TABLE 1.
Studies regarding mesenchymal stromal cell-conditioned medium for treating wounds in animal models.
| MSC source | Method of tissue extraction | MSC characterization | Preparation of MSC-CM | Model | Groups of treatments and via of administration | Follow-up (days) | Assessment | Main outcome | Other outcomes | |
| Chen et al., 2021 | Human subcutaneous adipose tissue samples | Liposuction | Flow cytometry (CD29+, CD90+, CD105+, CD31−, CD34−, CD45−). Osteogenic and adipogenic differentiation | MSCs of passage 3 at 90% confluence were used. CMs at different dilutions (100, 50, and 25%) were collected. Electrospun fibrous scaffolds formed by emulsion electrospinning; an ADSC-CM loaded micro-nano polylactic acid (PLA) electrospun fiber (MPF) was developed | Murine. Full-thickness excisional wounds, 15 mm × 15 mm, on dorsal surface | Wounds were covered with an electrospun membrane - Control - MPF alone - MPF loaded with 100% AT-MSC-CM - MPF loaded with 50% AT-MSC-CM - MPF loaded with 25% ATMSC-CM (n = 3/group) |
15 | Macroscopic appearance (photography), histology (HE, MT), IHC, qRT-PCR | MPF loaded with MSC-CM accelerated wound healing and prevented abnormal scar formation, promoting angiogenesis without adversely affecting epidermal cells | MPF loaded with MSC-CM inhibited ECM deposition, including Col I and Col III, and decreased α-SMA expression, showing an inhibitory effect on fibroblast differentiation. Groups with less collagen deposition and smaller scar area showed more VEGF expression and a faster healing rate. The wound area of MPF loaded with 100% AT-MSC-CM was only 34.9% of that of the control group |
| Zhou et al., 2020 | Human dental pulp isolated from periodontally compromised teeth (P-DP-MSCs) and healthy teeth (H-DP-MSCs) | Teeth extracting | Flow cytometry (CD90+, CD105+, CD146+, CD31−, CD34−, CD45−). Osteogenic, adipogenic, and chondrogenic differentiation | MSCs of passage 4 at 90% confluence were used. CM was collected and, it was used to obtain EVs | Murine. Full-thickness excisional wounds, on the dorsal surface | 100 μl subcutaneously injected at 4 sites around the wound (25 μl per site) - Control - P-DP-MSC-CM EVs - H-DP-MSC- CM EVs (n = 10/group) |
14 | Macroscopic appearance (photography), microscopic appearance (newly formed vessels), histology (HE), IHC (VEGF, CD31) | Wound closure was markedly accelerated by DP-MSC-CM EVs and P-DP-MSC-CM EVs. P-DP-MSC-CM EV group closed faster than H-DP-MSC-CM EV group (p < 0.05) | DP-MSC-CM extracellular vesicle-treated wounds had a lower level of scar formation and enhanced vessel formation in the wound sites. No adverse events were observed |
| Sabzevari et al., 2020 | Wharton’s jelly from human umbilical cords | Cesarean section | Flow cytometry (CD44+, CD73+, CD90+, CD105+, CD14−, CD31−, CD34−, CD45−, HLA-DR−). Osteogenic, adipogenic, and chondrogenic differentiation | MSC of passages 4–6 at 70% confluence were used. WJ-MSCs were transfected with a recombinant construct encoding hCAP-18/LL-37 gene. Next, the CM of the transfected cells (LL-37-MSCs) was harvested, and its concentrate was formulated in a sodium alginate (SA)/gelatine (G) hydrogel | Murine. Full-thickness circular excision wound, 20 mm, below the skull | The hydrogel was placed on the wound and covered by a wound dressing - PBS (control) - SA/G-PBS group (the SA/G-2/8 hydrogel contained only PBS) - SA/G-V-CM group (the SA/G-2/8 hydrogel contained only CM vehicle) - SA/G-LL-37-CM group (the SA/G-2/8 hydrogel contained LL-37-CM) (n = 3/group) |
