Designer self-assembling peptide scaffold stimulates pre-osteoblast attachment, spreading and proliferation.
A new peptide scaffold was made by mixing pure RADA16 (Ac-RADARADARADARADA-CONH2) and designer peptide RGDA16 (Ac-RADARGDARADARGDA-CONH2) solutions, and investigate any effect on attachment, spreading and proliferation of pre-osteoblast (MC3T3-E1). The peptides, RADA16 and RGDA16, were custom-synthesized. They were solubilized in deionized water at a concentration of 10 mg/ml (1% w/v), the RGDA16 peptide solution was mixed 1:1 with RADA16 solution and a new peptide solution RGDAmix was produced. The RGDAmix and RADA16 solution were directly loaded in 96-well plates and cover slips, and two different peptide scaffolds were formed with the addition of maintenance medium (alpha-MEM) in several minutes. About 1.0 x 10(4) MC3T3-E1 cells were seeded on each hydrogel scaffold, and then the cell morphological changes were observed using a fluorescence microscope at 1 h, 3 h and 24 h timepoint, respectively. Cell attachment was evaluated 1 h, 3 h and 24 h after cell seeding and cell proliferation was determined 4d, 7d and 14d after cell seeding. The RGDAmix scaffold significantly promoted the initial cell attachment compared with the RADA16 scaffold. MC3T3-E1 cells adhered and spread well on both scaffolds, however, cells spread better on the RGDAmix scaffold than on the RADA16 scaffold. Cell proliferation was greatly stimulated when cultured on RGDAmix scaffold. The RGD sequence contained peptide scaffold RGDAmix significantly enhances MC3T3-E1 cells attachment, spreading and proliferation.
Zhang
Feng
F
Department of Oral and Maxillofacial Surgery, The Affiliated Stomatology Hospital, College of Medicine, Zhejiang University, 395# Yan'an Road, Hangzhou 310006, People's Republic of China.
Shi
Geng-Sheng
GS
Ren
Ling-Fei
LF
Hu
Fei-Qing
FQ
Li
Sheng-Lai
SL
Xie
Zhi-Jian
ZJ
D000818:Animals; D002448:Cell Adhesion / Q000502:physiology; D018929:Cell Culture Techniques / Q000379:methods*; D002460:Cell Line; D002465:Cell Movement / Q000502:physiology; D049109:Cell Proliferation; D002478:Cells, Cultured; D015195:Drug Design; D005109:Extracellular Matrix / Q000737:chemistry*; D051379:Mice; D046912:Multiprotein Complexes / Q000737:chemistry / Q000648:ultrastructure; D010006:Osteoblasts / Q000166:cytology* / Q000502:physiology*; D010012:Osteogenesis / Q000502:physiology*; D010455:Peptides / Q000737:chemistry*; D023822:Tissue Engineering / Q000379:methods*
D046912:Multiprotein Complexes; D010455:Peptides