Piezo1-driven mechanotransduction as a key regulator of cartilage degradation in early osteoarthritis.

Osteoarthritis (OA) is a prevalent degenerative disease characterized by pain and cartilage damage in its later stages, while early OA is marked by the loss of cartilage's mechanical function. Recent studies suggest that Piezo1, a mechanotransducer, may contribute to cartilage degradation under abnormal physical stress. This study investigates the mechanism by which Piezo1 mediates the loss of cartilage's mechanical properties. Using rat chondrocytes cultured in a 3D in vitro model, we found that fluid flow-induced physical stress activates constitutively expressed Piezo1, leading to increased catabolic activity and apoptosis, which, in turn, disrupts the matrix structure. Ex vivo cartilage experiments further demonstrated that the mechanical stress-induced loss of cartilage's physical properties (approximately 10% reduction in relaxation modulus) is mediated by Piezo1 and depends on cell viability. Notably, Piezo1 agonists alone did not alter the mechanical behavior of cartilage tissue. In vivo, using an OA rat model induced by anterior cruciate ligament transection, we observed cartilage integrity degradation and loss of mechanical properties, which were partially mitigated by Piezo1 inhibition. RNA sequencing revealed significant modulation of the PI3K signaling and matrix regulation pathways. Collectively, this study demonstrates that Piezo1-mediated catabolic activity in chondrocytes is a key driver of the loss of cartilage's mechanical function during the relaxation phase.

Emerging signs of Alzheimer-like tau hyperphosphorylation and neuroinflammation in the brain post recovery from COVID-19.

Coronavirus disease 2019 (COVID-19) has been suggested to increase the risk of memory decline and Alzheimer's disease (AD), the main cause of dementia in the elderly. However, direct evidence about whether COVID-19 induces AD-like neuropathological changes in the brain, especially post recovery from acute infection, is still lacking. Here, using postmortem human brain samples, we found abnormal accumulation of hyperphosphorylated tau protein in the hippocampus and medial entorhinal cortex within 4-13 months post clinically recovery from acute COVID-19, together with prolonged activation of glia cells and increases in inflammatory factors, even though no SARS-COV-2 invasion was detected in these regions. By contrast, COVID-19 did not change beta-amyloid deposition and hippocampal neuron number, and had limited effects on AD-related pathological phenotypes in olfactory circuits including olfactory bulb, anterior olfactory nucleus, olfactory tubercle, piriform cortex and lateral entorhinal cortex. These results provide neuropathological evidences linking COVID-19 with prognostic increase of risk for AD.

Regenerated Fat Induced by a Decellularized Adipose Matrix Can Survive Long-term In Vivo.

Decellularized adipose matrix (DAM) is considered to be the most potential biological scaffold for soft tissue repair and reconstruction, as it is able to induce the regeneration of adipose tissue in situ in adulthood. But how does this adipose tissue regeneration happen and develop in vivo? Is it the same as the original autologous one? Temporary existence or long-term survival? These are the key questions that will determine the future applications of DAM. In this study, we investigated the composition, structure and biomechanical properties of DAM before implanting it into the subcutaneous back of immunodeficient mice. The entire regeneration process in vivo was closely monitored histologically from 3 days to 1 year after implantation, including fat regeneration, vascular growth, inflammatory responses, and matrix degradation and remodeling. Transcriptome sequencing was used to analyze the difference in gene expression between regenerated fat and autologous fat at different periods. The results showed that the DAM-induced regenerated fat first appeared at 1w and remained stable over 6m, indicating remarkable similarity to autologous fat at the later stages of implantation. And about (18.3 ± 29.3) % of the regenerated adipocytes were still viable after one year. The process of adipogenesis was enhanced by the decrease in inflammatory infiltration and proceeded in parallel with angiogenesis. STATEMENT OF SIGNIFICANCE: : The decellularized adipose matrix (DAM) is the only biological scaffold that can spontaneously generate adipocytes in vivo without the need to add exogenous cells. However, in the previous studies, the longest DAM-related animal experiments were about 3 months. The different stages and characteristics of DAM implantation cannot be fully captured. Comprehensive preclinical researches on the initiation, characteristics, and long-term outcomes of DAM-induced adipose tissue regeneration in adulthood is crucial. In this study, we closely observed various aspects of the entire process in vivo from 3 days to 1 year after implantation including fat regeneration, vascular growth, inflammatory reactions as well as matrix degradation and remodeling. The thorough research will contribute to the understanding of stability and dynamic remodeling of DAM regeneration models.

