Stem Cells Translational Medicine
·
2021
·Vol. 10
·期 5
·781-796
·
IF 6.94
Xu Haixia,Wang Chengqiang,Liu Chun,Peng Ziyue,Li Jianjun,Jin Yanglei,Wang Yihan,Guo Jiasong,Zhu Lixin
Steroid-induced osteonecrosis of the femoral head (ONFH) is characterized by decreased osteogenesis, angiogenesis, and increased adipogenesis. While bone tissue engineering has been widely investigated to treat ONFH, its therapeutic effects remain unsatisfactory. Therefore, further studies are required to determine optimal osteogenesis, angiogenesis and adipogenesis in the necrotic area of the femoral head. In our study, we developed a carboxymethyl chitosan/alginate/bone marrow mesenchymal stem cell/endothelial progenitor cell (CMC/ALG/BMSC/EPC) composite implant, and evaluated its ability to repair steroid-induced ONFH. Our in vitro studies showed that BMSC and EPC coculture displayed enhanced osteogenic and angiogenic differentiation. When compared with single BMSC cultures, adipogenic differentiation in coculture systems was reduced. We also fabricated a three-dimensional (3D) CMC/ALG scaffold for loading cells, using a lyophilization approach, and confirmed its good cell compatibility characteristics, that is, high porosity, low cytotoxicity and favorable cell adhesion. 3D coculture of BMSCs and EPCs also promoted secretion of osteogenic and angiogenic factors. Then, we established an rabbit model of steroid-induced ONFH. The CMC/ALG/BMSC/EPC composite implant was transplanted into the bone tunnel of the rabbit femoral head after core decompression (CD) surgery. Twelve weeks later, radiographical and histological analyses revealed CMC/ALG/BMSC/EPC composite implants had facilitated the repair of steroid-induced ONFH, by promoting osteogenesis and angiogenesis, and reducing adipogenesis when compared with CD, CMC/ALG, CMC/ALG/BMSC and CMC/ALG/EPC groups. Thus, our data show that cotransplantation of BMSCs and EPCs in 3D scaffolds is beneficial in treating steroid-induced ONFH.
研究领域:
组织工程
·
骨科
·
再生医学
✓ 已验证
DOI
PubMed
高影响因子
JOURNAL OF ANIMAL SCIENCE
·
2020
·Vol. 98
·期 2
·
IF 2.092
Wang Zhaobin,Li Jia,Wang Yu,Wang Lei,Yin Yuebang,Yin Lanmei,Yang Huansheng,Yin Yulong
Vitamin A (VA) is an important nutrient for weaning piglets. It plays a significant role in the normal formation, development, and maintenance of epithelial cells. Previous studies have shown that VA supplements could improve the host’s intestinal barrier function. Therefore, we hypothesized that VA supplements can affect intestinal function in weaned piglets by regulating intestinal stem cells. Thirty-two 21-d-old weaned [(Yorkshire × Landrace) × Duroc] piglets with an average weight of 8.34 ± 0.13 kg were randomly divided into 4 treatment groups, with 1) 2 mg/kg (control), 2) 4 mg/kg, 3) 8 mg/kg, and 4) 16 mg/kg doses of VA, respectively. The experiment lasted for 14 d. Weaned piglets were given ad libitum access to food and water during the test. The ADG (linear, P = 0.020) and G:F (linear, P = 0.005) of the piglets were found to increase significantly from days 8 to 14. The Lgr5+ gene expression (P = 0.012) in the jejunum mucosa of the 16 mg/kg VA group was increased. The jejunum villus height (P = 0.027) and villi surface area (P = 0.035) were significantly increased in the 4 mg/kg VA treatment group. The crypt depth increased significantly in the 4 and 8 mg/kg VA treatment groups (quadratic, P = 0.043), and the ratios of villus height to crypt depth significantly increased in the 16 mg/kg VA group (quadratic, P = 0.015). The maltase (P = 0.032), sucrose (P = 0.041), and alkaline phosphatase activity (linear, P = 0.024) were significantly increased when further supplemented with 4 mg/kg VA. Slc2a2 mRNA abundance was significantly increased in the 2 mg/kg VA group (linear, P = 0.024). Moreover, the budding rates, buddings number per organoid, and Chromogranin A and Muc2 expression of piglet intestinal organoids were significantly reduced (P < 0.05) by VA and its metabolites (retinoic acid). Compared with the control group, the expression of Spp1 and Trop2 increased. These results indicated that VA may increase the stemness of intestinal stem cell in vitro. This study suggested that VA could affect growth performance and intestinal function by regulating intestinal stem cells in the jejunum of weaned piglets.
