scholarly journals Human Infrapatellar Fat Pad Mesenchymal Stem Cells Show Immunomodulatory Exosomal Signatures

2021 ◽  
Author(s):  
Dimitrios Kouroupis ◽  
Lee D Kaplan ◽  
Thomas M Best
Keyword(s):  
Stem Cells ◽  
Mesenchymal Stem Cells ◽  
Macrophage Polarization ◽  
Viable Cell ◽  
Infrapatellar Fat Pad ◽  
Therapeutic Effects ◽  
Fat Pad ◽  
Proliferation Response ◽  
Transduction Signal ◽  
Inflammatory Molecules

Abstract Within the human knee infrapatellar fat pad (IFP) and synovium, resident synoviocytes and macrophages contribute to the onset and progression of inflammatory joint diseases. Our hypothesis is that IFP-derived mesenchymal stem cells (IFP-MSC) robust immunomodulatory therapeutic effects are largely exerted via their exosomal (IFP-MSC EXOs) secretome by attenuating synoviocyte and macrophage pro-inflammatory activation. IFP-MSC EXOs showed distinct miRNA and protein immunomodulatory profiles. Reactome analysis of 24 miRNAs highly present in exosomes showed their involvement in the regulation of six gene groups, including immune system. Exosomes were enriched for immunomodulatory and reparative proteins that are involved in positive regulation of cell proliferation, response to stimulus, signal transduction, signal receptor activity, and protein phosphorylation. Stimulated synoviocytes or macrophages exposed to IFP-MSC EXOs demonstrated significantly reduced proliferation, altered inflammation-related molecular profiles, and reduced secretion of pro-inflammatory molecules compared to stimulated alone. In an acute synovial/IFP inflammation rat model, IFP-MSC EXOs therapeutic treatment resulted in robust macrophage polarization towards an anti-inflammatory therapeutic M2 phenotype within the synovium/IFP tissues. Based on these findings, we propose a viable cell-free alternative to MSC-based therapeutics as an alternative approach to treating synovitis and IFP fibrosis.

CD10/Neprilysin Enrichment in Infrapatellar Fat Pad–Derived Mesenchymal Stem Cells Under Regulatory-Compliant Conditions: Implications for Efficient Synovitis and Fat Pad Fibrosis Reversal

2020 ◽  
Vol 48 (8) ◽  
pp. 2013-2027 ◽  
Author(s):  
Dimitrios Kouroupis ◽  
Annie C. Bowles ◽  
Thomas M. Best ◽  
Lee D. Kaplan ◽  
Diego Correa
Keyword(s):  
Stem Cells ◽  
Mesenchymal Stem Cells ◽  
Ex Vivo ◽  
Infrapatellar Fat Pad ◽  
Therapeutic Effects ◽  
Fat Pad ◽  
In Vitro Proliferation ◽  
Differentiation Potential