Decellularized adipose-derived matrix from Superficial layers of abdominal adipose tissue exhibits superior capacity of adipogenesis compared to deep layers.

The adipogenic property of decellularized adipose-derived matrix (DAM) varies widely across reports, making it difficult to make a horizontal comparison between reports and posing challenges for the stable clinical translation of DAM. It is possibly due to differences in donor characteristics, but the exact relationship remains unclear. Despite extensive research on the differences between superficial and deep layers of abdominal subcutaneous fat, a main donor of DAM, little is known about their extracellular matrix (ECM) which is promising in regenerative medicine. In this study, we first confirmed the distinct compositional profiles and adipogenic potential between superficial and deep DAM (S-DAM and D-DAM). Both in vitro and in vivo assays confirmed superior adipogenic induction potential in S-DAM over D-DAM. Total amounts of ECM proteins like collagen and laminin were similar, however, the predominant types differed, with collagen I dominating S-DAM and collagen XIV prevailing in D-DAM. S-DAM was enriched with mitochondrial and immunological proteins, whereas D-DAM featured more neuronal, vascular, muscular, and endocrine-related proteins. More proteins involved in mRNA processing were found in D-DAM, with Protein-Protein Interaction (PPI) analysis revealing HNRNPA2B1, HNRNPA1, and HNRNPC as the most tightly interacting members. These findings not only deepen our comprehension of the structural and functional heterogeneity of adipose tissues but also become one of the reason for the large variability between batches of DAM products, providing guidance for constructing more efficient and stable bio-scaffolds.

Hydrophilic Components as Key Active Ingredients in Adipose-Derived Matrix Bioscaffolds for Inducing Fat Regeneration.

Decellularized adipose-derived matrix (DAM) has emerged as a promising biomaterial for soft tissue reconstruction. However, due to a lack of research on its complex composition, the understanding of the key components in DAM remains limited, leading to inconsistent adipogenic properties and challenges in optimizing preparation methods purposefully. In this study, it is proposed for the first time that DAM comprises two distinct components: hydrophilic (H-DAM) and lipophilic (L-DAM), each with markedly different effects on fat regeneration. It is confirmed that H-DAM is the key component for inducing fat regeneration due to its enhanced cell-cell and cell-scaffold interactions, primarily mediated by the Hedgehog signaling pathway. In contrast, L-DAM exhibits poor cell adhesion and contains more antigenic components, leading to a higher immunoinflammatory response and reduced adipogenesis. In addition, it is found that intracellular proteins, which are more abundant in H-DAM, can be retained as beneficial components due to their hydrophilicity, contrary to the conventional view that they shall be removed. Accordingly, a purified bioscaffold with unprecedented efficacy is proposed for fat regeneration and reduced immunogenicity. This finding provides insights for developing scaffolds for fat regeneration and promotes the realization of xenotransplantation.

Dominant activities of fear engram cells in the dorsal dentate gyrus underlie fear generalization in mice.

Over-generalized fear is a maladaptive response to harmless stimuli or situations characteristic of posttraumatic stress disorder (PTSD) and other anxiety disorders. The dorsal dentate gyrus (dDG) contains engram cells that play a crucial role in accurate memory retrieval. However, the coordination mechanism of neuronal subpopulations within the dDG network during fear generalization is not well understood. Here, with the Tet-off system combined with immunostaining and two-photon calcium imaging, we report that dDG fear engram cells labeled in the conditioned context constitutes a significantly higher proportion of dDG neurons activated in a similar context where mice show generalized fear. The activation of these dDG fear engram cells encoding the conditioned context is both sufficient and necessary for inducing fear generalization in the similar context. Activities of mossy cells in the ventral dentate gyrus (vMCs) are significantly suppressed in mice showing fear generalization in a similar context, and activating the vMCs-dDG pathway suppresses generalized but not conditioned fear. Finally, modifying fear memory engrams in the dDG with "safety" signals effectively rescues fear generalization. These findings reveal that the competitive advantage of dDG engram cells underlies fear generalization, which can be rescued by activating the vMCs-dDG pathway or modifying fear memory engrams, and provide novel insights into the dDG network as the neuronal basis of fear generalization.

