Resting-State Functional Connectivity Between Centromedial Amygdala and Insula as Related to Somatic Symptoms in Depressed Patients: A Preliminary Study.

OBJECTIVE: Somatic symptoms are prevalent in patients with depression. The centromedial amygdala (CMA) is a key brain region that mediates autonomic and somatic responses. Abnormal function in the CMA may contribute to the development of somatic symptoms in depressed patients. METHODS: We compared the resting-state functional connectivity (RSFC) based on the seed of the left and right CMA between 37 patients with depression and 30 healthy controls. The severity of depressive and somatic symptoms was assessed using the Hamilton Depression Rating Scale (HDRS) and the 15-item somatic symptom severity scale of the Patient Health Questionnaire (PHQ-15). Correlation analysis was performed to investigate the relationship between the RSFC and clinical variables (HDRS and PHQ-15) in depressed patients. RESULTS: Compared with healthy controls, patients with depression exhibited decreased RSFC between the CMA and insula, and superior temporal gyrus. In addition, functional connectivity between the left CMA and left insula was negatively correlated with PHQ-15 (r = -0.348, p = .037) in depressed patients. No significant relation was found between the RSFC and HDRS in depressed patients. CONCLUSIONS: Functional connectivity between the CMA and insula is reduced in depressive patients, which is associated with the severity of somatic symptoms. Our findings may provide a potential neural substrate to interpret the co-occurrence of depression with somatic symptoms.

Sustained Release of a Peptide-Based Matrix Metalloproteinase-2 Inhibitor to Attenuate Adverse Cardiac Remodeling and Improve Cardiac Function Following Myocardial Infarction.

Following myocardial infarction (MI), degradation of extracellular matrix (ECM) by upregulated matrix metalloproteinases (MMPs) especially MMP-2 decreases tissue mechanical properties, leading to cardiac function deterioration. Attenuation of cardiac ECM degradation at the early stage of MI has the potential to preserve tissue mechanical properties, resulting in cardiac function increase. Yet the strategy for efficiently preventing cardiac ECM degradation remains to be established. Current preclinical approaches have shown limited efficacy because of low drug dosage allocated to the heart tissue, dose-limiting side effects, and cardiac fibrosis. To address these limitations, we have developed a MMP-2 inhibitor delivery system that can be specifically delivered into infarcted hearts at early stage of MI to efficiently prevent MMP-2-mediated ECM degradation. The system was based on an injectable, degradable, fast gelation, and thermosensitive hydrogel, and a MMP-2 specific inhibitor, peptide CTTHWGFTLC (CTT). The use of fast gelation hydrogel allowed to completely retain CTT in the heart tissue. The system was able to release low molecular weight CTT over 4 weeks possibly due to the strong hydrogen bonding between the hydrogel and CTT. The release kinetics was modulated by amount of CTT loaded into the hydrogel, and using chondroitin sulfate and heparin that can interact with CTT and the hydrogel. Both glycosaminoglycans augmented CTT release, while heparin more greatly accelerated the release. After it was injected into the infarcted hearts for 4 weeks, the released CTT efficiently prevented cardiac ECM degradation as it not only increased tissue thickness but also preserved collagen composition similar to that in the normal heart tissue. In addition, the delivery system significantly improved cardiac function. Importantly, the delivery system did not induce cardiac fibrosis. These results demonstrate that the developed MMP-2 inhibitor delivery system has potential to efficiently reduce adverse myocardial remodeling and improve cardiac function.

Modulation of wound healing and scar formation by MG53 protein-mediated cell membrane repair.