Background: Synovitis and infrapatellar fat pad (IFP) fibrosis participate in various conditions of the knee. Substance P (SP), a neurotransmitter secreted within those structures and historically associated with nociception, also modulates local neurogenic inflammatory and fibrotic responses. Exposure of IFP mesenchymal stem cells (IFP-MSCs) to a proinflammatory/profibrotic environment (ex vivo priming with TNFα, IFNγ, and CTGF) induces their expression of CD10/neprilysin, effectively degrading SP in vitro and in vivo. Purpose/Hypothesis: The purpose was to test the therapeutic effects of IFP-MSCs processed under regulatory-compliant protocols, comparing them side-by-side with standard fetal bovine serum (FBS)–grown cells. The hypothesis was that when processed under such protocols, IFP-MSCs do not require ex vivo priming to acquire a CD10-rich phenotype efficiently degrading SP and reversing synovitis and IFP fibrosis. Study Design: Controlled laboratory study. Methods: Human IFP-MSCs were processed in FBS or either of 2 alternative conditions—regulatory-compliant pooled human platelet lysate (hPL) and chemically reinforced medium (Ch-R)—and then subjected to proinflammatory/profibrotic priming with TNFα, IFNγ, and CTGF. Cells were assessed for in vitro proliferation, stemness, immunophenotype, differentiation potential, transcriptional and secretory profiles, and SP degradation. Based on a rat model of acute synovitis and IFP fibrosis, the in vivo efficacy of cells degrading SP plus reversing structural signs of inflammation and fibrosis was assessed. Results: When compared with FBS, IFP-MSCs processed with either hPL or Ch-R exhibited a CD10High phenotype and showed enhanced proliferation, differentiation, and immunomodulatory transcriptional and secretory profiles (amplified by priming). Both methods recapitulated and augmented the secretion of growth factors seen with FBS plus priming, with some differences between them. Functionally, in vitro SP degradation was more efficient in hPL and Ch-R, confirmed upon intra-articular injection in vivo where CD10-rich IFP-MSCs also dramatically reversed signs of synovitis and IFP fibrosis even without priming or at significantly lower cell doses. Conclusion: hPL and Ch-R formulations can effectively replace FBS plus priming to induce specific therapeutic attributes in IFP-MSCs. The resulting fine-tuned, regulatory-compliant, cell-based product has potential future utilization as a novel minimally invasive cell therapy for the treatment of synovitis and IFP fibrosis. Clinical Relevance: The therapeutic enhancement of IFP-MSCs manufactured under regulatory-compliant conditions suggests that such a strategy could accelerate the time from preclinical to clinical phases. The therapeutic efficacy obtained at lower MSC numbers than currently needed and the avoidance of cell priming for efficient results could have a significant effect on the design of clinical protocols to potentially treat conditions involving synovitis and IFP fibrosis.

scholarly journals Recent Advance in Source, Property, Differentiation, and Applications of Infrapatellar Fat Pad Adipose-Derived Stem Cells

2020 ◽  
Vol 2020 ◽  
pp. 1-14 ◽  
Author(s):  
Yu-chen Zhong ◽  
Shi-chun Wang ◽  
Yin-he Han ◽  
Yu Wen
Keyword(s):  
Stem Cells ◽  
Mesenchymal Stem Cells ◽  
Regenerative Medicine ◽  
3D Culture ◽  
Infrapatellar Fat Pad ◽  
Adipose Derived Stem Cells ◽  
Fat Pad ◽  
Stem Cell Source ◽  
Marrow Mesenchymal Stem Cells ◽  
And Function

Infrapatellar fat pad (IPFP) can be easily obtained during knee surgery, which avoids the damage to patients for obtaining IPFP. Infrapatellar fat pad adipose-derived stem cells (IPFP-ASCs) are also called infrapatellar fat pad mesenchymal stem cells (IPFP-MSCs) because the morphology of IPFP-ASCs is similar to that of bone marrow mesenchymal stem cells (BM-MSCs). IPFP-ASCs are attracting more and more attention due to their characteristics suitable to regenerative medicine such as strong proliferation and differentiation, anti-inflammation, antiaging, secreting cytokines, multipotential capacity, and 3D culture. IPFP-ASCs can repair articular cartilage and relieve the pain caused by osteoarthritis, so most of IPFP-related review articles focus on osteoarthritis. This article reviews the anatomy and function of IPFP, as well as the discovery, amplification, multipotential capacity, and application of IPFP-ASCs in order to explain why IPFP-ASC is a superior stem cell source in regenerative medicine.

scholarly journals Canine mesenchymal stem cells from synovium have a higher chondrogenic potential than those from infrapatellar fat pad, adipose tissue, and bone marrow

PLoS ONE ◽  
2018 ◽  
Vol 13 (8) ◽  
pp. e0202922 ◽  
Author(s):  
Akari Sasaki ◽  
Mitsuru Mizuno ◽  
Nobutake Ozeki ◽  
Hisako Katano ◽  
Koji Otabe ◽  
...  
Keyword(s):  
Stem Cells ◽  
Bone Marrow ◽  
Mesenchymal Stem Cells ◽  
Adipose Tissue ◽  
Infrapatellar Fat Pad ◽  
Fat Pad ◽  
Chondrogenic Potential

scholarly journals The infrapatellar fat pad from diseased joints inhibits chondrogenesis of mesenchymal stem cells

