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.

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

doi:10.3390/pharmaceutics12060538

Chien-Ming Hsieh, Hsiu-O Ho + Show 5 more

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https://doi.org/10.3390/pharmaceutics12060538

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Abstract

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 (aECM) 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 aECM with no presence of nuclei of cellular remnants. aECM hydrogel was fabricated by digesting aECM 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, aECMHCl 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 aECMHCl,25 (aECMHCl/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 ECMHCl, HA and ECMHCl/SC at higher concentrations (>0.05%) induced statistically significant proliferation. Diabetic wound healing study and histological examinations revealed that ECMHCl/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 ECMHCl/SC composite hydrogel for healing and regeneration of diabetic wounds.

Highlights

Extracellular matrix (ECM) is the largest component of normal skin and extracellular matrix (ECM) synthesis is crucial for wound healing [1]
Our studies found that the amount of DNA remaining was 42.95 ± 0.73 ng/mg in dry porcine skin treated with 30% formic acid for 72 h
The GAG content of decellularized porcine skin was significantly lower than that in the Phosphate buffered saline (PBS) group, it still remained at 5.50 ± 0.16 μg/mg dry weight (36.96% ± 1.08%) in the treatment with 30% formic acid solution for 72 h

Summary

Introduction

Extracellular matrix (ECM) is the largest component of normal skin and ECM synthesis is crucial for wound healing [1]. The methods can effectively remove cellular materials and preserve the collagen components but they might disrupt the ECM structure [8], a length incubation was necessitated [9] or with impractical device to assist [10]. Chemical methods by acidic and alkaline treatments can solubilize the cellular cytoplasmic components, remove nucleic acids (for instance, DNA), preserve the structure and function of the native ECM and simultaneously disinfect the material through entering microorganisms and oxidizing microbial enzymes [11]. A simple formic acid treatment able to remove most of the cellular contents and preserve the highest ECM contents in the decellularized porcine menisci was reported by Chen et al [12]. It was inevitable to develop simple, effective and optimized decellularization techniques to obtain ECM from porcine skin for wound-healing medical applications

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