Biological evaluation of human degenerated nucleus pulposus cells in functionalized self-assembling peptide nanofiber hydrogel scaffold.
Nucleus pulposus (NP) tissue engineering has been proposed as a novel biological treatment for early-stage intervertebral disc degeneration. In this study, a novel functional self-assembling peptide PKP was first designed by linking the short functional motif of bone morphogenetic protein-7 (BMP7) to the C-terminal of RADA16-I, and another new functional self-assembling peptide was obtained by mixing RKP with RADA16-I. Then, the biocompatibilities and bioactivities of RKP and RAD-RKP for human degenerated nucleus pulposus cells (hNPCs) were studied in vitro. Atomic force microscopy and scanning electron microscopy (SEM) confirmed that both RKP and RAD-RKP could self-assemble into three-dimensional (3D) nanofiber hydrogel scaffolds in a culture medium at 37°C. After the hNPCs were cultured in 3D scaffolds, both RKP and RAD-RKP exhibited reliable attachment and extremely low cytotoxicities (<14%), which were verified by SEM and cytotoxity assays, respectively. Our results also showed that the functional-based scaffolds could increase the proliferation and migration of hNPCs after 7 days compared with culture plates and pure RADA16-I. Quantitative real-time polymerase chain reaction demonstrated that the expressions of collagen II α1, Sox-9, and aggrecan were upregulated, while collagen I α1 was downregulated by functional-based scaffolds after 28 days. Furthermore, we also confirmed that RAD-RKP exhibited a higher hNPC proliferation, migration, and expression of Sox-9 and aggrecan compared with pure RKP. Therefore, the results of this study indicated that the BMP7 short motif-designed functional self-assembling peptide nanofiber hydrogels could be used as excellent scaffolds in NP tissue engineering, and RAD-RKP might have further potential application in human mild degenerated NP tissue regeneration.
1 The Third Affiliated Hospital of Southern Medical University , Guangzhou, People's Republic of China .
Hui
H
Tao
Yan
Y
Zhang
Chao-Feng
CF
Wang
Chao
C
Zhang
Xiu-Mei
XM
Wang
De-Li
DL
Wang
Xue-Dong
XD
Bai
Tian-Yong
TY
Wen
Hong-Kui
HK
Xin
Jian-Hong
JH
Wu
Yue
Y
Liu
Qin
Q
He
Dike
D
Ruan
D000595:Amino Acid Sequence; D002465:Cell Movement / Q000187:drug effects; D049109:Cell Proliferation / Q000187:drug effects; D002470:Cell Survival / Q000187:drug effects; D002478:Cells, Cultured; D002942:Circular Dichroism; D005260:Female; D005786:Gene Expression Regulation / Q000187:drug effects; D006801:Humans; D020136:Hydrogel, Polyethylene Glycol Dimethacrylate / Q000737:chemistry*; D007403:Intervertebral Disc / Q000187:drug effects / Q000473:pathology*; D055959:Intervertebral Disc Degeneration / Q000235:genetics / Q000473:pathology*; D008875:Middle Aged; D008958:Models, Molecular; D008969:Molecular Sequence Data; D057139:Nanofibers / Q000737:chemistry*; D010455:Peptides / Q000138:chemical synthesis / Q000737:chemistry* / Q000494:pharmacology; D054457:Tissue Scaffolds / Q000737:chemistry*
D010455:Peptides; D020136:Hydrogel, Polyethylene Glycol Dimethacrylate