Intra-Articular Platelet-Rich Plasma Combined With Hyaluronic Acid Injection for Knee Osteoarthritis Is Superior to Platelet-Rich Plasma or Hyaluronic Acid Alone in Inhibiting Inflammation and Improving Pain and Function.

PURPOSE: To evaluate the effectiveness and explore the therapeutic mechanisms of platelet-rich plasma (PRP) combined with hyaluronic acid (HA) as a treatment for knee osteoarthritis (KOA). METHODS: In total, 122 knees were randomly divided into HA (34 knees), PRP (40 knees), and PRP+HA (48 knees) groups. Platelet densities in whole blood and PRP were examined using Wright-Giemsa staining. Visual analogue scale, Lequesne, Western Ontario and McMaster Universities Osteoarthritis Index, Lysholm scores, and postoperative complications were evaluated. High-frequency color Doppler imaging was used to observe the synovium and cartilage. Enzyme-linked immunosorbent assays were used to quantify interleukin-1ß, tumor necrosis factor-α, matrix metalloproteinase-3, and tissue inhibitor of metalloproteinase-1 levels in synovial fluid. RESULTS: The platelet density in PRP was 5.13-times that in whole blood (P = .002). At 24 months, pain and function scores in the PRP+HA group were better than those in the HA-alone and PRP-alone groups (Ppain = .000; Pfunction = .000). At 6 and 12 months, synovial hyperplasia in the PRP and PRP+HA groups was improved (P < .05). After 6 and 12 months, the synovial peak systolic velocity, synovial end-diastolic velocity, systolic/diastolic ratio, and resistance index were improved in the PRP+HA group (P < .05). Complications were greatest in the PRP group (P = .008). After 6 and 12 months, interleukin-1ß, tumor necrosis factor-α, matrix metalloproteinase-3, and tissue inhibitor of metalloproteinase-1 in the PRP and PRP+HA groups decreased (P < .05), with more apparent inhibition in the PRP+HA group (P < .05). CONCLUSIONS: PRP combined with HA is more effective than PRP or HA alone at inhibiting synovial inflammation and can effectively improve pain and function and reduce adverse reactions. Its mechanism involves changes in the synovium and cytokine content. LEVEL OF EVIDENCE: Level II, Prospective cohort study.

Osteoblastic differentiation of stem cells induced by graphene oxide-hydroxyapatite-alginate hydrogel composites and construction of tissue-engineered bone.

This study aimed to investigate the effect of graphene oxide (GO)-hydroxyapatite (HA)-sodium alginate (SA) composite application in the field of bone tissue engineering. Four scaffold groups were established (SA-HA, SA-HA-0.8%GO, SA-HA-1.0%GO and SA-HA-1.2%GO) and mixed with bone marrow mesenchymal stem cells (BMSCs). Hydrogel viscosity was measured at room temperature, and after freeze-drying and Fourier-transform infrared spectroscopy (FTIR) and X-ray diffraction (XRD) to detect substance crystallinity, the printability of each hydrogel type was measured with a printing grid. Scanning electron microscopy (SEM) was used to observe the internal microstructure of the scaffolds and to evaluate the growth and proliferation of cells on the scaffold. A hollow cylinder was printed to compare the forming effect of the hydrogel bioinks, and cell-hydrogel composites were implanted under the skin of nude mice to observe the effect of the hydrogels on osteogenesis in vivo. Increased GO concentrations led to reduced scaffold degradation rates, increased viscosity, increased printability, increased mechanical properties, increased scaffold porosity and increased cell proliferation rates. In vivo experiments showed that hematoxylin and eosin (HE) staining, Alizarin red staining, alkaline phosphatase staining and collagen type I immunohistochemical staining increased as the implantation time increased. These results demonstrate that GO composites have high printability as bioinks and can be used for bioprinting of bone by altering the ratio of the different components.

Development and Application of a No-Clog Surgical Suction Tip Using 3D Printing Technology.