Cell membrane repair is an important aspect of physiology, and disruption of this process can result in pathophysiology in a number of different tissues, including wound healing, chronic ulcer and scarring. We have previously identified a novel tripartite motif family protein, MG53, as an essential component of the cell membrane repair machinery. Here we report the functional role of MG53 in the modulation of wound healing and scarring. Although MG53 is absent from keratinocytes and fibroblasts, remarkable defects in skin architecture and collagen overproduction are observed in mg53(-/-) mice, and these animals display delayed wound healing and abnormal scarring. Recombinant human MG53 (rhMG53) protein, encapsulated in a hydrogel formulation, facilitates wound healing and prevents scarring in rodent models of dermal injuries. An in vitro study shows that rhMG53 protects against acute injury to keratinocytes and facilitates the migration of fibroblasts in response to scratch wounding. During fibrotic remodeling, rhMG53 interferes with TGF-ß-dependent activation of myofibroblast differentiation. The resulting down-regulation of α smooth muscle actin and extracellular matrix proteins contributes to reduced scarring. Overall, these studies establish a trifunctional role for MG53 as a facilitator of rapid injury repair, a mediator of cell migration, and a modulator of myofibroblast differentiation during wound healing. Targeting the functional interaction between MG53 and TGF-ß signaling may present a potentially effective means for promoting scarless wound healing.

[Detection of Adulteration in Milk Powder with Starch Near Infrared].

Three China trademarks of milk powder called Mengniu, Yili, Wandashan were taken as testing samples. Each of them mixed varied amount of starch in different gradient, which were consisted of 32 adulterated milk powder samples mixed with starch, was taken as standard samples for constructing predicted model. To those 32 samples, the reflecting spectrum characteristics in middle wave of near infrared spectrum with Near Infrared Spectrum Analyzer (Micro NIR 1700) produced by JDSU Ltd. USA were collected for five repeats in five different days. The time span was nearly two months. Firstly, we build the model used the reflecting spectrum characteristics of those samples with biomimetic pattern recognition (BPR) arithmetic to do the qualitative analysis. The analysis included the reliability of testing result and stability of the model. When we took ninety percent as the evaluation threshold of testing result of CAR (Correct Acceptance Rate) and CRR (Correct Rejection Rate), the lowest starch content of adulterate milk powder in all tested samples which the tested result were bigger than that abovementioned threshold was designated CAR threshold (CAR-T) and CRR threshold (CRR-T). CAR means the correct rate of accepting a sample which is belong to itself, CRR means correct rate of refusing to accept a sample which is not belong to itself. The results were shown that, when we constructed a model based on the near infrared spectrum data from each of three China trademark milk powders, respectively, if we constructed a model with infrared spectrum data tested in a same day, both the CAR-T and CRR-T of adulterate starch content of a sample can reach 0.1% in predicting the remainder infrared spectrum data tested within a same day. The three China trademarks of milk powder had the same result. In addition, when we ignored the trademarks, put the spectrum data of adulterate milk powder samples mixed with the same content of starch of three China trademarks milk powder together to construct a model, the CAR-T of mixed starch content of a sample may reach 0.1%, the CRR-T can reach 1%, if the model construction and predicting were performed with near infrared spectrum data tested in a same day. However, the CAR-T can just stably reach up to 5% and the CRR-T have the same result, if the model construction and predicting were crossly performed with mixed near infrared spectrum data tested in different days. Furthermore, the correct recognizing threshold mixed starch of a sample can stably reach up to 1% and the CAR-T can reach 5%, if the model construction was based on near infrared spectrum data combined the previous four days to predict the output of the another day. On the other hand, we also engaged quantitative analysis to the starch content in milk power with two kinds of arithmetic (PLSR, LS-SVR). In contrast with the testing outputs, the reliability of both the CAR-T and CRR-T in qualitative analysis was further validated.

Periostin modulates myofibroblast differentiation during full-thickness cutaneous wound repair.

