scholarly journals Mitochondria transfer enhances proliferation, migration, and osteogenic differentiation of bone marrow mesenchymal stem cell and promotes bone defect healing

2020 ◽  
Author(s):  
Yusi Guo ◽  
Xiaopei Chi ◽  
Yifan Wang ◽  
Boon Chin Heng ◽  
Yan Wei ◽  
...  
Keyword(s):  
Bone Marrow ◽  
Stem Cell ◽  
Bone Defect ◽  
Atp Production ◽  
Bone Defect Repair ◽  
Mitochondria Transfer ◽  
Defect Repair

Abstract Background: Bone marrow-derived mesenchymal stem cells (BMSCs) transplantation is considered a promising therapeutic approach for bone defect repair. However, during the transplantation procedure, the functions and viability of BMSCs may be impaired due to extended durations of in vitro culture, aging and disease conditions of patients. Inspired by spontaneous intercellular mitochondria transfer that naturally occurs within injured tissues to rescue cellular or tissue function, we investigated whether artificial mitochondria transfer into pre-transplant BMSCs in vitro could improve cellular function and enhance their therapeutic effects on bone defect repair in situ . Methods: Mitochondria were isolated from donor BMSCs and transferred into recipient BMSCs of the same batch and passage. Subsequently, changes in proliferative capacity and cell senescence were evaluated by live cell imaging, Cell Counting Kit-8 assay, cell cycle analysis, Ki67 staining, qPCR and Western Blot analysis of C-myc expression, and β-galactosidase staining. Migration ability was evaluated by the transwell migration assay, wound scratch healing and cell motility tests. Alakine phosphatase (ALP) staining, Alizarin Red staining, combined qPCR and Western Blot analyses of Runx2 and BMP2 were performed to elucidate the effects of mitochondria transfer on the osteogenic potential of BMSCs in vitro . After that, in vivo experiments were performed by transplanting mitochondria-recipient BMSCs into a rat cranial critical-size bone defect model. Micro CT scanning and histological analysis were conducted at 4 and 8 weeks after transplantation to evaluate osteogenesis in situ . Finally, in order to establish the correlation between cellular behavioral changes and aerobic metabolism, OXPHOS (oxidative phosphorylation) and ATP production were assessed and inhibition of aerobic respiration by oligomycin was performed. Results: Mitochondria-recipient BMSCs exhibited significantly enhanced proliferation and migration, and increased osteogenesis upon osteogenic induction. The in vivo results showed more new bone formation after transplantation of mitochondria-recipient BMSCs in situ . Increased OXPHOS activity and ATP production were observed, which upon inhibition by oligomycin attenuated the enhancement of proliferation, migration and osteogenic differentiation induced by mitochondria transfer. Conclusions: Mitochondria transfer is a feasible technique to enhance BMSCs function in vitro and promote bone defect repair in situ through the up-regulation of aerobic metabolism. The results indicated that mitochondria transfer may be a novel promising technique for optimizing stem cell therapeutic function.

scholarly journals Mitochondria transfer enhances bone marrow mesenchymal stem cell functions and promotes bone defect healing

2020 ◽  
Author(s):  
Yusi Guo ◽  
Xiaopei Chi ◽  
Yifan Wang ◽  
Boon Chin Heng ◽  
Yan Wei ◽  
...  
Keyword(s):  
Bone Marrow ◽  
Stem Cell ◽  
Bone Defect ◽  
Atp Production ◽  
Bone Defect Repair ◽  
Mitochondria Transfer ◽  
Defect Repair

