Ethanolic extract from Sophora moorcroftiana inhibit cell proliferation and alter the mechanical properties of human cervical cancer.

BACKGROUND: Cervical cancer is one of the most common gynecological malignancies. Previous studies have shown that the ethanol extract of Sophora moorcroftiana seeds (EESMS) possesses an antiproliferative effect on several tumors in vitro. Therefore, in this study, we assessed the impact of EESMS on human cervical carcinoma (HeLa) cell proliferation. METHODS: The proliferation and apoptotic effects of HeLa cells treated with EESMS were evaluated using 3-[4,5-dimethylthiazol-2-yl]-2,5 diphenyl tetrazolium bromide assay, dual acridine orange/ethidium bromide double staining, flow cytometry, and western blotting. Single-cell level atomic force microscopy (AFM) was conducted to detect the mechanical properties of HeLa cells, and proteomics and bioinformatics methods were used to elucidate the molecular mechanisms of EESMS. RESULTS: EESMS treatment inhibited HeLa cell proliferation by blocking the G0/G1 phase, increasing the expression of Caspase-3 and affecting its mechanical properties, and the EESMS indicated no significant inhibitory effect on mouse fibroblasts L929 cell line. In total, 218 differentially expressed proteins were identified using two-dimensional electrophoresis, and eight differentially expressed proteins were successfully identified using matrix-assisted laser desorption/ionization-time-of-flight mass spectrometry. The differentially expressed proteins were involved in various cellular and biological processes. CONCLUSION: This study provides a perspective on how cells change through biomechanics and a further theoretical foundation for the future application of Sophora moorcroftiana as a novel low-toxicity chemotherapy medication for treating human cervical cancer.

In vitro differentiation of human induced pluripotent stem cells into temporomandibular joint disc like cells.

Temporomandibular joint discs (TMJ discs) are unable to repair themselves in disease states, while induced stem cell differentiation is a common method to repair tissue defects. Nowadays, kinds of stem cells are attempted for tissue regeneration of TMJ disc, but these methods have several downsides, which limit their wide application. The proliferation and differentiation ability of human induced pluripotent stem cells (hiPSC) provides a new research direction for TMJ disc tissue regeneration. In this study, we investigated the feasibility of induced differentiation of hiPSC into TMJ disc cells in vitro and the differentiation efficiency of different methods to clarify the possibility and conditions of hiPSC application in TMJ disc tissue engineering. We collected sheep TMJ disc cells cultures for adding in hiPSC culture environment and treated hiPSC by both direct induction and Transwell co-culture for 7 days, 14 days and 21 days. The secretion of extracellular matrix in TMJ disc cells was detected by Sirius Red and Safranin O staining. Collagen Ⅰ and Collagen Ⅱ were qualitatively detected by immunohistochemical staining. The expression of extracellular matrix genes (type I collagen (COL1A1), type II collagen(COL2), glycosaminoglycan (GAG)), chondrogenic differentiation gene SOX9 and pluripotency gene OCT4 were detected by RT-qPCR. Our results showed that hiPSC had the ability to differentiate to TMJ disc cells by direct induction in TMJ disc cell culture medium and by Transwell co-culture method. The highest degree of differentiation was observed after 14 days of direct induction, while Transwell co-culture showed significant differentiation at different times and with different major directions. Meanwhile, Transwell co-culture not only differentiates hiPSC but also promotes the growth and proliferation of TMJ disc cells. Our study is valuable to investigate the possibility of differentiation of hiPSC toward TMJ disc cells and to determine the time of differentiation. It provides new ideas for the selection of seed cells for TMJ disc tissue engineering.

Comparative effectiveness of hyaluronic acid, platelet-rich plasma, and platelet-rich fibrin in treating temporomandibular disorders: a systematic review and network meta-analysis.