BACKGROUND Clogging of the suction tip frequently occurs during orthopedic surgery. We developed a novel anti-clog suction tip using 3D printing technology to improve orthopedic surgery efficiency. MATERIAL AND METHODS We studied the root causes of obstructions in suction tips currently employed in orthopedic surgery during actual surgical cases. CAD software and 3D printer was used to design, modify, and print the novel suction tip. The frequency of clogging, the frequency of replacement of the suction tip, the time lost in replacing suction tips or connecting tubes, surgical duration, intraoperative surgical blood loss, and the satisfaction scores for the suction tips as rated by the surgeons were compared between the novel suction tip and the conventional suction tip. Comparisons were made first in laboratory experiments using a simulant liquid and then during total hip replacement surgeries. RESULTS In the simulant liquid experiments, the novel suction tips showed significantly reduced frequency of complete clogging and decreased time spent removing all fluid in comparison to the conventional suction tips (p<0.05). In the clinical trials, the novel suction tips exhibited significantly reduced frequency of complete clogging, shorter surgical duration, and reduced intraoperative surgical blood loss compared to the conventional suction tips (p<0.05). Surgeon satisfaction scores were higher for the novel tips than for the conventional tips (p<0.05). CONCLUSIONS Our surgeon-designed and -produced surgical suction tip utilizing 3D desktop printing technology was highly effective in resolving the problem of clogged suction tips during orthopedic surgery and yielded high surgeon satisfaction.

Bio-electrospraying is a safe technology for delivering human adipose-derived stem cells.

Bio-electrospraying (BES) is a technique for directly jetting living cells that has significant implications for tissue engineering and regenerative medicine. However, the effect of BES on human adipose-derived stem cells (hASCs) remains unknown. Here, we show that an hASC suspension was successfully electrosprayed via a continuous, stable and linearly directed electrospray at 10 kV and at 3 ml/h. Morphological observations and Trypan Blue and CCK-8 assays revealed that the cells remained viable and proliferated at a rate similar to that of the controls (0 kV). However, at 20 kV, BES became unstable and cell viability was reduced. Moreover, hASCs electrosprayed at 10 kV retained their multilineage potential, successfully differentiating into chondrogenic, osteogenic and neurogenic lineages. Thus, BES does not significantly affect cell morphology, viability or multipotency.

DCBLD1 Overexpression Is Associated With a Poor Prognosis in Head and Neck Squamous Cell Carcinoma.

Background: DCBLD1 is highly expressed in several kinds of cancer and plays a potential prognostic factor. However, the prognostic value and immune infiltration in head and neck squamous cell carcinoma remain unclear and need further research. Materials and Methods: DCBLD1 expression and clinical information were obtained from the Cancer Genome Atlas (TCGA) database. The mRNA level in cell lines (SCC25 and CAL27) and gingival fibroblasts were detected using quantitative PCR. Cox regression analysis was used to evaluate the prognostic values of DCBLD1 and clinical data in HNSCC. A nomogram was also established to predict the impact of DCBLD1 on prognosis based on Cox multivariate results. The methylation level of DCBLD1 in HNSC and its prognosis were analyzed in UALACN and MethSurv. Finally, the potential biological functions of DCBLD1 were investigated using gene set enrichment analysis (GSEA) and single-sample GSEA (ssGSEA). Results: The mRNA and protein expression levels of DCBLD1 were highly expressed in HNSCC tissue and cell lines. The Cox analyses demonstrate that highly expressed DCBLD1 is an independent prognosis marker (p < 0.05). ROC curve analysis showed the performance of DCBLD1 (area under the ROC curve: 0.948, sensitivity: 93.2%, specificity: 84.7%). The methylation was increased in HNSCC patients compared with normal subjects (p < 0.05) and was associated with poor prognosis at sites cg27642470 and cg21104965. Additionally, DCBLD1 expression is poorly associated with immune cell infiltration and immunological checkpoints PD-L1 and TIM-3. Conclusion: In head and neck squamous cell carcinoma, DCBLD1 is overexpressed, associated with poor patient prognosis. The detailed underlying mechanism merits further research.