Abstract Background: Bone marrow-derived mesenchymal stem cells (BMSCs) transplantation is considered a promising therapeutic approach for bone defect repair. However, during the transplantation procedure, the functions and viability of BMSCs may be impaired due to extended durations of in vitro culture, aging and disease conditions of patients. Inspired by spontaneous intercellular mitochondria transfer that naturally occurs within injured tissues to rescue cellular or tissue function, we investigated whether artificial mitochondria transfer into pre-transplant BMSCs in vitro could improve cellular function and enhance their therapeutic effects on bone defect repair in situ. Methods: First, mitochondria were isolated from donor BMSCs and transferred into recipient BMSCs of the same passage. Afterwards, changes in proliferative capability was evaluated by Cell Counting Kit-8, Ki67 staining, etc., while Transwell, wound scratch healing and cell motility tests were conducted to determine migration ability. Then, alkaline phosphatase (ALP) staining, Alizarin Red staining, combined with qPCR and Western Blot experiments of Runx2 and BMP2 were performed to elucidate the effect of mitochondria transfer on the osteogenic potential of BMSCs in vitro. After that, the in vivo experiments were completed by transplanting mitochondria-recipient BMSCs into a rat cranial critical-size bone defect model. Micro CT scanning and histological analysis were conducted 4 weeks and 8 weeks after transplantation to evaluate the osteogenesis effect in situ. Finally, in order to discover the potential connection between cellular behavioral changes and aerobic metabolism, OXPHOS (oxidative phosphorylation) and ATP production were assessed and inhibition of aerobic respiration by oligomycin was proceeded. Results: Mitochondria-recipient BMSCs exhibited significantly enhanced proliferation and migration, and increased osteogenic differentiation upon osteogenic induction. The in vivo results showed more new bone formation after transplantation of mitochondria-recipient BMSCs in situ. Increased OXPHOS activity and ATP production were further observed, whereas the inhibition of which impaired the enhancement of proliferation, migration and osteogenic differentiation induced by mitochondria transfer. Conclusions: Mitochondria transfer is a feasible technique to enhance BMSCs function in vitro and promote bone defect repair in situ through the up-regulation of aerobic metabolism. The results indicated that mitochondria transfer may be a novel promising technique for optimizing stem cell function.

Ectopic Human Mesenchymal Stem Cell-Coated Scaffolds Create An In Vivo Leukemia Niche in NOD/SCID Mice.

Blood ◽  
2009 ◽  
Vol 114 (22) ◽  
pp. 559-559
Author(s):  
Sarah Rivkah Vaiselbuh ◽  
Morris Edelman ◽  
Jeffrey Michael Lipton ◽  
Johnson M. Liu
Keyword(s):  
Stem Cells ◽  
Bone Marrow ◽  
Stem Cell ◽  
Stem Cell Niche ◽  
Scid Mice ◽  
Cxcr4 Antagonist ◽  
Cell Niche

Abstract Abstract 559 Introduction: Different cellular components of the normal hematopoietic niche have been identified. However, the niche for malignant hematopoiesis remains to be elucidated. Recent work of other groups has suggested that hematopoietic stem cells (HSC) within the bone marrow anchor themselves in place by attaching to osteoblasts and/or vascular sinusoid endothelial cells. We have recently identified mesenchymal stem cells (MSC) as niche-maker cells and found a crucial role of the SDF-1/CXCR4 axis in this process. Stromal Derived Factor-1 (SDF-1/CXCL12) regulates stem cell trafficking and the cell cycle via its receptor CXCR4. Methods: Polyurethane scaffolds, coated in vitro with human bone marrow MSC, were implanted subcutaneously in non-irradiated NOD/SCID mice. CD34+ HSC or primary AML cells (from a leukapheresis product) were injected either in situ or retro-orbitally in the mice and analyzed for engraftment. The mice were treated twice per week with in situ injections of SDF-1, AMD3100 (a CXCR4 antagonist) or PBS (control). After 2 to 4 weeks, the scaffolds were processed and evaluated for cell survival in the mesenchymal niche by immunohistochemistry. Results: We created in vitro MSC-coated scaffolds that retained inoculated AML cells in the presence of SDF-1, while AML cells seeded on empty scaffolds were not retained. In vivo in NOD/SCID mice, the MSC-coated scaffolds, in the presence of SDF-1 enabled homing of both in situ injected normal CD34+ HSC and retroorbital- or in situ injected primary human AML cells. The scaffolds were vascularized and showed osteoclasts and adipocytes present, suggestive of an ectopic human bone marrow microenvironment in the murine host. Finally, the SDF-1-treated scaffolds showed proliferation of the MSC stromal layer with multiple adherent AML cells, while in the AMD3100-treated scaffolds the stromal lining was thin and disrupted at several points, leaving AML cells free floating in proximity. The PBS-treated control-scaffold showed a thin single cell MSC stromal layer without disruption, with few AML cells attached. Conclusion: The preliminary data of this functional ectopic human microenvironment in NOD/SCID mice suggest that AMD3100 (a CXCR4 antagonist) can disrupt the stem cell niche by modulation of the mesenchymal stromal. Further studies are needed to define the role of mesenchymal stem cells in maintaining the hematopoietic/leukemic stem cell niche in vivo. In Vivo Leukemia Stem Cell Niche: (A) Empty polyurethane scaffold. (B)Vascularization in SQ implanted MSC-coated scaffold (s) niche in NOD/SCID mice. (C) DAB Peroxidase (brown) human CD45 positive nests of AML cells (arrows) 1 week after direct in situ AML injection. (D) Human CD45 positive myeloid cells adhere to MSC in vivo (arrows). Disclosures: No relevant conflicts of interest to declare.

