Supercritical Carbon Dioxide-decellularized Porcine Acellular Dermal Matrix combined with Autologous Adipose-derived Stem Cells: Its Role in Accelerated Diabetic Wound Healing

Ping-Ruey Chou; Yun-Nan Lin; Sheng-Hua Wu; Sin-Daw Lin; Periasamy Srinivasan; Dar-Jen Hsieh; Shu-Hung Huang

doi:10.7150/ijms.41155

Supercritical Carbon Dioxide-decellularized Porcine Acellular Dermal Matrix combined with Autologous Adipose-derived Stem Cells: Its Role in Accelerated Diabetic Wound Healing

Int J Med Sci. 2020 Feb 4;17(3):354-367. doi: 10.7150/ijms.41155. eCollection 2020.

Authors

Ping-Ruey Chou¹, Yun-Nan Lin², Sheng-Hua Wu^{3

4

5}, Sin-Daw Lin², Periasamy Srinivasan⁶, Dar-Jen Hsieh⁶, Shu-Hung Huang^{2

7

8}

Affiliations

¹ School of Medicine, College of Medicine, Kaohsiung Medical University, Kaohsiung 807, Taiwan.
² Division of Plastic Surgery, Department of Surgery, Kaohsiung Medical University Hospital, Kaohsiung 807, Taiwan.
³ Department of Anesthesiology, School of Medicine, College of Medicine, Kaohsiung Medical University, Kaohsiung 807, Taiwan.
⁴ Department of Anesthesiology, Kaohsiung Medical University Hospital, Kaohsiung 807, Taiwan.
⁵ Department of Anesthesiology, Kaohsiung Municipal Ta-Tung Hospital, Kaohsiung 807, Taiwan.
⁶ Center of Research and Development, ACRO Biomedical Co., Ltd. Kaohsiung 821, Taiwan.
⁷ Department of Surgery, School of Medicine, College of Medicine, Kaohsiung Medical University, Kaohsiung 807, Taiwan.
⁸ Regeneration Medicine and Cell Therapy Research Center, Kaohsiung Medical University, Kaohsiung 807, Taiwan.

Abstract

Diabetes mellitus (DM) causes impaired wound healing by affecting one or more of the biological mechanisms of hemostasis, inflammation, proliferation, and remodeling and a large number of cell types, extracellular components, growth factors, and cytokines. Interventions targeted toward these mechanisms might accelerate the wound healing process. To evaluate the wound healing efficacy of supercritical carbon dioxide (scCO₂)-decellularized porcine acellular dermal matrix (ADM) combined with autologous adipose-derived stem cells (ASCs) in streptozotocin (STZ)-induced DM rats. DM was induced by injecting rats with STZ; dorsal full-thickness skin (5 × 5 cm²) was created and treated with and without ASCs-scCO₂-treated ADM to evaluate the wound healing rate through histological examination, fluorescence microscopic observation, and immunohistochemical analysis. In the present study, complete decellularization of the porcine dermal matrix was achieved through scCO₂. Isolation of ASCs was conducted and evaluated using CD29⁺/CD31^-/CD45^-/CD90⁺ markers in flow cytometry, which indicated that more than 90% of cells were ASCs. The percentage of cells labeled with CD29⁺ and CD90⁺ was found to be 97.50% and 99.69%, respectively. The wound healing rate increased in all groups relative to the group with the DM wound without treatment. DM wound treated with ADM-ASCs showed significantly higher (p < 0.01) wound healing rate than DM wound without treatment. ADM-ASC-treated rats showed significantly increased epidermal growth factor, Ki67, and prolyl 4-hydroxylase and significantly decreased CD45 compared with the group with the DM wound without treatment. The intervention comprising ADM decellularized from porcine skin by using scCO₂ and ASCs was proven to improve diabetic wound healing. ADM-ASCs had a positive effect on epidermal regeneration, anti-inflammation, collagen production and processing, and cell proliferation; thus, it accelerated wound healing.

Keywords: acellular dermal matrix; autologous adipose-derived stem cells; diabetes mellitus; supercritical carbon dioxide; wound healing.

MeSH terms

Acellular Dermis / drug effects*
Adipocytes / cytology*
Animals
Carbon Dioxide / chemistry*
Carbon Dioxide / pharmacology
Cells, Cultured
Immunohistochemistry
Male
Microscopy, Fluorescence
Rats
Rats, Wistar
Stem Cells / cytology*
Stem Cells / drug effects
Swine
Wound Healing / drug effects

Substances

Carbon Dioxide