Nanostructured Lipid Carrier Gel Formulation of Recombinant Human Thrombomodulin Improve Diabetic Wound Healing by Topical Administration.

Hsiu-Ching Yu,Shang-Wen Lin,Hua-Lin Wu,Shu-Jhen Jheng,Yuan-Shuo Hsueh,Jui-Chen Tsai,Yan-Jye Shyong

doi:10.3390/pharmaceutics13091386

Abstract

Recombinant human thrombomodulin (rhTM), an angiogenesis factor, has been demonstrated to stimulate cell proliferation, keratinocyte migration and wound healing. The objective of this study was to develop nanostructured lipid carrier (NLC) formulations encapsulating rhTM for promoting chronic wound healing. RhTM-loaded NLCs were prepared and characterized. Encapsulation efficiency was more than 92%. The rate of rhTM release from different NLC formulations was influenced by their lipid compositions and was sustained for more than 72 h. Studies on diabetic mouse wound model suggested that rhTM-NLC 1.2 µg accelerated wound healing and was similar to recombinant human epidermal growth factor-NLC (rhEGF-NLC) 20 µg. By incorporating 0.085% carbopol (a highly crosslinked polyacrylic acid polymer) into rhTM NLC, the NLC-gel presented similar particle characteristics, and demonstrated physical stability, sustained release property and stability within 12 weeks. Both rhTM NLC and rhTM NLC-gel improved wound healing of diabetic mice and cell migration of human epidermal keratinocyte cell line (HaCaT) significantly. In comparison with rhTM solution, plasma concentrations of rhTM post applications of NLC and NLC-gel formulations were lower and more sustained in 24 h. The developed rhTM NLC and rhTM NLC-gel formulations are easy to prepare, stable and convenient to apply to the wound with reduced systemic exposure, which may warrant potential delivery systems for the care of chronic wound patients.

Highlights

Zeta potential of nanostructured lipid carrier (NLC) formulated with ATO5 had higher absolute value than NLC formulated with Geleol, and NLC formulated with higher ratio of oils showed more negative potential in comparison with lipids (Table S1 and Figure S1)
NLCs formulated with ratio of lipid to oil 5:5 (1:1) exhibited smallest and more consistent particle size around 200 nm and polydispersity index (PDI) of 0.2–0.3, and were selected for further studies
The importance of using growth factors on wound healing is rapidly increasing; howset of experiments substantiated that incorporation of carbopol with Recombinant human thrombomodulin (rhTM) NLC did not ever, having high concentration of these growth factors entering the circulation may cause affect its efficacy

Summary

Introduction

The wound healing process is a physiological dynamic interaction which involves several types of cells, tissues or proteins, such as parenchymal cells, blood cells, extracellular matrix, cytokines networks and growth factors, aiming at repairing the damaged cutaneous tissues [1,2,3]. Wound healing events can be divided into four phases: hemostasis, inflammatory, proliferative and remodeling phases [1,2,3,4,5,6]. The hemostasis phase leads to vasoconstriction and platelets aggregation at the site of injury to prevent exsanguination. The inflammatory phase is represented by a complex series of molecular signals

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Conclusion