The matricellular protein periostin is expressed in the skin. Although periostin has been hypothesized to contribute to dermal homeostasis and repair, this has not been directly tested. To assess the contribution of periostin to dermal healing, 6 mm full-thickness excisional wounds were created in the skin of periostin-knockout and wild-type, sex-matched control mice. In wild-type mice, periostin was potently induced 5-7 days after wounding. In the absence of periostin, day 7 wounds showed a significant reduction in myofibroblasts, as visualized by expression of α-smooth muscle actin (α-SMA) within the granulation tissue. Delivery of recombinant human periostin by electrospun collagen scaffolds restored α-SMA expression. Isolated wild-type and knockout dermal fibroblasts did not differ in in vitro assays of adhesion or migration; however, in 3D culture, periostin-knockout fibroblasts showed a significantly reduced ability to contract a collagen matrix, and adopted a dendritic phenotype. Recombinant periostin restored the defects in cell morphology and matrix contraction displayed by periostin-deficient fibroblasts in a manner that was sensitive to a neutralizing anti-ß1-integrin and to the FAK and Src inhibitor PP2. We propose that periostin promotes wound contraction by facilitating myofibroblast differentiation and contraction.

Oxygenated Scaffolds for Pancreatic Endocrine Differentiation from Induced Pluripotent Stem Cells.

A 3D microenvironment is known to endorse pancreatic islet development from human induced pluripotent stem cells (iPSCs). However, oxygen supply becomes a limiting factor in a scaffold culture. In this study, oxygen-releasing biomaterials are fabricated and an oxygenated scaffold culture platform is developed to offer a better oxygen supply during 3D iPSC pancreatic differentiation. It is found that the oxygenation does not alter the scaffold's mechanical properties. The in situ oxygenation improves oxygen tension within the scaffolds. The unique 3D differentiation system enables the generation of islet organoids with enhanced expression of islet signature genes and proteins. Additionally, it is discovered that the oxygenation at the early stage of differentiation has more profound impacts on islet development from iPSCs. More C-peptide+ /MAFA+ ß and glucagon+ /MAFB+ α cells formed in the iPSC-derived islet organoids generated under oxygenated conditions, suggesting enhanced maturation of the organoids. Furthermore, the oxygenated 3D cultures improve islet organoids' sensitivity to glucose for insulin secretion. It is herein demonstrated that the oxygenated scaffold culture empowers iPSC islet differentiation to generate clinically relevant tissues for diabetes research and treatment.

Electrospun acetalated dextran scaffolds for temporal release of therapeutics.

Electrospun acetalated dextran (Ac-DEX) scaffolds were fabricated to encapsulate resiquimod, an immunomodulatory toll-like-receptor (TLR) agonist. Ac-DEX has been used to fabricate scaffolds for sustained and temporal delivery of therapeutics because it has tunable degradation rates that are dependent on its synthesis reaction time or the molecular weight of dextran. Additionally, as opposed to commonly electrospun polyesters that shift the local pH upon degradation, the degradation products of Ac-DEX are pH-neutral: dextran, an alcohol, and the metabolic byproduct acetone. Formulations of Ac-DEX with two different degradation rates were used in this study. The effects of electrospinning conditions on the scaffold size and morphology were examined as well as fibroblast adhesion as imaged with fluorescence microcopy and scanning electron microscopy. Macrophage (MΦ) viability further indicates that the scaffolds are cytocompatible. Also, the controlled release profiles of resiquimod from loaded scaffolds and nitric oxide (NO) production by MΦ incubated with these scaffolds show the potential for Ac-DEX scaffolds to be used to temporally and efficiently deliver therapeutics. Overall, we present a novel scaffold that can have tunable and unique drug release rates for tissue engineering, drug delivery, immunomodulation, and wound healing applications.

Thermosensitive and antioxidant wound dressings capable of adaptively regulating TGFß pathways promote diabetic wound healing.