研究领域:
动物营养学
·
分子生物学
·
兽医学
✓ 已验证
DOI
PubMed
高影响因子
Biomedical Materials
·
2022
·Vol. 17
·期 2
·024105
·
IF 4.103
Yuheng Cao,Haobo He,Kaili Cao,Yongyang Liu,Dehua Huang,Tuanwei Li,Guangcun Chen
Controllable regulation of stem cell differentiation is a critical concern in stem cell-based regenerative medicine. In particular, there are still great challenges in controlling the directional differentiation of neural stem cells (NSCs) into neurons. Herein, we developed a novel linear-branched poly(β-amino esters) (S4-TMPTA-BDA-DT, STBD) through a two-step reaction. The synthesized linear-branched polymers possess multiple positively charged amine terminus and degradable intermolecular ester bonds, thus endowing them with excellent properties such as high gene load, efficient gene delivery, and effective gene release and transcription in cells. In the mCherry transfection test, a high transfection efficiency of approximately 70% was achieved in primary NSCs after a single transfection. Moreover, STBD also showed high biocompatibility to NSCs without disturbing their viability and neural differentiation. With the high gene delivery property, STBD is capable of delivering siRNA (shSOX9) expression plasmid into NSCs to significantly interfere with the expression of SOX9, thus enhancing the neuronal differentiation and maturation of NSCs. The STBD/DNA nano-polyplex represents a powerful non-viral approach of gene delivery for manipulating the differentiation of stem cells, showing broad application prospects in NSC-based regenerative therapy for treating neurodegenerative diseases.
研究领域:
再生医学
·
基因治疗
·
干细胞生物学
·
神经科学
✓ 已验证
DOI
PubMed
高影响因子
TOXICOLOGY IN VITRO
·
2021
·
IF 3.5
Meiting Wang,Jiaxian Wang,Alex Y.P. Tsui,Zhaomin Li,Yizhe Zhang,Qi Zhao,Hongyan Xing,Xijie Wang
The awareness of the long-term toxicities of cancer survivors after chemotherapy treatment has been gradually strengthened as the population of cancer survivors grows. Generally, chemotherapy-induced peripheral neurotoxicity (CIPN) is studied by animal models which are not only expensive and time-consuming, but also species-specific differences. The generation of human induced pluripotent stem cells (hiPSCs) and differentiation of peripheral neurons have provided an in vitro model to elucidate the risk of CIPN. Here, we developed a drug-induced peripheral neurotoxicity model using hiPSC-derived peripheral neurons (hiPSC-PNs) to study the mechanisms of different chemotherapeutic agents on neuronal viability using LDH assay, a cell apoptosis assay determined by caspase 3/7 activation, neurite outgrowth, ion channel expression and neurotransmitter release following treatment of cisplatin, bortezomib, ixabepilone, or pomalidomide. Our data showed that the multiple endpoints of the hiPSC-PNs model had different sensitivity to various chemotherapeutic agents. Furthermore, the chemotherapeutics separated cell viability from the decrease in neurite lengthand changed levels of ion channels and neurotransmitters to a certain extent. Thus, we study the mechanisms of peripheral neurotoxicity induced by chemotherapeutic agents through changes in these indicators.
研究领域:
肿瘤学
·
神经科学
·
干细胞生物学
·
毒理学
✓ 已验证
DOI
PubMed
高影响因子
Cellular Reprogramming
·
2018
·Vol. 20
·期 2
·99-106
·
IF 1.43
Mengya Zhao,Lin Li,Jundong Zhou,Xueyuan Cui,Qingmei Tian,Yaqing Jin,Yimin Zhu
Cancer stem cells (CSCs) are responsible for cancer initiating, recurrence, and drug resistance. Discovery of novel biomarkers for CSCs is helpful for early diagnosis and prognosis. Lung cancer stem cells (LCSCs) were closely related to the occurrence and development of lung cancer. In our study, the important role of miR-2861 in maintaining the stemness of LCSCs was investigated. The LCSC differentiation model was established through introducing serum into the medium of H460 spheres. miR-2861 expression was significantly higher in LCSCs no matter compared to the differentiation cells or normal cells. HDAC5 expression was positively correlated with miR-2861 in LCSCs, and knockdown of miR-2861 decreased the expression of HDAC5, which implied that HDAC5 may be involved in the differentiation of LCSCs mediated by miR-2861. The role of HDAC5 in the regulation of LCSC differentiation was further verified by the inhibitory effect of LMK-235 on the phosphorylation of ERK1/2, which was recognized as the regulator of CSC differentiation. Our study provided a better understanding of miR-2861 and HDAC5 axis in maintaining the stemness of LCSCs and laid a foundation for molecular targeted therapy.
