scholarly journals Induction of Articular Chondrogenesis by Chitosan/Hyaluronic-Acid-Based Biomimetic Matrices Using Human Adipose-Derived Stem Cells

2019 ◽  
Vol 20 (18) ◽  
pp. 4487 ◽  
Author(s):  
Yijiang Huang ◽  
Daniel Seitz ◽  
Fabian König ◽  
Peter E. Müller ◽  
Volkmar Jansson ◽  
...  

Cartilage repair using tissue engineering is the most advanced clinical application in regenerative medicine, yet available solutions remain unsuccessful in reconstructing native cartilage in its proprietary form and function. Previous investigations have suggested that the combination of specific bioactive elements combined with a natural polymer could generate carrier matrices that enhance activities of seeded stem cells and possibly induce the desired matrix formation. The present study sought to clarify this by assessing whether a chitosan-hyaluronic-acid-based biomimetic matrix in conjunction with adipose-derived stem cells could support articular hyaline cartilage formation in relation to a standard chitosan-based construct. By assessing cellular development, matrix formation, and key gene/protein expressions during in vitro cultivation utilizing quantitative gene and immunofluorescent assays, results showed that chitosan with hyaluronic acid provides a suitable environment that supports stem cell differentiation towards cartilage matrix producing chondrocytes. However, on the molecular gene expression level, it has become apparent that, without combinations of morphogens, in the chondrogenic medium, hyaluronic acid with chitosan has a very limited capacity to stimulate and maintain stem cells in an articular chondrogenic state, suggesting that cocktails of various growth factors are one of the key features to regenerate articular cartilage, clinically.

2014 ◽  
Vol 26 (1) ◽  
pp. 211 ◽  
Author(s):  
R. A. C. Rabel ◽  
L. Osterbur ◽  
A. Maki ◽  
J. Lewis ◽  
M. B. W. Wheeler

There is a great need for bioengineered cartilage because of the lack of medical or surgical therapies to improve articular cartilage healing. We hypothesised that porcine adipose-derived stem cells (pASC) can be induced to undergo chondrogenic differentiation within hyaluronic acid (HA) hydrogels. The objective of this study was to develop UV-curable pASC-laden HA hydrogels aimed at application in cartilage tissue engineering. HA was treated with glycidyl methacrylate (GM) to allow chemical gelation of the polymer upon exposure to UV light. 2% HAGM hydrogel was obtained by mixing HAGM with chondrogenic medium consisting of TGFβ, ascorbic acid, ITS+ premix (insulin, transferrin, selenous acid; Cat. No. 354352, BD Biosciences, Franklin Lakes, NJ), sodium pyruvate, and dexamethasone. Passage three-pASC were resuspended in 2% HAGM hydrogel with 2 × 107 cells mL–1. Twelve-and-one-half (12.5)-μL droplets (micromasses) of this suspension containing 250 000 pASC were placed in 24-well culture plates and incubated for 2 h at 37°C and 5% CO2 to allow for cell attachment. Subsequently, the cell-laden hydrogels were cured with ~10 mW cm–2 365-nm UV light for 10 min, covered with 500 μL of chondrogenic medium, and cultured for up to 11 days at 37°C and 5% CO2. Additionally, pASC micromasses were cultured in chondrogenic medium without loading on 2% HAGM hydrogels as positive controls, and in non-chondrogenic DMEM as negative controls. Samples were collected at 4, 7, and 11 days in to culture for cryopreservation (for immunohistochemistry; IHC) and dimethylmethylene blue (DMMB) assay. IHC on day 11 of culture demonstrated the expression of cartilage specific proteins type-II collagen and aggrecan. On the basis of data from the DMMB assay, chondrogenic differentiation of pASC-laden micromasses in positive controls and 2% HAGM treatments were not different (P > 0.05). This indicates that ASC can produce cartilage equally well under both conditions, supporting the idea that HAGM may be used as a matrix for cartilage formation in vitro and possibly in vivo. In conclusion, using a micromass cell culture system, we demonstrated that 2% HAGM hydrogels support proliferation and chondrogenic differentiation of pASC. Further experiments testing different concentrations of HAGM and UV exposure levels, and larger sample numbers are warranted to further improve this procedure.