DSCR1-1 attenuates osteoarthritis-associated chondrocyte injury by regulating the CREB1/ALDH2/Wnt/ß-catenin axis: An in vitro and in vivo study.

The progression of osteoarthritis (OA) includes the initial inflammation, subsequent degradation of the extracellular matrix (ECM), and chondrocyte apoptosis. Down syndrome candidate region 1 (DSCR1) is a stress-responsive gene and expresses in varied types of cells, including chondrocytes. Bioinformatics analysis of GSE103416 and GSE104739 datasets showed higher DSCR1 expression in the inflamed cartilage tissues and chondrocytes of OA. DSCR1 had two major isoforms, isoform 1 (DSCR1-1) and isoform 4 (DSCR1-4). We found that DSCR1-1 had a faster (in vitro) and higher expression (in vivo) response to OA compared to DSCR1-4. IL-1ß-induced apoptosis, inflammation, and ECM degradation in chondrocytes were attenuated by DSCR1-1 overexpression. DSCR1-1 triggered the phosphorylation of cAMP response element-binding 1 (CREB1) at 133 serine sites by decreasing calcineurin activity. Moreover, activated CREB1 moved into the cell nucleus and combined in the promoter regions of aldehyde dehydrogenase 2 (ALDH2), thus enhancing its gene transcription. ALDH2 could recover Wnt/ß-catenin signaling transduction by enhancing phosphorylation of ß-catenin at 33/37 serine sites and inhibiting the migration of ß-catenin protein from the cellular matrix to the nucleus. In vivo, adenoviruses (1 × 108 PFU) overexpressing DSCR1-1 were injected into the articular cavity of C57BL/6 mice with medial meniscus surgery-induced OA, and it showed that DSCR1-1 overexpression ameliorated cartilage injury. Collectively, our study demonstrates that DSCR1-1 may be a potential therapeutic target of OA.

Effects of nanographene oxide on adipose-derived stem cell cryopreservation.

Cryoinjury mitigation is key in cell cryopreservation. Here, we aimed to assess the effectiveness of nanographene oxide (nano-GO) for improving cryoprotectant agents (CPAs) in human adipose stem cell (hADSC) cryopreservation. For in vitro experiments, nano-GO (5 µg/mL) was added to the CPAs in the control, and passage (P) 2 hADSCs were collected and cryopreserved for around two weeks. We compared cytotoxicity, cell viability, immunophenotypes, proliferation, cell apoptosis, and tri-lineage differentiation. In vivo, studies used lipoaspirate to create non-enriched or hADSC-enriched fat tissues by combining it with PBS or hADSCs cryopreserved with the aforementioned CPAs. Each nude mouse received a 0.3 mL subcutaneous injection of the graft. At 12 weeks, the grafts were harvested. Histology, adipocyte-associated genes and protein, vascular density and angiogenic cytokines, macrophage infiltration, and inflammatory cytokines were analyzed. Nano-GO CPA contributed to increased cell viability, improved cell recovery, and lowered levels of early apoptosis. Nano GO at concentrations of 0.01-100 µg/mL caused no cytotoxicity to hADSCs. The absence of nano GOs in the intracellular compartments of the cells was confirmed by transmission electron microscopy. The fat grafts from the CPA-GO group showed more viable adipocytes and significantly increased angiogenesis compared to the PBS and CPA-C groups. Adding hADSCs from the CPA-GO group to the graft reduced macrophage infiltration and MCP-1 expression. Nano-GO plays an anti-apoptotic role in the cryopreservation of hADSCs, which could improve the survival of transplanted fat tissues, possibly via improved angiogenesis and lower inflammatory response in the transplanted adipose tissue.

Predictive value of thrombocytopenia for bloodstream infection in patients with sepsis and septic shock.

BACKGROUND: Thrombocytopenia is common in patients with sepsis and septic shock. AIM: To analyse the decrease in the number of platelets for predicting bloodstream infection in patients with sepsis and septic shock in the intensive care unit. METHODS: A retrospective analysis of patients admitted with sepsis and septic shock in Xingtai People Hospital was revisited. Patient population characteristics and laboratory data were collected for analysis. RESULTS: The study group consisted of 85 (39%) inpatients with bloodstream infection, and the control group consisted of 133 (61%) with negative results or contamination. The percentage decline in platelet counts (PPCs) in patients positive for pathogens [57.1 (41.3-74.6)] was distinctly higher than that in the control group [18.2 (5.1-43.1)] (P < 0.001), whereas the PPCs were not significantly different among those with gram-positive bacteraemia, gram-negative bacteraemia, and fungal infection. Using receiver operating characteristic curves, the area under the curve of the platelet drop rate was 0.839 (95%CI: 0.783-0.895). CONCLUSION: The percentage decline in platelet counts is sensitive in predicting bloodstream infection in patients with sepsis and septic shock. However, it cannot identify gram-positive bacteraemia, gram-negative bacteraemia, and fungal infection.

