Effect of HBO therapy on adipose-derived stem cells,fibroblasts and co-cultures: In vitro study of oxidative stress,angiogenic potential and production of pro-inflammatory growth factors in co-cultures 1

Abstract

BACKGROUND:

A key moderator of wound healing is oxygen. Wound healing is a dynamic and carefully orchestrated process involving blood cells, cytokines, parenchymal cells (i.e. fibroblasts and mesenchymal stem cells) and extracellular matrix reorganization. Human adipose derived stem cells as well as human fibroblasts produce soluble factors, exhibit diverse effects on inflammation and anti inflammation response and are involved in wound healing processes.

Hyperbaric oxygen therapy is an effective adjunct treatment for ischemic disorders such as chronic infection or chronic wounds. In vitro effects of hyperbaric oxygen therapy on human cells were presented in many studies except for those on mono- and co-cultures of human adipose derived stem cells and fibroblasts.

OBJECTIVE:

The aim of this study was to investigate the effects of hyperbaric oxygen therapy on mono- and co-cultures of human adipose derived stem cells and fibroblasts.

METHODS:

Mono- and co-cultures from human adipose derived stem cells and fibroblasts were established. These cultures were exposed to hyperbaric oxygen therapy every 24 h for five consecutive days. Measuring experiments were performed on the first, third and fifth day. Therapy effects on the expression of VEGF, IL 6 and reactive oxygen species were investigated.

RESULTS:

After exposure to hyperbaric oxygen, cell culturess showed a significant increase in the expression of VEGF after 3 and 5 days. All cultures showed significantly reduced formation of reactive oxygen species throughout the experiments. The expression of IL-6 decreased during the experiment in mono-cultures of human adipose derived stem cells and co-cultures. In contrast, mono-cultures of human skin fibroblasts showed an overall significantly increased expression of IL-6.

CONCLUSIONS:

Hyperbaric oxygen therapy leads to immunmodulatory and proangiogenetic effects in a wound-like enviroment of adipose derived stem cells and fibroblasts.

Keywords

HBO therapy adipose derived stem cells cell oxygenation cell co-culture angiogenesis hyperbaric oxygen

Get full access to this article

View all access options for this article.

References

Sen

. Wound healing essentials: let there be oxygen. Wound Repair Regen. 2009;17(1):1–18.

Greif

, et al. Supplemental perioperative oxygen to reduce the incidence of surgical-wound infection. N Engl J Med. 2000;342(3):161–7.

Phillips

. Hyperbaric oxygen therapy: a therapy in search of a disease? Dermatol Surg. 2000;26(12):1167–69.

Tibbles

, Edelsberg

. Hyperbaric-oxygen therapy. N Engl J Med. 1996;334(25):1642–8.

Dunnill

, et al. Reactive oxygen species (ROS) and wound healing: the functional role of ROS and emerging ROS-modulating technologies for augmentation of the healing process. Int Wound J. 2017;14(1):89–96.

Klopfleisch

, Jung

. The pathology of the foreign body reaction against biomaterials. J Biomed Mater Res. A. 2017;105(3):927–40.

Krüger-Genge

, Blocki

, Franke

, Jung

. Vascular Endothelial Cell Biology: An Update. Int J Mol Sci. 2019;20(18):4411. 10.3390/ijms20184411

Sheikh

, et al. Effect of hyperoxia on vascular endothelial growth factor levels in a wound model. Arch Surg. 2000;135(11):1293–7.

Sureda

, et al. Antioxidant Response of Chronic Wounds to Hyperbaric Oxygen Therapy. PLoS One. 2016;11(9):e0163371.

10.

Francis

, et al. Hyperbaric Oxygen Inhibits Reperfusion-Induced Neutrophil Polarization and Adhesion Via Plasmin-Mediated VEGF Release. Plast Reconstr Surg Glob Open. 2017;5(9):e1497.

11.

Ersson

, et al. Cytokine response after acute hyperbaric exposure in the rat. Undersea Hyperb Med. 1998;25(4):217–21.

12.

Sun

, et al. Hyperbaric oxygen preconditioning attenuates postoperative cognitive impairment in aged rats. Neuroreport. 2014;25(9):718–24.

13.

Dimitrijevich

, et al. Effect of hyperbaric oxygen on human skin cells in culture and in human dermal and skin equivalents. Wound Repair Regen. 1999;7(1):53–64.

14.

Kang

, et al. Effect of hyperbaric oxygen on the growth factor profile of fibroblasts. Arch Facial Plast Surg. 2004;6(1):31–5.

15.

Qian

, et al. Protective Role of Adipose-Derived Stem Cells in Staphylococcus aureus-Induced Lung Injury is Mediated by RegIIIgamma Secretion. Stem Cells. 2016;34(7):1947–56.

16.

Pilehvar-Soltanahmadi

, et al. Cytoprotection, proliferation and epidermal differentiation of adipose tissue-derived stem cells on emu oil based electrospun nanofibrous mat. Exp Cell Res. 2017;357(2):192–201.

17.

