Table 2.
Study characteristics and outcomes.
| Author | Journal | Year | Cell source | Target disease | Study type | Cells management | Bioengineering method for MSCs paracrine mediators | Measurement instrument |
|---|---|---|---|---|---|---|---|---|
| Chang J | Bone Res | 2015 | C57BL/6 mice | Bone fracture | In-vitro | BMSC CM and Mφs cell contract co-culture | BMSCs intrinsic bio-activation: the supernatants from BMSCs cultures were collected and stored at −80 °C until used as conditioned medium. | Scratch assay, BMSCs migration assay, IL-6 ELISA assay, cell growth assay, Gene expression by RT-PCR, Western blot |
| Cosenza et al., 2017 | Sci Rep | 2017 | C57BL/6 mice | Osteoarthritis | In-vitro and in-vivo |
In-vitro: BMSC Exos and Mφs In-vivo (arthritis model): IA injections of BM-MSCs, MPs or Exos. |
BMSCs intrinsic bio-activation: BMSC-CM was centrifuged at 300 g for 10 min to eliminate cells and 2,500 g for 25 min to remove debris and apoptotic bodies. For MP isolation, CM was centrifuged at 18,000 g for 1 h in polyallomer tubes; the pellet was then suspended in PBS and submitted to a second round of centrifugation. For Exos, supernatant from MP fraction was filtered on 0.22 μm porous membrane and centrifuged at 100,000 g for 2 h. | Flow cytometry analysis, Bone parameter analyses, Confocal laser scanning microscopy, Histological analysis |
| Lo Sicco | Stem Cells Transl Med | 2017 | Human-ADSCs; C57BL/6 mice–Mφs | Muscle damage | In-vitro and in-vivo |
In-vitro: ADSC EVs and Mφs In-vivo (muscle injury model): IL injection of ADSCs EVs |
ADSCs extrinsic modification: ADSC EVs were isolated from normoxic- and hypoxic-conditioned media by differential centrifugation at 300 g for 10 min, 2,000 g for 20 min, 10,000 g for 30 min at 4°C to eliminate cells and debris | Protein quantification and immunoblot analysis, Flow cytometry analysis, qRT-PCR, Immunofluorescence analysis, Histology and morphometric analysis |
| Hyvärinen K | Front Immunol | 2018 | Human | / | In-vitro | BMSCs or BMSC-EVs and Mφs co-culture | BMSCs intrinsic bio-activation: BMSCs media were collected and centrifuged at 2,000 g for 10 min to remove cell debris. The supernatant was ultracentrifuged with Optima™ MAX-XP Ultracentrifuge (Beckman Coulter) at 100,000 g 1.5 h +4°C with MLA-50 rotor (k-factor = 92, Beckman Coulter), and the pelleted EVs were combined. For the second EV collection, the cell starvation was continued in 200 ml α-MEM at 37°C, 5% CO2 for 2 days followed by replication of EV centrifugation steps. | Flow cytometry analysis, cytokine (IL-10, IL-23, IL-22) and LMs measurements |
| Zhang S | Biomaterials | 2018 | Human embryonic stem cell derived MSCs | Osteoarthritis | In-vivo | Sprague-Dawley rat osteochondral defect model: IA injection of MSCs-Exos | BMSCs intrinsic bio-activation: MSCs were grown in a chemically defined medium for 3 days and exosomes were purified from the CM. | Histology and immunohistochemistry, Multiplex cytokine assay (IL-1β, IL-6, TNF-β) |
| Chamberlain CS | Stem Cells | 2019 | Human | Tendon injury | In-vitro and in-vivo |
In-vitro: BMSC-EVs and Mφs; In-vivo (Foxn1nu mouse model of Achilles tendon injury): IL injection of BMSCs, CD14+ Mφs or EEMs |
