3D printed carboxymethyl cellulose scaffolds for autologous growth factors delivery in wound healing.

This work aims to use carboxymethyl cellulose (CMC) as main structural and functional component of 3D printed scaffolds for healing of diabetic wounds. Differently from previous inks involving small contents in CMC, herein sterile (steam-heated) concentrated CMC solely dispersions (10-20%w/v) were screened regarding printability and fidelity properties. CMC (15%w/v)-citric acid inks showed excellent self-healing rheological properties and stability during storage. CMC scaffolds loaded with platelet rich plasma (PRP) sustained the release of relevant growth factors. CMC scaffolds both with and without PRP promoted angiogenesis in ovo, stem cell migration in vitro, and wound healing in a diabetic model in vivo. Transparent CMC scaffolds allowed direct monitoring of bilateral full-thickness wounds created in rat dorsum. CMC scaffolds facilitated re-epithelialization, granulation, and angiogenesis in full-thickness skin defects, and the performance was improved when combined with PRP. Overall, CMC is pointed out as outstanding component of active dressings for diabetic wounds.

Local delivery of hydrogel encapsulated vascular endothelial growth factor for the prevention of medication-related osteonecrosis of the jaw.

The anti-angiogenic effects of bisphosphonates have been hypothesized as one of the major etiologic factors in the development of medication-related osteonecrosis of the jaw (MRONJ), a severe debilitating condition with limited treatment options. This study evaluated the potential of a gelatine-hyaluronic acid hydrogel loaded with the angiogenic growth factor, vascular endothelial growth factor (VEGF), as a local delivery system to aid in maintaining vascularization in a bisphosphonate-treated (Zoledronic Acid) rodent maxillary extraction defect. Healing was assessed four weeks after implantation of the VEGF-hydrogel into extraction sockets. Gross examination and histological assessment showed that total osteonecrosis and inflammatory infiltrate was significantly reduced in the presence of VEGF. Also, total vascularity and specifically neovascularization, was significantly improved in animals that received VEGF hydrogel. Gene expression of vascular, inflammatory and bone specific markers within the defect area were also significantly altered in the presence of VEGF. Furthermore, plasma cytokine levels were assessed to determine the systemic effect of locally delivered VEGF and showed similar outcomes. In conclusion, the use of locally delivered VEGF within healing extraction sockets assists bone healing and prevents MRONJ via a pro-angiogenic and immunomodulatory mechanism.

A Structural Overview of Vascular Endothelial Growth Factors Pharmacological Ligands: From Macromolecules to Designed Peptidomimetics.

The vascular endothelial growth factor (VEGF) family of cytokines plays a key role in vasculogenesis, angiogenesis, and lymphangiogenesis. VEGF-A is the main member of this family, alongside placental growth factor (PlGF), VEGF-B/C/D in mammals, and VEGF-E/F in other organisms. To study the activities of these growth factors under physiological and pathological conditions, resulting in therapeutic applications in cancer and age-related macular degeneration, blocking ligands have been developed. These have mostly been large biomolecules like antibodies. Ligands with high affinities, at least in the nanomolar range, and accurate structural data from X-ray crystallography and NMR spectroscopy have been described. They constitute the main focus of this overview, which evidences similarities and differences in their binding modes. For VEGF-A ligands, and to a limited extent also for PlGF, a transition is now observed towards developing smaller ligands like nanobodies and peptides. These include unnatural amino acids and chemical modifications for designed and improved properties, such as serum stability and greater affinity. However, this review also highlights the scarcity of such small molecular entities and the striking lack of small organic molecule ligands. It also shows the gap between the rather large array of ligands targeting VEGF-A and the general absence of ligands binding other VEGF members, besides some antibodies. Future developments in these directions are expected in the upcoming years, and the study of these growth factors and their promising therapeutic applications will be welcomed.

Probing the helical stability in a VEGF-mimetic peptide.