Impaired neuronal macroautophagy in the prelimbic cortex contributes to comorbid anxiety-like behaviors in rats with chronic neuropathic pain.

A large proportion of patients with chronic pain experience co-morbid anxiety. The medial prefrontal cortex (mPFC) is proposed to underlie this comorbidity, but the molecular and neuronal mechanisms are not fully understood. Here, we reported that impaired neuronal macroautophagy in the prelimbic cortical (PrL) subregion of the mPFC paralleled the occurrence of anxiety-like behaviors in rats with chronic spared nerve injury (SNI). Intriguingly, such macroautophagy impairment was mainly observed in a FOS/c-Fos+ neuronal subpopulation in the PrL. Chemogenetic inactivation of this comorbid anxiety-related neuronal ensemble relieved pain-induced anxiety-like behaviors. Rescuing macroautophagy impairment in this neuronal ensemble relieved chronic pain-associated anxiety and mechanical allodynia and restored synaptic homeostasis at the molecular level. By contrast, artificial disruption of macroautophagy induced early-onset co-morbid anxiety in neuropathic rats, but not general anxiety in normal rats. Taken together, our work identifies causal linkage between PrL neuronal macroautophagy dysfunction and comorbid anxiety in neuropathic pain and provides novel insights into the role of PrL by differentiating its contribution in pain-induced comorbid anxiety from its modulation over general anxiety-like behaviors.Abbreviation: AAV: adeno-associated viruses; ACC: anterior cingulate cortex; ATG5: autophagy related 5; ATG7: autophagy related 7; ATG12: autophagy related 12; CAMK2/CaMKII: calcium/calmodulin-dependent protein kinase II; CNO: clozapine-N-oxide; CQ: chloroquine; DIA: data independent acquisition; DIO: double floxed inverse orf; DLG4/PSD-95: discs large MAGUK scaffold protein 4; Dox: doxycycline; GABA: γ-aminobutyric acid; GFP: green fluorescent protein; GO: gene ontology; Gi: inhibitory guanine nucleotide-binding proteins; HsCHRM4/M4D: human cholinergic receptor muscarinic 4; HsSYN: human synapsin; KEGG: Kyoto encyclopedia of genes and genomes; LAMP1: lysosomal-associated membrane protein 1; LC3-II: PE conjugated microtubule-associated protein 1 light chain3; MAP1LC3/LC3: microtubule-associated protein 1 light chain 3; mPFC: medial prefrontal cortex; P2A: 2A self-cleaving peptide; PPI: protein-protein interaction networks; PrL: prelimbic cortex; RBFOX3/NeuN: RNA binding protein, fox-1 homolog (C. elegans) 3; rtTA: reverse tetracycline-transactivator; SDS-PAGE: sodium dodecylsulfate-polyacrylamide gel electrophoresis; SHANK3: SH3 and multiple ankyrin repeat domains 3; SLC1A1/EAAC1: solute carrier family 1 (neuronal/epithelial high affinity glutamate transporter, systemXag), member 1; SNAP23: synaptosomal-associated protein 23; SNI:spared nerve injury; SQSTM1/p62: sequestosome 1; SYT3: synaptotagmin 3; TRE: tetracycline-responsive element; TRE3G: third-generation tetracycline-responsive element.

Study on the Photocatalytic Properties of Metal-Organic Framework-Derived C-, N-Co-Doped ZnO.

In this study, C- and N-co-doped ZnO photocatalysts were prepared through pyrolysis using metal-organic frameworks (MOFs) as precursor materials. The crystal structure, morphology, and surface chemical composition of the samples were characterised via X-ray diffraction (XRD), scanning electron microscopy (SEM), and X-ray photoelectron spectroscopy (XPS). Their activities in photocatalytic reactions were also evaluated through photocatalytic experiments. The results show that C-, N-co-doped ZnO has a high specific surface area, which is favourable for a photocatalytic reaction. Meanwhile, C-, N-doping can effectively modulate the energy band structure of ZnO, broaden its light absorption range, and improve the separation efficiency of photogenerated electron-hole pairs. The photocatalytic experiments show that the C/N-ZnO-500 samples, which have the optimal photocatalytic performances, have improved performances of 50% and 35%, respectively, compared with those of the blank control group and the ZIF-8 samples. The preparation of ZnO materials with a morphology change and doping using metal frameworks as precursors provides a new idea for designing efficient photocatalysts.