研究领域:
肿瘤学
·
分子生物学
·
癌症研究
✓ 已验证
DOI
PubMed
高影响因子
INTERNATIONAL JOURNAL OF BIOCHEMISTRY & CELL BIOLOGY
·
2018
·
IF 3.247
Chao Zhong,Zhenhuang Chen,Xiaocui Luo,Cuicui Wang,Hui Jiang,Jianzhong Shao,Minxin Guan,Liquan Huang,Xiao Huang,Jinfu Wang
Inner ear hair cells are mechanoreceptors responsible for hearing. Pathogenic defects of hair cell-specific genes are one of the major causes of deafness. The BarH class homeobox gene Barhl1 is a deafness gene expressed in developing hair cells, yet the role of Barhl1 during hair cell development remains poorly understood. In the present study, we first established an in vitro differentiation system to efficiently obtain mouse embryonic stem cell (mESC)-derived hair cell-like cells. Subsequently, a mESC line carrying a targeted disruption of Barhl1 was generated using CRISPR/Cas9 technology and subjected to the established in vitro hair cell differentiation protocol. Targeted disruption of Barhl1 does not affect the induction of mESCs toward early primitive ectoderm-like (EPL) cells and otic progenitors but strongly inhibits the differentiation of hair cell-like cells. Using RNA-sequencing and bioinformatics, we further unravel the molecular mechanism underlying Barhl1 -mediated hair cell development. Our data demonstrate the essential role of Barhl1 during hair cell development and provide a basis for the treatment of Barhl1 mutation-based deafness.
研究领域:
发育生物学
·
干细胞研究
·
基因组学
·
听觉科学
✓ 已验证
DOI
PubMed
高影响因子
Frontiers in Bioengineering and Biotechnology
·
2022
·
IF 6.064
Javad Harati,Xuelian Tao,Hosein Shahsavarani,Ping Du,Massimiliano Galluzzi,Kun Liu,Zhen Zhang,Peter Shaw,Mohammad Ali Shokrgozar,Haobo Pan,Peng-Yuan Wang
Polydopamine (PDA) is a biocompatible cell-adhesive polymer with versatile applications in biomedical devices. Previous studies have shown that PDA coating could improve cell adhesion and differentiation of human mesenchymal stem cells (hMSCs). However, there is still a knowledge gap in the effect of PDA-mediated protein adsorption on the epigenetic status of MSCs. This work used gelatin-coated cell culture surfaces with and without PDA underlayer (Gel and PDA-Gel) to culture and differentiate primary human adipose-derived stem cells (hASCs). The properties of these two substrates were significantly different, which, in combination with a variation in extracellular matrix (ECM) protein bioactivity, regulated cell adhesion and migration. hASCs reduced focal adhesions by downregulating the expression of integrins such as αV, α1, α2, and β1 on the PDA-Gel compared to the Gel substrate. Interestingly, the ratio of H3K27me3 to H3K27me3+H3K4me3 was decreased, but this only occurred for upregulation of AGG and BMP4 genes during chondrogenic differentiation. This result implies that the PDA-Gel surface positively affects the chondrogenic, but not adipogenic and osteogenic, differentiation. In conclusion, for the first time, this study demonstrates the sequential effects of PDA coating on the biophysical property of adsorbed protein and then focal adhesions and differentiation of hMSCs through epigenetic regulation. This study sheds light on PDA-mediated mechanotransduction.