2017 ◽  
Vol 42 (3) ◽  
pp. 1037-1050 ◽  
Author(s):  
Guangxin Huang ◽  
Yan Kang ◽  
Zhiyu Huang ◽  
Zhiqi Zhang ◽  
Fangang Meng ◽  
...  

Background/Aims: Long noncoding RNAs (lncRNAs) play important roles in stem cell differentiation. However, their role in osteogenesis of human adipose-derived stem cells (ASCs), a promising cell source for bone regeneration, remains unknown. Here, we investigated the expression profile and potential roles of lncRNAs in osteogenic differentiation of human ASCs. Methods: Human ASCs were induced to differentiate into osteoblasts in vitro, and the expression profiles of lncRNAs and mRNAs in undifferentiated and osteogenic differentiated ASCs were obtained by microarray. Bioinformatics analyses including subgroup analysis, gene ontology analysis, pathway analysis and co-expression network analysis were performed. The function of lncRNA H19 was determined by in vitro knockdown and overexpression. Quantitative reverse transcription polymerase chain reaction was utilized to examine the expression of selected genes. Results: We identified 1,460 upregulated and 1,112 downregulated lncRNAs in osteogenic differentiated human ASCs as compared with those of undifferentiated cells (Fold change ≥ 2.0, P < 0.05). Among these, 94 antisense lncRNAs, 85 enhancer-like lncRNAs and 160 lincRNAs were further recognized. We used 12 lncRNAs and 157 mRNAs to comprise a coding-non-coding gene expression network. Additionally, silencing of H19 caused a significantly increase in expression of osteogenesis-related genes, including ALPL and RUNX2, while a decrease was observed after H19 overexpression. Conclusion: This study revealed for the first time the global expression profile of lncRNAs involved in osteogenic differentiation of human ASCs and provided a foundation for future investigations of lncRNA regulation of human ASC osteogenesis.


2017 ◽  
Vol 16 (4) ◽  
pp. 4095-4100 ◽  
Author(s):  
Jiayan Guo ◽  
Shu Guo ◽  
Yuxin Wang ◽  
Yanqiu Yu

2012 ◽  
Vol 27 (7) ◽  
pp. 471-476 ◽  
Author(s):  
Silvana Gaiba ◽  
Lucimar Pereira de França ◽  
Jerônimo Pereira de França ◽  
Lydia Masako Ferreira

PURPOSE: There is a growing scientific interest in the plasticity and therapeutic potential of adipose-derived stem cells (ASCs), which are multipotent and abundant in adipose tissue and can differentiate in vitro into multiple lineages, including adipocytes, chondrocytes, osteoblasts, neural cells, endothelial cells and cardiomyocytes. The aim of this study was to isolate, cultivate and identify ASCs. METHODS: Human adipose precursor cells were obtained from subcutaneous abdominal tissue. Recently dispersed cells were separated by density centrifugation gradient, cultured and then analyzed. RESULTS: Human ASCs were able to replicate in our culture conditions. The cells maintained their phenotypes throughout the studied period on different passages confirming they suitability for in vitro cultivation. We also induced their adipogenic, osteogenic and chondrogenic differentiation, verifying their mesenchymal stem cells potentiality in vitro. Flow cytometry results showed that these cells expressed CD73, CD90 and CD105, (mesenchymal stem-cells markers), contrasting with the lack of expression of CD16, CD34 and CD45 (hematopoietic cells markers). CONCLUSION: It was possible to isolate human adipose-derived stem cells by in vitro cultivation without adipogenic induction, maintaining their functional integrity and high proliferation levels. The cells demonstrated adipogenic, osteogenic and chondrogenic differentiation potential in vitro.