The analysis of the forces governing helix formation and stability in peptides and proteins has attracted considerable interest in order to shed light on folding mechanism. We analyzed the role of hydrophobic interaction, steric hindrance and chain length on i, i + 3 position in QK peptide, a VEGF mimetic helical peptide. We focused on position 10 of QK, occupied by a leucine, as previous studies highlighted the key role of the Leu7-Leu10 interaction in modulating the helix formation and inducing an unusual thermodynamic stability. Leu10 has been replaced by hydrophobic amino acids with different side-chain length, hydrophobicity and steric hindrance. Ten peptides were, hence, synthesized and analyzed combining circular dichroism, calorimetry and NMR spectroscopy. We found that helical content and thermal stability of peptide QK changed when Leu10 was replaced. Interestingly, we observed that the changes in the helical content and thermal stability were not always correlated and they depend on the type of interaction (strength and geometry) that could be established between Leu7 and the residue in position 10.

Snake venom vascular endothelial growth factors (svVEGFs): Unravelling their molecular structure, functions, and research potential.

Vascular endothelial growth factor (VEGF) is a key regulator of angiogenesis, a physiological process characterized by the formation of new vessels from a preexisting endothelium. VEGF has also been implicated in pathologic states, such as neoplasias, intraocular neovascular disorders, among other conditions. VEGFs are distributed in seven different families: VEGF-A, B, C, D, and PIGF (placental growth factor), which are identified in mammals; VEGF-E, which are encountered in viruses; and VEGF-F or svVEGF (snake venom VEGF) described in snake venoms. This is the pioneer review of svVEGF family, exploring its distribution among the snake venoms, molecular structure, main functions, and potential applications.

Microfluidic-assisted fabrication of reverse micelle/PLGA hybrid microspheres for sustained vascular endothelial growth factor delivery.

In this study, poly (d, l-lactide-co-glycolide) (PLGA) composite microspheres containing anhydrous reverse micelle (R.M.) dipalmitoylphosphatidylcholine (DPPC) nanoparticles loaded vascular endothelial growth factor (VEGF) were produced using microfluidic platforms. The VEGF-loaded R.M. nanoparticles (VRM) were achieved by initial self-assembly and subsequent lipid inversion of the DPPC vesicles. The fabricated VRMs were encapsulated into the PLGA matrix by flow-focusing geometry microfluidic platforms. The encapsulation efficiency, in vitro release profile, and the bioactivity of the produced composite microspheres were investigated. The release study showed that VEGF was slowly released from the PLGA composite microspheres over 28 days with a reduced initial burst (18 â€¯± â€¯4.17% in the first 24 H). The VEGF stability during encapsulation and release period was also investigated, and the results indicated that encapsulated VEGF was well preserved. Also, the bioactivity assay of the PLGA composite microspheres on human umbilical vein endothelial cells was confirmed that the encapsulated VEGF was utterly active. The present monodisperse and controllable VEGF-loaded microspheres with reproducible manner could be widely used in tissue engineering and therapeutic applications.

Hydrogen/deuterium exchange mass spectrometry with improved electrochemical reduction enables comprehensive epitope mapping of a therapeutic antibody to the cysteine-knot containing vascular endothelial growth factor.

Hydrogen/deuterium exchange mass spectrometry (HDX-MS) has become a popular method for analysis of the conformational dynamics and interactions of proteins. Disulfide-bonded proteins, however, present a challenge to HDX-MS as they require efficient disulfide bond reduction prior to enzymatic proteolysis. Electrochemical reduction (ER) provides an attractive solution to tackle disulfide-bonded proteins that are resistant to conventional chemical reduction during HDX-MS. However, ER-enabled HDX-MS has been limited by technical challenges including partial unwanted protein oxidation side-reactions, incompatibility with certain buffer components and most importantly, a lack of overall method robustness. In this study, we have sought to address these challenges. We perform a systematic screening of the compatibility of ER to buffers commonly used in HDX-MS samples by using a reliable and simple system suitability test (SST). Furthermore, we demonstrate the benefits of a new design of the electrochemical cell (EC) for ER-enabled HDX-MS, which include a) high repeatability and robustness over large sample batches without the need for electrode polishing and b) high reduction efficiency of disulfide-bonded proteins without unwanted oxidation side-reactions. We show the real-world applicability of the optimized ER-enabled HDX-MS workflow by performing an epitope mapping of a Fab fragment of a therapeutic monoclonal antibody (mAb) to the cysteine knot-containing vascular endothelial growth factor (VEGF). The results allow us to comprehensively map sites in VEGF involved in mAb binding. Overall, our findings show how ER and HDX-MS can be combined to enable analysis of the conformation and interactions of challenging disulfide-rich proteins.

