Tailoring Polymerization Controllability and Dispersity Through a Photoswitchable Catalyst Strategy.

Modulating on-demand polymerization is a challenge in synthetic macromolecules. Herein, tailoring polymerization controllability and dispersity during single-electron transfer mediated living radical polymerization (SET-LRP) of methyl methacrylate (MMA) is achieved. Hexaarylbiimidazole (HABI) is employed as a photoswitchable catalyst, allowing reversible control of catalytic activity between an active and inactive state. In the presence of HABI and with the light on (active state), control SET-LRP of MMA follows first-order kinetics, resulting in polymers with a narrow molecular weight distribution. In contrast, polymerization responds to light and reverts to their original uncontrolled state with light off (inactive state). Therefore, repeatable resetting polymerization can be easily performed. The key to photomodulating dispersity is to use an efficient molecular switch to tailor the breadths of dispersity. Besides, the mechanism of HABI-mediated SET-LRP with switchable ability is proposed.

Upregulation of mmu_circ_0001066 attenuates the inhibitory effect of bisphosphonates on osteoclastogenesis.

OBJECTIVE: Medication-related osteonecrosis of the jaw (MRONJ) is the main adverse side effect of bisphosphonates (BPs), mainly owing to the inhibitory effect of BPs on osteoclastogenesis. CircRNAs were identified to be an important factor in regulating cellular processes. The aim of this study was to explore the effect of mmu_circ_0001066 on BP-inhibited osteoclastogenesis. MATERIALS AND METHODS: The expression of MRONJ-related miRNA in RANKL-induced RAW264.7 cells treated with BP was analyzed using qRT-PCR analysis. Bioinformatics techniques were applied to screen potential circRNAs. Tartrate-resistant acid phosphatase (TRAP) staining and bone resorption assays were used to examine the effect of mmu_circ_0001066 on osteoclastogenesis. Bioinformatics analysis, luciferase reporter assays, and Western blotting assays were performed to investigate the underlying mechanism. RESULTS: Four MRONJ-related miRNAs were upregulated in BP-treated RAW264.7 cells, and the expression of mmu_circ_0001066 was negatively correlated with those of MRONJ-related miRNAs. Furthermore, the upregulation of mmu_circ_0001066 partially attenuated the inhibitory effect of BP on osteoclastogenesis in RAW264.7 cells. Mechanistically, upregulated miR-16 suppressed osteoclastogenesis and miR-16 inhibitor increased osteoclastogenesis. Furthermore, we have identified that miR-16 is a downstream effector of mmu_circ_0001066. CONCLUSION: Our results suggest that mmu_circ_0001066 played an important role in the BP-mediated suppression of osteoclastogenesis, which lays a foundation for identifying mmu_circ_0001066 as a potential biomarker for MRONJ.

Regenerable-Catalyst-Aided, Opened to Air and Sunlight-Driven "CuAAC&ATRP" Concurrent Reaction for Sequence-Controlled Copolymer.

An ideal stimuli-responsive controlled/living radical polymerization should have the ability to manipulate the reaction through spatiotemporal "on/off" controls, achieving the polymerization under fully open conditions and allowing for precise control over macromolecular architecture with defined molecular weights and monomer sequence. In this contribution, the photo (sunlight)-induced electron transfer atom transfer radical-polymerization (PET-ATRP) can be realized to be reversibly activated and deactivated under fully open conditions utilizing one-component copper(II) thioxanthone carboxylate as multifunctional photocatalyst and oxygen scavenger. The polymerization behaviors are investigated, presenting controlled features with first-order kinetics and linear relationships between molecular weights and monomer conversions. More importantly, "CuAAC&ATRP" concurrent reaction combining PET-ATRP, photodriven deoxygenation, and photoactivated CuAAC click reaction is successfully employed to synthesize the sequence-defined multiblock functional copolymers, in which the iterative monomer additions can be easily manipulated under fully open conditions.

Integrating Sugar and Dopamine into One Polymer: Controlled Synthesis and Robust Surface Modification.