2021 ◽  
Vol 30 ◽  
pp. 096368972110354
Author(s):  
Eun-Jung Yoon ◽  
Hye Rim Seong ◽  
Jangbeen Kyung ◽  
Dajeong Kim ◽  
Sangryong Park ◽  
...  

Stamina-enhancing effects of human adipose derived stem cells (hADSCs) were investigated in young Sprague-Dawley rats. Ten-day-old male rats were transplanted intravenously (IV) or intracerebroventricularly (ICV) with hADSCs (1 × 106 cells/rat), and physical activity was measured by locomotor activity and rota-rod performance at post-natal day (PND) 14, 20, 30, and 40, as well as a forced swimming test at PND 41. hADSCs injection increased the moving time in locomotor activity, the latency in rota-rod performance, and the maximum swimming time. For the improvement of physical activity, ICV transplantation was superior to IV injection. In biochemical analyses, ICV transplantation of hADSCs markedly reduced serum creatine phosphokinase, lactate dehydrogenase, alanine transaminase, and muscular lipid peroxidation, the markers for muscular and hepatic injuries, despite the reduction in muscular glycogen and serum triglycerides as energy sources. Notably, hADSCs secreted brain-derived neurotrophic factor (BDNF) and nerve growth factor in vitro, and increased the level of BDNF in the brain and muscles in vivo. The results indicate that hADSCs enhance physical activity including stamina not only by attenuating tissue injury, but also by strengthening the muscles via production of BDNF.


2019 ◽  
Vol 103 (3) ◽  
pp. 925-934
Author(s):  
Eкaterina Vackova ◽  
Darko Bosnakovski ◽  
Bodil Bjørndal ◽  
Penka Yonkova ◽  
Natalia Grigorova ◽  
...  

2012 ◽  
Vol 32 (8) ◽  
pp. 1255-1263 ◽  
Author(s):  
Chengcheng Ying ◽  
Wanli Hu ◽  
Bei Cheng ◽  
Xinmin Zheng ◽  
Shiwen Li

2009 ◽  
Vol 1239 ◽  
Author(s):  
Karla Brammer ◽  
Seunghan Oh ◽  
Sungho Jin

AbstractTwo important goals in stem cell research are to control the cell proliferation without differentiation, and also to direct the differentiation into a specific cell lineage when desired. Recent studies indicate that the nanostructures substantially influence the stem cell behavior. It is well known that mesenchymal stem cells (MSCs) are multipotent stem cells that can differentiate into stromal lineages such as adipocyte, chondrocyte, fibroblast, myocyte, and osteoblast cell types. By examining the cellular behavior of MSCs cultured in vitro on nanostructures, some understanding of the effects that the nanostructures have on the stem cell’s response has been obtained. Here we demonstrate that TiO2 nanotubes produced by anodization on Ti implant surface can regulate human mesenchymal stem cell (hMSC) differentiation towards an osteoblast lineage in the absence of osteogenic inducing factors. Altering the dimensions of nanotubular-shaped titanium oxide surface structures independently allowed either augmented human mesenchymal stem cell (hMSC) adhesion at smaller diameter levels or a specific differentiation of hMSCs into osteoblasts using only the geometric cues. Small (˜30 nm diameter) nanotubes promoted adhesion without noticeable differentiation, while larger (˜70 - 100 nm diameter) nanotubes elicited a dramatic, ˜10 fold stem cell elongation, which induced cytoskeletal stress and selective differentiation into osteoblast-like cells, offering a promising nanotechnology-based route for novel orthopaedics-related hMSC treatments. The fact that a guided and preferential osteogenic differentiation of stem cells can be achieved using substrate nanotopography alone without using potentially toxic, differentiation-inducing chemical agents is significant, which can be useful for future development of novel and enhanced stem cell control and therapeutic implant development.


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