Vascular endothelial growth factor-loaded poly-lactic-co-glycolic acid nanoparticles with controlled release protect the dopaminergic neurons in Parkinson's rats.

Vascular endothelial growth factor (VEGF) had neuroprotective effects on dopaminergic (DA) neurons. In order to overcome the gastrointestinal digestion and bioaccessibility, VEGF was encapsulated with poly-lactic-co-glycolic acid nanospheres (NS) in order to prevent the VEGF degradation until its release. The caudal administration of VEGF and NS encapsulated VEGF at different doses (1.0, 10.0, and 100.0 ng/ml) on the rats with Parkinson's disease lesion was evaluated. Intravenous injected VEGF at the dose of 1 ng/ml displayed the strongest neuroprotective effect than other groups as well as the stereotaxic group. The NS encapsulated with VEGF can pass through blood-brain barrier and protect the DA neurons. There was no significant difference between intravenous injection method and stereotaxic method, while the first method is simpler and convenient. Injection of NS encapsulated with VEGF may become a valuable neurorescuing therapeutic approach for Parkinson's disease.

Biomimetic proteoglycan nanoparticles for growth factor immobilization and delivery.

The delivery of growth factors is often challenging due to their short half-life, low stability, and rapid deactivation. In native tissues, the sulfated residual of glycosaminoglycan (GAG) polymer chains of proteoglycans immobilizes growth factors through the proteoglycans'/proteins' complexation with nanoscale organization. These biological assemblies can influence growth factor-cell surface receptor interactions, cell differentiation, cell-cell signaling, and mechanical properties of the tissues. Here, we introduce a facile procedure to prepare novel biomimetic proteoglycan nanocarriers, based on naturally derived polymers, for the immobilization and controlled release of growth factors. We developed polyelectrolyte complex nanoparticles (PCNs) as growth factor nanocarriers, which mimic the dimensions, chemical composition, and growth factor immobilization of proteoglycans in native tissues. PCNs were prepared by a polymer-polymer pair reaction method and characterized for physicochemical properties. Fourier transform infrared spectroscopy (FTIR) analysis indicated that complexation occurred through electrostatic interactions. Transmission electron microscopy (TEM) results showed that the nanocarriers had a diameter of 60 ± 11 nm and 91 ± 33 nm for dermatan sulfate sodium salt-poly-l-lysine (DS-PLL) and gum tragacanth-poly-l-lysine (GT-PLL) complexes, respectively. The colloidal nanoparticles were stable due to their negative zeta potential, i.e.-25 ± 4 mV for DS-PLL and -18 ± 3.5 mV for GT-PLL. Cytocompatibility of PCNs in contact with human bone marrow stromal cells (HS-5) was confirmed through a live/dead assay and metabolic activity measurement. In addition, vascular endothelial growth factor (VEGF) was used to evaluate the ability of PCNs to stabilize growth factors. The capability of PCNs to preserve VEGF activity for up to 21 days was confirmed by analyzing the metabolic and mitogenic characteristics of human umbilical vein endothelial cells (HUVECs). Our results demonstrated the potential applications of these nanoparticles in therapeutic delivery for tissue regeneration applications.

Bioengineering a pre-vascularized pouch for subsequent islet transplantation using VEGF-loaded polylactide capsules.