Glycopolymers attached to a surface possess the ability to bind to certain carbohydrate binding proteins in a highly specific manner, and because of this, the fabrication of glycopolymer-modified surfaces has evolved as an effective route toward bioresponsive systems. Poly(N-3,4-dihydroxybenzenethyl methacrylamide-co-2-(methacrylamido) glucopyranose) copolymers, containing sugar and catechol functionalities, are for the first time successfully prepared in a well-controlled manner via room temperature single-electron transfer initiation and propagation through radical addition fragmentation chain transfer technique. The polymerization behavior is investigated and it presents controlled features with first-order kinetics and linear relationships between molecular weights and monomer conversions. Moreover, the copolymers are used to modify different types of surfaces (silicon, steel, and plastic), the properties of the surfaces and the specific lectin-binding abilities are investigated by a combination of water contact angle, Fourier transform infrared spectroscopy (FT-IR), X-ray photoelectron spectra, scanning electron microscopy with energy dispersive X-ray (SEM/EDX), atomic force microscopy, and confocal microscope measurements.

Controlled synthesis and self-assembly of dopamine-containing copolymer for honeycomb-like porous hybrid particles.

Dopamine-containing monomers, N-3,4-dihydroxybenzenethyl methacrylamide (DMA) and dimethylaminoethyl methacrylate (DMAEMA), are successfully copolymerized in a well-controlled manner via ambient temperature single-electron transfer initiation and propagation through the radical addition fragmentation chain transfer (SET-RAFT) method. The controlled behaviors of the copolymerization are confirmed by the first-order kinetic plots, the linear relationships between molecular weights, and the monomer conversions while keeping relatively narrow molecular weight distribution (Mw/Mn ≤ 1.45). Moreover, biomimetic self-assembly of poly(N-3,4-dihydroxybenzenethyl methacrylamide-co-dimethylaminoethyl methacrylate) PDMA-co-PDMAEMA and inorganic particles are employed to prepare tunable honeycomb-like porous hybrid particles (HPHPs) by regulating the predesigned chemical composition. In addition, the inorganic sacrificial templates are successfully selective etched for the formation of porous organic materials.

Inflammation Can Be a High-Risk Factor for Mucosal Nonunion of MRONJ by Regulating SIRT1 Signaling When Treated with an Oncologic Dose of Zoledronate.

Purpose: Zoledronate (ZA) stands as a highly effective antiresorptive agent known to trigger medication-related osteonecrosis of the jaw (MRONJ). Its clinical dosages primarily encompass those used for oncologic and osteoporosis treatments. While inflammation is recognized as a potential disruptor of mucosal healing processes associated with ZA, prior research has overlooked the influence of varying ZA dosages on tissue adaptability. Therefore, a deeper understanding of the specific mechanisms by which inflammation exacerbates ZA-induced MRONJ, particularly when inflammation acts as a risk factor, remains crucial. Methods: Cell proliferation and migration of human oral keratinocytes (HOK) was analyzed after treatment with different doses of ZA and/or lipopolysaccharide (LPS) to assess their possible effect on mucosal healing of extraction wounds. Mouse periodontitis models were established using LPS, and histological changes in extraction wounds were observed after the administration of oncologic dose ZA. Hematoxylin and eosin (HE) staining and immunofluorescence were used to evaluate mucosal healing. Results: In vitro, LPS did not exacerbate the effects of osteoporosis therapeutic dose of ZA on the proliferation and migration of HOK cells, while aggravated these with the oncologic dose of ZA treatment by inducing mitochondrial dysfunction and oxidative stress via regulating SIRT1 expression. Furthermore, SIRT1 overexpression can alleviate this process. In vivo, local injection of LPS increased the nonunion of mucous membranes in MRONJ and decreased the expression of SIRT1, PGC-1α, and MnSOD. Conclusion: Inflammation aggravates oncologic dose of ZA-induced mitochondrial dysfunction and oxidative stress via a SIRT1-dependent pathway, enhancing the risk of impaired mucosal healing in MRONJ. Our study implies that inflammation becomes a critical risk factor for MRONJ development at higher ZA concentrations. Elucidating the mechanisms of inflammation as a risk factor for mucosal non-healing in MRONJ could inform the development of SIRT1-targeted therapies.