Various therapies have been utilized for treating diabetic wounds, yet current regiments do not simultaneously address the key intrinsic causes of slow wound healing, i.e., abnormal skin cell functions (particularly migration), delayed angiogenesis, and chronic inflammation. To address this clinical gap, we develop a wound dressing that contains a peptide-based TGFß receptor II inhibitor (PTßR2I), and a thermosensitive and reactive oxygen species (ROS)-scavenging hydrogel. The wound dressing can quickly solidify on the diabetic wounds following administration. The released PTßR2I inhibits the TGFß1/p38 pathway, leading to improved cell migration and angiogenesis, and decreased inflammation. Meanwhile, the PTßR2I does not interfere with the TGFß1/Smad2/3 pathway that is required to regulate myofibroblasts, a critical cell type for wound healing. The hydrogel's ability to scavenge ROS in diabetic wounds further decreases inflammation. Single-dose application of the wound dressing significantly accelerates wound healing with complete wound closure after 14 days. Overall, using wound dressings capable of adaptively modulating TGFß pathways provides a new strategy for diabetic wound treatment.

Co-Delivery of Bioengineered Exosomes and Oxygen for Treating Critical Limb Ischemia in Diabetic Mice.

Diabetic patients with critical limb ischemia face a high rate of limb amputation. Regeneration of the vasculature and skeletal muscles can salvage diseased limbs. Therapy using stem cell-derived exosomes that contain multiple proangiogenic and promyogenic factors represents a promising strategy. Yet the therapeutic efficacy is not optimal because exosomes alone cannot efficiently rescue and recruit endothelial and skeletal muscle cells and restore their functions under hyperglycemic and ischemic conditions. To address these limitations, we fabricated ischemic-limb-targeting stem cell-derived exosomes and oxygen-releasing nanoparticles and codelivered them in order to recruit endothelial and skeletal muscle cells, improve cell survival under ischemia before vasculature is established, and restore cell morphogenic function under high glucose and ischemic conditions. The exosomes and oxygen-releasing nanoparticles, delivered by intravenous injection, specifically accumulated in the ischemic limbs. Following 4 weeks of delivery, the exosomes and released oxygen synergistically stimulated angiogenesis and muscle regeneration without inducing substantial inflammation and reactive oxygen species overproduction. Our work demonstrates that codelivery of exosomes and oxygen is a promising treatment solution for saving diabetic ischemic limbs.

Increased 10-year cardiovascular disease risk in depressed patients with coexisting subclinical hypothyroidism.

Purpose: The prevalence of depressive disorder (DD) and subclinical hypothyroidism (SH) was almost twofold higher in women compared with men, both of which are confirmed to be related to cardiovascular disease (CVD) risk. The current study aimed to identify the prevalence of CVD risk factors and evaluate the 10-year CVD risk in female depressed patients with and without comorbid SH. Methods: We recruited 1744 female inpatients with a diagnosis of DD. Venous blood samples were taken from all patients for lipid and thyroid hormones. Framingham Risk Score (FRS) was used to estimate the 10-year CVD risk. Results: Female depressed patients with SH had increased BMI, higher Hamilton Anxiety Scale (HAMA) scores, higher LDL-C, TC, UA, and a higher 10-year CVD risk than euthyroid DD groups. Serum TSH levels and HAMA scores were critical predictive variables for 10-year CVD risk in female depressed patients with comorbid SH. Conclusion: Our study suggests that female depressed patients with SH have a high 10-year CVD risk. Serum TSH levels and HAMA scores may be helpful to predict cardiovascular risk in female patients with SH. The increased CVD risk in female depressed patients with comorbid SH requires more attention from researchers and clinicians.

DNA methylation-mediated Rbpjk suppression protects against fracture nonunion caused by systemic inflammation.