21 | Macroscopic appearance (photography), histology (HE, MT), qRT-PCR | WJ-MSC-CM hydrogel effectively accelerated wound contraction and promoted wound healing, even more in the SA/G-LL-37-CM group | Blood vessels were higher in SA/G-LL-37-CM group, showing the early angiogenesis profile with higher VEGF-A levels. This group also showed the best collagen arrangement, thickness, and density |
| Joseph et al., 2020 | Caprine, canine, and guinea pig bone marrow | Aspiration needle from the iliac crest | Flow cytometry (CD 73+, CD90+, CD 105+, CD34−). Chondrogenic, osteogenic, and adipogenic differentiation | MSCs of passage 3 at 80% confluence were used. CM was collected and then lyophilized by the freeze−drying method and was vacuum sealed and stored at 4°C. A serum−free DMEM medium was processed for use as a control | Guinea pig. Full-thickness excisional skin wounds, 20 mm × 20 mm, on the dorsum on either side of the midline | 100 μl of formulated respective CM was applied topically with sterile applicator over the wounds one per week - Control (laminin gel+DMEM) - Canine CM (laminin gel+canine MSC−CM) - Caprine CM (laminin gel+caprine MSC−CM) |
28 | Macroscopic appearance (photography), histology (HE) | MSC−CM accelerates excision wound healing compared with control (p < 0.05) | Surface epithelium, neovascularization, and collagen depositions improved more in MSC-CM-treated group than in controls (p < 0.05). Allogeneic and xenogenic application of CM significantly improved wound healing quality, with minimal scar formation. Better healing rate was observed in the allogeneic group |
| - Guinea pig CM (laminin gel+guinea pig MSC−CM) (n = 6/group) | ||||||||||
| He et al., 2020 | Human amnion | Amniocentesis | Flow cytometry (CD73, CD45, CD31, CD105, CD44, CD34, CD90, CD29, SSEA-3, SSEA-4, EP-CAM, HLA-DR). Chondrogenic, osteogenic, and adipogenic differentiation | MSCs of passage 2 at 90% confluence were used. CM was collected | Murine. Full-thickness excisional skin wounds, 10 mm, on the dorsum | 50 μl subcutaneous injected into the surrounding tissues of the wound bed at four sites - PBS (control) - MSC-CM - Murine LOXL2 (5 μg, Sino Biologicals, China) (n = 4/group) |
12 | Macroscopic appearance (photography), histology (HE, TC) | Wound sizes were significantly reduced in the LOXL2 and MSC-CM groups compared with controls (p < 0.05) | MSC-CM and LOXL2 enhanced wound healing. Epidermis of the MSC-CM and LOXL2 group mice resembled normal skin and the keratinocytes were well organized and tightly arranged. These treatments also significantly reduced fibrosis and improved keratinocyte proliferation. No adverse events were reported |
| Ahangar et al., 2020 | Human bone marrow | Aspiration | − | MSCs of passage 1 at 70% confluence were used. CM was collected | Murine. Full-thickness excisional skin wounds, 6 mm2, on dorsal surface | 100 μl was injected at four sites into the margins of each wound MSC-CM - DMEM (n = 8/group) |
14 | Macroscopic appearance (photography), histology (HE, TC), IHC, qRT-PCR | MSC-CM improved wound healing, showing a significant reduction in average wound area and wound width compared with DMEM-treated group (p < 0.05) | At day 7 post-wounding, wounds in MSC-CM-treated mice were 90% re-epithelialized compared with 78% in control group. MSC-CM also decreased inflammatory response, increased endothelial cell number and angiogenesis (showed in an increased numbers of CD31-positive |
| endothelial cells), and increased both collagen I and III expression | ||||||||||
