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Hydroxyapatite-Based Colloidal Gels Facilitate the Proliferation and Migration of Chondrocytes and the Adhesion of Umbilical Cord Mesenchymal Stem Cells.

ABSTRACT: Collective movement of cells that have been delivered on biomaterials for transplantation purposes would be a desirable attribute that would promote wound healing, cell proliferation, and eventual growth and regeneration of damaged organs. We hypothesized that colloidal gels made from hydroxyapatite (HA) and poly(D,L-lactic-co-glycolic acid) (PLGA) particles will be conducive to the growth and migration of porcine chondrocytes, will allow the adhesion of human umbilical cord mesenchymal stem cells, and will have negligible effects on the cell cycle of these cells. Then, we performed experiments designed to assess the viability and migratory properties of porcine chondrocytes studded on nanosized HA/PLGA particles. Our experiments show that porcine chondrocytes migrated in and around a hydroxyapatite-based biomaterial that could be described as a colloidal gel. Cells in the colloidal gel demonstrated unidirectional movement. Cells were seen to be extending lamellae and were followed by other cells.

SUBMITTER: Jamal SA

PROVIDER: S-EPMC4897378 | biostudies-literature | 2014

REPOSITORIES: biostudies-literature

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Hydroxyapatite-Based Colloidal Gels Facilitate the Proliferation and Migration of Chondrocytes and the Adhesion of Umbilical Cord Mesenchymal Stem Cells.

Jamal Syed A SA Ye Qiang Q

International scholarly research notices 20141229

Collective movement of cells that have been delivered on biomaterials for transplantation purposes would be a desirable attribute that would promote wound healing, cell proliferation, and eventual growth and regeneration of damaged organs. We hypothesized that colloidal gels made from hydroxyapatite (HA) and poly(D,L-lactic-co-glycolic acid) (PLGA) particles will be conducive to the growth and migration of porcine chondrocytes, will allow the adhesion of human umbilical cord mesenchymal stem cel ...[more]

PMID: 27382607

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Project description:BACKGROUND:Mesenchymal stem cells (MSCs) are known to home to injured and inflamed regions via the bloodstream to assist in tissue regeneration in response to signals of cellular damage. However, the factors and mechanisms that affect their transendothelial migration are still unclear. In this study, the mechanisms involved in interleukin-1? (IL-1?) enhancing the transendothelial migration of MSCs were investigated. METHODS:Immunofluorescence staining and Western blotting were used to observe IL-1?-induced CXC chemokine receptor 3 (CXCR3) expression on MSCs. Quantitative real-time PCR and ELISA were used to demonstrate IL-1? upregulated both chemokine (C-X-C motif) ligand 9 (CXCL9) mRNA and CXCL9 ligand secretion in human umbilical vein endothelial cells (HUVECs). Monolayer co-cultivation, agarose drop chemotaxis, and transwell assay were conducted to investigate the chemotaxis invasion and transendothelial migration ability of IL-1?-induced MSCs in response to CXCL9. RESULTS:In this study, our immunofluorescence staining showed that IL-1? induces CXCR3 expression on MSCs. This result was confirmed by Western blotting. Following pretreatment with protein synthesis inhibitor cycloheximide, we found that IL-1? induced CXCR3 on the surface of MSCs via protein synthesis pathway. Quantitative real-time PCR and ELISA validated that IL-1? upregulated both CXCL9 mRNA and CXCL9 ligand secretion in HUVECs. In response to CXCL9, chemotaxis invasion and transendothelial migration ability were increased in IL-1?-stimulated MSCs. In addition, we pretreated MSCs with CXCR3 antagonist AMG-487 and p38 MAPK inhibitor SB203580 to confirm CXCR3-CXCL9 interaction and the role of CXCR3 in IL-1?-induced chemotaxis invasion and transendothelial migration. CONCLUSION:We found that IL-1? induces the expression of CXCR3 through p38 MAPK signaling and that IL-1? also enhances CXCL9 ligand secretion in HUVECs. These results indicated that IL-1? promotes the transendothelial migration of MSCs through CXCR3-CXCL9 axis. The implication of the finding could enhance the efficacy of MSCs homing to target sites.