OBJECTIVE: This study aims to compare the efficacy of intra-articular injections of hyaluronic acid (HA), platelet-rich plasma (PRP), and platelet-rich fibrin (PRF) for treating temporomandibular disorders (TMDs) and summarize their mechanisms of action. METHODS: Randomized controlled trials (RCTs) published until November 13, 2021, were identified using electronic and manual searches. Each study was evaluated for the risk of bias using the Cochrane risk of bias tool. The studies found via searches were categorized by follow-up time (1, 3, or 6 months). Evidence quality was graded according to the GRADE system. RESULTS: Twelve RCTs were included that involved 421 patients with TMD. The network meta-analysis showed that all treatment groups improved compared to the placebo groups in terms of pain and maximal mouth opening (MMO). For pain evaluated via the visual analog scale, PRF exhibited better analgesic effects than PRP or HA after 1 and 3 months. PRP appeared to be more effective than PRF was after 6 months but there were no statistically significant differences between the two. For MMO, the effect of PRP was superior to those of PRF and HA after 1 month. However, after 3 and 6 months, PRF provided more encouraging results in improving MMO. CONCLUSION: PRP and PRF exhibited similar short-term efficacy in treating TMD, while PRF was more advantageous in terms of long-term efficacy. Therefore, PRF was recommended for treating TMD.

Effects of glucose concentration and oxygen partial pressure on the respiratory metabolism of sheep temporomandibular joint disc cells.

Temporomandibular joint (TMJ) disc degeneration is a common disease characterized by a decrease in metabolic function. The present study aimed to investigate the pathogenesis of TMJ disc degeneration by analyzing the effects of oxygen and glucose concentrations on metabolism in a simulated TMJ disc cell growth environment. Cell samples were divided into 10 groups and cultured in different nutritional environments, including 21 and 2% O2 partial pressures and various glucose concentrations (0, 0.5, 3, 5.5 and 22.5 mmol/l). Cell proliferation, extracellular matrix content, mitochondrial function, and cell metabolism were subsequently analyzed. The results demonstrated that hypoxia and a low glucose concentration inhibited cell growth, and low glucose concentration inhibited extracellular matrix synthesis and adenosine 5'-monophosphate-activated protein kinase expression. Hypoxic conditions also induced a compensatory increase in the number of mitochondria, whereas mitochondrial deformation and swelling were observed in the absence of glucose. According to this study, the primary metabolic pathway of TMJ disc cells is glycolysis. It was concluded that hypoxic conditions and normal glucose concentrations are needed for the growth of TMJ disc cells. Glucose is necessary to ensure cell survival, extracellular matrix synthesis and mitochondrial function. Glucose deficiency may be related to disc degeneration, aging and disease mechanisms.

Comparative effectiveness of low-level laser therapy with different wavelengths and transcutaneous electric nerve stimulation in the treatment of pain caused by temporomandibular disorders: A systematic review and network meta-analysis.

OBJECTIVE: To assess the efficacy of low-level laser therapy (LLLT) with different wavelengths and transcutaneous electric nerve stimulation (TENS) and explore the optimal wavelength range of laser application in the treatment of pain caused by temporomandibular disorders (TMD). METHODS: An electronic search on PubMed, Cochrane Library, Embase, Scopus and Web of Science was undertaken to identify the randomised clinical trials (RCTs) published from database inception to 16 April 2021, aiming to compare the effects of LLLT with different wavelengths (632.8-672 nm, 780-904 nm, and 910-1100 nm) or TENS or placebo group on TMD patients pain reduction. In addition, manual search of the studies was performed. The reviewers assessed the risk of bias of individual studies with the Cochrane risk of bias tool and excluded the RCTs with a high risk of bias in any field. Meanwhile, the reviewers, after performing the network meta-analysis, assessed the quality of evidence, which contributed to network estimate via the GRADE framework. RESULTS: Twenty-seven RCTs with 969 patients with TMD were included. In the meta-analysis, all treatment groups showed an overall improvement in pain scores, when compared with the placebo group. LLLT with wavelength ranging from 910 nm to 1100 nm produced more pain relief in the visual analogue scale (VAS) immediately after treatment [mean difference (MD) = 4.68, 95% confidence interval (CI):(3.08,6.28)]. After one-month follow-up, LLLT with wavelength ranging from 910 nm to 1100 nm also showed superior pain-relieving effects [MD = 3.61, 95% CI: (-1.77, 8.99)]. However, no significant difference was observed. Based on the SUCRA ranking, L3 ranked first immediately after treatment and 1 month later. CONCLUSION: The results of the meta-analysis showed the LLLT had better short-term efficacy than TENS in the treatment of pain caused by TMD. Better results can be achieved with higher wavelengths. Therefore, we recommended to treat TMD using LLLT with wavelength ranging from 910 nm to 1100 nm.