The effectiveness of cell transplantation can be improved by optimization of the transplantation site. For some types of cells that form highly oxygen-demanding tissue, e.g., pancreatic islets, a successful engraftment depends on immediate and sufficient blood supply. This critical point can be avoided when cells are transplanted into a bioengineered pre-vascularized cavity which can be formed using a polymer scaffold. In our study, we tested surface-modified poly(lactide-co-caprolactone) (PLCL) capsular scaffolds containing the pro-angiogenic factor VEGF. After each modification step (i.e., amination and heparinization), the surface properties and morphology of scaffolds were characterized by ATR-FTIR and XPS spectroscopy, and by SEM and AFM. All modifications preserved the gross capsule morphology and maintained the open pore structure. Optimized aminolysis conditions decreased the Mw of PLCL only up to 10% while generating a sufficient number of NH2 groups required for the covalent immobilization of heparin. The heparin layer served as a VEGF reservoir with an in vitro VEGF release for at least four weeks. In vivo studies revealed that to obtain highly vascularized PLCL capsules (a) the optimal VEGF dose for the capsule was 50 µg and (b) the implantation time was four weeks when implanted into the greater omentum of Lewis rats; dense fibrous tissue accompanied by vessels completely infiltrated the scaffold and created sparse granulation tissue within the internal cavity of the capsule. The prepared pre-vascularized pouch enabled the islet graft survival and functioning for at least 50 days after islet transplantation. The proposed construct can be used to create a reliable pre-vascularized pouch for cell transplantation.

Multifunctional fibrous scaffolds for bone regeneration with enhanced vascularization.

Due to the structural similarity to the extracellular matrix of human tissue and the ultra-high surface area-to-volume ratio, three dimensional electrospun fibrous structures have been increasingly used as tissue engineering scaffolds. Given that successful bone regeneration requires both good osteogenesis and vascularization, producing scaffolds that have both osteogenic and angiogenic potential is highly desirable. In this investigation, tricomponent fibrous scaffolds simultaneously incorporated with recombinant human vein endothelial growth factor (rhVEGF), recombinant human bone morphogenetic protein-2 (rhBMP-2) and bioactive calcium phosphate (Ca-P) nanoparticles are produced through a novel multi-source multi-power electrospinning method, and sequential growth factor release with a quick rhVEGF release and a steady rhBMP-2 release is achieved. The enhanced human umbilical vein endothelial cell (HUVEC) migration and tube formation, and up-regulated human bone marrow derived mesenchymal stem cell (hBMSC) osteogenic differentiation and mineralization demonstrate that tricomponent scaffolds have balanced angiogenic-osteogenic properties in vitro. 8 weeks after the scaffold implantation into the cranial defects of mice, obvious new bone regeneration and newly formed capillaries are observed in tricomponent scaffolds, suggesting that the tricomponent scaffolds enhance osteogenesis in vivo with required vascularization, which shows the great potential of the tricomponent scaffolds in bone tissue regeneration.

Dual-functional gold nanoparticles with antimicrobial and proangiogenic activities improve the healing of multidrug-resistant bacteria-infected wounds in diabetic mice.

Gold nanoparticles (Au NPs) are conjugated with the vascular endothelial growth factor-A165 (VEGF-A165) and (11-mercaptoundecyl)-N,N,N-trimethylammonium (11-MTA) cation to form dual-functional gold nanoparticles (11-MTA/VEGF-Au NPs) that possess antimicrobial and proangiogenic activities for wound healing in diabetic (db/db) mice. VEGF-A165 is a popular proangiogenic growth factor that stimulates multiple components in the wound-healing cascade. On the other hand, 11-MTA possesses antibacterial activity and can be bound to Au NPs easily through Au-S bonding. We have found that the surface density of VEGF-A165 plays a vital role in promoting the proliferation, migration, and tube formation of human umbilical vein endothelial cells. 11-MTA tethered on the VEGF-modified Au NPs enables the nanocomposites (i.e., 11-MTA/VEGF-Au NPs) to exhibit a strong antimicrobial activity against multidrug-resistant bacteria [methicillin-resistant S. aureus (MRSA)]. The minimal inhibition concentration of 11-MTA/VEGF-Au NPs is ∼450-fold lower than that of 11-MTA, revealing their high antibacterial efficiency. 11-MTA/VEGF-Au NPs exhibit high biocompatibility. 11-MTA/VEGF-Au NPs as dressing materials to treat MRSA-infected wounds in diabetic mice not only show strong in vivo bactericidal activities but also enhance the healing process of the formation of collagen fibers and epithelialization. Our results show that dual-functional 11-MTA/VEGF-Au NPs are promising agents for clinical applications like treating chronic wound infections.