Comparison of the quality of Nurse-Led palliative care with standard medical care during six months in 405 patients with Parkinson's disease and burdens of their Caregivers: A retrospective study at a single center in China.

BACKGROUND: Palliative care is mainly used to improve the quality of life of patients with chronic diseases by addressing their medical conditions and psychological problems. End-stage Parkinson's disease (PD) is also a progressive disease like cancer and could be managed by palliative care. This study was conducted at a single center in China and aimed to compare the quality of nurse-led palliative care with standard medical care during six months in 405 patients with Parkinson's disease (PPD) and their caregivers using the Chinese version of the 39-item Parkinson's Disease Questionnaire (PDQ-39) and the Chinese Zarit Burden Interview (ZBI) scale. METHODS: PPD (stage 2-5) received nurse-led palliative care (NP cohort, 103 patients; 103 caregivers) or neurologist-led standard care (NS cohort, 134 patients; 134 caregivers), or primary care practitioner-led usual care (PS cohort, 168 patients; 168 caregivers) for six months. RESULTS: Before the health professional-led care (BN), the PDQ-39 score of PPD was 68 (71-64) and their caregivers had 54.86 ± 7.64 a ZBI scale. After 6-months of the health professional-led care (AN), the PDQ-39 score of PPD and a ZBI scale of their caregivers decreased for the NP cohort as compared to those of BN condition and those of patients in the NS and PS cohorts at AN condition (p < 0.001 for all). CONCLUSIONS: The quality of life of PPD must be improved and the burden on their caregivers must be relieved. Nurse-led palliative care successfully improved the quality of life of PPD and reduced their caregiver burden.

Harnessing fine fibers in decellularized adipose-derived matrix for enhanced adipose regeneration.

Decellularized Adipose-Derived Matrix (DAM) has the function of inducing adipogenesis, but the distribution of adipogenesis is uneven. We found for the first time that DAM contains two structural components: The tough fibers DAM (T-DAM) and the fine fibers DAM (F-DAM). T-DAM was a dense vortex structure composed of a large number of coarse fibers, characterized by myoblast-related proteins, which cannot achieve fat regeneration and forms a typical "adipose-free zone". While the F-DAM was a loose structure consisting of uniform fine fibers, has more matrix-related proteins and adipose-related proteins. It can not only better promote the adhesion and proliferation of adipose stem cells in vitro, but also achieve the regeneration of adipose tissue in vivo earlier and better, with a uniform range of adipogenesis. The F-DAM is the main and effective kind of DAM to initiate adipose tissue regeneration, which can be picked out by ultrasound fragmentation.

Transcriptomic and machine learning analyses identify hub genes of metabolism and host immune response that are associated with the progression of breast capsular contracture.

Capsular contracture is a prevalent and severe complication that affects the postoperative outcomes of patients who receive silicone breast implants. At present, prosthesis replacement is the major treatment for capsular contracture after both breast augmentation procedures and breast reconstruction following breast cancer surgery. However, the mechanism(s) underlying breast capsular contracture remains unclear. This study aimed to identify the biological features of breast capsular contracture and reveal the potential underlying mechanism using RNA sequencing. Sample tissues from 12 female patients (15 breast capsules) were divided into low capsular contracture (LCC) and high capsular contracture (HCC) groups based on the Baker grades. Subsequently, 41 lipid metabolism-related genes were identified through enrichment analysis, and three of these genes were identified as candidate genes by SVM-RFE and LASSO algorithms. We then compared the proportions of the 22 types of immune cells between the LCC and HCC groups using a CIBERSORT analysis and explored the correlation between the candidate hub features and immune cells. Notably, PRKAR2B was positively correlated with the differentially clustered immune cells, which were M1 macrophages and follicular helper T cells (area under the ROC = 0.786). In addition, the expression of PRKAR2B at the mRNA or protein level was lower in the HCC group than in the LCC group. Potential molecular mechanisms were identified based on the expression levels in the high and low PRKAR2B groups. Our findings indicate that PRKAR2B is a novel diagnostic biomarker for breast capsular contracture and might also influence the grade and progression of capsular contracture.

