Recombinant human growth hormone expression in E.coli Origami (DE3) hosts and preparing a topical formulation of liposomes containing recombinant human growth hormone for skin regeneration and repair

Abstract

Introduction: Topical administration of recombinant human growth hormone (rhGH) is an interesting strategy for therapeutic purposes. We have formulated and characterized a topical rhGH-loaded liposomal formulation (rhGH-Lip) and evaluated its safety, biological activity, and protective role against UVB-induced skin damage. Materials and Methods: The formulations’ size, zeta potential and PDI were determined using dynamic light scattering. The morphological evaluations were perfomrd usig TEM. The stability of hGH was determined using SEC-HPLC, SDS-PAGE and circular dichroism. The safety and biological activity of rhGH-Lip was evaluated on BaF3-hGHR, HaCaT and HFFF-2 cell lines using MTT assay. The protective role of hGH on skin cells against UVB damage was assessed by flowcytometry. Anti-UVB effects and skin hydration by rhGH-Lip were studied in nude mice. Type I collagen, elastin and P53 expression in nude mice skin following UVB exposure was assessed by real-time PCR. Results: The rhGH-Lip had an average size and zeta potential of 63 nm and -33 mV, respectively, with 70% encapsulation efficiency. The formulation was stable at 4 ºC for at least one year. The SDS-PAGE and circular dichroism results showed no structural alterations in rhGH upon encapsulation. In Vitro, studies in HaCaT, HFFF-2, and Ba/F3-rhGHR cell lines confirmed the safety and biological activity of rhGH-Lip. In addition, fow cytmometry analysis revealed that rhGH can increase cell viability and reduce UVB-induced apoptosis in HaCaT and HFFF-2 cell lines. Treatment of cells with rhGH also increased the mRNA expression of collagen and elastin genes in HFFF-2 cells. Franz diffusion cell study showed increased rhGH skin permeation compared to free rhGH. Animal studies in nude mice showed that liposomal rhGH prevented UVB-induced epidermal hyperplasia, angiogenesis, wrinkle formation, and collagen loss, as well as improving skin moisture. Conclusion: The results of this study show that rhGH-Lip is a stable, safe, and effective skin delivery system and has potential as an anti-wrinkle formulation for topical application. This study also provides a new method for the topical delivery of proteins and merits further investigation.