Identification and characterization of two novel vascular endothelial growth factor genes in Litopenaeus vannamei.

Vascular endothelial growth factor (VEGF) signaling pathway induces endothelial cell proliferation, promotes cell migration, and inhibits apoptosis. Although three VEGF and two VEGF receptor genes have been identified in Litopenaeus vannamei and demonstrated their roles in WSSV infection, another two novel VEGF genes (LvVEGF4, LvVEGF5) were isolated and their involvements in the WSSV infection of shrimp were studied in the present study. The deduced amino acid sequences of both LvVEGF4 and LvVEGF5 contained a signal peptide, a typical PDGF/VEGF domain and a cysteine knot motif (CXCXCX). Tissue distribution analysis showed that LvVEGF4 was predominantly expressed in gill and hemocytes, while LvVEGF5 was mainly detected in hemocytes and intestine. WSSV infection could cause up-regulation of the transcriptional levels of LvVEGF4 and LvVEGF5. Their functions were studied by double-strand RNA interference. The results showed that knock-down of LvVEGF4 and LvVEGF5 led to a decrease of the viral copy number in WSSV infected shrimp. Yeast two-hybrid analysis showed that both LvVEGF4 and LvVEGF5 could interact with LvVEGFR1 rather than LvVEGFR2. In addition, knock-down of LvVEGF4 and LvVEGF5 could reduce the expressional levels of downstream genes FAK and PI3K. The present study provides new clues in demonstrating that the VEGF signaling pathway is involved in the process of WSSV infection in shrimp.

Vascular endothelial growth factor immobilized on mussel-inspired three-dimensional bilayered scaffold for artificial vascular graft application: In vitro and in vivo evaluations.

Currently, there is a great clinical demand for biocompatible and robust tissue-engineered tubular scaffolds for use as artificial vascular graft materials. Despite considerable research on vascular scaffolds, there has still been only limited development of scaffold materials possessing both sufficient mechanical strengths and biological effects for vascular application. In this work, we designed a mechanically robust, bilayered scaffold and manufactured it by combining electrospinning (ELSP) and three-dimensional (3D) printing techniques. This material was coated with polydopamine (PDA) and vascular endothelial growth factor (VEGF) was grafted directly on the scaffold surface to induce potent angiogenic activity. We confirmed that the coated-PDA layer was evenly deposited on the bare polycaprolactone (PCL) scaffold and could enable abundant VEGF immobilization with enhanced hydrophilicity. The VEGF immobilized porous tubular scaffold was well prepared without mechanical weakness induced by surface modification steps. During in vitro and in vivo testing, VEGF immobilized scaffolds elicited markedly enhanced vascular cell proliferation and angiogenic differentiation, as compared to non-treated groups. These results demonstrate that the developed scaffolds may represent an innovative paradigm in vascular tissue engineering by inducing angiogenesis as a means of remodeling and healing vascular defects for use in restorative procedures.

One-pot construction of a twice-condensed pDNA polyplex system for peripheral nerve crush injury therapy.

