-vesicles (GUVs) composed of 1,2-dioleoyl-sn-glycero-3-phosphocholine (DOPC), 2-[[(E,2S,3R)-1,3-dihydroxy-2-(octadecanoylamino) octadec-4-enyl]peroxy-hydroxyphosphoryl]oxyethyl-trimethylazanium (sphingomyelin, SM) and (3β)-cholest-5-en-3-ol (cholesterol, CHOL) (1:1:1 M ratio), which present liquid-order (L

Polyglycerol sebacate (PGS) scaffolds obtained using a leaching technique were modified with iodine-doped polypyrrole (PPy-I) in a plasma reactor in order to study the effect of exposure time on the cell viability of hDPSCs. SEM analysis showed the formation and growth of PPy-I particles as the exposure time was increased, while FTIR and XPS analysis revealed the presence of -NH- and N+ groups in the chemical composition of the surfaces, relating to the increase in the amount of PPY-I particles. The water contact angle measurements showed an increase in the scaffold’s hydrophilicity with greater exposure times which was also attributed to the rising of PPy-I particles. It was also observed that PPy-I promotes the rigidity of the treated PGS scaffolds. when in direct contact with treated PGS scaffolds, cell viability improved with respect to non-treated scaffolds, however only at shorter time exposures. Extracts of plasma-treated PGS scaffolds showed high cytotoxicity as the time exposure to plasma treatment was increased.

Highly sensitive, flexible sensors that can be manufactured with minimum environmental footprint and be seamlessly integrated into wearable devices are required for real-time tracking of complex human movement, gestures, and health conditions. This study reports on how biodegradation can be used to enhance the sensitivity and electromechanical performance of piezoresistive sensors. Poly(glycerol sebacate) (PGS) elastomeric porous sensor was synthesized and blended with multiwall carbon nanotubes (MWCNTs) and sodium chloride (NaCl). Because of their unique porous characteristics, a single linear behavior over a large range of pressures (≤8 kPa) and an increase in their sensitivity from 0.12 ± 0.03 kPa

Dual meshes are often preferred in the treatment of umbilical and incisional hernias where the abdominal wall defect is large. These meshes are generally composed of either two nonabsorbable layers or a nonabsorbable layer combined with an absorbable one that degrades within the body upon healing of the defect. The most crucial point in the design of a dual mesh is to produce the respective layers based on the structure and requirements of the recipient site. We herein developed a dual mesh that consists of two layers: a nanofibrous layer made of poly (glycerol sebacate)/poly (caprolactone) (PGS/PCL) to support the healing of the abdominal wall defect and a nondegradable, nonadhesive smooth layer made of polycarbonateurethane (PU) with suitable properties to avoid the adhesion of the viscera to the mesh. To prepare the double-sided structure, PGS/PCL was directly electrospun onto the PU film. This processing approach provided a final product with well-integrated layers as observed by a scanning electron microscope. Tensile test performed at the dry state of the samples showed that the dual mesh has the ability to elongate seven times more as compared with the commercially available counterparts, mimicking the native tissue properties. The degradation test carried out at physiological conditions revealed that PGS started to degrade within the first 15 days. in vitro studies with human umbilical vein endothelial cells demonstrated the double function of the meshes, in which PU layer did not allow cell adhesion, whereas PGS/PCL layer has the ability to support cell adhesion and proliferation. Therefore, the material developed in this study has the potential to be an alternative to the existing hernia mesh products.

Poly(glycerol sebacate) (PGS) is a new biodegradable polymer with good biocompatibility used in many fields of biomedicine and drug delivery. Sunitinib‐loaded PGS/gelatine nanoparticles were prepared by the de‐solvation method for retinal delivery and treatment of diabetic retinopathy. The nanoparticles were characterised by Fourier‐transform infrared and differential scanning calorimetry. The effects of different formulation variables including drug‐to‐carrier ratio, gelatine‐to‐PGS ratio, and glycerine‐to‐sebacate ratio were assessed on the encapsulation efficiency (EE%), particle size, release efficiency (RE), and zeta potential of the nanoparticles. The in vitro cytotoxicity of PGS/gelatine nanoparticles was studied on L929 cells. Draize test on rabbit eyes was also done to investigate the possible allergic reactions caused by the polymer. Glycerine/sebacic acid was the most effective parameter on the EE and RE. Gelatine‐to‐PGS ratio had the most considerable effect on the particle size while the RE was more affected by the glycerine/sebacic acid ratio. The optimised formulation (S(1) G(0.7) D(21.2)) exhibited a particle size of 282 nm, 34.6% EE, zeta potential of −8.9 mV, and RE% of about 27.3% for drug over 228 h. The 3‐(4,5‐dimethylthuazol‐2‐yl)‐2,5‐diphenyltetrazolium bromide assay indicated PGS/gelatine nanoparticles were not cytotoxic and sunitinib‐loaded nanoparticles were not toxic at concentrations