Menaquinone-4 prevents medication-related osteonecrosis of the jaw through the SIRT1 signaling-mediated inhibition of cellular metabolic stresses-induced osteoblast apoptosis.

Long-term usage of bisphosphonates, especially zoledronic acid (ZA), induces osteogenesis disorders and medication-related osteonecrosis of the jaw (MRONJ) in patients, thereby contributing to the destruction of bone remodeling and the continuous progression of osteonecrosis. Menaquinone-4 (MK-4), a specific vitamin K2 isoform converted by the mevalonate (MVA) pathway in vivo, exerts the promotion of bone formation, whereas ZA administration suppresses this pathway and results in endogenous MK-4 deficiency. However, no study has evaluated whether exogenous MK-4 supplementation can prevent ZA-induced MRONJ. Here we showed that MK-4 pretreatment partially ameliorated mucosal nonunion and bone sequestration among ZA-treated MRONJ mouse models. Moreover, MK-4 promoted bone regeneration and inhibited osteoblast apoptosis in vivo. Consistently, MK-4 downregulated ZA-induced osteoblast apoptosis in MC3T3-E1 cells and suppressed the levels of cellular metabolic stresses, including oxidative stress, endoplasmic reticulum stress, mitochondrial dysfunction, and DNA damage, which were accompanied by elevated sirtuin 1 (SIRT1) expression. Notably, EX527, an inhibitor of the SIRT1 signaling pathway, abolished the inhibitory effects of MK-4 on ZA-induced cell metabolic stresses and osteoblast damage. Combined with experimental evidences from MRONJ mouse models and MC3T3-E1 cells, our findings suggested that MK-4 prevents ZA-induced MRONJ by inhibiting osteoblast apoptosis through suppression of cellular metabolic stresses in a SIRT1-dependent manner. The results provide a novel translational direction for the clinical application of MK-4 for preventing MRONJ.

Sex difference in the morbidity and pain response with stage 0 of medication-related osteonecrosis of the jaws.

OBJECTIVE: Medication-related osteonecrosis of the jaws (MRONJ) is a potentially severe complication associated with antiresorptive or antiangiogenic therapies. Prior studies, including our own clinical data, have indicated a higher incidence of MRONJ among women compare to men. However, robust evidence establishing a relationship between sex and the prevalence of MRONJ is lacking. METHODS: We conducted a meta-analysis and utilized murine models to investigate potential sex-based differences in the morbidity associated with MRONJ. RESULTS: Our results revealed no significant difference in the incidence of MRONJ between the sexes when using exposed necrotic bone as a diagnostic criterion. However, a histological examination of the murine models identified the presence of stage 0 MRONJ. Notably, pain assessments across all groups revealed that male mice with stage 0 MRONJ displayed less severe pain symptoms than their female counterparts. CONCLUSIONS: Our findings suggested that sex does not contribute to the risk of developing MRONJ. However, considering that approximately 50% of stage 0 MRONJ cases progress to more advanced stages, the less pronounced pain in male patients might delay medical consultation and potentially lead to disease progression. Clinicians should be particularly vigilant about the subdued pain response in male patients with stage 0 MRONJ to prevent disease advancement.

Brain-derived neurotrophic factor promotes orthodontic tooth movement by alleviating periodontal ligament stem cell senescence.