Project description:BackgroundSevere injuries of the uterus may trigger uterine scar formation, ultimately leading to infertility or obstetrical complications. To date, few methods have adequately solved the problem of collagen deposition in uterine scars. Umbilical cord-derived mesenchymal stem cells (UC-MSCs) have shown great promise in clinical applications. The objective of this study was to investigate the effect of a scaffold/UC-MSCs construct on collagen degradation and functional regeneration in rat uterine scars following full-thickness excision of uterine walls.MethodsIn order to establish a rat model of uterine scars, the uterine wall of approximately 1.0 cm in length and 0.5 cm in width (one-third of the uterine circumference) was excised from each uterine horn. A total of 128 scarred uterine horns from 64 rats were randomly assigned to four groups, including a PBS group (n = 32 uterine horns), scaffold group (n = 32 uterine horns), UC-MSCs group (n = 32 uterine horns) and scaffold/UC-MSCs group (n = 32 uterine horns) to investigate the effect of different treatments on the structure and function of uterine scars. PBS, degradable collagen fibres, UC-MSCs or UC-MSCs mixed with gelatinous degradable collagen fibres were injected into four pre-marked points surrounding each uterine scar, respectively. At days 30 and 60 post-transplantation, a subset of rats (n = 8 uterine horns) from each group was euthanized and serial sections of uterine tissues containing the operative region were prepared. Haematoxylin-eosin staining, Masson's trichrome staining, and immunohistochemical staining for MMP-2, MMP-9, α-SMA and vWF were performed. Finally, another subset of rats (n = 16 uterine horns) from each group was mated with male rats at day 60 post-transplantation and euthanized 18 days after the presence of vaginal plugs to check numbers, sizes and weights of fetuses, as well as sites of implantation.ResultsThe scaffold/UC-MSCs group exhibited obvious collagen degradation compared with the other three groups. At day 60 post-transplantation, the number of MMP-9-positive cells in the scaffold/UC-MSCs group (25.96 ± 3.63) was significantly higher than that in the PBS group (8.19 ± 1.61, P < 0.01), the scaffold group (7.25 ± 2.17, P < 0.01) and the UC-MSCs group (8.31 ± 2.77, P < 0.01). The pregnancy rate in the scaffold/UC-MSCs group (10/16) was also significantly higher than that in the PBS group (2/16, P < 0.017), the scaffold group (1/16, P < 0.017) and the UC-MSCs group (3/16, P < 0.017).ConclusionsThe scaffold/UC-MSCs system facilitated collagen degradation in uterine scars via upregulation of MMP-9, which was secreted by transplanted UC-MSCs, and promoted regeneration of the endometrium, myometrium and blood vessels in uterine scars. Furthermore, the scaffold/UC-MSCs-treated uterine scars showed nearly complete restoration of receptive fertility.

Project description:UnlabelledTherapeutic applications of mesenchymal stem cells (MSCs) for treating various diseases have increased in recent years. To ensure that treatment is effective, an adequate MSC dosage should be determined before these cells are used for therapeutic purposes. To obtain a sufficient number of cells for therapeutic applications, MSCs must be expanded in long-term cell culture, which inevitably triggers cellular senescence. In this study, we investigated the surface markers of human umbilical cord blood-derived MSCs (hUCB-MSCs) associated with cellular senescence using fluorescence-activated cell sorting analysis and 242 cell surface-marker antibodies. Among these surface proteins, we selected the melanoma cell adhesion molecule (MCAM/CD146) for further study with the aim of validating observed expression differences and investigating the associated implications in hUCB-MSCs during cellular senescence. We observed that CD146 expression markedly decreased in hUCB-MSCs following prolonged in vitro expansion. Using preparative sorting, we found that hUCB-MSCs with high CD146 expression displayed high growth rates, multilineage differentiation, expression of stemness markers, and telomerase activity, as well as significantly lower expression of the senescence markers p16, p21, p53, and senescence-associated ?-galactosidase, compared with that observed in hUCB-MSCs with low-level CD146 expression. In contrast, CD146 downregulation with small interfering RNAs enhanced the senescence phenotype. In addition, CD146 suppression in hUCB-MSCs caused downregulation of other cellular senescence regulators, including Bmi-1, Id1, and Twist1. Collectively, our results suggest that CD146 regulates cellular senescence; thus, it could be used as a therapeutic marker to identify senescent hUCB-MSCs.SignificanceOne of the fundamental requirements for mesenchymal stem cell (MSC)-based therapies is the expansion of MSCs during long-term culture because a sufficient number of functional cells is required. However, long-term growth inevitably induces cellular senescence, which potentially causes poor clinical outcomes by inducing growth arrest and the loss of stem cell properties. Thus, the identification of markers for evaluating the status of MSC senescence during long-term culture may enhance the success of MSC-based therapy. This study provides strong evidence that CD146 is a novel and useful marker for predicting senescence in human umbilical cord blood-derived MSCs (hUCB-MSCs), and CD146 can potentially be applied in quality-control assessments of hUCB-MSC-based therapy.

Dataset Information

Hydroxyapatite-Based Colloidal Gels Facilitate the Proliferation and Migration of Chondrocytes and the Adhesion of Umbilical Cord Mesenchymal Stem Cells.

Publications

Hydroxyapatite-Based Colloidal Gels Facilitate the Proliferation and Migration of Chondrocytes and the Adhesion of Umbilical Cord Mesenchymal Stem Cells.

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