Effectiveness of a New Enzyme-Free Method for the Preparation of a Decellularized Adipose-Derived Matrix.

BACKGROUND: Decellularized adipose-derived matrix (DAM) represents a new alternative to tissue fillers. The function of DAM is closely associated with the decellularization technique used for its preparation. However, most techniques are time-consuming and expensive, and this might reduce the popularity of DAM. OBJECTIVES: The study aimed to investigate an enzyme-free adipose decellularization method and generate a DAM capable of adipose tissue regeneration. METHODS: DAMs prepared by the enzyme-free and Flynn's methods were compared and co-cultured with human adipose-derived stem cells (hADSCs) to investigate cytocompatibility. Adipose tissue formation was evaluated by injecting the DAMs into the backs of nude mice over 4 weeks. Samples were harvested for gross and perilipin immunohistochemistry analysis at 1 and 4 weeks. RESULTS: The enzyme-free method is effective for adipose decellularization because it removes adipocytes and preserves the microstructure. In vitro, the DAM made by the enzyme-free method could support the attachment, growth, proliferation, and differentiation of hADSCs, and promote the enhanced secretion of vascular endothelial growth factor by hADSCs; this DAM also induced the formation and maturity of adipocytes in vivo. CONCLUSIONS: This study describes a highly effective enzyme-free method for adipose tissue decellularization that also promotes adipocyte formation and adipose tissue volume stability in vitro and in vivo, resulting in a new alternative tissue filler.

Effect of radiation sterilization on the ability to induce adipose regeneration in vivo in decellularized adipose-derived matrix.

BACKGROUND: Decellularized adipose-derived matrix (DAM), a biological scaffold that can induce adipose regeneration. The balance between its sterilization efficiency and its ability to maintain in situ adipose regeneration should be considered in terminal sterilization. The purpose of this study was to investigate the effects of radiation sterilization of cobalt-60 (60 Co)with different doses on adipogenesis induced by different forms of DAM, so as to reduce radiation dose under the premise of safe and effective sterilization and ensure adipogenesis induced by DAM in vivo. METHODS: High dose (25 kGy) and low dose (5 kGy) radiation were used to sterilize freeze-dried and wet DAM, respectively. The sterilization efficiency, macro and micro characteristics, mechanical and mechanical properties of DAM were compared, and then implanted into the immunocompromised mice to evaluate the adipose regeneration. RESULTS: Under the two radiation doses, no microbial growth was found in the freeze-dried and wet DAM sterility tests, and no significant changes were observed in the macro and micro structures. In terms of mechanical properties, the elastic modulus of high dose freeze-dried DAM decreased significantly (p < 0.001). In vivo animal experiments, the freeze-dried DAM irradiated with high dose almost completely lost its function of adipogenesis in vivo. Although the wet DAM irradiated with high dose could induce fat regeneration in the early stage, the adipocyte deformation and atrophy appeared in the later stage. The freeze-dried and wet DAM after low dose irradiation was similar to the wet DAM without irradiation in the blank control, which could maintain excellent adipogenic and angiogenic functions in vivo. CONCLUSION: High dose 60 Co irradiation can completely destroy the ability of freeze-dried DAM to induce adipose regeneration in situ, while low dose irradiation (5 kGy) can effectively sterilize the DAM without damaging in vivo induced adipose regeneration. Radiation has more damage to freeze-dried DAM than wet DAM in adipogenesis properties.

Selection of Mechanical Fragmentation Methods Based on Enzyme-Free Preparation of Decellularized Adipose-Derived Matrix.

