Design of bio-scaffold conjugated with chitosan-PEG nano-carriers containing bio-macromolecules of Verbascum sinuatum L. to differentiate human adipose-derived stem cells into dermal keratinocytes

Abstract

Regenerative medicine and drug delivery systems provide promising approaches for the treatment of skin lesions. However, the design of engineered substrates containing therapeutic agents for cell proliferation and its differentiation into skin cells, with skin-like patterns, is the major challenge. Here, to overcome this problem, a hybrid scaffold conjugated with nanoparticles containing the extract of Verbascum sinuatum L. flowers (HE) was designed. To this end, (chitosan-PEG)-based nanocarriers (Chi-PEG) were first prepared in the volume ratios of 90:10, 80:20, 70:30, and 50:50 v/v. The results indicated that the 70:30 ratio possessed better physical/morphologic properties along with more suitable stability than other nanoparticles (encapsulation-efficiency:86.34 %, zeta-potential:21.2 mV, and PDI:0.30). Afterward, PCL-collagen biologic scaffold (PCL-Coll) were prepared by the lyophilization method, then conjugated with selected nanoparticles(Chi-PEG_70:30-HE). Notably, in addition to PCL-Coll/Chi-PEG-HE, two scaffolds of PCL-Coll and PCL-Coll/Chi-PEG were prepared to evaluate the role of conjugation in the release behavior of herbal bio-macromolecules. Based on the results, the conjugation process was led to a more stable release, compared to unconjugated nanoparticles. The mentioned process also created an integrated network along with better physicomechanical properties [modulus:12.31 MPa, tensile strength:4.44 MPa, smaller pore size(2 μm), and better swelling (100.27 %) with a symmetrical wettability on the surface]. PCL-Coll/Chi-PEG-HE scaffold was also resulted in higher expression levels of K10 and K14 keratinocytes with biomimetic patterns than PCL-Coll/Chi-PEG scaffold. This could be due to the active ingredients of V. sinuatum extract like alkaloids, flavonoids, and triterpenoids which imparts the wound healing (anti-inflammatory, anti-bacterial, anti-oxidant) properties to this scaffold. It seems that the use of bioactive materials like herbal extracts, in the form of encapsulated into polymeric nanocarriers, in the structure of engineered scaffolds can be a promising option for regenerating damaged skin without scarring. Hence, this study can provide innovative insights into the combination of two techniques of drug delivery and tissue engineering to design bio-scaffolds containing bioactive molecules with better therapeutic approaches.

Keywords: Bio-macromolecules; Bio-scaffold; Conjugation process; Drug delivery systems; Herbal extract-loaded nanoparticles; Regenerative medicine.