Challenging skeletal repairs are frequently seen in patients experiencing systemic inflammation. To tackle the complexity and heterogeneity of the skeletal repair process, we performed single-cell RNA sequencing and revealed that progenitor cells were one of the major lineages responsive to elevated inflammation and this response adversely affected progenitor differentiation by upregulation of Rbpjk in fracture nonunion. We then validated the interplay between inflammation (via constitutive activation of Ikk2, Ikk2ca) and Rbpjk specifically in progenitors by using genetic animal models. Focusing on epigenetic regulation, we identified Rbpjk as a direct target of Dnmt3b. Mechanistically, inflammation decreased Dnmt3b expression in progenitor cells, consequently leading to Rbpjk upregulation via hypomethylation within its promoter region. We also showed that Dnmt3b loss-of-function mice phenotypically recapitulated the fracture repair defects observed in Ikk2ca-transgenic mice, whereas Dnmt3b-transgenic mice alleviated fracture repair defects induced by Ikk2ca. Moreover, Rbpjk ablation restored fracture repair in both Ikk2ca mice and Dnmt3b loss-of-function mice. Altogether, this work elucidates a common mechanism involving a NF-κB/Dnmt3b/Rbpjk axis within the context of inflamed bone regeneration. Building on this mechanistic insight, we applied local treatment with epigenetically modified progenitor cells in a previously established mouse model of inflammation-mediated fracture nonunion and showed a functional restoration of bone regeneration under inflammatory conditions through an increase in progenitor differentiation potential.

Micropatterned thermoresponsive surfaces by polymerization of monomer crystals: modulating cellular morphology and cell-substrate interactions.

A novel and facile approach has been developed to create thermoresponsive surfaces with macroscale patterns together with microscale features. The surface patterns were formed by applying macroscale nucleation agent patterns onto saturated N-isopropylacrylamide monomer solution membranes to induce the divergent growth of needlelike monomer crystals; the patterned monomer crystals were then photopolymerized to form patterned thermoresponsive films. A series of analytical tools (i.e., scanning electron microscopy, profilometry, and contact angle measurement) were used to characterize the properties of the patterned films. Cell coculture on this patterned thermoresponsive films enables cell separation and sorting by modulating temperature- and topography-dependent cell-substrate interactions and cell morphology, respectively. This versatile technique allows the formation of various macroscale patterns with microscale features over large areas, and on most solid substrates, within minutes, all of this without the need for expensive equipment and facilities. Such patterned surfaces can act as both in vitro tumor models and separation platforms for cancer studies. This method can also be applied to other cell-based biological studies and clinical applications.

Placement of an elastic biodegradable cardiac patch on a subacute infarcted heart leads to cellularization with early developmental cardiomyocyte characteristics.

BACKGROUND: Placement of an elastic biodegradable patch onto a subacute myocardial infarct (MI) provides temporary elastic support that may act to effectively alter adverse left ventricular (LV) remodeling processes. METHODS: Two weeks after permanent left coronary ligation in Lewis rats, the infarcted anterior wall was covered with polyester urethane urea (MI + PEUU; n = 15) or expanded polytetrafluoroethylene (MI + ePTFE; n = 15) patches, or had no implantation (MI + sham; n = 12). Eight weeks after surgery, cardiac function and histology were assessed. RESULTS: The ventricular wall in the MI + ePTFE and MI + sham groups was composed of fibrous tissue, whereas PEUU implantation induced α-smooth muscle actin-positive muscle bundles coexpressing sarcomeric α-actinin and cardiac-specific troponin-T. This pattern of colocalization was also found in developing embryonic myocardium. Cardiac transcription factors Nkx-2.5 and GATA-4 were strongly expressed in the muscle bundles. In the MI + sham group, end-diastolic LV cavity area (EDA) increased and the percentage of fractional area change (%FAC) decreased. For ePTFE patched animals, both EDA and %FAC decreased. In contrast, with MI + PEUU patching, %FAC increased and EDA was maintained. With dobutamine-stress echocardiography, MI + PEUU patched LVs possessed contractile reserve significantly larger than the MI + sham group. CONCLUSIONS: MI + PEUU patch implantation onto subacute infarcted myocardium induced muscle cellularization with characteristics of early developmental cardiomyocytes as well as providing a functional reserve.

A thermosensitive hydrogel capable of releasing bFGF for enhanced differentiation of mesenchymal stem cell into cardiomyocyte-like cells under ischemic conditions.