Corrigendum: DCBLD1 Overexpression is associated with a poor prognosis in head and neck squamous cell carcinoma.

[This corrects the article DOI: 10.3389/fimmu.2022.939344.].

Clean version: Electrospun fibrinogen scaffolds from discarded blood for wound healing.

Immediate reutilization of discarded blood from surgery has not received much attention, leading to the waste of a large amount of autologous blood. We used a concentration gradient and high-voltage electrospinning technology to immediately prepare a scaffold material with high biological activity but without immunogenicity from autologous waste blood collected during surgery. Here, we fabricated and characterized a 90 mg/mL group, 110 mg/mL group, and 130 mg/mL group of fibrinogen (FBG) scaffolds. Analyses revealed that the FBG scaffolds had good film-forming properties and a clear fiber structure. in vitro cell viability experiments confirmed that the cells showed an increasing trend with increasing FBG concentrations. The cells grew well in the scaffold material and secreted more cell matrix. When human bone mesenchymal stem cells (hBMSCs) were cocultured with the scaffold material, the hBMSCs expressed osteogenic and chondrogenic biomarkers. The cellular scaffold complexes from the 3 groups were implanted in four full-thickness round wounds (Φ12 mm) on the dorsal back of each rat, the 130 mg/mL group showed a 90% reduction in wound size and the data compared to other groups were better at 14 day. These results suggest that electrospinning technology-based FBG scaffold materials derived from autologous waste blood may become an ideal tissue engineering scaffold and can be immediately used for autologous hemostasis, anti-adhesion films, and wound dressing in surgery.

Agarose composite hydrogel and PVA sacrificial materials for bioprinting large-scale, personalized face-like with nutrient networks.

Large, deep, complex, and severe tissue defects and deformities of the face are the problems encountered in clinical practice. Autologous tissue reconstruction or allograft face transplantation has been adopted but has problems such as blood supply difficulties, collateral damage, immune rejection, and ethical disputes. 3D bioprinting enables personalized tissue regeneration. However, simple hydrogels are prone to collapse during printing, are limited in size, and have poor shape and structure. The present study used three polysaccharide hydrogel composites of nanocellulose, agarose, and sodium alginate with seeded cells as bioinks and polyvinyl alcohol (PVA) as sacrificial material to construct the structures that did not collapse (characteristic parts, such as lips and nose). The nutrient network gradually formed a blood vessel-like structure. The hydrogels prepared using these three polysaccharides have great potential in the construction of personalized, complex, and vascularized tissue-engineered anatomical faces and provide a new strategy for autologous full face reconstruction.

Fabrication of multilayered electrospun poly(lactic-co-glycolic acid)/polyvinyl pyrrolidone + poly(ethylene oxide) scaffolds and biocompatibility evaluation.

Poly(lactic-co-glycolic acid)/polyvinyl pyrrolidone + poly(ethylene oxide) [PLGA/(PVP + PEO)] scaffolds with different polymer concentrations were fabricated using multilayered electrospinning, and their physicochemical properties and biocompatibility were examined to screen for scaffolds with excellent performance in tissue engineering (TE). PLGA solution (15% w/v) was used as the bottom solution, and a mixed solution of 12% w/v PVP + PEO was applied as the surface layer solution. The mass ratios of PVP vs. PEO in each 10 ml surface layer mixed solution were 1.08 g: 0.12 g; 0.96 g: 0.24 g; and 0.84 g: 0.36 g. Compared to the conventional electrospinning method used to fabricate the pure PVP + PEO (0.96 g: 0.24 g, Group A) scaffold and pure PLGA (Group E) scaffold, the multilayer electrospinning technique of alternating sprays of the bottom layer solution and the surface layer solution was adopted to fabricate multilayer nanofiber scaffolds, including PLGA/(PVP + PEO) (1.08 g: 0.12 g, Group B), PLGA/(PVP + PEO) (0.96 g: 0.24 g, Group C), and PLGA/(PVP + PEO) (0.84 g: 0.36 g, Group D). The morphology and characteristics of the five scaffolds were analyzed, and the biocompatibilities of the cell-scaffold composites were assessed through methods including Cell Counting Kit-8 (CCK8) analysis, 4',6-diamidino-2-phenylindole (DAPI) staining, and scanning electron microscopy. Therefore, with a PVP-to-PEO mass ratio of 0.96 g: 0.24 g, an optimal multilayer nanofiber scaffold was fabricated by the multilayer electrospinning technique. The excellent biocompatibility and mechanical properties of the scaffold were confirmed by in vitro experiments, which demonstrated the scaffold's promising application potential in the field of TE.