BMSCs intrinsic bio-activation: BMSCs CM was centrifuged using a Beckman Coulter Allegra X-15R centrifuge at 2,000 g at 4°C for 20 min. Clarified supernatant CM was then centrifuged in a Beckman Coulter Optima L-80 XP Ultracentrifuge at 100,000 g at 4°C for 2 h with a swinging bucket SW 28 rotor to pellet EVs. | Flow cytometry analysis, IHC/Immunofluorescence/Histology; Fractal analysis; Mechanical testing |
| Pacienza N | Mol Ther Methods Clin Dev | 2019 | Human–BMSCs; RAW 264.7–Mφs cell |
/ | In-vitro and in-vivo |
In-vitro: LPS in combination with Exos and Mφs; In-vivo (endotoxemia mouse model): IV injection of Exos |
BMSCs intrinsic bio-activation: BMSCs CM was applied directly at room temperature to a column containing the anion exchange resin (Q Sepharose Fast Flow, GE Healthcare, Chicago, IL, USA) that had been equilibrated with 50 mM NaCl in 50 mM phosphate buffer (pH 7.5). The column resin was washed with 100 mM NaCl in 50 mM phosphate buffer (pH 7.5) and then eluted with 500 mM NaCl in 50 mM phosphate buffer (pH 7.5). | qRT-PCR, Quantitation of TNF-a, IL-1β, and IL-6 by ELISA |
| Shi Z | J Transl Med | 2019 | Sprague-Dawley rats–BMSCs | Tendon injury | In-vivo | Sprague-Dawley rat patellar tendon defect model: injection of fibrin containing EVs | BMSCs CM was centrifuged sequentially at 300 g for 10 min followed by 2,000 g for 10 min to remove cellular debris. The supernatants were then ultracentrifuged at 100,000 g for 2 h to obtain a pellet containing the EVs, which was resuspended in 200 μL of PBS. EVs-enriched fraction was centrifuged at 1,500 g, 30 min with 100-kDa molecular weight cut off (MWCO) hollow fiber membrane (Millipore, Billerica, MA, USA). Then, EVs were passed through a 0.22-μm filter. | Histology and immunohistochemistry, Gene expression, Histological analysis |
| Li Y | J Cell Biochem | 2019 | C57BL/6 mice | Bone fracture | In-vitro and in-vivo | In-vitro: ADSCs and Mφs cell contract co-culture;In-vivo (a cylindrical bone defect model): IL injection of ADSCs | / | Immunohistochemistry, Western-blot analysis, RT-PCR, Enzyme-linked immunosorbent assay, micro-CT |
| Shen H | J Orthop Res | 2020 | Scleraxis-GFP tendon reporter mice or NF-κB-GFP-luciferase transgenic reporter mice–ADSCs Wild type FVB/NJ (FVB) –Mφs |
Tendon injury | In-vitro and in-vivo |
In-vitro: Exos and Mφs co-culture; In-vivo (NGL mouse model of Achilles tendon injury and repair): collagen sheet loaded with EVs from naÃ_ve ASCs or IFNγ-primed ASCs |
ADSCs intrinsic bio-activation: ADSCs were cultured in an EV collection medium (2% EV-free FBS in α-MEM) for 48 h. CM from ASC culture (150 ml from ~2.5 E+07 cells per isolation) with or without IFNγ pre-treatment was collected and centrifuged at 500 g for 10 min and 10,000 g for 30 min at 4°C to remove large vesicles. After passing through a 0.22 μm filter, the medium was further centrifuged at 100,000 g for 90 min at 4°C. | Mφs assays, NF-κB-Luciferase Imaging in-vivo, RT-PCR, Histology |
Both the methodology employed and the results obtained by each article are represented in this table. BMSCs, bone marrow stem cells; ADSCs, adipose tissue derived stem cells; Mφs, macrophages; Exos, exosomes; MPs, microparticles; EVs, extracellular vesicles; CM, conditioned medium; IA, intra-articularly; IL, intralesional; IV, intravenous; LMs, lipid mediators; EEMs, exosome-educated macrophages; LPS, lipopolysaccharide.