Persistence of fluorescent nanoparticle-labelled bone marrow mesenchymal stem cells in vitro and after intra-articular injection.
Abstract: Mesenchymal stem cells (MSCs) improve the osteoarthritis condition, but the fate of MSCs after intra-articular injection is unclear. We used fluorescent nanoparticles (quantum dots [QDs]) to track equine MSCs (QD-labelled MSCs [QD-MSCs]) in vivo after intra-articular injection into normal and osteoarthritic joints. One week after injection of QD-MSCs, unlabelled MSCs, or vehicle, we determined the presence of QD-MSCs in synovium and articular cartilage histologically. In vitro, we evaluated the persistence of QDs in MSCs and whether QDs affected proliferation, immunophenotype, or differentiation. In joints injected with QD-MSCs, labelled cells were identified on the synovial membrane and significantly less often on articular cartilage, without differences between normal and osteoarthritic joints. Joints injected with QD-MSCs and MSCs had increased synovial total nucleated cell count and protein compared with vehicle-injected joints. In vitro, QDs persisted in nonproliferating cells for up to 8 weeks (length of the study), but QD fluorescence was essentially absent from proliferating cells within two passages (approximately 3 to 5 days). QD labelling did not affect MSC differentiation into chondrocytes, adipocytes, and osteocytes. QD-MSCs had slightly different immunophenotype from control cells, but whether this was due to an effect of the QDs or to drift during culture is unknown. QD-MSCs can be visualized in histological sections 1 week after intra-articular injection and are more frequently found in the synovial membrane versus cartilage in both normal and osteoarthritic joints. QDs do not alter MSC viability and differentiation potential in vitro. However, QDs are not optimal markers for long-term tracking of MSCs, especially under proliferative conditions.
© 2018 John Wiley & Sons, Ltd.
Publication Date: 2019-01-23 PubMed ID: 30536848PubMed Central: PMC6393194DOI: 10.1002/term.2781Google Scholar: Lookup
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- Journal Article
- Research Support
- N.I.H.
- Extramural
- Research Support
- Non-U.S. Gov't
- Articular Cartilage
- Biomedical Research
- Biotechnology
- Cell Proliferation
- Cell Viability
- Equine Diseases
- Equine Health
- Fluorescence
- Histology
- Immunophenotyping
- In Vitro Research
- In Vivo
- Intra-Articular Injection
- Mesenchymal Cells
- Nanoparticles
- Osteoarthritis
- Stem Cells
- Synovial Fluid
- Veterinary Medicine
- Veterinary Research
Summary
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This research investigates the behavior of equine bone marrow mesenchymal stem cells (MSCs) labelled with fluorescent nanoparticles, or quantum dots (QDs), in vivo and in vitro after being injected into both healthy and osteoarthritic joints. It provides insights into the QDs’ impact on MSC proliferation, immunophenotype, and differentiation, concluding that while QDs are practical for short-term tracking of MSCs, they are not suitable for long-term tracking, particularly in proliferative conditions.
Methodology and Objectives
- This research aimed to determine the properties and fate of equine mesenchymal stem cells (MSCs) labeled with fluorescent nanoparticles (quantum dots) when injected into both normal and osteoarthritic joints.
- Researchers aimed to study whether the QDs would affect the proliferation, immune characteristics, or the ability of MSCs to change into other forms, a process known as differentiation.
Processes
- The researchers injected QD-labelled MSCs into normal and osteoarthritic joints and observed their positions in the synovium (the soft tissue that lines the non-cartilaginous surfaces within joints) and articular cartilage.
- They also observed how QDs influenced the activity of MSCs in a controlled lab environment to evaluate their persistence.
- Finally, they compared the effect on the synovial fluid when joints are injected with QDs, MSCs, or a vehicle.
- In vitro, the persistence of QDs in MSCs was analyzed and the possible effects of QDs on proliferation, immunophenotype or differentiation were evaluated.
Findings
- The MSCs labelled with QDs were identified in the synovial membrane considerably more frequently than the articular cartilage, regardless of whether or not the joint was osteoarthritic.
- The presence of QD-MSCs and MSCs increased the total count of nucleated cells and protein in the synovial fluid when compared to joints injected with only the vehicle.
- The QDs remained in non-dividing cells for the duration of the study (which lasted eight weeks), but were basically absent from dividing cells within two times of cell division (approximately 3 to 5 days).
- The labelling of MSCs with QDs did not seem to alter the cells’ potential to differentiate into other types of cells or affect their viability.
- The immunophenotype was slightly different in QD-MSCs compared to control cells, however the cause is unknown: it could be an effect of the QDs or possibly due to drift during the culturing process.
Conclusion
- Results indicate that QD-labelled MSCs can be visualized in tissue samples a week after being injected into a joint and are typically found more frequently in the synovial membrane than in the cartilage, regardless of whether or not the joint is osteoarthritic.
- While QDs do not influence the viability and differentiation potential of MSCs, they are not an ideal tool for long-term tracking of MSCs due to their tendency to not persist in cells that are actively dividing.
Cite This Article
APA
Grady ST, Britton L, Hinrichs K, Nixon AJ, Watts AE.
(2019).
Persistence of fluorescent nanoparticle-labelled bone marrow mesenchymal stem cells in vitro and after intra-articular injection.
J Tissue Eng Regen Med, 13(2), 191-202.
https://doi.org/10.1002/term.2781 Publication
Researcher Affiliations
- Department of Veterinary Physiology and Pharmacology, Texas A&M University, College Station, Texas, USA.
- Department of Veterinary Physiology and Pharmacology, Texas A&M University, College Station, Texas, USA.
- Department of Veterinary Physiology and Pharmacology, Texas A&M University, College Station, Texas, USA.
- Department of Large Animal Clinical Sciences, Texas A&M University, College Station, Texas, USA.
- Clinical Sciences, Cornell University, Ithaca, New York.
- Department of Large Animal Clinical Sciences, Texas A&M University, College Station, Texas, USA.
MeSH Terms
- Allografts
- Animals
- Bone Marrow Cells / metabolism
- Bone Marrow Cells / pathology
- Horse Diseases / metabolism
- Horse Diseases / pathology
- Horse Diseases / therapy
- Horses
- Joints / metabolism
- Joints / pathology
- Mesenchymal Stem Cell Transplantation
- Mesenchymal Stem Cells / metabolism
- Mesenchymal Stem Cells / pathology
- Osteoarthritis / metabolism
- Osteoarthritis / pathology
- Osteoarthritis / therapy
- Quantum Dots / chemistry
Grant Funding
- T32 RR007059 / NCRR NIH HHS
- F32 AR057299 / NIAMS NIH HHS
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