| Rong et al., 2019 | ASC: sticks of antlers from sika deer hU-MSC: human umbilical cords | hU-MSC: Cesarean sections | − | MSCs of passage 3 at 80% confluence were used. CM was collected | Murine. Full-thickness skin excisional wounds, 8 mm in diameter, on the dorsal surface | MSC-CM was mixed with the hydrogel. 200 μl of each group was pipetted onto the wound every 2 days - DMEM - EGF - MSC-CM - ASC-CM (n = 8/group) |
16 | Photography, histology (HE), IHC | ASC-CM group accelerated wound healing compared with other treatments. At 16 days, wounds in ASC-CM group were completely closed, while other groups showed different sizes of unhealed wounds (DMEM, 6.14 ± 4.1 mm2; EGF, 1.79 ± 3.2 mm2; hU-MSC-CM, 0.61 ± 2.3 mm2, p < 0.05) | ASC-CM group had the thickest dermis containing the highest number of cutaneous appendages and the highest vessel numbers. The ASC-CM treatment significantly upregulated the expression ratios of Col3A1/Col1A2, TGF-β3/TGF-β1, MMP1/TIMP1 and MMP3/TIMP1 |
| Robert et al., 2019 | Human skin | Face-lifting | − | MSCs of passage 6 at 90% confluence were used. CM was collected | Murine. Full-thickness excisional wound, 6 mm in diameter, on dorsum surface | CM was directly applied to the wounds or embedded within hydrogels - Control, no treatment (n = 12) - Carrageenan hydrogel (n = 9) - Carrageenan hydrogel embedded with MSC-CM (n = 10) - Polyvinyl alcohol hydrogel (n = 10) - Polyvinyl alcohol hydrogel-embedded with MSC-CM (n = 11) - Only MSC-CM (n = 8) |
14 | Macroscopic appearance (photography), histology (HE) | All groups showed successfully repaired and closed wounds, although the animals treated with CM embedded in PVA (or only with PVA) displayed slightly larger wounds (p > 0.05) | Improvements in wound closure were not observed, but MSC-CM increased angiogenesis, independently of the hydrogel used |
| Raj et al., 2019 | Human umbilical cord | Umbilical cord dissection | Flow cytometry (CD13+, CD29+, CD44+, CD90+, CD10−, CD14−, CD34−, CD117−) | MSCs were transduced with a lentiviral vector (green fluorescence protein tagged). MSCs of passage 3–4 were used. CM was collected. Wound dressing patches: impregnated of aloe verapolycaprolactone (AV/PCL) nanoscaffolds with hWJSCs or hWJSC−CM were also created | Murine. Full-thickness excisional wound, 8 mm, on the back | 100 μl of: - PBS with 1 × 106 MSCs - MSC-CM - PBS with 1 × 106 fibroblast - Fibroblast-CM - UCM - PBS with 1 × 106 MSCs+AV/PCL - MSC-CM+AV/PCL - PBS with 1 × 106 fibroblast + AV/PCL - Fibroblast-CM+AV/PCL - PBS+AV/PCL - Untreated group (n = 9/group) |
28 | Macroscopic appearance (photography), histology (HE), IHC, qRT−PCR | The dermal thickness of both MSCs+AV/PCL (290.55 μm) and MSC−CM+AV/PCL (338.3 μm) treatment groups was significantly greater than that of the controls (PBS+AV/PCL, 193.51 μm; UCM, 266.55 μm; fibroblast+AV/PCL, 235.29 μm; fibroblast−CM+ AV/PCL, 227.31 μm) | CD31 marker showed strong positive signals in the MSC-CM group and in the MSCs group compared with untreated controls |
| Im et al., 2019 | Human (type is not specified) | Purchased from Lonza (Basel, Switzerland) | − | MSCs of passages 6–12 were used. CM from the MSCs treated with or without gold–iron nanoparticles was collected | Murine. Full-thickness excisional wound, 20 mm × 20 mm, on the back | Daily injection of CM (200 μl/wound) was for 4 days - MSC-CM passage 6 - MSC-CM passage 12 - MSC-CM passage 12 with gold–iron nanoparticles (n = 4/group) |