Restoration of the dentition in a patient with a history of bruxism and amelogenesis imperfecta: A clinical report.

The report describes a 27-year-old female patient with amelogenesis imperfecta (AI) accompanied by nocturnal bruxism, who was treated with a combination of occlusal splint and full-mouth fixed prosthetic rehabilitation through follow-ups within 2 years. Soft splint protection, regular follow-up, and monitoring of carries are guaranteed to maintain a long-term curative effect.

Effects of different energy density low-level laser therapies for temporomandibular joint disorders patients: a systematic review and network meta-analysis of parallel randomized controlled trials.

PURPOSE: To compare the effects of different energy density LLLT on TMD patients' pain reduction. METHODS: Reviewers searched RCTs published in Embase, PubMed and The Cochrane Library before 1 April 2020. Reviewers included parallelRCTs that compared the effects of LLLT with placebo or different energy density LLLT for TMD patients' pain reduction. Reviewers did network meta-analysis and evaluated the quality of evidence using the GRADE process. RESULTS: From 352 studies, reviewers included 16 RCTs. Immediately after treatment, the d1 laser therapy (energy density ranging from 0 to 10 J/cm2) ranked first. The d1 laser therapy showed more pain reduction than placebo ( MD = 2.49, 95% CI ranging from 1.28 to 3.71). The quality of "d1 vs p" comparison was assessed as "moderate" quality. A month after treatment, the d1 laser therapy also performed better than placebo (MD = 1.69, 95%CI = [-0.78, 4.16]). The quality of secondary outcome was assessed as "low" quality. CONCLUSIONS: For clinical application, d1 laser therapy (energy density ranging from 0 to 10 J/cm2) is recommended for short-term pain management of TMD patients (moderate quality evidence). A month after treatment, the d1 laser therapy also performed better than placebo and other laser groups but the result didn't reach the point of statistical significance (low quality evidence). TRIAL REGISTRATION: PROSPERO-CRD42018118313.

Nanomorphological and mechanical reconstruction of mesenchymal stem cells during early apoptosis detected by atomic force microscopy.

Stem cell apoptosis exists widely in embryonic development, tissue regeneration, repair, aging and pathophysiology of disease. The molecular mechanism of stem cell apoptosis has been extensively investigated. However, alterations in biomechanics and nanomorphology have rarely been studied. Therefore, an apoptosis model was established for bone marrow mesenchymal stem cells (BMSCs) and the reconstruction of the mechanical properties and nanomorphology of the cells were investigated in detail. Atomic force microscopy (AFM), scanning electron microscopy (SEM), laser scanning confocal microscopy (LSCM), flow cytometry and Cell Counting Kit-8 analysis were applied to assess the cellular elasticity modulus, geometry, nanomorphology, cell surface ultrastructure, biological viability and early apoptotic signals (phosphatidylserine, PS). The results indicated that the cellular elastic modulus and volume significantly decreased, whereas the cell surface roughness obviously increased during the first 3 h of cytochalasin B (CB) treatment. Moreover, these alterations preceded the exposure of biological apoptotic signal PS. These findings suggested that cellular mechanical damage is connected with the apoptosis of BMSCs, and the alterations in mechanics and nanomorphology may be a sensitive index to detect alterations in cell viability during apoptosis. The results contribute to further understanding of apoptosis from the perspective of cell mechanics.