A thermosensitive hydrogel capable of differentiating mesenchymal stem cells (MSCs) into cardiomyocyte-like cells was synthesized. The hydrogel was based on N-isopropylacrylamide (NIPAAm), N-acryloxysuccinimide, acrylic acid, and hydroxyethyl methacrylate-poly(trimethylene carbonate). The hydrogel was highly flexible at body temperature with breaking strain >1000% and Young's modulus 45 kPa. When MSCs were encapsulated in the hydrogel and cultured under normal culture conditions (10% FBS and 21% O(2)), the cells differentiated into cardiomyocyte-like cells. However, the differentiation was retarded, and even diminished, under low nutrient and low oxygen conditions, which are typical of the infarcted heart. We hypothesized that enhancing MSC survival under low nutrient and low oxygen conditions would restore the differentiation. To enhance cell survival, a pro-survival growth factor (bFGF) was loaded in the hydrogel. bFGF was able to sustainedly release from the hydrogel for 21 days. Under the low nutrient and low oxygen conditions (1% O(2) and 1% FBS), bFGF enhanced MSC survival and differentiation in the hydrogel. After 14 days of culture, survival of 70.5% of MSCs remained in the bFGF-loaded hydrogel, while only 4.9% of MSCs remained in the hydrogel without bFGF. The differentiation toward cardiomyocyte-like cells was completely inhibited at 1% FBS and 1% oxygen. Loading bFGF in the hydrogel restored the differentiation, as confirmed by the expression of cardiac markers at both the gene (MEF2C and CACNA1c) and protein (cTnI and connexin 43) levels. bFGF loading also up-regulated the paracrine effect of MSCs. VEGF expression was significantly increased in the bFGF-loaded hydrogel. These results demonstrate that the developed bFGF-loaded hydrogel may potentially be used to deliver MSCs into hearts for regeneration of heart tissue.

Creating 3D angiogenic growth factor gradients in fibrous constructs to guide fast angiogenesis.

Fast angiogenesis in 3D fibrous constructs that mimic the morphology of the extracellular matrix remains challenging due to limited porosity in the densely packed constructs. We investigated whether mimicking the in vivo chemotaxis microenvironment for native blood vessel formation would stimulate angiogenesis in the fibrous constructs. The chemotaxis microenvironment was created by introducing 3D angiogenic growth factor gradients into the constructs. We have developed a technique that can quickly fabricate (∼40 min) such 3D gradients by simultaneously electrospinning polycaprolactone (PCL) fibers, encapsulating gradient amount of bFGF (stabilized by heparin) into poly(lactide-co-glycolide) (PLGA) microspheres, and electrospraying the microspheres into PCL fibers. Gradient formation was confirmed by fluorescence microscopy. Gradients with different steepnesses were obtained by modulating the initial concentration of the bFGF solution. All of the constructs were able to sustainedly release bioactive bFGF over a 28 day period. The release kinetics was dependent on the bFGF loading and steepness of the gradient. In vitro cell migration study demonstrated that bFGF gradients significantly increased the depth of cell migration. To assess the efficacy of bFGF gradients in inducing angiogenesis, we implanted constructs subcutaneously using mouse model. bFGF gradients significantly promoted cell penetration into the constructs. After 10 days of implantation, a high density of mature blood vessels (positive to both CD31 and α-SMA) were formed in the constructs. Vessel density was increased with the increase in steepness of the bFGF gradient. These gradient constructs may have potential to engineer vascularized tissues for various applications.

[Early treatment of atelectasis by bronchoscopy in craniotomy patients].