研究领域:
生物医学工程
·
干细胞生物学
·
组织工程
·
表观遗传学
✓ 已验证
DOI
PubMed
高影响因子
Journal of Nanomaterials
·
2022
·
IF 3.791
Yifan Wang,Xin Wu,Wenjing Yang,Pei Feng,Wei Tan,Youwen Deng,Cijun Shuai
Magnesium ion (Mg2+)-based materials are known to exert osteogenic effects that can be enhanced by the bioelectrical properties of magnetic fields. In this study, we examined the effect of a medium-strength static magnetic field (SMF), combined with a Mg2+-containing medium, on the proliferation and osteogenic differentiation of mouse bone marrow mesenchymal stem cells (BMSCs). Mouse BMSCs were divided into a control group, 7.5 mM Mg2+ group, 15 mT SMF group, and 7.5 mM Mg2+ plus 15 mT SMF group. Osteoblast proliferation was measured using a Cell Counting Kit-8 assay, whereas osteogenic differentiation was detected using alkaline phosphatase (ALP) staining and western blot analysis, respectively. The number and size of calcium nodules were determined using Alizarin Red staining. Compared with those in the control group, the ALP activity, calcium nodule formation, and osteogenic protein expression were promoted in other groups. In particular, Mg2+-SMF had a significant effect after 7 days of intervention and more effectively promoted BMSC differentiation and proliferation than either Mg2+ or the SMF alone, suggesting that Mg2+-SMF synergistically contributed to osteogenic differentiation and cell proliferation. To examine their roles in bone differentiation, the Magt1 and Creb1 genes were silenced in BMSCs, and the findings indicated that the synergistic intervention with Mg2+ and magnetic fields might exert osteogenic effects via the MAGT1 channel and CREB1 protein. This study provides an experimental basis for a potential Mg2+-SMF synergistic artificial bone material that could be clinically applied in the treatment of bone defects.
研究领域:
生物医学工程
·
干细胞生物学
·
骨科
·
材料科学
✓ 已验证
DOI
高影响因子
Nanoscale
·
2020
·Vol. 12
·期 18
·10106-10116
·
IF 6.895
Kaipeng Wang,Qian Wu,Xichao Wang,Guohai Liang,Anli Yang,Jinming Li
Finding a method to control and detect the differentiation of stem cells in real time remains a challenge for regenerative medicine. Here we developed the multifunctional upconversion nanoparticle (UCNP) approach for both near-infrared (NIR) control and the real-time detection of osteogenic differentiation in mesenchymal stem cells (MSCs). We first synthesized Tm/Er doped core–shell UCNPs (NaYF4:Yb/Tm/Er@NaYF4), and the core–shell UCNPs were coated with mesoporous silica for drug loading and installing photomechanical azobenzene (azo). Then the Arg-Gly-Asp (RGD) peptide and the matrix metalloproteinase 13 (MMP13) sensitive peptide-black hole quencher-3 group (CGPLGVRGK-BHQ-3) were conjugated on the surface of UCNPs for cell targeting and detection of cell differentiation. The final multifunctional UCNPs are called UCNP@mSiO2-azo-peptide-BHQ-3. The drug icariin (ICA), which can induce the osteogenic differentiation of MSCs, was loaded into UCNP@mSiO2-azo-peptide-BHQ-3 to form the UCNP nanocomplexes. ICA could be released from UCNP nanocomplexes in a NIR-controlled manner that is based on the transformation of the trans-isomer of azo into the cis isomer under the upconverted UV and visible light. Meanwhile, UCNP@mSiO2-azo-peptide-BHQ-3 could also be used as a nanoprobe to detect the activity of the MMP13 enzyme by enzyme digestion and UCNP fluorescence recovery. By detecting MMP13, which is produced by osteogenic differentiation, a real-time detection of cell differentiation in living differentiated MSCs could be achieved using UCNP nanoprobes. Thus, the multifunctional UCNPs combined the control of cell differentiation with the real-time detection of cell differentiation in MSCs, which makes them a powerful tool for regulating and detecting the differentiation of MSCs in regenerative medicine.
