A Novel Composite Hydrogel Composed of Formic Acid-Decellularized Pepsin-Soluble Extracellular Matrix Hydrogel and Sacchachitin Hydrogel as Wound Dressing to Synergistically Accelerate Diabetic Wound Healing

oleh: Chien-Ming Hsieh, Weu Wang, Ying-Hsuan Chen, Pu-Sheng Wei, Yu-Hsuan Liu, Ming-Thau Sheu, Hsiu-O Ho

Format: Article
Diterbitkan: MDPI AG 2020-06-01

Deskripsi

Extracellular matrix (ECM) hydrogel can create a favorable regenerative microenvironment and act as a promising dressing for accelerating the healing of diabetic wound. In this study, a simple and effective decellularization technique was developed and optimized to obtain acellular extracellular matrix (<i>a</i>ECM) from porcine skin. It was found that decellularization at 30% formic acid for 72 h effectively decellularized porcine skin while retaining >75% collagen and ~37% GAG in the <i>a</i>ECM with no presence of nuclei of cellular remnants. <i>a</i>ECM hydrogel was fabricated by digesting <i>a</i>ECM with pepsin in various acidic solutions (0.1 N HCl, glycolic acid (GA) and 2-pyrrolidone-5-carboxylic acid (PCA)) and then treated with a pH-controlled neutralization and temperature-controlled gelation procedure. Based on physical characterizations, including SDS-PAGE, rheological analysis and SEM analysis, <i>a</i>ECM<sub>HCl</sub> hydrogels fabricated at 25 mg/mL in 0.1 N HCl were selected. Four polymeric ECM-mimic hydrogels, including sacchachitin (SC), hyaluronic acid (HA) and chitosan (CS) and three composite hydrogels of combining SC either with <i>a</i>ECM<sub>HCl,25</sub> (<i>a</i>ECM<sub>HCl</sub>/SC), HA (HA/SC) or CS (SC/CS) were prepared and evaluated for WS-1 cell viability and wound-healing effectiveness. Cell viability study confirmed that no hydrogel dressings possessed any toxicity at all concentrations examined and ECM<sub>HCl</sub>, HA and ECM<sub>HCl</sub>/SC at higher concentrations (>0.05%) induced statistically significant proliferation. Diabetic wound healing study and histological examinations revealed that ECM<sub>HCl</sub>/SC hydrogel was observed to synergistically accelerate wound healing and ultimately stimulated the growth of hair follicles and sweat glands in the healing wound indicating the wound had healed as functional tissues. The results support the great potential of this newly produced ECM<sub>HCl</sub>/SC composite hydrogel for healing and regeneration of diabetic wounds.