Orthodontic treatment in older adults is more difficult than in younger adults, partially due to delayed osteogenesis caused by senescence of human periodontal ligament stem cells (hPDLSCs). The production of brain-derived neurotrophic factor (BDNF) which regulates the differentiation and survival of stem cells decreases with age. We aimed to investigate the relationship between BDNF and hPDLSC senescence and its effects on orthodontic tooth movement (OTM). We constructed mouse OTM models using orthodontic nickel­titanium springs and compared the responses of wild-type (WT) and BDNF+/- mice with or without addition of exogenous BDNF. In vitro, hPDLSCs subjected to the mechanical stretch were used to simulate the cell stretch environment during OTM. We extracted periodontal ligament cells from WT and BDNF+/- mice to evaluate their senescence-related indicators. The application of orthodontic force increased BDNF expression in the periodontium of WT mice, while the mechanical stretch increased BDNF expression in hPDLSCs. Osteogenesis-related indicators, including RUNX2 and ALP decreased and cellular senescence-related indicators such as p16, p53 and ß-galactosidase increased in BDNF+/- mice periodontium. Furthermore, periodontal ligament cells extracted from BDNF+/- mice exhibited more senescent compared with cells from WT mice. Application of exogenous BDNF decreased the expression of senescence-related indicators in hPDLSCs by inhibiting Notch3, thereby promoting osteogenic differentiation. Periodontal injection of BDNF decreased the expression of senescence-related indicators in periodontium of aged WT mice. In conclusion, our study showed that BDNF promotes osteogenesis during OTM by alleviating hPDLSCs senescence, paving a new path for future research and clinical applications.

Eldecalcitol effectively prevents alveolar bone loss by partially improving Th17/Treg cell balance in diabetes-associated periodontitis.

Background: Diabetes-associated periodontitis (DPD) is an inflammatory and destructive disease of periodontal tissues in the diabetic population. The disease is manifested as more severe periodontal destruction and is more difficult to treat when compared with periodontitis (PD). Eldecalcitol (ELD) is a novel active vitamin D3 analog; however, little clinical evidence is available on its role on improving PD and DPD, and its specific mechanisms remain unclear. In this study, we evaluated the preventative effects of ELD toward PD and DPD and explored its underlying molecular mechanisms. Methods: Experimental PD and DPD mouse models were established by ligation combined with lipopolysaccharide (LPS) from Porphyromonas gingivalis injection in C57BL/6J and C57BLKS/J Iar- + Leprdb/+Leprdb (db/db) mice, respectively. Simultaneously, ELD (0.25 µg/kg) was orally administered to mice via an intragastric method. Micro-computed tomography (CT), hematoxylin-eosin (HE) staining, immunohistochemistry (IHC), and tartrate-resistant acid phosphatase (TRAP) staining were used to evaluate alveolar bone alterations in vivo. Flow cytometry, immunofluorescence, and real-time polymerase chain reaction (qRT-PCR) were also used to examine gene expression and probe systemic and local changes in Treg and Th17 cell numbers. Additionally, western blotting and immunofluorescence staining were used to examine changes in STAT3/STAT5 signaling. Results: Micro-CT and HE staining showed that the DPD group had higher alveolar bone loss when compared with the PD group. After applying ELD, alveolar bone loss decreased significantly in both PD and DPD groups, and particularly evident in the DPD group. IHC and TRAP staining also showed that ELD promoted osteoblast activity while inhibiting the number of osteoclasts, and after ELD treatment, the receptor activator of nuclear factor-κB ligand (RANKL) to osteoprotegerin (OPG) ratio decreased. More importantly, this decreasing trend was more obvious in the DPD group. Flow cytometry and qRT-PCR also showed that the systemic Th17/Treg imbalance in PD and DPD groups was partially resolved when animals were supplemented with ELD, while immunofluorescence staining and qRT-PCR data showed the Th17/Treg imbalance was partially resolved in the alveolar bone of both ELD supplemented groups. Western blotting and immunofluorescence staining showed increased p-STAT5 and decreased p-STAT3 levels after ELD application. Conclusion: ELD exerted preventative effects toward PD and DPD by partially rectifying Th17/Treg cell imbalance via STAT3/STAT5 signaling. More importantly, given the severity of DPD, we found ELD was more advantageous in preventing DPD.

Responses of nitrogen and phosphorus resorption of understory plants to microscale soil nutrient hetero-geneity in Chinese fir plantation.