14 | Macroscopic appearance (photography), histology (HE), IHC, qRT-PCR | All wounds were almost closed with similar appearance in all groups | Increased CD31 expression was observed in MSC- CM passage 12 with gold–iron nanoparticles group compared with the passage 12 without gold–iron nano- particles. SM-α expression did not exhibit differences between groups. The relative amount of involucrin and laminin was higher in MSC-CM passage 6 and MSC-CM passage 12 with gold–iron nanoparticles compared with MSC-CM passage 12 |
| Yuan et al., 2018 | Human adipose tissue | − | Flow cytometry (CD49+, CD73+, CD90+, CD105+, CD34−). Adipogenic and chondrogenic differentiation | MSCs of passages 2–5 were used. Cells were cultured in the presence of the profibrogenic cytokine TGF-β1. CM was collected | Murine. Full-thickness skin wounds, 6 mm punch, on the back | Wounds were applied with 0, 10, 50, and 100% MSC-CM* or left untreated (n = 7/group) | 14 | Macroscopic appearance (photography) | MSC-CM treatment significantly accelerated wound healing (p < 0.05) | Compared with the non-treated wounds and 0% MSC-CM group, 100% MSC-CM treatment accelerated wound healing on the seventh day after wounding (ratio of untreated wound area: 100% in untreated group, 100% in 0% MSC-CM, and 50% in 100% MSC-CM) |
| Payushina et al., 2018 | Murine bone marrow from tibia and femur | − | Flow cytometry (CD73+, CD90+). Osteogenic and adipogenic differentiation | MSCs of passage 2 were used. CM was collected | Murine. Full-thickness skin wounds, 20 mm in diameter, on the back | 100 μl MSC-CM or control CM was injected into the wound bed each other day five times (n = not specified) | 14 | Clinical examination, histology | MSC-CM group showed greater number of blood vessels (53.20 ± 2.58 blood vessels per field of view) compared with controls (37.20 ± 4.73), p < 0.05 | MSC-CM group showed less intensive inflammation and more complete epithelialization compared with controls |
| Park et al., 2018 | Human amniotic fluid | Amniocentesis | Flow cytometry (CD13+, CD29+, CD44+, CD71+, CD90+, CD120a+, CD31−, CD106−, CD15−, CD33−, CD34−, CD45−). Adipogenic, osteogenic, and chondrogenic differentiation | MSCs of passage 12 at 70–100% confluence were used. They were supplemented with selenium and bFGF. CM was collected | Murine. Full-thickness skin wounds, 8 mm, side of the midline | Vehicle, CM supplemented with selenium (−/s), CM supplemented with bFGF (b/−), or CM supplemented with bot selenium and bFGF (b/s), was topically applied to the induced wounds in a volume of 20 μl (n = 10/group) | 11 | Macroscopic appearance (photography), histology (HE, 3,3′-diaminobenzidine tetrahydrochloride staining), IHC | Treatment with CM (b/s) resulted in complete wound closure, while not completed closure was observed in the other groups | The CM (−/s) group exhibited better recovery than the CM (b/−) group. The CM (b/s) group showed the thickest epidermis region and the highest expression of involucrin. Smad2, AKT-MEK1/2-ERK, and NFκB signaling pathways were more effectively activated by CM (−/s) than by CM (b/−), and their |
| highest activation was seen when treated with CM (b/s) | ||||||||||
| Tarcisia et al., 2017 | Adipose tissue | − | Flow cytometry positive for (CD73+, CD90+, CD34−). Osteogenic, chondrogenic, and adipogenic differentiation | MSCs of passage 3 were used. CM was then collected | Murine. Full-thickness excisional skin wounds, 20 mm long and 5 mm depth, on the back | The four cuts of each rat were treated randomly with MSC-CM (100%), complete culture medium, basal medium, and without treatment (control). The treatment was only done once after the rat skin injury (n = 30/group) | 28 | Macroscopic appearance (photography), histology (HE, MT). | Wounds treated with MSC-CM showed improvement in wound healing process | MSC-CM showed greater collagen density, angiogenesis, ratio, and length of epithelialization than the other groups (p < 0.05) |