Effects of dynamic radial tensile stress on fibrocartilage differentiation of bone marrow mesenchymal stem cells.

BACKGROUND: Uniaxial/biaxial tensile stress has been employed to induce chondrocyte differentiation of mesenchymal stem cells. However, the effects of radial tensile stimuli on differentiation of MSCs into fibrocartilage remain unclear. RESULTS: It was found that induced bone marrow mesenchymal stem cells (BMSCs) were not only similar to TMJ disc cells in morphology, but also could synthesize type I collagen (Col I), a small amount of type II collagen (Col II) and glycosaminoglycans (GAGs). The synthesis of Col I significantly increased while that of Col II gradually decreased with increasing tensile strength. The ratio of Col I to Col II was 1.8 to 1 and 2 to 1 in the 10% and 15% stretching groups, respectively. The gene expression of Col I and GAGs was significantly upregulated, whereas that of Col II was downregulated. However, the higher tensile stimulation (15%) promoted the synthesis of α-smooth muscle actin (α-SMA). Too much α-SMA is not conducive to constructing engineered tissue. CONCLUSION: Therefore, the 10% radial tensile stimulus was the optimal strength for inducing the BMSCs to differentiate into fibrochondrocytes of the temporomandibular joint (TMJ) disc. This work provided a novel approach for inducing BMSCs to differentiate into fibrochondrocytes.

A novel zinc oxide eugenol modified by polyhexamethylene biguanide: Physical and antimicrobial properties.

This study was to prepare and screen a novel root canal sealing agent modified by polyhexamethylene biguanide (PHMB) that was in accordance with the ISO 6876:2001 standard and to study its physical and antimicrobial properties. The modified sealers were produced by mixing a certain amount of zinc oxide with eugenol containing different concentrations of PHMB (0.05, 0.1, 0.2, 0.4, 0.6 and 0.8%) at a ratio of 1:1 (w/v). The setting time, flow, film thickness, solubility and dimensional change after solidifying were assessed to screen out the modified sealing agents that the physical properties met the mentioned standards. The modified direct contact test (DCT) was used to evaluate the antimicrobial activity against Enterococcus faecalis. The results suggested that when the concentrations of PHMB were 0.05, 0.1 and 0.2%, the modified root canal sealers showed the best performance in physical and antimicrobial properties.

[Effects of staurosporine on the contraction of self-assembled constructs of goat temporomandibular joint disc cells].

OBJECTIVE: The effects of the staurosporine on contraction of self-assembled constructs and extracellular matrix syntheses of goat temporomandibular joint discs were investigated. METHODS: Goat temporomandibular joint disc cells were isolated and cultured to P3, and 5.5×106 cells were combined with different concentrations of staurosporine (0, 0.1, 1, 10, 100 nmol·L⁻¹) in agarose wells and cultured for one week. The samples were frozen and sectioned. Safranin-O,  Picro-sirius red and immunohistochemical staining were performed to observe the distributions of the extracellular matrix and the expression of alpha-smooth muscle actin (α-SMA). Enzyme linked immunosorbent assay (ELISA) and Blyscan kits were utilized to quan--titatively detect the contents of type Ⅰ collagen (ColⅠ) and glycosaminoglycans (GAGs). RESULTS: Each group of goat temporo-mandibular joint disc cells in the agarose wells were gathered to self-assemble into a disc-shaped base for 4 hours and then to gradually contract into a round shape. The Picro-sirius red staining was strong and indicated collagen distribution. The Safranin-O staining observed GAGs throughout the entire construct. The expression of ColⅠ was strongly posi-tive in the staurosporine groups; however, the expression of α-SMA was weak. ColⅠ and GAGs contents in the stau-rosporine groups were greater than that of the control group, especially in the 10 nmol·L⁻¹ group (P<0.01). CONCLUSIONS: Staurosporine has a certain effect on the shrinkage of self-assembled constructs; however, such effect is not prominent. Staurosporine contributes to the construction synthesis of extracellular matrix.