研究领域:
再生医学
·
纳米技术
·
干细胞生物学
·
生物医学工程
✓ 已验证
DOI
PubMed
高影响因子
Nanoscale
·
2015
·Vol. 8
·期 4
·1897-1904
·
IF 7.394
Weibo Guo,Shu Wang,Xin Yu,Jichuan Qiu,Jianhua Li,Wei Tang,Zhou Li,Xiaoning Mou,Hong Liu,Zhonglin Wang
The cell–material interface is one of the most important considerations in designing a high-performance tissue engineering scaffold because the surface of the scaffold can determine the fate of stem cells. A conductive surface is required for a scaffold to direct stem cells toward neural differentiation. However, most conductive polymers are toxic and not amenable to biological degradation, which restricts the design of neural tissue engineering scaffolds. In this study, we used a bioactive three-dimensional (3D) porcine acellular dermal matrix (PADM), which is mainly composed of type I collagen, as a basic material and successfully assembled a layer of reduced graphene oxide (rGO) nanosheets on the surface of the PADM channels to obtain a porous 3D, biodegradable, conductive and biocompatible PADM–rGO hybrid neural tissue engineering scaffold. Compared with the PADM scaffold, assembling the rGO into the scaffold did not induce a significant change in the microstructure but endowed the PADM–rGO hybrid scaffold with good conductivity. A comparison of the neural differentiation of rat bone-marrow-derived mesenchymal stem cells (MSCs) was performed by culturing the MSCs on PADM and PADM–rGO scaffolds in neuronal culture medium, followed by the determination of gene expression and immunofluorescence staining. The results of both the gene expression and protein level assessments suggest that the rGO-assembled PADM scaffold may promote the differentiation of MSCs into neuronal cells with higher protein and gene expression levels after 7 days under neural differentiation conditions. This study demonstrated that the PADM–rGO hybrid scaffold is a promising scaffold for neural tissue engineering; this scaffold can not only support the growth of MSCs at a high proliferation rate but also enhance the differentiation of MSCs into neural cells.
研究领域:
组织工程
·
神经科学
·
生物材料科学
✓ 已验证
DOI
PubMed
高影响因子
JOURNAL OF BIOACTIVE AND COMPATIBLE POLYMERS
·
2019
·
IF 1.976
Zengjie Fan,Yingying Nie,Zizi Chen,Xuzhuzi Xie,Xiaozhu Liao,Yuan Wei
Temperature-responsive hydrogel culture system is considered as an ideal platform for cell sheet harvest, but its complex preparation methods and harsh reaction conditions limit its application. Inspired by the marine mussels, a biomimetic method presented here is to construct a novel temperature-responsive hydrogel culture system for stem cell sheet harvest. The tissue culture polystyrene is first modified with polydopamine coating, and then amine-terminated poly(N-isopropylacrylamide) is grafted onto the coating via the Schiff base or Michael addition reaction to construct the temperature-sensitive hydrogel culture system. Then, bone marrow stromal cells are cultured on the culture system to construct cell sheets. The prepared culture system shows significant temperature-sensitive property with the grafted concentrations of poly(N-isopropylacrylamide) ranging from 0.5 to 1 g/L. Meanwhile, the constructed culture system has low cytotoxicity and facilitates the stem cell adhesion, proliferation, and cell sheet formation at 37°C. When the culture system is placed in a 20°C environment, the cell sheet can be completely detached from the surface of tissue culture polystyrene without being treated with any enzymes. More importantly, the cell morphology, cell sheet thickness, and the fibril structure of the associated proteins are similar to the cells cultured on the tissue culture polystyrene without modification. The biomimetic, simple, inexpensive, and environmentally friendly preparation of the culture system enables it to be used for the harvest of cell sheet and even applied to tissue engineering for tissue regeneration.
研究领域:
材料科学
·
生物材料
·
组织工程
·
细胞生物学
✓ 已验证
DOI
高影响因子
Cell Reports
·
2022
·
IF 9.995
Ruipeng Zhuge,Chao Wang,Jie Wang,Shuyu Yu,Liming Liao,Xiaofeng Zheng
Summary YAP1 functions in lineage differentiation of pluripotent embryonic stem cells (ESCs); however, the detailed mechanisms underlying the regulation of YAP1 activity during ESC differentiation remain elusive. Here, we report that hCINAP serves as a negative regulator of YAP1 during ESC fate decisions. The expression of mCINAP, the murine homolog of hCINAP, is downregulated during the differentiation process of murine ESC (mESC) ectoderm lineage, leading to liquid-liquid phase separation (LLPS) of NEDD4 and activation of YAP1. Mechanistically, hCINAP interacts with and prevents NEDD4 from forming cytoplasmic condensates that compartmentalize YAP1 and its kinase NLK, facilitating YAP1 phosphorylation at Ser128 and promoting YAP1 activation. mCINAP depletion leads to the formation of NEDD4 condensates and YAP1 activation, which impedes endoderm differentiation of mESCs. Our study shows that hCINAP is a vital regulator of YAP1 activity and is essential for stem cell fate decisions, which provides mechanistic insight into early embryogenesis.
研究领域:
干细胞生物学
·
细胞信号传导
·
发育生物学
✓ 已验证
DOI
PubMed
高影响因子
危险品化学品经营许可证（带存储）