Non-viral vector gene delivery is generally limited by its potential toxicity problems, poor transfection abilities, serum stability, or relatively complex construction processes of modified polyplexes. Thus, we develop an efficient and stable polyplex system through convenient construction methods. Here, polyethyleneimine (PEI) 1.8 kDa and glutaraldehyde (GA) are used to construct a novel twice-condensed pDNA polyplex system using a one-pot construction method, including pH-responsive C[double bond, length as m-dash]N linkages by which different PEI molecules on one single polyplex can link with each other. In this system, smaller particle sizes, higher zeta potentials and better serum stabilities are achieved without PEGylation or other chemical modifications using lyophobic segments, but via pH-responsive linkages that ensure the escape of nucleic acids. This polyplex system is used to deliver the pDNA of vascular endothelial growth factor (VEGF) whose half-life period in vivo is only around 30 minutes. Compared with polyplexes prepared using PEI 25 kDa, cells and rats treated with twice-condensed VEGF pDNA polyplexes express significantly more VEGF or myelin basic protein (MBP), and this new polyplex system showed fewer adverse effects in vitro and in vivo. In addition, revascularization and neurogenesis are also discovered in the rat sciatic nerve crush injury model.

VEGF (Vascular Endothelial Growth Factor) and Fibrotic Lung Disease.

Interstitial lung disease (ILD) encompasses a group of heterogeneous diseases characterised by varying degrees of aberrant inflammation and fibrosis of the lung parenchyma. This may occur in isolation, such as in idiopathic pulmonary fibrosis (IPF) or as part of a wider disease process affecting multiple organs, such as in systemic sclerosis. Anti-Vascular Endothelial Growth Factor (anti-VEGF) therapy is one component of an existing broad-spectrum therapeutic option in IPF (nintedanib) and may become part of the emerging therapeutic strategy for other ILDs in the future. This article describes our current understanding of VEGF biology in normal lung homeostasis and how changes in its bioavailability may contribute the pathogenesis of ILD. The complexity of VEGF biology is particularly highlighted with an emphasis on the potential non-vascular, non-angiogenic roles for VEGF in the lung, in both health and disease.

Molecularly Imprinted Nanocavities Capable of Ligand-Binding Domain and Size/Shape Recognition for Selective Discrimination of Vascular Endothelial Growth Factor Isoforms.

Vascular endothelial growth factor 165 (VEGF165) is known to be predominantly expressed in the first stage of vascularization; therefore, the detection of VEGF165 is important in the stage diagnosis of cancers. Molecularly imprinted nanocavities, capable of the selective discrimination of VEGF165 from other VEGF isoforms, were prepared by surface-initiated atom transfer radical polymerization. VEGF165 was immobilized on a gold-coated glass substrate by anchored heparin moieties, where the immobilized heparin was able to capture VEGF165 by binding with the heparin-binding domain (HBD) on VEGF165. Molecular imprinting was conducted on the immobilized VEGF165 by using methacrylic acid (MAA) as a functional monomer to interact with basic amino acids outside of the HBD of VEGF165 by electrostatic interaction. After the removal of VEGF165 from the obtained polymer thin layer (ca. 7 nm), VEGF165-imprinted nanocavities remained, in which the heparin moiety and MAA residues were located in suitable positions for VEGF165 recognition. The molecularly imprinted polymer (MIP) thin layer showed a binding affinity for VEGF165 (dissociation constant: 3.4 nM) that was ten times higher than that of the substrate before polymerization (heparin-immobilized substrate). A much lower binding affinity for VEGF121, which contains no heparin-binding domain, was observed. Moreover, the MIP thin layer distinguished VEGF165 from VEGF189, which possesses a larger molecular size than VEGF165, an amino acid sequence homology of 87%, and contains HBDs, whereas the heparin-immobilized substrate showed almost no selectivity. These results suggested that the heparin moiety within the nanocavity provided HBD selectivity and the polymer matrix composed of the molecularly imprinted nanocavity provided size/shape selectivity, which resulted in the highly selective discrimination of VEGF isoforms.

Characterization of two types of vascular endothelial growth factor from Litopenaeus vannamei and their involvements during WSSV infection.