Synthesizing Functional Biomacromolecular Wet Adhesives with Typical Gel-Sol Transition and Shear-Thinning Features.

Functional wet adhesives have become increasingly important in the field of biomedicine. Herein, modified chitosan (CS) biomacromolecule is synthesized as tissue wet adhesives and hemostatic materials. In a typical case, after getting through two steps of organic reactions, catechol and lysine groups are grafted onto the chitosan backbone, enabling for the successful preparation of one kind of novel biomacromolecule of chitosan-catechol-lysine (CHIC-Lys). The as-prepared CHIC-Lys biomacromolecule shows improved wet adhesion strength, concentration-dependent gel-sol transition feature when shear stress is cycled between low stress (0.2 Pa) and high stress (2 Pa) at room temperature, and obvious shear-thinning feature under wide concentration ranges, along with good biocompatibility, comparable with traditional CS. Based on these obvious characteristics, CHIC-Lys is successfully coated onto the surface of a syringe needle and the decorated needle shows considerable hemostatic effect in a test of rats venous bleeding. Overall, the as-synthesized CHIC-Lys biomacromolecule exhibits considerable application potential in the biomedical field.

Mechanical, nanomorphological and biological reconstruction of early­stage apoptosis in HeLa cells induced by cytochalasin B.

There is a growing interest in the fact that mechanical signals may be as important as biological signals in evaluating cell viability. To investigate the alterations in biomechanics, nanomorphology and biological apoptotic signals during early apoptosis, an apoptosis model was established for cervical cancer HeLa cells induced by cytochalasin B (CB). The cellular mechanical properties, geometry, morphology and expression of key apoptotic proteins were systematically analyzed. The findings indicated a marked decline in cellular elastic modulus and volume and a considerable increase in surface roughness occurring prior to the activation of biological apoptosis signals (such as phosphatidylserine exposure or activation of CD95/Fas). Moreover, the depolymerization of filamentous actin aggravated the intracellular crowding degree, which induced the redistribution of different­sized protein molecules and protrusions across the cell membrane arising from excluded volume interactions. Statistical analysis revealed that the disassembly of the actin cytoskeleton was negatively correlated with the cellular elastic modulus and volume, but was positively correlated with surface roughness and CD95/Fas activation. The results of the present study suggest that compared with biological signals, mechanical and geometrical reconstruction is more sensitive during apoptosis and the increase in cell surface roughness arises from the redistribution of biophysical molecules. These results contribute to our in­depth understanding of the apoptosis mechanisms of cancer cells mediated by cytochalasin B.

[Topography and mechanical property of goat temporomandibular joint disc cells].

OBJECTIVE: This study is performed to investigate the cell topographies and biomechanical properties of two different types of temporomandibular joint (TMJ) discs from goats by using JPK Nano Wizard 3 biological atomic force microscopy (AFM). This process provides a guideline for selecting seed cells for TMJ disc tissue engineering. METHODS: TMJ disc cells from primary goats were cultured by monolayer culture method. AFM was used to contact scan the topographies of the two types of TMJ disc cells under physiological environment. Approximately 20 chondrocyte-like and fibroblast-like cells were selected randomly to plot the force-versus-distance curves of the cytoplasm and nucleus. Young's modulus and adhesion were analyzed by JPK Data Processing. RESULTS: The triangle-shapednucleus of the chondrocyte-like cell occupied a large portion of the cell. Cytoskeleton was arranged dendritically on the surface. Pseudopodia were extended from cell edges. The spindle-shaped nucleus of the fibroblast-like cell occupied a significantly larger region compared with the cytoplasmic region. Cytoskeleton was arranged regularly. Cell edges were smooth with less pseudopodia extended. No difference was found in the surface roughness between the two types of cells. According to the force-versus-distance curves, the Young's moduli of the two types of cells were not statistically different (P>0.05), but differences were found in the cytoplasmic regions (P=0.047). No statistical difference was found in the adhesions between the two types of cells (P>0.05). CONCLUSION: The AFM topography and curves were compared and analyzed. The two types of TMJ disc cells exhibited significantly different topographies, but only slight difference in their mechanical abilities.