[Study on the correlation between the mandibular masticatory muscle movement and sleep tooth wear].

OBJECTIVE: To investigate the correlation between the clinical diagnostic criteria of sleep bruxism and the frequency of mandibular movements during sleep. METHODS: Video polysomnography was used to record 20 healthy adults with at least one of the following clinical symptoms and signs: 1) report of frequent tooth grinding; 2) tooth wear and dentin exposure with at least three occlusal surfaces; 3) masticatory muscle symptoms in the morning; 4) masseter muscle hypertrophy. The rhythmic masticatory muscle activity (RMMA) and isolated tonic activity were scored to compare the correlations with clinical symptoms and signs. Finally, the incidence of temporomandibular disorders (TMD) was investigated in patients with isolated tonic and RMMA subjects. RESULTS: Among the 20 subjects, RMMA events were observed (5.8±3.1) times·h⁻¹ and isolated tonic episodes were observed (2.1±0.9) times·h⁻¹. The frequency of RMMA events was significantly greater in the patients with acoustic molars than in those without (P<0.05). Similarly, the frequency of RMMA events was significantly greater in the patients with tooth attrition than in those without (P<0.05). However, no difference was observed between the occurrence of RMMA and the symptoms of masticatory muscles or masseter hypertrophy in the morning. The incidence of TMD was significantly higher in the patients with RMMA than in the isolated tonic patients. CONCLUSIONS: The clinical symptoms and signs often used to diagnose sleep bruxism are different clinical and physiological mandibular movements during sleep. RMMA during sleep can reflect the occurrence of tooth attrition and the high risk of TMD.

DLX2 activates Wnt1 transcription and mediates Wnt/ß-catenin signal to promote osteogenic differentiation of hBMSCs.

OBJECTIVES: The molocular mechanism underlying human bone marrow mesenchymal stem cells (hBMSCs) differentiation remains to be further elucidated. DLX2 has been confirmed to accelerate osteogenic differentiation which is one member of Distal-less family genes. However, how DLX2 regulates in osteogenic differentiation is still unclear. METHODS: The hBMSCs were isolated and identified by the antigen CD29, CD4, CD90 through flow cytometry. DLX2 expression level, molecules related signaling pathways and transcriptional markers in osteogenesis were examined by western blot and real time-PCR. Osteogenic state was weighed by the ALP Detection Kit and Alizarin red S staining. The combination between DLX2 and WNT1 was detected by Chromatin immunoprecipitation (CHIP) assay. RESULTS: The results showed that in the process of osteoblast differentiation, DLX2 was up-regulated accompanied with osteogenic transcriptional factor. DLX2 elevated cellular alkaline phosphatase activity, accelerated BMSC mineralization along with up-regulation of osteogenic-related gene expression. Besides, DLX2 is a transcription factor of WNT1, which activated the Wnt/ß-Catenin signaling pathway resulting in osteogenic differentiation. Whereas, the inhibitor of ß-Catenin FH535 restrained enhanced osteogenic capability induced by DLX2. CONCLUSIONS: Taken together, these results suggest that by up-regulation of Wnt/ß-Catenin signaling, DLX2 accelerated human osteogenic differentiation.