We compared the interspecific differences in leaf nutrient resorption of two dominant understory species (Lophatherum gracile and Oplimenus unulatifolius), and analyzed the correlations between the intraspecific efficiency of leaf nutrient resorption and nutrient properties of soil and leaves in Chinese fir plantation. The results showed high soil nutrient heterogeneity in Chinese fir plantation. Soil inorganic nitrogen content and available phosphorus content varied from 8.58 to 65.29 mg·kg-1 and from 2.43 to 15.20 mg·kg-1 in the Chinese fir plantation, respectively. The soil inorganic nitrogen content in O. undulatifolius community was 1.4 times higher than that in L. gra-cile community, but there was no significant difference in soil available phosphorus content between the two communities. Both leaf nitrogen and phosphorus resorption efficiency of O. unulatifolius was significantly lower than that of L. gracile under the three measurement bases of leaf dry weight, leaf area, and lignin content. Resorption efficiency in L. gracile community expressed on leaf dry weight was lower than that expressed on leaf area and lignin content, while resorption efficiency expressed on leaf area was the lowest in O. unulatifolius community. The intraspecific resorption efficiency was significantly correlated with leaf nutrient contents, but was less correlated with soil nutrient content, and only the nitrogen resorption efficiency of L. gracile had significant positive correlation with soil inorganic nitrogen content. The results indicated that there was significant difference in the leaf nutrient resorption efficiency between the two understory species. Soil nutrient heterogeneity exerted a weak effect on the intraspecific nutrient resorption, which might be attributed to high soil nutrient availability and potential disturbance from canopy litter in Chinese fir plantation.

High familiar faces have both eye recognition and holistic processing advantages.

People recognize familiar faces better than unfamiliar faces. However, it remains unknown whether familiarity affects part-based and/or holistic processing. Wang et al., Frontiers in Psychology, 6, 559 (2015), Vision Research, 157, 89-96 (2019) found both enhanced part-based and holistic processing in eye relative to mouth regions (i.e., in a region-selective manner) for own-race and own-species faces, i.e., faces with more experience. Here, we examined the role of face familiarity in eyes (part-based, region-selective) and holistic processing. Face familiarity was tested at three levels: high-familiar (faces of students from the same department and the same class who attended almost all courses together), low-familiar (faces of students from the same department but different classes who attended some courses together), and unfamiliar (faces of schoolmates from different departments who seldom attended the same courses). Using the old/new task in Experiment 1, we found that participants recognized eyes of high-familiar faces better than low-familiar and unfamiliar ones, while similar performance was observed for mouths, indicating a region-selective, eyes familiarity effect. Using the "Perceptual field" paradigm in Experiment 2, we observed a stronger inversion effect for high-familiar faces, a weaker inversion effect for low-familiar faces, but a non-significant inversion effect for unfamiliar faces, indicating that face familiarity plays a role in holistic processing. Taken together, our results suggest that familiarity, like other experience-based variables (e.g., race and species), can improve both eye processing and holistic processing.

New monoterpenes, diterpenes, and lignans from Abies recurvata.

From the aerial part of Abies recurvata, 62 miscellaneous chemical constituents were isolated including 6 new and 56 known ones. The new compounds comprised three monoterpenes, two diterpenes, and one lignan. Their chemical structures were characterized on the basis of various spectroscopic techniques. Dihydrodehydrodiconiferyl alcohol (35) showed the strongest inhibitory effects against lipopolysaccharide-induced nitric oxide production in RAW 264.7 cells with an IC50 value of 66.4 µM.

A review: Development and application of surface molecularly imprinted polymers toward amino acids, peptides, and proteins.