Prophage-encoded Escherichia coli O157:H7 transcription factor (TF), PchE, inhibits biofilm formation and attachment to cultured epithelial cells by reducing curli fimbriae expression and increasing flagella expression. To identify pchE regulators that might be used in intervention strategies to reduce environmental persistence or host infections, we performed a computational search of O157:H7 strain PA20 pchE promoter sequences for binding sites used by known TFs. A common site shared by MarA/SoxS/Rob TFs was identified and the typical MarA/Rob inducers, salicylate and decanoate, were tested for biofilm and motility effects. Sodium salicylate, a proven biofilm inhibitor, but not sodium decanoate, strongly reduced O157:H7 biofilms by a pchE-independent mechanism. Both salicylate and decanoate enhanced O157:H7 motility dependent on pchE using media and incubation temperatures optimum for culturing human epithelial cells. However, induction of pchE by salicylate did not activate the SOS response. MarA/SoxS/Rob inducers provide new potential agents for controlling O157:H7 interactions with the host and its persistence in the environment. IMPORTANCE There is a need to develop E. coli serotype O157:H7 nonantibiotic interventions that do not precipitate the release and activation of virulence factor-encoded prophage and transferrable genetic elements. One method is to stimulate existing regulatory pathways that repress bacterial persistence and virulence genes. Here we show that certain inducers of MarA and Rob have that ability, working through both pchE-dependent and pschE-independent pathways.

Preparing a micropatterned elastomer film with characteristics that can simulate the mechanical properties, anisotropy, and electroactivity of natural myocardial tissues is crucial in cardiac tissue engineering after myocardial infarction (MI). Therefore, in this study, we developed several elastomeric films with a surface micropattern based on poly (glycerol sebacate) (PGS) and graphene (Gr). These films have sufficient mechanical strength (0.6 ± 0.1–3.2 ± 0.08 MPa) to withstand heartbeats, and the micropatterned structure also satisfies the natural myocardium anisotropy in the transverse and vertical. Moreover, Gr makes these films conductive (up to 5.80 × 10−7 S m−1), which is necessary for the conduction of electrical signals between cardiomyocytes and the cardiac tissue. Furthermore, they have good cytocompatibility and can promote cell proliferation in H9c2 rat cardiomyocyte cell lines. In vivo test results indicate that these films have good biocompatibility. Notably, a film with 1 wt% Gr content (PGS–Gr1) significantly affects the recovery of myocardial function in rats after MI. This film effectively decreased the infarct size and degree of myocardial fibrosis and reduced collagen deposition. Echocardiographic evaluation showed that after treatment with this film, the left ventricular internal dimension (LVID) in systole and LVID in diastole of rats exhibited a significant downward trend, whereas the fractional shortening and ejection fraction were significantly increased compared with the control group. These data indicate that this electroactive micropatterned anisotropic elastomer film can be applied in cardiac tissue engineering.

The mycobacterial membrane protein large 3 (MmpL3) transporter is essential and required for shuttling the lipid trehalose monomycolate (TMM), a precursor of mycolic acid (MA)-containing trehalose dimycolate (TDM) and mycolyl arabinogalactan peptidoglycan (mAGP), in Mycobacterium species, including Mycobacterium tuberculosis and Mycobacterium smegmatis. However, the mechanism that MmpL3 uses to facilitate the transport of fatty acids and lipidic elements to the mycobacterial cell wall remains elusive. Here, we report 7 structures of the M. smegmatis MmpL3 transporter in its unbound state and in complex with trehalose 6-decanoate (T6D) or TMM using single-particle cryo-electron microscopy (cryo-EM) and X-ray crystallography. Combined with calculated results from molecular dynamics (MD) and target MD simulations, we reveal a lipid transport mechanism that involves a coupled movement of the periplasmic domain and transmembrane helices of the MmpL3 transporter that facilitates the shuttling of lipids to the mycobacterial cell wall., Mycobacterial membrane protein Large 3 (MmpL3) is a transporter required for shuttling trehalose monomycolate. Structures of M. smegmatis MmpL3 with and without substrate reveal the mechanism by which MmpL3 transports this essential precursor of lipids for the mycobacterial cell wall.