| Tarcisia et al., 2017 | Adipose tissue | − | Flow cytometry positive for (CD73+, CD90+, CD34−). Osteogenic, chondrogenic, and adipogenic differentiation | MSCs of passage 3 were used. CM was then collected | Murine. Full-thickness excisional skin wounds, 20 mm long and 5 mm depth, on the back | The four cuts of each rat were treated randomly with MSC-CM (100%), complete culture medium, basal medium, and without treatment (control). The treatment was only done once after the rat skin injury (n = 30/group) | 28 | Macroscopic appearance (photography), histology (HE, MT). | Wounds treated with MSC-CM showed improvement in wound healing process | MSC-CM showed greater collagen density, angiogenesis, ratio, and length of epithelialization than the other groups (p < 0.05) |
| Lee et al., 2017 | Adipose tissue | − | − | Ell3 expression was suppressed using siRNA transfection in MSCs. CM harvested from MSCs transfected with siNS or siEll3 was collected | Murine. Full-thickness excisional skin wound, back | 100 μl of CM prepared from siNS- or siEll3-transfected MSCs or serum-free media was applied to the wound (n = not specified) | 7 | Macroscopic appearance (photography), histology (HE), IHC. | Skin wounds treated with siEll3 CM recovered to a lesser extent than those treated with serum-free media | siEll3 CM could not enhance the wound healing rate, whereas siNS CM significantly promoted wound repair |
| Du et al., 2017 | Rabbit bone marrow | − | − | MSCs of passage 2 were used. MSCs were cultured in normoxic (Nc) and chemical hypoxic conditioning by adding CoCl2 (Cc). Decellularization was conducted and extracellular matrices (ECM) cell-free were constructed | Murine. Full-thickness excisional skin wound, 7 mm, on the back | No treated group (control), mice treated with Nc-ECM sheets or Cc-ECM sheets (n = 16/group) | 7 | Macroscopic appearance (photography), histology (HE, MT, picrosirius red) | All the Cc-ECM-treated wounds completely healed on day 7, while Nc-ECM-treated wounds healed about 85.0 ± 8.6%, and non-treated wounds only healed 69.8 ± 9.6% | No inflammatory signs or visible indication of necrosis or other adverse events were observed |
| Dong et al., 2017 | Human umbilical cord | − | − | MSCs were transduced with human Wnt7a cDNA retroviral vector. MSCs of passage 3 at 50–60% confluence were used. Passage-3 MSCs were. CM from MSCs (MSC-CM) and from MSC with Wnt7a (Wnt-CM) was collected | Murine. Full-thickness excisional wound, 10 mm, on the back | 100 μl of Wnt-CM, MSC-CM, or DMEM was subcutaneously injected at multiple points into the wound area (n = 3/group) | 14 | Macroscopic appearance (photography), histology (HE, MT) | Wnt-CM significantly enhanced the closure rates in comparison with MSC-CM and DMEM (91.5 vs. 76.3 vs. 65.1%, respectively) | Wnt-CM accelerated migration of HFs into the wound area, had a thicker epidermis with more organized cell layers, and showed regeneration of more hair follicles. Increased expression of the α-SMA, collagen I, and collagen III was also observed in Wnt-CM group |