OBJECTIVE: To investigate the safety and effects of early bronchoscopy on atelectasis of the ventilation patients, whom had experienced craniotomy for severe cranial trauma and hemorrhage. METHODS: Fifty-five patients suffered from severe cranial trauma and hemorrhage with Glascow coma scores (GCS) less than 8 complicated by atelectasis after craniotomy were early given sputum suction by bronchoscope via extratracheal intubation and broncho-alveolar lavage (BAL) during tracheal intubation and mechanical ventilation. During the treatment, patients' consciousness, vital signs and arterial blood gas were closely monitored. The relevant data, before, during (5, 10, and 25 minutes), bronchoscopy treatment completed and 30 minutes after bronchoscopy, were recorded and analyzed. RESULTS: Eighty-two time of bronchoscopies and 111 time of local BALs in 55 patients were completed and were effective for atelectasis. The patient's GCS (5.6±2.5 vs. 5.4±2.6, P>0.05), heart rate (HR), respiratory rate (RR), systolic blood pressure (SBP), blood oxygenous saturation (SaO(2)) were not deteriorated during bronchoscopy. Compared with pre-bronchoscopy, the HR and SBP decreased (HR: 88.2±14.2 bpm vs. 98.2±18.3 bpm, SBP: 110.6±18.2 mm Hg vs. 118.4±18.5 mm Hg, both P<0.05), and SaO(2) increased (0.982±0.022 vs. 0.945±0.035, P<0.05), pH, arterial partial pressure of oxygen (PaO(2)) and arterial partial pressure of carbon dioxide (PaCO(2)) had no significant changes during bronchoscopy. There was obviously increased in PaO(2) (84.5±14.4 mm Hg, 81.6±18.2 mm Hg vs. 76.2±15.4 mm Hg, both P<0.05), and decreased in PaCO(2) (27.0±12.8 mm Hg, 29.3±18.2 mm Hg vs. 36.5±11.6 mm Hg, both P<0.05) respectively, significantly decreased in alveolar arterial pressure of oxygen difference [P ((A-a))O(2)] at 10 minutes and 25 minutes, and at the time bronchoscopy treatment completed and the time 30 minutes after compared with before bronchoscopy (36.1±4.7 mm Hg, 32.4±6.2 mm Hg, 32.5±5.2 mm Hg, 31.2±7.2 mm Hg vs. 38.5±5.6 mm Hg, all P<0.05). All patients had not encounter side effects related with bronchoscopy and ventilation. CONCLUSION: The bronchoscope via extratracheal intubation for sputum suction and BAL were safe and effective treatment to the patients suffered from severe cranial trauma or hemorrhage complicated by atelectasis after craniotomy during mechanical ventilation, without obvious changes of the vital signs.

Delivery of VEGF and delta-like 4 to synergistically regenerate capillaries and arterioles in ischemic limbs.

Vascularization of the poorly vascularized limbs affected by critical limb ischemia (CLI) is necessary to salvage the limbs and avoid amputation. Effective vascularization requires forming not only capillaries, but also arterioles and vessel branching. These processes rely on the survival, migration and morphogenesis of endothelial cells in the ischemic limbs. Yet endothelial cell functions are impaired by the upregulated TGFß. Herein, we developed an injectable hydrogel-based drug release system capable of delivering both VEGF and Dll4 to synergistically restore endothelial cellular functions, leading to accelerated formation of capillaries, arterioles and vessel branching. In vitro, the Dll4 and VEGF synergistically promoted the human arterial endothelial cell (HAEC) survival, migration, and formation of filopodial structure, lumens, and branches under the elevated TGFß1 condition mimicking that of the ischemic limbs. The synergistic effect was resulted from activating VEGFR2, Notch-1 and Erk1/2 signaling pathways. After delivering the Dll4 and VEGF via an injectable and thermosensitive hydrogel to the ischemic mouse hindlimbs, 95% of blood perfusion was restored at day 14, significantly higher than delivery of Dll4 or VEGF only. The released Dll4 and VEGF significantly increased density of capillaries and arterioles, vessel branching point density, and proliferating cell density. Besides, the delivery of Dll4 and VEGF stimulated skeletal muscle regeneration and improved muscle function. Overall, the developed hydrogel-based Dll4 and VEGF delivery system promoted ischemic limb vascularization and muscle regeneration. STATEMENT OF SIGNIFICANCE: Effective vascularization of the poorly vascularized limbs affected by critical limb ischemia (CLI) requires forming not only capillaries, but also arterioles and vessel branching. These processes rely on the survival, migration and morphogenesis of endothelial cells. Yet endothelial cell functions are impaired by the upregulated TGFß in the ischemic limbs. Herein, we developed an injectable hydrogel-based drug release system capable of delivering both VEGF and Dll4 to synergistically restore endothelial cell functions, leading to accelerated formation of capillaries, arterioles and vessel branching.