BACKGROUND: The decellularized adipose-derived matrix (DAM) has emerged as a promising biomaterial for inducing adipose tissue regeneration. Various methods have been employed to produce DAM, among which the enzyme-free method is a relatively recent preparation technique. The mechanical fragmentation step plays a crucial role in determining the efficacy of the enzyme-free preparation. METHODS: The adipose tissue underwent fragmentation through the application of ultrasonication, homogenization, and freeze ball milling. This study compared the central temperature of the mixture immediately following crushing, the quantity of oil obtained after centrifugation, and the thickness of the middle layer. Fluorescence staining was utilized to compare the residual cell activity of the broken fat in the middle layer, while electron microscopy was employed to assess the integrity and properties of the adipocytes among the three methods. The primary products obtained through the three methods were subsequently subjected to processing using the enzyme-free method DAM. The assessment of degreasing and denucleation of DAM was conducted through HE staining, oil red staining, and determination of DNA residues. Subsequently, the ultrasonication-DAM (U-DAM) and homogenation-DAM (H-DAM) were implanted bilaterally on the back of immunocompromised mice, and a comparative analysis of their adipogenic and angiogenic effects in vivo was performed. RESULTS: Oil discharge following ultrasonication and homogenization was significantly higher compared to that observed after freeze ball milling (p < 0.001), despite the latter exhibiting the lowest center temperature (p < 0.001). The middle layer was found to be thinnest after ultrasonication (p < 0.001), and most of the remaining cells were observed to be dead following fragmentation. Except for DAM obtained through freeze ball milling, DAM obtained through ultrasonication and homogenization could be completely denucleated and degreased. In the in vivo experiment, the first adipocytes were observed in U-DAM as early as 1 week after implantation, but not in H-DAM. After 8 weeks, a significant number of adipocytes were regenerated in both groups, but the U-DAM group demonstrated a more efficient adipose regeneration than in H-DAM (p = 0.0057). CONCLUSIONS: Ultrasonication and homogenization are effective mechanical fragmentation methods for breaking down adipocytes at the initial stage, enabling the production of DAM through an enzyme-free method that facilitates successful regeneration of adipose tissues in vivo. Furthermore, the enzyme-free method, which is based on the ultrasonication pre-fragmentation approach, exhibits superior performance in terms of denucleation, degreasing, and the removal of non-adipocyte matrix components, thereby resulting in the highest in vivo adipogenic induction efficiency.

A Bibliometric Analysis of Research on Decellularized Matrix for Two Decades.

The articles and reviews in the field of decellularized extracellular matrix (dECM) from 2001 to 2021 were retrieved and extracted from the Web of Science Core Collection. The R package Bibliometrix, CiteSpace, VOSviewer, and the online BIBLIOMETRC platform were utilized for bibliometric analysis, including specific characteristics of annual publications, influential countries/regions, core journals, leading institutions, keywords, key references, cocited authors, journals and institutions, cooperation, and historical direct citations. Our study concluded core references that fueled the development of dECM and highlighted current research directions, hotpots, and trends. From 2001 to 2021, 3,046 publications were retrieved in total, including 2,700 articles and 349 reviews. The United States (n = 895) produced the majority of publications, and the University of Pittsburgh (n = 318) published most productions. Biomaterials were identified as the most productive and influential journal in the dECM field considering the number of publications (n = 194), and total citations (n = 15,694). Immunomodulation, bioreactors, aging, three-dimensional (3D) bioprinting, bone tissue engineering, bioink, hydrogel, biomaterials, and regeneration were the latest high-frequency keywords, indicating the emerging frontiers of dECM. In the field, decellularization techniques lay the foundation. Orthotopic transplantation of recellularized dECM and induction of specific cell differentiation promoted the bursts of research. The 3D bioprinting and hydrogel based on dECM were extensively studied in recent years. The present study provided developmental trajectories, current research status, global collaboration patterns, hotpots, and trending topics of dECM. Decellularization techniques, tissue engineering to regenerate organs, and improvements in application are the major themes over the past two decades. Impact Statement The review article is significant because decellularized extracellular matrix (dECM), which derived from biological tissues and removal of immunogenic cells, is characterized by safety, biocompatibility, and low in toxicity. Showing great application prospects, dECM has been applied in multiple scenarios of tissue repairment and reconstruction, among the most popular topics in tissue engineering. Thus, analyzing and concluding the development, current condition and future trends are of great significance. Comparing to conventional review, this review article systemically and comprehensively concluded the historical development, current status, and research trending topics. Thus, it allows scholars to get a rapid overview of the dECM field, and plan research directions.

GuiLu-ErXian Glue extract promotes mesenchymal stem cells (MSC)-Induced chondrogenesis via exosomes release and delays aging in the MSC senescence process.