2015 ◽  
Vol 23 ◽  
pp. A145-A146 ◽  
Author(s):  
W. Wei ◽  
R. Rudjito ◽  
N. Fahy ◽  
K.P. Bos ◽  
J.A. Verhaar ◽  
...  
Keyword(s):  
Stem Cells ◽  
Mesenchymal Stem Cells ◽  
Infrapatellar Fat Pad ◽  
Fat Pad

scholarly journals Mechanism of mesenchymal stem cells in spinal cord injury repair through macrophage polarization

2021 ◽  
Vol 11 (1) ◽  
Author(s):  
Nan An ◽  
Jiaxu Yang ◽  
Hequn Wang ◽  
Shengfeng Sun ◽  
Hao Wu ◽  
...  
Keyword(s):  
Spinal Cord ◽  
Stem Cells ◽  
Spinal Cord Injury ◽  
Mesenchymal Stem Cells ◽  
Clinical Medicine ◽  
Macrophage Polarization ◽  
Modern Medicine ◽  
Therapeutic Effects ◽  
Inflammatory Reactions ◽  
Cord Injury

AbstractTreatment and rehabilitation of spinal cord injury (SCI) is a major problem in clinical medicine. Modern medicine has achieved minimal progress in improving the functions of injured nerves in patients with SCI, mainly due to the complex pathophysiological changes that present after injury. Inflammatory reactions occurring after SCI are related to various functions of immune cells over time at different injury sites. Macrophages are important mediators of inflammatory reactions and are divided into two different subtypes (M1 and M2), which play important roles at different times after SCI. Mesenchymal stem cells (MSCs) are characterized by multi-differentiation and immunoregulatory potentials, and different treatments can have different effects on macrophage polarization. MSC transplantation has become a promising method for eliminating nerve injury caused by SCI and can help repair injured nerve tissues. Therapeutic effects are related to the induced formation of specific immune microenvironments, caused by influencing macrophage polarization, controlling the consequences of secondary injury after SCI, and assisting with function recovery. Herein, we review the mechanisms whereby MSCs affect macrophage-induced specific immune microenvironments, and discuss potential avenues of investigation for improving SCI treatment.

scholarly journals Gelactin-9 Mediates the Therapeutic Effect of Mesenchymal Stem Cells on Experimental Endotoxemia

2020 ◽  
Author(s):  
Yiming Zhao ◽  
Dingding Yu ◽  
Hongda Wang ◽  
Wang Jin ◽  
Xiang Li ◽  
...  
Keyword(s):  
Stem Cells ◽  
Flow Cytometry ◽  
Mesenchymal Stem Cells ◽  
Survival Rate ◽  
Inflammatory Mediators ◽  
Macrophage Polarization ◽  
Therapeutic Effects ◽  
Dependent Manner ◽  
Experimental Endotoxemia ◽  
Ifn Γ

