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Home / Equine Research Bank / Stem Cells Int / Insulin Enhances the In Vitro Osteogenic Capacity of Flexor ...
Stem cells international2019; 2019; 1602751; doi: 10.1155/2019/1602751

Insulin Enhances the In Vitro Osteogenic Capacity of Flexor Tendon-Derived Progenitor Cells.

Abstract: There is increased incidence of tendon disorders and decreased tendon healing capacity in people with diabetes mellitus (DM). Recent studies have also suggested pathological ossification in repair tendon of people with DM. Therefore, the objective of this study is to investigate the effects of insulin supplementation, an important pathophysiologic stimulus of DM, on tendon progenitor cell (TPC) proliferation and in vitro osteogenic capacity. Passage 3 TPCs were isolated from collagenase-digested, equine superficial digital flexor tendons. TPC proliferation was measured via MTT assay after 3 days of monolayer culture in medium supplemented with 0, 0.007, 0.07, and 0.7 nM insulin. In vitro osteogenic capacity of TPCs (Alizarin Red staining, osteogenic mRNA expression, and alkaline phosphatase bioactivity) was assessed with 0, 0.07, and 0.7 nM insulin-supplemented osteogenic induction medium. Insulin (0.7 nM) significantly increased TPC proliferation after 3 days of monolayer culture. Alizarin Red staining of insulin-treated TPC osteogenic cultures demonstrated robust cell aggregation and mineralized matrix secretion, in a dose-dependent manner. Runx2, alkaline phosphatase, and Osteonectin mRNA and alkaline phosphatase bioactivity of insulin-treated TPC cultures were significantly higher at day 14 of osteogenesis compared to untreated controls. Addition of picropodophyllin (PPP), a selective IGF-I receptor inhibitor, mitigated the increased osteogenic capacity of TPCs, indicating that IGF-I signaling plays an important role. Our findings indicate that hyperinsulinemia may alter TPC phenotype and subsequently impact the quality of repair tendon tissue.
Copyright © 2019 Sushmitha S. Durgam et al.
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Publication Date: 2019-12-27 PubMed ID: 31949435PubMed Central: PMC6948345DOI: 10.1155/2019/1602751Google Scholar: Lookup
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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.

The research aims to investigate how adding insulin can affect the proliferative and osteogenic ability of tendon progenitor cells (TPCs), a process which has implications on tendon healing, especially in diabetics.

Understanding the Context

  • Diabetes Mellitus (DM) patients often suffer from tendon disorders and exhibit reduced tendon healing capacity. Some studies have also suggested an abnormality known as pathological ossification in the repaired tendons of DM patients.
  • The research was focused on the effects of insulin, a vital element in DM pathology, on the rate of proliferation and osteogenic (bone forming) capacity of TPCs.

Methodology

  • Third passage TPCs were isolated from the superficial digital flexor tendons of horses, treated with collagenase.
  • The isolated TPCs were cultured in mediums supplemented with different concentrations of insulin. They used four distinct concentrations: 0, 0.007, 0.07, and 0.7 nM.
  • The proliferation of TPCs was measured after 3 days of culturing, using an MTT assay.
  • The ability of TPCs to form bone (osteogenic capacity) was assessed using Alizarin Red staining, osteogenic mRNA expression tests, and testing for alkaline phosphatase bioactivity, with the medium being supplemented with either 0, 0.07, or 0.7 nM insulin.

Results

  • The highest dose of insulin used (0.7 nM) showed a significant increase in TPC proliferation after 3 days of culture.
  • Insulin-treated TPCs showed remarkable cell aggregation and secretion of the mineralized matrix in a dose-dependent manner, as evidenced by Alizarin Red staining.
  • Gene expressions of Runx2, alkaline phosphatase, and Osteonectin mRNA in insulin-treated TPCs were significantly higher on day 14 of osteogenesis compared to those left untreated. Alkaline phosphatase bioactivity followed the same pattern.
  • They discovered that the addition of Picropodophyllin (PPP), an inhibitor of IGF-I receptors, decreased the osteogenic capacity of TPCs, indicating the role of IGF-I signaling in this process.

Conclusion

  • The research outcomes suggest that high levels of insulin, known as hyperinsulinemia, could alter the normal functioning and characteristics of TPCs and subsequently affect the quality of the repaired tendon tissue.

Cite This Article

APA
Durgam SS, Altmann NN, Coughlin HE, Rollins A, Hostnik LD. (2019). Insulin Enhances the In Vitro Osteogenic Capacity of Flexor Tendon-Derived Progenitor Cells. Stem Cells Int, 2019, 1602751. https://doi.org/10.1155/2019/1602751

Publication

Stem cells international
ISSN: 1687-966X
NlmUniqueID: 101535822
Country: United States
Language: English
Volume: 2019
Pages: 1602751
PII: 1602751

Researcher Affiliations

Durgam, Sushmitha S
  • Department of Veterinary Clinical Sciences, College of Veterinary Medicine, The Ohio State University, 601 Vernon L. Tharp Street, Columbus, OH, 43210, USA.
Altmann, Nadine N
  • Department of Veterinary Clinical Sciences, College of Veterinary Medicine, The Ohio State University, 601 Vernon L. Tharp Street, Columbus, OH, 43210, USA.
Coughlin, Haley E
  • Department of Veterinary Clinical Sciences, College of Veterinary Medicine, The Ohio State University, 601 Vernon L. Tharp Street, Columbus, OH, 43210, USA.
Rollins, Audrey
  • Department of Veterinary Clinical Sciences, College of Veterinary Medicine, The Ohio State University, 601 Vernon L. Tharp Street, Columbus, OH, 43210, USA.
Hostnik, Laura D
  • Department of Veterinary Clinical Sciences, College of Veterinary Medicine, The Ohio State University, 601 Vernon L. Tharp Street, Columbus, OH, 43210, USA.

Conflict of Interest Statement

The authors declare that they have no conflicts of interest.

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