A novel dimeric thymosin beta 4 with enhanced activities accelerates the rate of wound healing.

ABSTRACT: Thymosin beta 4 (T?4) is a peptide with 43 amino acids that is critical for repair and remodeling tissues on the skin, eye, heart, and neural system following injury. To fully realize its utility as a treatment for disease caused by injury, the authors constructed a cost-effective novel T?4 dimer and demonstrated that it was better able to accelerate tissue repair than native T?4.A prokaryotic vector harboring two complete T?4 genes with a short linker was constructed and expressed in Escherichia coli. A pilot-scale fermentation (10 L) was performed to produce engineered bacteria and the T?4 dimer was purified by one-step hydrophobic interaction chromatography. The activities of the T?4 dimer to promote endothelial cell proliferation, migration, and sprouting were assessed by tetramethylbenzidine (methylthiazol tetrazolium), trans-well, scratch, and tube formation assays. The ability to accelerate dermal healing was assessed on rats.After fermentation, the T?4 dimer accounted for about 30% of all the bacteria proteins. The purity of the T?4 dimer reached 98% after hydrophobic interaction chromatography purification. An average of 562.4 mg/L T?4 dimer was acquired using a 10 L fermenter. In each assay, the dimeric T?4 exhibited enhanced activities compared with native T?4. Notably, the ability of the dimeric T?4 to promote cell migration was almost two times higher than that of T?4. The rate of dermal healing in the dimeric T?4-treated rats was approximately 1 day faster than with native T?4-treated rats.The dimeric T?4 exhibited enhanced activity on wound healing than native T?4, and the purification process was simple and cost-effective. This data could be of significant benefit for the high pain and morbidity associated with chronic wounds disease. A better strategy to develop T?4 as a treatment for other diseases caused by injuries such as heart attack, neurotrophic keratitis, and multiple sclerosis was also described.

SUBMITTER: Xu TJ 

PROVIDER: S-EPMC3792846 | biostudies-literature | 2013

REPOSITORIES: biostudies-literature