ETHNOPHARMACOLOGICAL RELEVANCE: The treatment of osteoarthritis (OA) patients is a challenging problem. Mesenchymal stem cells (MSCs) are multipotent cells and play key roles in regenerative medicine for cartilage degeneration. GuiLu-ErXian Glue (GLEXG) is an herbal remedy widely used in traditional Chinese medicine to treat joint pain and disability in elderly OA patients. However, the mechanisms of how GLEXG affects MSCs-induced chondrogensis remains to be elucidated. AIM OF THE STUDY: The aim of this study was to investigate the effects of GLEXG on MSC-derived chondrogenesis, both in vitro and in vivo and its potential mechanisms. METHODS: Using human MSC (hMSCs) as in vitro model, the effects of HPLC-profiled GLEXG water extract on chondrogenic differentiation were investigated by 3D spheroid cultures under chondrogenesis-inducing medium (CIM) condition. The chondrogenesis process was evaluated by measuring the sphere sizes, chondrogenesis-related genes expression by reverse transcription real-time PCR that targeted type II/X collagens, SOX9, aggrecan, and protein expression by immunostaining. Anti-TGF-ß1 neutralization antibody was used for mechanistic study. Mono-iodoacetate (MIA) induced OA joint was used to evaluate the effects of GLEXG on in vivo model. MSCs-derived exosomes were purified for proteomics study and senescence process was evaluated by cumulative population doublings and senescence-associated ß-Galactosidase staining. RESULTS: The results showed that GLEXG enhanced hMSCs chondrogenesis and upregulated RNA expression of type II/X collagen, SOX9 and aggrecan at 0.1 µg/mL, 0.3 µg/mL in vitro. In vivo, GLEXG at the dose of 0.3 µg intraarticular (i.a.) injection rescued the MIA-induced cartilage defect. Proteomics and ingenuity pathway analysis obtained from MSCs-released exosomes suggested that senescence pathway was less activated in GLEXG group than in vehicle group. Besides, GLEXG was able to increase cumulative population doubling and delayed hMSCs senescence process after four passages in cultures. CONCLUSION: we conclude that GLEXG promotes in vitro MSC-induced chondrogenesis possibly via exosomes release and delays aging in the MSC senescence process and that treatment with GLEXG (0.3 µg, i.a.) rescued cartilage defects in rat OA knee model.

Interrater agreement between children's self-reported and their mothers' proxy-reported dental anxiety: a Chinese cross-sectional study.

BACKGROUND: Children's dental anxiety is common in dental clinics. This study aimed to determine the interrater agreement between children's self-reported and their mothers' proxy-reported dental anxiety and its affecting factors. METHODS: In this cross-sectional study performed in a dental clinic, primary school students and their mothers were assessed for enrollment eligibility. The Modified Dental Anxiety Scale plus Facial Image Scale (MDAS-FIS) was employed to test both the children's self-reported and their mothers' proxy-reported dental anxiety independently. The interrater agreement was analyzed using percentage agreement and the linear weighted kappa (k) coefficient. Factors affecting children's dental anxiety were analyzed using univariate and multivariate logistic regression models. RESULTS: One hundred children and their mothers were enrolled. The median ages of the children and mothers were 8.5 and 40.0 years old, respectively, and 38.0% (38/100) of the children were female. The scores of children's self-reported dental anxiety were significantly higher than their mothers' proxy-reported dental anxiety (MDAS-Questions 1-5, all p < 0.05); moreover, there was no agreement between the two groups in terms of all anxiety hierarchies (kappa coefficient = 0.028, p = 0.593). In the univariate model, a total of seven factors (age, gender, maternal anxiety, number of dental visits, mother's presence or absence, oral health status, and having siblings or not) were involved for analysis, and age [every 1-year increase, odds ratio (OR) = 0.661, 95% confidence interval (CI) = 0.514-0.850, p = 0.001], several dental visits (every 1 visit increase, OR = 0.409, 95% CI = 0.190-0.880, p = 0.022), and mother presence (OR = 0.286, 95% CI = 0.114-0.714, p = 0.007) were affecting factors. In the multivariate model, only age (every 1 year increase) and maternal presence were associated with 0.697-fold (95% CI = 0.535-0.908, p = 0.007) and 0.362-fold (95% CI = 0.135-0.967, p = 0.043) decreases in the risk of children's dental anxiety during dental visits and treatment, respectively. CONCLUSION: There was no significant agreement between elementary school students' self-reported dental anxiety and mothers' proxy ratings of children's dental anxiety, which suggests that self-reported dental anxiety by children should be encouraged and adopted, and the mother's presence during dental visits is strongly recommended.