Vascular endothelial growth factors (VEGFs) are important signaling proteins in VEGF signaling pathway which play key roles in inducing endothelial cell proliferation, migration, angiogenesis, vascular permeability, inhibition of apoptosis and virus infection. In the present study, we isolated and characterized two VEGF genes, LvVEGF1 and LvVEGF2 from Litopenaeus vannamei. The deduced amino acid sequences of both LvVEGF1 and LvVEGF2 contained a signal peptide, a typical PDGF/VEGF domain and a cysteine knot motif (CXCXC). Tissue distribution analysis showed that LvVEGF1 was predominantly expressed in lymphoid organ (Oka) while LvVEGF2 was mainly detected in gill and hemocytes. The transcriptional levels of LvVEGF1 in Oka and LvVEGF2 in gill or hemocytes were apparently up-regulated during WSSV infection. Double-stranded RNA interference was used for further functional studies. The data showed that silencing of LvVEGF1 and LvVEGF2 caused a decrease of the copy numbers of the virus in WSSV infected shrimp and a reduction of the cumulative mortality rate of shrimp during WSSV infection. The present study indicated that LvVEGF1 and LvVEGF2 might facilitate WSSV infection, which provided new evidence to understand the function of VEGF signaling pathway during WSSV infection in shrimp.

The cellular response to vascular endothelial growth factors requires co-ordinated signal transduction, trafficking and proteolysis.

VEGFs (vascular endothelial growth factors) are a family of conserved disulfide-linked soluble secretory glycoproteins found in higher eukaryotes. VEGFs mediate a wide range of responses in different tissues including metabolic homoeostasis, cell proliferation, migration and tubulogenesis. Such responses are initiated by VEGF binding to soluble and membrane-bound VEGFRs (VEGF receptor tyrosine kinases) and co-receptors. VEGF and receptor splice isoform diversity further enhances complexity of membrane protein assembly and function in signal transduction pathways that control multiple cellular responses. Different signal transduction pathways are simultaneously activated by VEGFR-VEGF complexes with membrane trafficking along the endosome-lysosome network further modulating signal output from multiple enzymatic events associated with such pathways. Balancing VEGFR-VEGF signal transduction with trafficking and proteolysis is essential in controlling the intensity and duration of different intracellular signalling events. Dysfunction in VEGF-regulated signal transduction is important in chronic disease states including cancer, atherosclerosis and blindness. This family of growth factors and receptors is an important model system for understanding human disease pathology and developing new therapeutics for treating such ailments.

3D-printed dimethyloxallyl glycine delivery scaffolds to improve angiogenesis and osteogenesis.

Angiogenesis-osteogenesis coupling processes are vital in bone tissue engineering. Normal biomaterials implanted in bone defects have issues in the sufficient formation of blood vessels, especially in the central part. Single delivery of vascular endothelial growth factors (VEGF) to foci in previous studies did not show satisfactory results due to low loading doses, a short protein half-life and low efficiency. Development of a hypoxia-mimicking microenvironment for cells by local prolyl-4-hydroxylase inhibitor release, which can stabilize hypoxia-inducible factor 1α (HIF-1α) expression, is an alternative method. The aim of this study was to design a dimethyloxallyl glycine (DMOG) delivering scaffold composed of mesoporous bioactive glasses and poly(3-hydroxybutyrate-co-3-hydroxyhexanoate) polymers (MPHS scaffolds), so as to investigate whether the sustained release of DMOG promotes local angiogenesis and bone healing. The morphology and microstructure of composite scaffolds were characterized. The DMOG release patterns from scaffolds loaded with different DMOG dosages were evaluated, and the effects of DMOG delivery on human bone marrow stromal cell (hBMSC) adhesion, viability, proliferation, osteogenic differentiation and angiogenic-relative gene expressions with scaffolds were also investigated. In vivo studies were carried out to observe vascular formations and new bone ingrowth with DMOG-loaded scaffolds. The results showed that DMOG could be released in a sustained manner over 4 weeks from MPHS scaffolds and obviously enhance the angiogenesis and osteogenesis in the defects. Microfil perfusion showed a significantly increased formation of vessels in the defects with DMOG delivery. Furthermore, micro-CT imaging and fluorescence labeling indicated larger areas of bone formation for DMOG-loaded scaffolds. It is concluded that MPHS-DMOG scaffolds are promising for enhancing bone healing of osseous defects.