Nguyen

, et al. Stromal vascular fraction: A regenerative reality? Part Current concepts and review of the literature. J Plast Reconstr Aesthet Surg. 2016;69(2):170–9.

18.

Guo

, et al. Stromal vascular fraction: A regenerative reality? Part Mechanisms of regenerative action. J Plast Reconstr Aesthet Surg. 2016;69(2):180–8.

19.

Haubner

, et al. A Co-Culture Model of Fibroblasts and Adipose Tissue-Derived Stem Cells Reveals New Insights into Impaired Wound Healing After Radiotherapy. Int J Mol Sci. 2015;16(11):25947–58.

20.

Felthaus

, et al. Adipose tissue-derived stem cells from affected and unaffected areas in patients with multiple symmetric lipomatosis show differential regulation of mTOR pathway genes. Clin Hemorheol Microcirc. 2018;69(1-2):141–51.

21.

Sheridan

, Shank

. Hyperbaric oxygen treatment: a brief overview of a controversial topic. J Trauma. 1999;47(2):426–35.

22.

Zhao

, et al. Effect of hyperbaric oxygen and growth factors on rabbit ear ischemic ulcers. Arch Surg. 1994;129(10):1043–9.

23.

Park

, et al. Adipose-derived stem cells and their secretory factors as a promising therapy for skin aging. Dermatol Surg. 2008;34(10):1323–6.

24.

Hsu

, et al. The effect of exogenous histone H1 on rat adipose-derived stem cell proliferation, migration, and osteogenic differentiation in vitro. J Cell Physiol. 2012;227(10):3417–25.

25.

Kappy

, et al. Human adipose-derived stem cell treatment modulates cellular protection in both in vitro and in vivo traumatic brain injury models. J Trauma Acute Care Surg. 2018;84(5):745–51.

26.

Seo

, et al. Neuroprotective effect of secreted factors from human adipose stem cells in a rat stroke model. Neurol Res. 2017;39(12):1114–24.

27.

Tra

, et al. Hyperbaric oxygen treatment of tissue-engineered mucosa enhances secretion of angiogenic factors in vitro. Tissue Eng Part A. 2014;20(9-10):1523–30.

28.

Park

, Lee

, Son

. Uniaxial cyclic strain stimulates cell proliferation and secretion of interleukin-6 and vascular endothelial growth factor of human dermal fibroblasts seeded on chitosan scaffolds. J Biomed Mater Res A. 2014;102(7):2268–76.

29.

Namiki

, et al. Hypoxia induces vascular endothelial growth factor in cultured human endothelial cells. J Biol Chem. 1995;270(52):31189–95.

30.

Brogi

, et al. Hypoxia-induced paracrine regulation of vascular endothelial growth factor receptor expression. J Clin Invest. 1996;97(2):469–76.

31.

Lin

, et al. Essential involvement of IL-6 in the skin wound-healing process as evidenced by delayed wound healing in IL-6-deficient mice. J Leukoc Biol. 2003;73(6):713–21.

32.

Johnson

, Wilgus

. Vascular Endothelial Growth Factor and Angiogenesis in the Regulation of Cutaneous Wound Repair. Adv Wound Care (New Rochelle). 2014;3(10):647–61.

33.

Duncan

, Berman

. Stimulation of collagen and glycosaminoglycan production in cultured human adult dermal fibroblasts by recombinant human interleukin 6. J Invest Dermatol. 1991;97(4):686–92.

34.

Kim

, et al. Wound healing effect of adipose-derived stem cells: a critical role of secretory factors on human dermal fibroblasts. J Dermatol Sci. 2007;48(1):15–24.

35.

Castilla

, Liu

, Velazquez

. Oxygen: Implications for Wound Healing. Adv Wound Care (New Rochelle). 2012;1(6):225–30.

36.

Hopf

, et al. Hyperoxia and angiogenesis. Wound Repair Regen. 2005;13(6):558–64.

37.

Fosen

, Thom

. Hyperbaric oxygen, vasculogenic stem cells, and wound healing. Antioxid Redox Signal. 2014;21(11):1634–47.

38.

Francis

, Baynosa

. Ischaemia-reperfusion injury and hyperbaric oxygen pathways: a review of cellular mechanisms. Diving Hyperb Med. 2017;47(2):110–17.

39.

Ganguly

, et al. Hyperbaric oxygen enhances apoptosis in hematopoietic cells. Apoptosis. 2002;7(6):499–510.

40.

Chen

, et al. Apoptosis of T-leukemia and B-myeloma cancer cells induced by hyperbaric oxygen increased phosphorylation of p38 MAPK. Leuk Res. 2007;31(6):805–15.

41.

Granowitz

, et al. Hyperbaric oxygen inhibits benign and malignant human mammary epithelial cell proliferation. Anticancer Res. 2005;25(6b):3833–42.

42.

Tandara

, Mustoe

. Oxygen in wound healing–more than a nutrient. World J Surg. 2004;28(3):294–300.

43.

Sen

, et al. Oxidant-induced vascular endothelial growth factor expression in human keratinocytes and cutaneous wound healing. J Biol Chem. 2002;277(36):33284–90.