The interactions of mono-rhamnolipids (mono-RLs) with model membranes were investigated through a biomimetic approach using phospholipid-based liposomes immobilized on a gold substrate and also by the multiparametric surface plasmon resonance (MP-SPR) technique. Biotinylated liposomes were bound onto an SPR gold chip surface coated with a streptavidin layer. The resulting MP-SPR signal proved the efficient binding of the liposomes. The thickness of the liposome layer calculated by modeling the MP-SPR signal was about 80 nm, which matched the average diameter of the liposomes. The mono-RL binding to the film of the phospholipid liposomes was monitored by SPR and the morphological changes of the liposome layer were assessed by modeling the SPR signal. We demonstrated the capacity of the MP-SPR technique to characterize the different steps of the liposome architecture evolution, i.e., from a monolayer of phospholipid liposomes to a single phospholipid bilayer induced by the interaction with mono-RLs. Further washing treatment with Triton X-100 detergent left a monolayer of phospholipid on the surface. As a possible practical application, our method based on a biomimetic membrane coupled to an SPR measurement proved to be a robust and sensitive analytical tool for the detection of mono-RLs with a limit of detection of 2 μg mL-1.

Plasma surface modification is one of the new methods for improving the surface properties of the scaffold and accelerating tissue regeneration. The aim of this study was to create poly glycerol sebacate/poly lactic acid (PGS/PLA) composite scaffold by electrospun method and modified the scaffold by oxygen plasma for use as a vascular graft. Plasma surface modified PGS/PLA scaffold morphology study showed relatively uniform fibers with an average diameter of 637 ± 149.4 nm and porosity of 82%. The mechanical evaluation of the PGS/PLA scaffold showed properties close to the natural vessels. Atomic force microscopy images exhibited an increase in the roughness of the scaffold after plasma surface modification; however, hemocompatibility studies revealed that it had no adverse effect on blood compatibility. Wettability studies revealed the superhydrophilic property of the modified scaffold (contact angle near to zero). Besides, the human umbilical vein endothelial cells proliferation and adhesion were improved significantly. Obtaining mechanical properties near to the natural vessels due to the suitable composition and significant improvement in blood compatibility and cell growth make the modified PGS/PLA composite a suitable candidate for vascular tissue regeneration.

In the cardiovascular system, heart valves and vessels are subjected to continuous cyclic mechanical loadings due to the pulsatile nature of blood flow. Hence, in leveraging tissue engineering (TE) strategies to regenerate such a system, the candidate scaffold should not only be biocompatible with the desired biodegradation rate, but it should also be mechanically competent to provide a supportive structure for facilitating stem cells retention, growth, and differentiation. To this end, herein, we introduced a novel scaffold composed of poly(glycerol-sebacate) (PGS) and polyurethane (PU), which comprises of two layers: an electrospun pure PU layer beneath another electrospun PGS/PU layer with a different ratio of PGS to PU (3:2, 1:1, 2:3 Wt:Wt). The electrospun PGS/PU-PU scaffold was mechanically competent and showed intended hydrophilicity and a good biodegradation rate. Moreover, the PGS/PU-PU scaffold indicated cell viability and proliferation within ten days of in vitro cell culture and upon 7 day vascular endothelial growth factor (VEGF) stimulation, supported endothelial differentiation of mesenchymal stem cells by significant overexpression of platelet-endothelial cell adhesion molecule-1, von Willebrand factor, and VEGF receptor 2. The results of this study could be implemented in cardiovascular TE strategies when regeneration of blood vessel or heart valve is desired.

Pseudomonas aeruginosa is an opportunistic pathogen that uses malonate among its many carbon sources. We recently reported that, when grown in blood from trauma patients, P. aeruginosa expression of malonate utilization genes was upregulated. In this study, we explored the role of malonate utilization and its contribution to P. aeruginosa virulence. We grew P. aeruginosa strain PA14 in M9 minimal medium containing malonate (MM9) or glycerol (GM9) as a sole carbon source and assessed the effect of the growth on quorum sensing, virulence factors, and antibiotic resistance. Growth of PA14 in MM9, compared to GM9, reduced the production of elastases, rhamnolipids, and pyoverdine; enhanced the production of pyocyanin and catalase; and increased its sensitivity to norfloxacin. Growth in MM9 decreased extracellular levels of N-acylhomoserine lactone autoinducers, an effect likely associated with increased pH of the culture medium; but had little effect on extracellular levels of PQS. At 18 hr of growth in MM9, PA14 formed biofilm-like structures or aggregates that were associated with biomineralization, which was related to increased pH of the culture medium. These results suggest that malonate significantly impacts P. aeruginosa pathogenesis by influencing the quorum sensing systems, the production of virulence factors, biofilm formation, and antibiotic resistance.

