Open AccessEffect of freeze drying on stability, thermo-responsive characteristics, and in vivo wound healing of erythropoietin-loaded trimethyl chitosan/glycerophosphate hydrogel
Author(s) -
Vajihe Akbari,
Mahboubeh Rezazadeh,
Mohsen Minayian,
Marjan Amirian,
Azadeh Moghadas,
Ardeshir Talebi
Publication year - 2018
Publication title -
research in pharmaceutical sciences
Language(s) - English
Resource type - Journals
SCImago Journal Rank - 0.685
H-Index - 29
eISSN - 1735-9414
pISSN - 1735-5362
DOI - 10.4103/1735-5362.245959
Subject(s) - bioadhesive , mucositis , chitosan , chemistry , erythropoietin , in vivo , wound healing , freeze drying , chromatography , pharmacology , medicine , biochemistry , surgery , polymer , toxicity , microbiology and biotechnology , organic chemistry , biology
Erythropoietin (EPO) was successfully incorporated into a bioadhesive thermosensitive hydrogel based on trimethyl chitosan (TMC)/β-glycerophosphate (GP) for prevention and treatment of oral mucositis in cancerous patients. The aim of the present study was to evaluate the effect of freeze drying on thermo-responsive property of the hydrogel and structural stability of the loaded protein. The freeze-dried EPO-loaded hydrogel were characterized using various methods. Gelation property by rheological analysis, EPO aggregation in formulations by sodium dodecyl sulfate polyacrylamide gel electrophoresis (SDS-PAGE), protein secondary structure by far ultraviolet-circular dichroism (CD), and the antigenic activity of EPO with ELISA techniques. The healing effects of the freeze-dried formulation was also investigated in Sprague-Dawley rats with chemotherapy-induced mucositis and compared with freshly prepared mixture. Finally, the retention time of the gel in the oral cavity was assessed in healthy volunteers. SDS-PAGE, CD, and ELISA confirmed the stability of conformational structure of loaded and released EPO. Severity of mucositis was markedly reduced in animals treated with freeze-dried EPO hydrogel; whereas the group received normal saline did not show any significant healing. EPO salvia level was decreased rapidly following EPO solution compared to the gel application. Approximately, 40% of EPO was maintained on the buccal areas in patients receiving the hydrogel system after 30 min. Therefore, the TMC/GP could preserve EPO stability after freeze drying and has the potential in the treatment of oral mucositis and other oral or subcutaneous wounds.