Impact of Hyperglycemia and Low Oxygen Tension on Adipose-Derived Stem Cells Compared with Dermal Fibroblasts and Keratinocytes: Importance for Wound Healing in Type 2 Diabetes

PLoS One. 2016 Dec 19;11(12):e0168058. doi: 10.1371/journal.pone.0168058. eCollection 2016.

Abstract

Aim: Adipose-derived stem cells (ASC) are currently proposed for wound healing in those with type 2 diabetes mellitus (T2DM). Therefore, this study investigated the impact of diabetes on adipose tissue in relation to ASC isolation, proliferation, and growth factor release and the impact of hyperglycemia and low oxygen tension (found in diabetic wounds) on dermal fibroblasts, keratinocytes, and ASC in vitro.

Methods: Different sequences of hypoxia and hyperglycemia were applied in vitro to ASC from nondiabetic (n = 8) or T2DM patients (n = 4) to study cell survival, proliferation, and growth factor release. Comparisons of dermal fibroblasts (n = 8) and keratinocytes (primary lineage) were made.

Results: No significant difference of isolation and proliferation capacities was found in ASC from nondiabetic and diabetic humans. Hypoxia and hyperglycemia did not impact cell viability and proliferation. Keratinocyte Growth Factor release was significantly lower in diabetic ASC than in nondiabetic ASC group in each condition, while Vascular Endothelial Growth Factor release was not affected by the diabetic origin. Nondiabetic ASC exposition to hypoxia (0.1% oxygen) combined with hyperglycemia (25mM glucose), resulted in a significant increase in VEGF secretion (+64%, p<0.05) with no deleterious impact on KGF release in comparison to physiological conditions (5% oxygen and 5 mM glucose). Stromal cell-Derived Factor-1α (-93%, p<0.001) and KGF (-20%, p<0.05) secretion by DF decreased in these conditions.

Conclusions: A better profile of growth factor secretion (regarding wound healing) was found in vitro for ASC in hyperglycemia coupled with hypoxia in comparison to dermal fibroblasts and keratinocytes. Interestingly, ASC from T2DM donors demonstrated cellular growth rates and survival (in hypoxia and hyperglycemic conditions) similar to those of healthy ASC (from normoglycemic donors); however, KGF secretion was significantly depleted in ASC obtained from T2DM patients. This study demonstrated the impact of diabetes on ASC for regenerative medicine and wound healing.

Publication types

  • Comparative Study

MeSH terms

  • Adipose Tissue / cytology*
  • Adipose Tissue / metabolism
  • Adult
  • Aged
  • Cell Hypoxia
  • Cell Proliferation
  • Cell Survival
  • Cells, Cultured
  • Diabetes Mellitus, Type 2 / complications
  • Diabetes Mellitus, Type 2 / metabolism
  • Diabetes Mellitus, Type 2 / pathology*
  • Female
  • Fibroblasts / cytology*
  • Fibroblasts / metabolism
  • Glucose / pharmacology*
  • Humans
  • Intercellular Signaling Peptides and Proteins / metabolism
  • Keratinocytes / cytology*
  • Keratinocytes / metabolism
  • Male
  • Middle Aged
  • Stem Cells / cytology*
  • Stem Cells / metabolism
  • Wound Healing
  • Young Adult

Substances

  • Intercellular Signaling Peptides and Proteins
  • Glucose

Grants and funding

This work was supported by public grants from the Fond pour la Recherche Scientifique (FRS-FNRS) and from the Feédeération Wallonie-Bruxelles (Action de Recherche Concerteée). The Fond pour la Recherche Scientifique provided salaries for Aurore Lafosse. The Action de Recherche Concerteée provided support to acquire the research materials. The authors have no financial interest to declare in relation to the content of this article. Novadip Biosciences did not fund the study in terms of study design, data collection and analysis, decision to publish, or preparation of the manuscript. Denis Dufrane is co-founder of Novadip Biosciences which provides a salary for the position of Chief Scientific Officer; however, Novadip Biosciences did not have any additional role in the study design, data collection and analysis, decision to publish, or preparation of the manuscript. There are no commercially relevant declarations (relating to employment, consultancy, patents, products in development, or marketed products, etc.) in relation this study.