scholarly journals In Vivo and in Vitro Studies of the Effect of Chitosan on the Bone Defect Repair Process

2016 ◽  
Vol 16 (2) ◽  
pp. 171-179 ◽  
Author(s):  
I. V. Zudina ◽  
◽  
A. P. Vedyaeva ◽  
P. V. Ivanov ◽  
A. F. A. Alzubaidi ◽  
...  
Keyword(s):  
Bone Defect ◽  
Repair Process ◽  
In Vitro Studies ◽  
Bone Defect Repair ◽  
Defect Repair

scholarly journals Fabrication and Application of Novel Porous Scaffold in Situ-Loaded Graphene Oxide and Osteogenic Peptide by Cryogenic 3D Printing for Repairing Critical-Sized Bone Defect

Molecules ◽  
2019 ◽  
Vol 24 (9) ◽  
pp. 1669 ◽  
Author(s):  
Zhang ◽  
Wang ◽  
Fu ◽  
Ye ◽  
Wang ◽  
...  
Keyword(s):  
Biological Activity ◽  
Graphene Oxide ◽  
Stem Cell ◽  
3D Printing ◽  
Mesenchymal Stem Cell ◽  
Bone Defect ◽  
The Novel

Osteogenic peptides have been reported as highly effective in directing mesenchymal stem cell osteogenic differentiation in vitro and bone formation in vivo. Therefore, developing novel biomaterials for the controlled delivery of osteogenic peptides in scaffolds without lowering the peptide’s biological activity is highly desirable. To repair a critical-sized bone defect to efficiently achieve personalized bone regeneration, a novel bioactive poly(lactic-co-glycolic acid) (PLGA)/β-tricalcium phosphate (β-TCP) composite scaffold, in which graphene oxide (GO) and bone morphogenetic protein (BMP)-2-like peptide were loaded in situ (PTG/P), was produced by an original cryogenic 3D printing method. The scaffolds were mechanically comparable to human cancellous bone and hierarchically porous. The incorporation of GO further improved the scaffold wettability and mechanical strength. The in situ loaded peptides retained a high level of biological activity for an extended time, and the loading of GO in the scaffold further tuned the peptide release so that it was more sustained. Our in vitro study showed that the PTG/P scaffold promoted rat bone marrow-derived mesenchymal stem cell ingrowth into the scaffold and enhanced osteogenic differentiation. Moreover, the in vivo study indicated that the novel PTG/P scaffold with sustained delivery of the peptide could significantly promote bone regeneration in a critical bone defect. Thus, the novel bioactive PTG/P scaffold with a customized shape, improved mechanical strength, sustainable peptide delivery, and excellent osteogenic ability has great potential in bone tissue regeneration.

scholarly journals Tuning filament composition and microstructure of 3D-printed bioceramic scaffolds facilitate bone defect regeneration and repair