| Dong et al., 2017 | Human adipose tissues | − | − | Coleman adipose tissue was mechanically emulsified to obtain ECM/SVF-gel. For SVF preparation, the Coleman adipose tissue was digested. Supernatant from ECM/SVF-gel (gel-CM), Coleman adipose tissue (adi-CM), and SVF (SVF-CM) culture were collected to obtain CM | Murine. Full-thickness excisional skin wound, 8 mm, on the back | 100 ml PBS (control), gel-CM, adi-CM, or SVF-CM was injected into the wounds (n = 15/group) | 14 | Macroscopic appearance (photography), histology (HE, MT), ELISA | CM treatments resulted in a significant upregulation of collagen production compared with controls, revealing that the Gel-CM-group had the highest production of collagen | Higher expression of bFGF, EGF, and TGF-b in Gel-CM was observed |
| Mehanna et al., 2015 | Murine bone marrow | Needle flushing | Flow cytometry (CD44+, CD45−) | MSCs of passage 3 were used, and CM was collected | Murine. Full-thickness square-shaped skin wound (35 mm × 35 mm), on the back | Topical application - Untreated group (control) - Fibrin glue only group - Fibrin+MSCs group - Fibrin +MSC-CM group (n = 13/group) |
35 | Macroscopic appearance (photography), functional parameters (TEWL, SCH, tensile strength assessment), histology (HE, MT), IHC | Wounds’ size in fibrin+MSCs and fibrin+MSC-CM groups showed significant decrease in comparison with only fibrin and controls | CD68+ macrophages infiltrating granulation tissue were considerably higher in fibrin+MSC and fibrin+CM groups. SCH and tensile strength were higher, while TEWL was lower in both fibrin+MSC and fibrin+CM than in only fibrin and control group |
| Tam et al., 2014 | Wharton’s jelly from human umbilical cord | Cutting umbilical cords | Flow cytometry (CD13+, CD29+, CD44+, CD90+, CD10−, CD14−, CD34, CD117) | MSCs of passage 3–4 were used. It was a constructed wound dressing patch made up of an aloe vera−PCL (AV/PCL) nanoscaffold impregnated with WJ-MSCs or its CM | Murine. Full-thickness skin wounds, 8 mm, on dorsal region | - MSCs+AV/PC - MSC-CM+AV/PCL - Fibroblast+ AV/PCL - Fibroblast-CM+AV/PCL - PBS+ AV/PCL - Untreated (n = 9/group) |
28 | Macroscopic appearance (photography), histology (HE and MT), IF, WB, qRT-PCR | MSCs+AV/PCL and MSC-CM+AV/PCL treatment arms showed faster wound closure compared with other groups (p < 0.05) | MSCs+AV/PCL and MSC-CM+AV/PCL groups showed higher numbers of sebaceous glands, hair follicles, cellularity, and vasculature compared with other groups |
| Sun et al., 2014 | Murine abdominal subcutaneous adipose tissue | Incision | Flow cytometry (CD29+, CD90+, CD105+, CD34−). Osteogenic and adipogenic differentiation | MSCs of passage 3 at 90% confluence were used. Hypoxic microenvironment was generated using a disposable oxygen-absorbed and CO2 generator. CM was collected | Murine. Full-thickness skin incisions, 30 mm in diameter, on the back | Topical application - Concentrated hypoxic MSC-CM - Hypoxic MSC-CM - Serum medium (n = 10/group) |
21 | Macroscopic appearance (photography), histology (HE) | The average healing time was lower in concentrated hypoxic MSC-CM than in hypoxic MSC-CM and control group (16.2 ± 0.98 vs. 17.7 ± 0.78 vs. 21.3 ± 1.10 days) | Wound closure after treatment with concentrated hypoxic MSC-CM showed well-organized epidermis, thick cuticular layer, and increased collagen content than the other groups |
| Jun et al., 2014 | Human amniotic fluid | Amniocentesis | Immunofluorescence. Adipogenic and osteogenic differentiation | MSCs at 70% confluence were used. Cells were cultured in normoxic (nor) and hypoxic (hypo) conditions. CM was collected | Murine. Full-thickness excisional skin wound, 2 mm full thickness, on each side of the midline | 100 μl topically applied - MSC-hypoCM - MSC-norCM - DMEM (n = 10/group) |