The goal of this study was to develop a scaffold for the generation of an artificial intestine that specifically mimics the architecture and biomechanical properties of the native small intestine, and to evaluate the scaffold in vitro and in vivo. Scaffolds mimicking the microarchitecture of native intestine were fabricated from poly(glycerol sebacate) (PGS) with a thickness of 647 μm (±241 μm) and villus height of 340 μm (±29.5 μm). The scaffolds showed excellent biological properties, as 71.4% (±7.2%) and 58.7% (±12.7%) mass remained after 5 weeks of in vitro exposure to control and digestive media, respectively. Tensile properties of the scaffolds approached those of native porcine intestine and scaffolds maintained their mechanical properties over 6 weeks based on rheometer measurements. Scaffolds accommodated intestinal epithelial stem cells and demonstrated maintenance of size and microarchitecture after 12 weeks of omental implantation in mice. There was an expected amount of inflammation, but less tissue infiltration and tissue formation than anticipated. In conclusion, we developed novel scaffolds using PGS that mimic the microarchitecture and mechanical properties of native intestine with promise for use in artificial intestine for individuals with short bowel syndrome. IMPACT STATEMENT: This study is significant because it demonstrates an attempt to design a scaffold specifically for small intestine using a novel fabrication method, resulting in an architecture that resembles intestinal villi. In addition, we use the versatile polymer poly(glycerol sebacate) (PGS) for artificial intestine, which has tunable mechanical and degradation properties that can be harnessed for further fine-tuning of scaffold design. Moreover, the utilization of PGS allows for future development of growth factor and drug delivery from the scaffolds to promote artificial intestine formation.

Tracheal reconstruction remains a major surgical challenge, mainly owing to the scarce of resilient hollow grafts with identifiable vascular pedicle in humans. In this study, we developed a three-layer, elastomeric, trachea-like composite made of poly glycerol sebacate (PGS) and polycaprolactone (PCL), which presented appropriate resilient property, timely degradation and interconnected pores. C shape PCL rings fabricated with selective laser sintering (SLS) techniques are regularly positioned around porous PGS tubes and fixed by PCL electrospinning sheath. Such an elastomeric composite underwent host remodeling including rapid vascularization and tissue infiltration after fascia wrapping. With degrading of PGS, C rings well incorporated into growing fascia and lead to the formation of pedicled tracheal grafts, which attributes to the strong and resilient properties of generated hollow grafts thus enabled orthotopic transplantation in segmental tracheal defect. Progressive remodeling on such vascularized and mechanically stable grafts resulted in epithelium regeneration on luminal side as well as production of adequate amount of collagen and elastin, which warrantees the air passage during breathing. Future study employing large animal models more representative of human tracheal regeneration is warranted before clinical translation. Using fast degrading PGS combined with PCL rings represents a philosophical shift from the prevailing focus on tough grafts in airway reconstruction and may impact regenerative medicine in general.

The use of fibrous scaffolds for tissue repair or regeneration is advantageous for its microstructure similar to that of the native ECM. Aligned fibrous scaffold, in particular, can be used to manipulate cell alignment and hence the microstructure of the resultant tissue. In our previous study, nanofibers consisting of solely poly(glycerol sebacate) (PGS) have been successfully fabricated using core-shell coaxial electrospinning followed by curing and subsequent shell removal. When we tried to fabricate aligned PGS fibrous membranes by collecting the electrospun fibers on a rapidly rotating drum, however, loss of fibrous structure was observed upon curing. This might be due to the broken fibers that were collected under tension; the core PGS prepolymer that melts at high temperature could leak from the broken ends during curing. In this study, attempts were made to reduce the possibility of the fiber breakage. At each stage of preparation, fiber morphology was examined by SEM and fiber compositions were verified by Fourier transform infrared spectroscopy and differential scanning calorimetry. Mechanical properties of the aligned PGS fibrous membrane were evaluated by uniaxial tensile testing both in parallel and perpendicular to the principal fiber direction. SEM images showed that fibrous morphology was better preserved upon the adjustment of the shell composition and the rotational speed of the collector drum. The final PGS fibers remained to be aligned although the alignment was less strong than that of as-spun core-shell fibers. The aligned PGS fibrous membrane exhibited anisotropic mechanical properties with Young's modulus in parallel and perpendicular to the principal fiber direction being 0.98 ± 0.04 MPa and 0.52 ± 0.02 MPa, respectively. The aligned PGS fibrous membrane was capable of guiding the orientation of cultured cells and therefore has the potential to be used to fabricate structurally anisotropic tissue-engineered constructs.