2021 ◽  
Vol 8 (2) ◽  
Author(s):  
Yi Chen ◽  
Jiaping Huang ◽  
Jiamei Liu ◽  
Yingming Wei ◽  
Xianyan Yang ◽  
...  
Keyword(s):  
3D Printing ◽  
Bone Defect ◽  
Core Shell ◽  
Shell Layer ◽  
Ion Release ◽  
Component Distribution ◽  
Bone Defect Repair ◽  
Defect Repair

Abstract It is still a challenge to optimize the component distribution and microporous structures in scaffolds for tailoring biodegradation (ion releasing) and enhancing bone defect repair within an expected time stage. Herein, the core–shell-typed nonstoichiometric wollastonite (4% and 10% Mg-doping calcium silicate; CSiMg4, CSiMg10) macroporous scaffolds with microporous shells (adding ∼10 μm PS microspheres into shell-layer slurry) were fabricated via 3D printing. The initial mechanical properties and bio-dissolution (ion releasing) in vitro, and osteogenic capacity in vivo of the bioceramic scaffolds were evaluated systematically. It was shown that endowing high-density micropores in the sparingly dissolvable CSiMg10 or dissolvable CSiMg4 shell layer inevitably led to nearly 30% reduction of compressive strength, but such micropores could readily tune the ion release behaviour of the scaffolds (CSiMg4@CSiMg10 vs. CSiMg4@CSiMg10-p; CSiMg10@CSiMg4 vs. CSiMg10@CSiMg4-p). Based on the in rabbit femoral bone defect repair model, the 3D μCT reconstruction and histological observation demonstrated that the CSiMg4@CSiMg10-p scaffolds displayed markedly higher osteogenic capability than the other scaffolds after 12 weeks of implantation. It demonstrated that core–shell bioceramic 3D printing technique can be developed to fabricate single-phase or biphasic bioactive ceramic scaffolds with accurately tailored filament biodegradation for promoting bone defect regeneration and repair in some specific pathological conditions.

Enhanced Bone Regeneration and Bone Defect Repair Using Magnesium/Lithium-Co-Modified, Porous, Hydroxyapatite Composite Scaffolds

2020 ◽  
Vol 12 (1) ◽  
pp. 124-136
Author(s):  
Hai-Yan Zhao ◽  
Releken-Yeersheng ◽  
Ya-Yi Xia ◽  
Xing-Wen Han ◽  
Chao Zhang ◽  
...  
Keyword(s):  
Bone Regeneration ◽  
Bone Defect ◽  
Biological Degradation ◽  
Porous Hydroxyapatite ◽  
Bone Defect Repair ◽  
Hydroxyapatite Composite ◽  
Defect Repair ◽  
Osteogenic Effect

Background: Hydroxyapatite (HA) has been frequently used in clinic, but it is hard to be degraded, and insufficient in osteogenesis and angiogenesis. This study aimed to modify HA by doping magnesium/lithium (Mg/Li) and assess the Mg/LiHA scaffold's bone regeneration and bone defect repair effects. Materials and Methods: The biomaterial was identified using XRD, FTIR and SEM. The porosity, cell mediated degradation behavior and mechanical property were investigated. Meanwhile, cell proliferation and adhesion were also exploited. Finally, osteogenic effect of Mg/LiHA scaffold in vitro, and bone defect repair effect in vivo were researched. Results: The results suggested that low-content of Mg/Li incorporation did not influence on the structure of HA. The cells mediated degradation experiments indicated that Mg/Li doped HA could improve the biological degradation and release the Mg2+ and Li+ sustainedly. The compressive strength of Mg/LiHA scaffolds with 63% porosity reached to 3.9 MPa. Cells proliferation and adhesion experiments demonstrated that Mg/LiHA scaffolds were beneficial to cell growth and attachment. Furthermore, Mg/LiHA scaffolds increased ALP expression, calcium phosphate deposition and VEGF expression in vitro. The bone defect repair in vivo was enhanced by using Mg/LiHA scaffolds. Conclusion: Mg/Li-co-substituted HA could enhance bone regeneration and bone defect repair, and may be recommended to further research on bone defect repair.

Sign in / Sign up

Export Citation Format

Share Document