Rescuing Cardiac Cells and Improving Cardiac Function by Targeted Delivery of Oxygen-Releasing Nanoparticles after or Even before Acute Myocardial Infarction.

Myocardial infarction (MI) causes massive cell death due to restricted blood flow and oxygen deficiency. Rapid and sustained oxygen delivery following MI rescues cardiac cells and restores cardiac function. However, current oxygen-generating materials cannot be administered during acute MI stage without direct injection or suturing methods, both of which risk rupturing weakened heart tissue. Here, we present infarcted heart-targeting, oxygen-releasing nanoparticles capable of being delivered by intravenous injection at acute MI stage, and specifically accumulating in the infarcted heart. The nanoparticles can also be delivered before MI, then gather at the injured area after MI. We demonstrate that the nanoparticles, delivered either pre-MI or post-MI, enhance cardiac cell survival, stimulate angiogenesis, and suppress fibrosis without inducing substantial inflammation and reactive oxygen species overproduction. Our findings demonstrate that oxygen-delivering nanoparticles can provide a nonpharmacological solution to rescue the infarcted heart during acute MI and preserve heart function.

Sustained delivery of rhMG53 promotes diabetic wound healing and hair follicle development.

MG53 is an essential component of the cell membrane repair machinery, participating in the healing of dermal wounds. Here we develop a novel delivery system using recombinant human MG53 (rhMG53) protein and a reactive oxygen species (ROS)-scavenging gel to treat diabetic wounds. Mice with ablation of MG53 display defective hair follicle structure, and topical application of rhMG53 can promote hair growth in the mg53 -/- mice. Cell lineage tracing studies reveal a physiological function of MG53 in modulating the proliferation of hair follicle stem cells (HFSCs). We find that rhMG53 protects HFSCs from oxidative stress-induced apoptosis and stimulates differentiation of HSFCs into keratinocytes. The cytoprotective function of MG53 is mediated by STATs and MAPK signaling in HFSCs. The thermosensitive ROS-scavenging gel encapsulated with rhMG53 allows for sustained release of rhMG53 and promotes healing of chronic cutaneous wounds and hair follicle development in the db/db mice. These findings support the potential therapeutic value of using rhMG53 in combination with ROS-scavenging gel to treat diabetic wounds.

Differential roles of hypoxia inducible factor subunits in multipotential stromal cells under hypoxic condition.

Cell therapy with bone marrow multipotential stromal cells (MSCs) represents a promising approach to promote wound healing and tissue regeneration. MSCs expanded in vitro lose early progenitors with differentiation and therapeutic potentials under normoxic condition, whereas hypoxic condition promotes MSC self-renewal through preserving colony forming early progenitors and maintaining undifferentiated phenotypes. Hypoxia inducible factor (HIF) pathway is a crucial signaling pathway activated in hypoxic condition. We evaluated the roles of HIFs in MSC differentiation, colony formation, and paracrine activity under hypoxic condition. Hypoxic condition reversibly decreased osteogenic and adipogenic differentiation. Decrease of osteogenic differentiation depended on HIF pathway; whereas decrease of adipogenic differentiation depended on the activation of unfolded protein response (UPR), but not HIFs. Hypoxia-mediated increase of MSC colony formation was not HIF-dependent also. Hypoxic exposure increased secretion of VEGF, HGF, and basic FGF in a HIF-dependent manner. These findings suggest that HIF has a limited, but pivotal role in enhancing MSC self-renewal and growth factor secretions under hypoxic condition.