Molecularly imprinted polymers (MIPs) have received wide interests in the bioanalysis field as "artificial antibodies". They can mimic biological receptors by selectively recognizing and adsorbing target molecules owing to their specific affinity to the targets. Traditional MIPs obtained by bulk imprinting have some defects, including low adsorption capacity, poor site accessibility, restricted mass transfer, and irregular morphology, which limit their development. Surface molecularly imprinted polymers (SMIPs) show the features of large surface area, allow fast mass transfer, and have high adsorption capacity and efficiency. They have been intensively used in the research of amino acids, peptides, and proteins due to these advantages. In this review, we systematically summarize the preparation of SMIPs including components and polymerization strategies, and their applications focusing on amino acids, peptides, and proteins are discussed in detail. Finally, future trends and challenges for the design and development of SMIPs are described.

Retention durability of one-retainer versus two-retainer posterior RBFDPs after chewing simulation.

The clinical use of one-retainer RBFDPs in the anterior region has shown higher survival rates compared to conventional two-retainer RBFDPs. The motivation for this study was to assess the validity of this observation when extended to the posterior region. The aim was thus to evaluate different preparation and framework designs for replacing premolars, particularly one-retainer versus two-retainer designs, on the retention of monolithic zirconia posterior RBFDPs. Extracted caries-free human premolars and third molars were embedded in auto-curing resin to create models with an edentulous space of premolar width. Abutment teeth were prepared according to these six designs (n = 8 each): one or two upper retainers with narrow rests, one or two upper retainers with wide rests, and one or two-retainers with wide rests. RBFDPs were milled from monolithic zirconia (KATANA Zirconia HT), and were adhesively bonded using Panavia V5 with corresponding primers. After thermodynamic loading, the quasi-static tensile force required for failure was determined. Failure modes were evaluated using a microscope. Survival rates after thermodynamic loading were 75% for one group (one upper-molar retainer with narrow rest), 100% for the other groups. The debonding forces ranged from 310 ± 224 N (group one upper-molar retainer with narrow rest) to 927 ± 292 N (group two upper retainers with narrow rests). Two-retainer designs failed at significantly higher tensile forces than designs with one retainer (p ≤ 0.05). There were no significant differences between upper and lower designs, or rest widths. Although RBFDPs with two retainers withstood higher debonding forces, RBFDPs with one retainer and wide rest still have a high potential for clinical treatment because of the high forces required for their debonding.

MiR-152-5p suppresses osteogenic differentiation of mandible mesenchymal stem cells by regulating ATG14-mediated autophagy.

BACKGROUND: Osteoporosis affects the mandible resulting in bone loss. Though impairments are not life threatening, they affect a person's quality-of-life particularly vulnerable elderly. MicroRNAs (miRNAs) are novel regulatory factors that play an important role in regulating bone metabolism. Autophagy is evolutionarily conserved intracellular self-degradation process and is vital in the maintenance of both miRNA and bone homeostasis. However, the role of autophagy in the pathogenesis of miRNA regulating osteoporosis remains unclear. METHODS: In the study, we established a rat osteoporosis model induced by ovariectomy (OVX) and isolated mesenchymal stem cells from mandible (MMSCs-M). Several miRNAs were identified to regulate osteoporosis in some studies. qRT-PCR was applied to examine the expression of miRNA, autophagy and osteogenic differentiation-related genes. Western blotting assays were performed to detect the expression of autophagy and osteogenic differentiation proteins. Immunofluorescence and transmission electron microscope were used to verify the autophagy activity. Transfecting technology was used to enhance or suppress the expression of miR-152-5p which enable us to observe the relationship between miR-152-5p, autophagy and osteogenic differentiation. Additionally, the measurement of reactive oxygen species was used to investigate the mechanism of autophagy affecting osteogenic differentiation. RESULTS: We found an upregulated expression of miR-152-5p in MMSCs-M in OVX group. Downregulated autophagy-related gene, proteins and autophagosome were detected in vitro of OVX group compared with sham group. Moreover, downregulation of miR-152-5p promoted osteogenic differentiation of MMSCs-M as well as enhanced autophagy-related proteins in OVX group. Conversely, overexpression of miR-152-5p showed opposite effect in sham group. Meanwhile, we found Atg14 (autophagy-related protein homolog 14) was identified to be a direct target of miR-152-5p theoretically and functionally. In other words, we confirmed inhibition of miR-152-5p promoted the osteogenic differentiation via promoting ATG14-mediated autophagy. Furthermore, miR-152-5p/ATG14-mediated autophagy regulated osteogenic differentiation by reducing the endogenous ROS accumulation and maintaining cellular redox homeostasis. CONCLUSION: Our data suggest that miR-152-5p is the first identified to regulate osteogenic differentiation by directly targeting autophagy-related protein ATG14 and regulating oxidative stress and therapeutic inhibition of miR-152-5p may be an efficient anabolic strategy for osteoporosis.