Biofabrication of valentine-shaped heart with a composite hydrogel and sacrificial material.

3D bioprinting represents a potential solution for organs regeneration, however, the production of complex tissues and organs that are in large size, randomly shaped, hollow, and contain integrated pre-vascularization still faces multiple challenges. This study aimed to test the feasibility of our 3D printing scheme for the manufacturing of micro-fluid channel networks complex three-dimensional tissue structures. The reverse engineering software was used to design the CAD model and polyvinyl alcohol (PVA) was used as the sacrificial material to print the sacrificial stent use the bioprinter nozzle 1. Hydrogel composite H9c2 and human umbilical vein endothelial cells (HUVECs) were mixed with sodium alginate, agarose solution and platelet-rich plasma (PRP) as cellular bioink, which was extruded through nozzle 2 to deposit the internal pores of the sacrificial scaffold. The scaffold dissolved, change to a flexible, hollow and micro-fluid channel networks complex structure. The 3D-bioprinting technology can construct a micro-fluid channel networks valentine heart with a self-defined height and hollow in suitable mechanical properties. The cells proliferate and maintain their biological properties within the printed constructs. This study demonstrates that valentine heart-like constructs can be fabricated with 3D bioprinting using sacrificial and hydrogel materials.

Preparation of P3HB4HB/(Gelatin + PVA) Composite Scaffolds by Coaxial Electrospinning and Its Biocompatibility Evaluation.

This study was conducted to prepare coaxial electrospun scaffolds of P3HB4HB/(gelatin + PVA) with various concentration ratios with P3HB4HB as the core solution and gelatin + PVA mixture as the shell solution; the mass ratios of gelatin and PVA in each 10 mL shell mixture were 0.6 g : 0.2 g (Group A), 0.4 g : 0.4 g (Group B), and 0.2 g : 0.6 g (Group C). The results showed that the pore size, porosity, and cell proliferation rate of Group C were better than those of Groups A and B. The ascending order of the tensile strength and modulus of elasticity was Group A < Group B < Group C. The surface roughness was Group C > Group B > Group A. The osteogenic and chondrogenic-specific staining showed that Group C was stronger than Groups A and B. This study demonstrates that when the mass ratio of gelatin : PVA was 0.2 g : 0.6 g, a P3HB4HB/(gelatin + PVA) composite scaffold with a core-shell structure can be prepared, and the scaffold has good biocompatibility that it may be an ideal scaffold for tissue engineering.

PHB/PHBHHx scaffolds and human adipose-derived stem cells for cartilage tissue engineering.

The goal of this study was to investigate the potential of polyhydroxybutyrate (PHB)/poly(hydroxybutyrate-co-hydroxyhexanoate) (PHBHHx) (PHB/PHBHHx) to produce neocartilage upon seeding with differentiated human adipose-derived stem cells (hASCs). hASCs were grown on a three-dimensional PHB/PHBHHx scaffold in vitro with or without chondrogenic media for 14 days. Scanning electron microscopy showed that differentiated cells produced abundant extracellular matrices with increasing culture time. No cytotoxicity was observed by the live/dead cell viability assay. GAG and total collagen content in the differentiated cells increased significantly with in vitro culture time. After 14 days of in vitro culture, the differentiated cells grown on the (PHB/PHBHHx) scaffold (differentiated cells/(PHB/PHBHHx)) were implanted into the subcutaneous layer nude mice for 12 or 24 weeks, non-differentiated cells/(PHB/PHBHHx) were implanted as the control group. The differentiated cells/(PHB/PHBHHx) implants formed cartilage-like tissue after 24 weeks of implantation, and stained positive for collagen type II, safranin O, and toluidine blue. In addition, typical cartilage lacuna was observed, and there were no remnants of PHB/PHBHHx. Collagen type II was detected by Western blot at 12 and 24 weeks of implantation. In the control group, no cartilage formation was observed. This study demonstrated that PHB/PHBHHx is a suitable material for cartilage tissue engineering.