5 | Macroscopic appearance (photography), histology (HE), IHC | MSC-hypoCM significantly accelerated wound closure compared with DMEM and MSC-norCM groups (p < 0.05) | The skin structure of wounds treated by MSC-hypoCM was more similar to normal skin structure. TGF-β/SMAD2 and PI3K/AKT signal pathways were upregulated in AF-MSC-hypoCM |
| Fong et al., 2014 | Wharton’s jelly from human umbilical cord | Full-term delivery | Immunofluorescence (CD10+, CD13+, CD29+, CD44+, CD90+) | MSCs of passages 3–4 at 80% confluence were used. CM was then collected | Murine. Full-thickness excisional skin wound, 8 mm, on dorsum surface | 100 μl injected intraperitoneally - MSCs - MSC-CM - Fibroblast - Fibroblast-CM - UCM (n = 9/group) |
28 | Macroscopic appearance (photography), histology (HE), IHC | MSC and MSC-CM healing rates were greater compared with controls (p < 0.05) | Wounds treated with MSCs and MSC-CMs showed greater re-epithelialization, vascularity, cellular density, sebaceous gland, and hair follicle numbers compared with controls |
| Chen et al., 2014 | Human bone marrow | − | − | MSCs of passages 3–4 at 80% confluence were used. MSCs were cultured and expanded under normoxic or hypoxic conditions. CM was collected from the normoxic and hypoxic MSCs to yield norCM and hypoCM | Murine. Full-thickness excisional skin wounds, 18 mm, on dorsal surface | 100 μl of treatment topically applied to skin wounds and covered with dressings daily for the first 4 days - MSC-norCM - MSC-hypoCM - Vehicle (control) (n = 16/group) |
14 | Macroscopic appearance (photography), IHC, IF | HypoCM-treated mice showed smaller wound area compared with norCM groups and vehicle control (26.42 ± 62.48 vs. 37.92 ± 62.44 vs. 45.00 ± 61.97%, respectively) | MSC-hypoCM accelerated wound closure compared with norCM and vehicle control |
| Tamari et al., 2013 | Human bone marrow | − | − | MSCs purchased from LONZA | Murine. Full-thickness excisional skin wound, 8 mm, on the midline on the back | Injection administered at 4 spots around the wound - PBS - MSCs - MSC-CM (n = not specified) |
14 | Macroscopic appearance (photography), histology, ELISA | Epithelialization rate was higher in MSC and MSC-CM group compared with controls: non-epithelialized wound area after treatment was 42.64 ± 5.36% in the control group, 4.90 ± 2.36% in MSC group, and 5.74 ± 2.85% in MSC-CM group | MSC and MSC-CM accelerated wound healing. HA production in MSC and MSC-CM group was increased compared with controls |
| Lee et al., 2011 | hCB: human cord blood hESC: human embryo | − | Fluorescence-activated cell sorter (CD133+, KDR+) | MSCs of passages 5–8 at 80 confluence were used. CM was collected | Murine. Full-thickness skin wound, 12 mm, on dorsal surface | 200 μl of CM was subcutaneously injected around the wound site or applied topically - hESC-CM - hCB-CM - Control medium (n = 10/group) |
21 | Macroscopic appearance (photography), histology | Wound healing rate was higher in groups treated with CM (90, 70, and 40% in the hESC-CM, hCB-CM, and vehicle medium, respectively) | Wound closure rate in hESC-CM group was higher when it was used topically instead of subcutaneously |
| Heo et al., 2011 | Human adipose tissue | Elective surgery | − | MSCs of passages 2–5 were used. CM was collected. Immunoprecipitation of TNF-α was also used, and CM implemented TNF-α was collected | Murine. Full-thickness excisional skin wounds, 8 mm, on the dorsal surface | 20 μl topically applied on the wound bed daily - PBS - MSC-CM - MSC-CM with TNF-α (n = 8/group) |