This study explored the applicability, cellular efficacy, and osteogenic activities of porous nano-hydroxyapatite/Poly (glycerol sebacate)-grafted maleic anhydride (n-HA/PGS-g-M) composite scaffolds. Nuclear magnetic resonance (NMR) analyses indicated that approximately 43% of the hydroxide radicals in PGS were displaced by maleic anhydride. Resonance bands at 1036 cm−1 occurred in scaffolds containing nHA powders, and peak areas increased when n-HA weight increased in PGS-M-n-HA-0.4, PGS-M-n-HA-0.5, and PGS-M-n-HA-0.6 scaffolds. The n-HA/PGS-g-M composite scaffolds exhibited porous microstructure with average pore size of 150–300 µm in scanning electron microscopy (SEM) analysis. Differential scanning calorimetry (DSC) identified the glass transition temperature (Tg) as −25–30 °C, indicative of quality resilience. The modulus of compressibility increased when n-HA content increased. Interestingly, viability of human adipose-derived stem cells (hADSCs) in vitro and expression of the osteogenic related genes RUNX2, OCN, and COL1A1 was enhanced in the n-HA/PGS-g-M composite scaffolds compared to those factors observed in PGS-g-M scaffolds. Finally, simulated body fluid (SBF) tests indicated more apatite deposits on the surface of n-HA/PGS-g-M scaffolds compared to PGS-g-M scaffolds. Overall, porous n-HA/PGS-g-M composite scaffolds possessed acceptable biocompatibility and mechanical properties, and they stimulated hADSC cell proliferation and differentiation. Given these qualities, the composite scaffolds have potential applications in bone tissue engineering.

Autologous veins are the most widely used grafts for bypassing small arteries in coronary and peripheral arterial occlusive diseases. However, they have limited availability and cause donor-site morbidity. Here, we report a direct comparison of acellular biodegradable synthetic grafts and autologous veins as interposition grafts of rat carotid arteries, which is a good model for clinically relevant small arteries. Notably, extensive but transient infiltration of circulating monocytes at day 14 in synthetic grafts leads to a quickly-resolved inflammation and arterial-like tissue remodeling. The vein graft exhibits a similar inflammation phase except the prolonged presence of inflammatory monocytes. The walls of the remodeled synthetic graft contain many circumferentially aligned contractile non-proliferative smooth muscle cells (SMCs), collagen and elastin. In contrast, the walls of the vein grafts contain disorganized proliferating SMCs and thicken over time, suggesting the onset of stenosis. At 3 months, both grafts have a similar patency, extracellular matrix composition, and mechanical properties. Furthermore, synthetic grafts exhibit recruitment and re-orientation of newly synthesized collagen fibers upon mechanical loading. To our knowledge, this is the first demonstration of a biodegradable synthetic vascular graft with a performance similar to an autologous vein in small artery grafting.

© 2021 John Wiley & Sons Ltd., The effort to develop an effective and safe temporomandibular joint (TMJ) disc substitute has been one of the mainstreams of tissue engineering. Biodegradable customized scaffolds could approach safety and effectiveness to regenerate a new autologous disc, rather than using non-biodegradable materials. However, it is still technically challenging to mimic the biomechanical properties of the native disc with biodegradable polymers. In this study, new 3D tailored TMJ disc implants were developed: (1) Poly(glycerol sebacate) (PGS) scaffold reinforced with electrospun Poly(εcaprolactone) (PCL) fibers on the outer surface (PGS+PCL); (2) PCL and polyethylene glycol diacrylate (PEGDA) (PCL+PEGDA); and (3) PCL. The TMJ implants were tested in a randomized preclinical trial, conducted in 24 black Merino sheep TMJ, perfoming bilateral interventions. Histologic, imaging, and kinematics analysis was performed. No statistical changes were observed between the PGS+PCL disc and the control group. The PCL+PEGDA and PCL groups were associated with statistical changes in histology (p = 0.004 for articular cartilage mid-layer; p = 0.019 for structure changes and p = 0.017 for cell shape changes), imaging (p = 0.027 for global appreciation) and dangerous material fragmentation was observed. No biomaterial particles were observed in the multi-organ analysis in the different groups. The sheep confirmed to be a relevant animal model for TMJ disc surgery and regenerative approaches. The PCL and PCL+PEGDA discs presented a higher risk to increase degenerative changes, due to material fragmentation. None of the tested discs regenerate a new autologous disc, however, PGS+PCL was safe, demonstrated rapid resorption, and was capable to prevent condyle degenerative changes., This work was supported by European Regional Development Fund—Centro2020 through the Project reference: BIODISCUS (CENTRO‐01‐0247‐FEDER‐039969).