Enhanced osteogenesis of human mesenchymal stem cells by self-assembled peptide...
引用 | Wiley Online Library報導
Enhanced osteogenesis of human mesenchymal stem cells by self-assembled peptide hydrogel functionalized with glutamic acid templated peptides
Abstract 自組裝肽 (SAP) 水凝膠已被證明是用於三維細胞培養和刺激細胞遷移和分化為支架以及修復骨組織缺損的優良生物材料。在此，我們通過與短生物活性基序 O1 (EEGGC) 和 O2 (EEEEE) 的直接偶聯設計了一種 SAP 支架 KLD (KLDLKLDLKLDL)，其對成骨分化的生物活性先前已在不同濃度 (0.5%, 1 % 和 2%）。我們的目的是增強可注射的SAP水凝膠的成骨作用和生物礦化，使其具有受控的機械性能，從而使肽水凝膠也能夠被注射到骨缺損中。使用原子力顯微鏡 (AFM) 和掃描電子顯微鏡 (SEM) 觀察納米纖維肽支架的分子整合。評估 SAP 水凝膠的流變特性和降解曲線以確保 SAP 的穩定性。與純 KLD 支架相比，我們發現這些設計的生物活性肽支架顯著促進了 hMSCs 增殖，通過鹼性磷酸酶 (ALP) 活性、總鈣沉積的生化分析顯示。此外，通過實時聚合酶鏈反應 (PCR) 和免疫熒光分析確定的 ALP 活性、I 型膠原蛋白 (COL-1)、骨橋蛋白 (OP) 和骨鈣素 (OCN) 表達水平的關鍵成骨標誌物也顯著增加。向 KLD 添加谷氨酸殘基。我們證明了設計的 SAP 支架促進了 hMSCs 的增殖和成骨分化。我們的結果表明，這些設計的生物活性肽支架可能有助於促進骨組織再生。 Self-assembling peptide (SAP) hydrogel has been shown to be an excellent biological material for three-dimensional cell culture and stimulatie cell migration and differentiation into the scaffold, as well as for repairing bone tissue defects. Herein, we designed one of the SAP scaffolds KLD (KLDLKLDLKLDL) through direct coupling to short bioactive motif O1 (EEGGC) and O2 (EEEEE) of which bioactivity on osteogenic differentiation was previously demonstrated and self-assembled in different concentrations (0.5%, 1%, and 2%). Our aim was to enhance osteogenesis and biomineralization of injectable SAP hydrogels with controlled mechanical properties so that the peptide hydrogel also becomes capable of being injected to bone defects. The molecular integration of the nanofibrous peptide scaffolds was observed using atomic force microscopy (AFM) and scanning electron microscopy (SEM). The rheological properties and degradation profile of SAP hydrogels were evaluated to ensure stability of SAPs. Compared with pure KLD scaffold, we found that these designed bioactive peptide scaffolds significantly promoted hMSCs proliferation depicted by biochemical analysis of alkaline phosphatase (ALP) activity, total calcium deposition. Moreover, key osteogenic markers of ALP activity, collagen type I (COL-1), osteopontin (OP), and osteocalcin (OCN) expression levels determined by real-time polymerase chain reaction (PCR) and immunofluorescence analysis were also significantly increased with the addition of glutamic acid residues to KLD. We demonstrated that the designed SAP scaffolds promoted the proliferation and osteogenic differentiation of hMSCs. Our results suggest that these designed bioactive peptide scaffolds may be useful for promoting bone tissue regeneration.