Abstract Background: Endotoxemia, mediated by uncontrolled immunocytes activation toward Lipopolysaccharide, could deteriorate into severe septic shock, but with limited treatment effect. Mesenchymal stem cells (MSCs), with excellent immune regulatory capacities, have displayed potential in multiple inflammatory disease treatment. Gelactin-9 (Gal-9), a newly discovered immune checkpoint, has been demonstrated to mediate immunomodulatory effect of MSCs in vitro. However, its in vivo role in alleviating endotoxemia remains to be elucidated.Methods: MSCs (2.5×10⁵/ml) were obtained and stimulated with IFN-γ (20ng/ml) for 72 hours. Gal-9 expression on MSCs were measured by ELISA, RT-PCR, flow cytometry and immunofluorescence respectively. Then, experimental endotoxemia was induced by LPS injection (10mg/kg, i.p.), followed by the treatment with Gal-9-MSC (20ng/ml, 72 hours), MSCs and MSC+α-lactose (10.8mg/mL, 500ul, i.v.). Therapeutic effects of MSC-based treatments were assessed by monitoring murine sepsis score, survival rate, splenocyte proportion, phenotype polarization, inflammatory mediator levels and pathological manifestations. Furthermore, Gal-9 expression in multiple organs was also detected after administering the treatments.Results: It has been found that MSCs expressed Gal-9 and its level was increased in a dose-dependent manner after being stimulated by IFN-γ. Moreover, adoptive transferred of IFN-γ pre-stimulated MSCs into endotoxemia mice was found with relieved symptoms and increased survival rate. Flow cytometry analysis indicated that Gal-9-MSC could promote macrophage polarization to M2-subtype and increase Treg ratios in spleen. Further results also demonstrated that, Gal-9-mediated MSC therapy could assist in attenuating local and circulating pro-inflammatory mediators expression (TNF-α, IL-1β, IFN-γ and iNOS), but increasing anti-inflammatory mediators expression (T-SOD and IL-35). Additionally, after administrating Gal-9-MSC, it was also found there was a significant relief in pathological manifestations, and with a higher expression of Gal-9 in liver, kidney and lung homogenate.Conclusions: This study revealed that Gal-9 mediated therapeutic effects of MSCs in alleviating endotoxemia injury, which provides a novel idea for supplementing the research of MSC immunoregulatory mechanism, and offers an excellent candidate to be used in treatment of endotoxemia in the clinical settings.

scholarly journals Inability of Low Oxygen Tension to Induce Chondrogenesis in Human Infrapatellar Fat Pad Mesenchymal Stem Cells

2021 ◽  
Vol 9 ◽  
Author(s):  
Samia Rahman ◽  
Alexander R. A. Szojka ◽  
Yan Liang ◽  
Melanie Kunze ◽  
Victoria Goncalves ◽  
...  
Keyword(s):  
Stem Cells ◽  
Mesenchymal Stem Cells ◽  
Oxygen Tension ◽  
Endochondral Ossification ◽  
Infrapatellar Fat Pad ◽  
Low Oxygen ◽  
Fat Pad ◽  
Chondrogenic Medium ◽  
Low Oxygen Tension

ObjectiveArticular cartilage of the knee joint is avascular, exists under a low oxygen tension microenvironment, and does not self-heal when injured. Human infrapatellar fat pad-sourced mesenchymal stem cells (IFP-MSC) are an arthroscopically accessible source of mesenchymal stem cells (MSC) for the repair of articular cartilage defects. Human IFP-MSC exists physiologically under a low oxygen tension (i.e., 1–5%) microenvironment. Human bone marrow mesenchymal stem cells (BM-MSC) exist physiologically within a similar range of oxygen tension. A low oxygen tension of 2% spontaneously induced chondrogenesis in micromass pellets of human BM-MSC. However, this is yet to be demonstrated in human IFP-MSC or other adipose tissue-sourced MSC. In this study, we explored the potential of low oxygen tension at 2% to drive the in vitro chondrogenesis of IFP-MSC. We hypothesized that 2% O2 will induce stable chondrogenesis in human IFP-MSC without the risk of undergoing endochondral ossification at ectopic sites of implantation.MethodsMicromass pellets of human IFP-MSC were cultured under 2% O2 or 21% O2 (normal atmosphere O2) in the presence or absence of chondrogenic medium with transforming growth factor-β3 (TGFβ3) for 3 weeks. Following in vitro chondrogenesis, the resulting pellets were implanted in immunodeficient athymic nude mice for 3 weeks.ResultsA low oxygen tension of 2% was unable to induce chondrogenesis in human IFP-MSC. In contrast, chondrogenic medium with TGFβ3 induced in vitro chondrogenesis. All pellets were devoid of any evidence of undergoing endochondral ossification after subcutaneous implantation in athymic mice.

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