12 | Macroscopic appearance (photography), histology | MSC-CM with TNF-α accelerated wound closure compared with PBS MSC-CM | Number of blood vessel was the highest in MSC-CM with TNF-α group |
| Yoon et al., 2010 | Human amniotic fluid | Amniocentesis | Immunofluorescence (CD13+, CD29+, CD44+). Osteogenic, adipogenic, and chondrogenic differentiation | MSCs of passage 3 at 70% confluence were used. CM was collected | Murine. Full-thickness excisional skin wound, 2 mm, on each side of the midline. | 100 μl subcutaneous injection around the wound and topically applied on the wound bed | 8 | Macroscopic appearance (photography), histology (HE), IHC | MSC-CM accelerated wound closure compared with control | No difference in skin structure was observed between groups |
| - Control medium | ||||||||||
| - MSC-CM (n = 10/group) | ||||||||||
| Cho et al., 2010 | Human subcutaneous adipose tissue | Liposuction | Flow cytometry (CD90+, CD49d−) | MSCs of passage. TGF-β1-treated MSC-CM or non-treated MSC-CM was collected | Murine. Circular full-thickness skin wounds, 4-mm diameter, on the back | Intradermal injections (0.05 ml/point × 4 points) into the wound base twice per week | 10 | Macroscopic appearance (photography), histology (HE) | Wound size was reduced in both groups | TGF-β1-treated MSC-CM accelerated wound healing compared with MSC-CM |
| - Bactroban oint with MSC-CM - Bactroban oint with TGF-β1-treated MSC-CM (n = 3/group) |
||||||||||
| Templin et al., 2009 | Murine bone marrow | − | − | Retroviral gene transfer into lin- cells was performed, and DK mix cells were created. Cells and CM were then collected | Murine. Full-thickness skin wounds, 5-mm diameter, on dorsal surface | 200 μl subcutaneously injected around the wound at 8 different sites - MSC-CM - MSCs - PBS (n = 8/group) |
13 | Macroscopic appearance (photography), | MSC and MCS-CM accelerated wound healing compared with PBS (area non-epithelialized after follow-up: MSC = 25.9 ± 2.6%, MSC-CM = 25.1 ± 1.5%, PBS = 48.3 ± 4.7%) | Capillary density was higher in MSC-CM and MSC groups |
| Chen et al., 2008 | Murine bone marrow from femurs and tibia | − | q | MSCs in passage 3 at 80% confluence under hypoxic conditions were used | Murine. Full-thickness skin wounds, 6-mm diameter, on each side of midline | 100 ml (80 ml for subcutaneous injection around the wound and 20 ml for topical application on the wound bed) - MSC-CM - Fibroblast-CM - Vehicle (n = 5/group) |
14 | Macroscopic appearance (photography), histology (HE), IHC | MSC-CM significantly accelerated wound closure compared with fibroblast-CM or vehicle (wound closure percentage: 61 vs. 53 vs. 51%, respectively) | MSC-CM increased cell recruitment. |
ASCs, antler stem cells; AT, Adipose tissue-derived; bFGF, basic fibroblast growth factor; AF, amniotic fluid; BM, bone marrow; CM, conditioned medium; DMEM, Dulbecco’s modified Eagle medium; DP, dental pulp; ECM, extracellular matrix; EGF, epidermal growth factor; ELISA, enzyme-linked immunosorbent assay; EVs, extracellular vesicles; Exos, exosomes; HE, hematoxylin and eosin; hCB, human cord blood; hESC, human embryonic stem cell; IHC, immunohistochemistry; IF, immunofluorescence; MPF, micro-nano polylactic acid electrospun fiber; MSCs, mesenchymal stromal cells; MT, Masson’s trichrome; PBM, photobiomodulation; PBS, phosphate-buffered saline; qRT-PCR, real-time quantitative polymerase chain reaction; SVF, stromal vascular fraction; UCM, unconditioned medium; WB, Western blotting; WJ, Wharton’s jelly. *0, 10, 50, and 100% MSC-CM refers to the dilutions of the conditioned medium in DMEM/F12 with 2% FBS.