Effect of different ionic liquids and organic solvents on the structural and physicochemical properties of cellulose-protein complexes extracted from Se-enriched peanut leaves.

Abundant cellulose and insoluble protein were contained in the Se-enriched peanut leaf residue, a by-product from leaf protein extraction. Ionic liquids (ILs) were used to extract the cellulose-protein complexes (CPCs) from Se-enriched peanut leaf residue. The effects of various ILs as extractants and organic solvents as regenerant on the physicochemical properties of CPCs were compared. The results showed that the yield of CPCs and recovery yield of [AMIM]Cl (1-allyl-3-methylimidazole chloride) were better than those of [BMIM]Cl (1-butyl-3-methylimidazolium chloride). Simultaneously, it could be seen from the infrared absorption peaks and secondary structure fitting results that [BMIM]Cl seemed stronger than [AMIM]Cl in destroying the secondary structure of CPCs. Scanning electron microscope (SEM) showed that the CPCs extracted by [BMIM]Cl were lamellate with holes on the surface, and the CPCs extracted by [AMIM]Cl were rough, almost without holes on the surface. Furthermore, the transmittance and tensile strength of the film which contained BA-CPC ([BMIM]Cl as extractant and acetonitrile as regenerant) film were better than those contained AA-CPC ([AMIM]Cl as extractant and acetonitrile as regenerant) film, which might be mainly because the types of ILs and regenerants affect the particle size of CPCs, thereby influencing the mechanical properties of the film.

Polymer chimera of stapled oncolytic peptide coupled with anti-PD-L1 peptide boosts immunotherapy of colorectal cancer.

Rationale: Scarce tumor mutation burden and neoantigens create tremendous obstacles for an effective immunotherapy of colorectal cancer (CRC). Oncolytic peptides rise as a promising therapeutic approach that boosts tumor-specific immune responses by inducing antigenic substances. However, the clinical application of oncolytic peptides has been hindered because of structural instability, proteolytic degradation, and undesired toxicity when administered systemically. Methods: Based on wasp venom peptide, an optimized stapled oncolytic peptide MP9 was developed with rigid α-helix, protease-resistance, and CRC cell cytotoxicity. By incorporating four functional motifs that include D-peptidomimetic inhibitor of PD-L1, matrix metalloproteinase-2 (MMP-2) cleavable spacer, and MP9 with 4-arm PEG, a novel peptide-polymer conjugate (PEG-MP9-aPDL1) was obtained and identified as the most promising systemic delivery vehicle with PD-L1 targeting specificity and favorable pharmacokinetic properties. Results: We demonstrated that PEG-MP9-aPDL1-driven oncolysis induces a panel of immunogenic cell death (ICD)-relevant damage-associated molecular patterns (DAMPs) both in vitro and in vivo, which are key elements for immunotherapy with PD-L1 inhibitor. Further, PEG-MP9-aPDL1 exhibited prominent immunotherapeutic efficacy in a CRC mouse model characterized by tumor infiltration of CD8+ T cells and induction of cytotoxic lymphocytes (CTLs) in the spleens. Conclusion: Our findings suggest that PEG-MP9-aPDL1 is an all-in-one platform for oncolytic immunotherapy and immune checkpoint blockade (ICB).

Fucoidan-loaded nanofibrous scaffolds promote annulus fibrosus repair by ameliorating the inflammatory and oxidative microenvironments in degenerative intervertebral discs.

