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Stem cell research & therapy2017; 8(1); 120; doi: 10.1186/s13287-017-0577-3

Effect of bone marrow-derived mesenchymal stem cells and stem cell supernatant on equine corneal wound healing in vitro.

Abstract: We aimed to determine and compare the in vitro effects of autologous bone marrow-derived mesenchymal stem cells (BM-MSCs) and mesenchymal stem cell supernatant (MSC-Sp) on the wound healing capacity of equine corneal fibroblasts using a scratch assay. Bone marrow aspirates and eyes were collected from normal, euthanized horses with subsequent isolation and culture of BM-MSCs and corneal stromal cells. Corneal stromal cells were culture-expanded in the culture well of transwell plates and then treated with an autologous BM-MSC suspension (dose: 2.5 × 105/100 μL media with the BM-MSCs contained within the insert well), MSC-Sp solution, or naive culture media (control) for 72 h. A linear defect in confluent cell cultures was created (i.e., corneal scratch assay) to assess the cellular closure ("healing") over time. Three representative areas of the scratch in each culture were photographed at each time point and the scratch area was quantitated using image analysis software (ImageJ). Media from the scratches were analyzed for various growth factors using human enzyme-linked immunosorbent assay (ELISA) kits that crossreact with the horse. There was a significant percentage decrease in the scratch area remaining in the BM-MSC and MSC-Sp groups compared to the control group. There was also a significant percentage decrease in the scratch area remaining in the BM-MSC group compared to the MSC-Sp group at 36 h post-scratch and all time points thereafter. The concentration of transforming growth factor (TGF)-β1 in the media was significantly higher in the BM-MSC group compared to the control group. The significant decrease in scratch area in equine corneal fibroblast cultures treated with autologous BM-MSCs compared to MSC-Sp or control treatments suggests that BM-MSCs may substantially improve corneal wound healing in horses. MSC-Sp may also improve corneal wound healing given the significant decrease in scratch area compared to control treatments, and would be an immediately available and cost-effective treatment option.
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Publication Date: 2017-05-25 PubMed ID: 28545510PubMed Central: PMC5445363DOI: 10.1186/s13287-017-0577-3Google Scholar: Lookup
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  • Journal Article

Summary

This research summary has been generated with artificial intelligence and may contain errors and omissions. Refer to the original study to confirm details provided. Submit correction.

This research investigates the effect of bone marrow-derived stem cells and their surrounding fluid on the healing ability of horse cornea cells. The results reveal that both the stem cells and their fluid significantly improved the healing process in cell culture, hinting that they could be beneficial in treating corneal wounds in horses.

Research Methodology

  • The research team used the scratch assay technique to determine the healing impacts of bone marrow-derived mesenchymal stem cells (BM-MSCs) and mesenchymal stem cell supernatant (MSC-Sp) on horse corneal cells. These cells and bone marrow samples were obtained from healthy, euthanized horses.
  • The collected corneal stromal cells were expanded in a lab culture. These cells were then exposed to either a BM-MSC suspension, MSC-Sp solution, or untreated culture media (used as a control) for 72 hours.
  • To measure healing progress, the researchers created a linear defect or “scratch” in the confluent cell cultures. Images of the scratch were taken from three representative areas over time and the remaining scratch area was calculated using image analysis software.

Findings

  • The scratch assay revealed a significant decrease in the remaining scratch area in the samples treated with BM-MSCs and MSC-Sp when compared with the control. This implies an enhanced cell healing ability due to the presence of stem cells or their supernatant.
  • The BM-MSC treated samples showed an even more significant decrease in remaining scratch area compared to those treated with MSC-Sp after the 36-hour mark. This suggests that BM-MSCs are more beneficial in promoting wound healing processes.
  • The study also observed higher concentrations of the transforming growth factor (TGF)-β1 in the BM-MSC treated cultures compared to the control, possibly implicating the growth factor in the improved wound healing.

Implications

  • The significant reduction in the scratch area in corneal cell cultures treated with BM-MSCs suggests these stem cells could play a vital role in enhancing corneal wound healing in horses.
  • The MSC-Sp could also serve as an efficient treatment option for corneal wound healing due to its documented healing benefits, apart from being an immediately available and cost-effective treatment option.

Cite This Article

APA
Sherman AB, Gilger BC, Berglund AK, Schnabel LV. (2017). Effect of bone marrow-derived mesenchymal stem cells and stem cell supernatant on equine corneal wound healing in vitro. Stem Cell Res Ther, 8(1), 120. https://doi.org/10.1186/s13287-017-0577-3

Publication

Stem cell research & therapy
ISSN: 1757-6512
NlmUniqueID: 101527581
Country: England
Language: English
Volume: 8
Issue: 1
Pages: 120

Researcher Affiliations

Sherman, Amanda B
  • Department of Clinical Sciences, North Carolina State University College of Veterinary Medicine, Raleigh, NC, 27606, USA.
Gilger, Brian C
  • Department of Clinical Sciences, North Carolina State University College of Veterinary Medicine, Raleigh, NC, 27606, USA.
Berglund, Alix K
  • Department of Clinical Sciences, North Carolina State University College of Veterinary Medicine, Raleigh, NC, 27606, USA.
Schnabel, Lauren V
  • Department of Clinical Sciences, North Carolina State University College of Veterinary Medicine, Raleigh, NC, 27606, USA. lvschnab@ncsu.edu.

MeSH Terms

  • Animals
  • Biomarkers / metabolism
  • Bone Marrow Cells / cytology
  • Corneal Injuries / pathology
  • Corneal Injuries / therapy
  • Enzyme-Linked Immunosorbent Assay
  • Epidermal Growth Factor / metabolism
  • Female
  • Flow Cytometry
  • Horses
  • Male
  • Mesenchymal Stem Cells / cytology
  • Mesenchymal Stem Cells / metabolism
  • Transforming Growth Factor beta1 / metabolism
  • Wound Healing

Grant Funding

  • K08 AR060875 / NIAMS NIH HHS

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