Tissue engineering of the temporomandibular joint disc: current status and future trends.

INTRODUCTION: Temporomandibular joint disorders are extremely prevalent and there is no ideal treatment clinically for the moment. For severe cases, a discectomy often need to be performed, which will further result in the development of osteoarthritis. In the past thirty years, tissue engineering has provided a promising approach for the effective remedy of severe TMJ disease through the creation of viable, effective, and biological functional implants. METHODS: Although TMJ disc tissue engineering is still in early stage, unremitting efforts and some achievements have been made over the past decades. In this review, a comprehensive summary of the available literature on the progress and status in tissue engineering of the TMJ disc regarding cell sources, scaffolds, biochemical and biomechanical stimuli, and other prospects relative to this field is provided. RESULTS AND CONCLUSIONS: Even though research studies in this field are too few compared to other fibrocartilage (e.g., knee meniscus) and numerous, difficult tasks still exist, we believe that our ultimate goal of regenerating a biological implant whose histological, biochemical, and biomechanical properties parallel native TMJ discs for clinical therapy will be achieved in the near future.

[Effects of basic fibroblast growth factor on bone marrow mesenchymal stem cell differentiation into temporomandibular joint disc cells].

The present paper is aimed to observe the effects of basic fibroblast growth factor (bFGF) on bone marrow mesenchymal stem cell (BMSCs) differentiation. The bFGF was used to stimulate BMSCs and histology, immunohistochemistry and enzyme linked immunosorbent assay (ELISA) were used to examine the extracellular matrix produced by induced BMSCs, evaluated the feasibility of BMSCs being the seeding cells of temporomandibular joint (TMJ) disc tissue engineering. The results showed that having been induced with bFGF, the BMSCs could differentiate into fibroblast-like cells, which could synthesize GAG and collagen type I matrix. So it is feasible for BMSCs as seeding cells for engineered TMJ disc.

[Research progress of anticarious Chinese herbal].

Caries is a chronicly infectious disease which happens to the hard tissue of the teeth and is the most common dental disease of human. Many scholars have devoted to research anticarigenics drugs and the Chinese herbal was considerably concerned. The paper briefly introduces kinds of anticarigenics Chinese herbals, such as Turkish galls, TFA and Magnolia.

[Influence on mechanical properties and microstructure of nano-zirconia toughened alumina ceramics with nano-zirconia content].

OBJECTIVE: To investigate the mechanical properties and microstructure of nano -zirconia toughened alumina ceramics with variety of nano-zirconia content in centrifugal infiltrate casting processing of dental all-ceramic. METHODS: Composite powder with different ethanol-water ratio, obtained serosity from ball milling and centrifugal infiltrate cast processing of green, then sintered at 1 450 degrees C for 8 h. The physical and mechanical properties of the sintered sample after milling and polishing were tested. Microstructures of the surface and fracture of the sintered sample were investigated by SEM. RESULTS: The experimental results showed that there had statistical significience (P < 0.01) on static three-point flexure strength and Vickers Hardness in three kinds of different nano-zirconia content sintered sample. Fracture toughness of 20% group was different from other two groups, while 10% group had not difference from 30% group (P < 0.05). The mechanical properties of this ceramic with 20% nano-zirconia was the best of the three, the static three-point flexure strength was (433 +/- 19) MPa and fracture toughness was (7.50 +/- 0.56) MPa x min 1/2. The intra/inter structure, fracture of intragranular and intergranular on the surface and fracture of sintered sample in microstrucre was also found. CONCLUSION: Intra/inter structure has strengthen toughness in ceramics. It has better toughness with 20% nano-zirconia, is suitable dental all-ceramic restoratives.