Tissue engineering holds potential in the treatment of intervertebral disc degeneration (IDD). However, implantation of tissue engineered constructs may cause foreign body reaction and aggravate the inflammatory and oxidative microenvironment of the degenerative intervertebral disc (IVD). In order to ameliorate the adverse microenvironment of IDD, in this study, we prepared a biocompatible poly (ether carbonate urethane) urea (PECUU) nanofibrous scaffold loaded with fucoidan, a natural marine bioactive polysaccharide which has great anti-inflammatory and antioxidative functions. Compared with pure PECUU scaffold, the fucoidan-loaded PECUU nanofibrous scaffold (F-PECUU) decreased the gene and protein expression related to inflammation and the oxidative stress in the lipopolysaccharide (LPS) induced annulus fibrosus cells (AFCs) significantly (p<0.05). Especially, gene expression of Il 6 and Ptgs2 was decreased more than 50% in F-PECUU with 3.0 wt% fucoidan (HF-PECUU). Moreover, the gene and protein expression related to the degradation of extracellular matrix (ECM) were reduced in a fucoidan concentration-dependent manner significantly, with increased almost 3 times gene expression of Col1a1 and Acan in HF-PECUU. Further, in a 'box' defect model, HF-PECUU decreased the expression of COX-2 and deposited more ECM between scaffold layers when compared with pure PECUU. The disc height and nucleus pulposus hydration of repaired IVD reached up to 75% and 85% of those in the sham group. In addition, F-PECUU helped to maintain an integrate tissue structure with a similar compression modulus to that in sham group. Taken together, the F-PECUU nanofibrous scaffolds showed promising potential to promote AF repair in IDD treatment by ameliorating the harsh degenerative microenvironment. STATEMENT OF SIGNIFICANCE: Annulus fibrosus (AF) tissue engineering holds potential in the treatment of intervertebral disc degeneration (IDD), but is restricted by the inflammatory and oxidative microenvironment of degenerative disc. This study developed a biocompatible polyurethane scaffold (F-PECUU) loaded with fucoidan, a marine bioactive polysaccharide, for ameliorating IDD microenvironment and promoting disc regeneration. F-PECUU alleviated the inflammation and oxidative stress caused by lipopolysaccharide and prevented extracellular matrix (ECM) degradation in AF cells. In vivo, it promoted ECM deposition to maintain the height, water content and mechanical property of disc. This work has shown the potential of marine polysaccharides-containing functional scaffolds in IDD treatment by ameliorating the harsh microenvironment accompanied with disc degeneration.

Multifunctional Nanofibrous Scaffolds with Angle-Ply Microstructure and Co-Delivery Capacity Promote Partial Repair and Total Replacement of Intervertebral Disc.

There is an urgent clinical need for the treatment of annulus fibrosus (AF) impairment caused by intervertebral disc (IVD) degeneration or surgical injury. Although repairing injured AF through tissue engineering is promising, the approach is limited by the complicated angle-ply microstructure, inflammatory microenvironment, poor self-repairing ability of AF cells and deficient matrix production. In this study, electrospinning technology is used to construct aligned core-shell nanofibrous scaffolds loaded with transforming growth factor-ß3 (TGFß3) and ibuprofen (IBU), respectively. The results confirm that the rapid IBU release improves the inflammatory microenvironment, while sustained TGFß3 release enhances nascent extracellular matrix (ECM) formation. Biomaterials for clinical applications must repair local AF defects during herniectomy and enable AF regeneration during disc replacement, so a box defect model and total IVD replacement model in rat tail are constructed. The dual-drug delivering electrospun scaffolds are assembled into angle-ply structure to form a highly biomimetic AF that is implanted into the box defect or used to replace the disc. In two animal models, it is found that biomimetic scaffolds with good anti-inflammatory ability enhance ECM formation and maintain the mechanical properties of IVD. Findings from this study demonstrate that the multifunctional nanofibrous scaffolds provide inspirations for IVD repair.