Self-assembling short-peptide hydrogel for three-dimensional culture of rabbit articular chondrocytes in vitro
- VernacularTitle:自组装短肽水凝胶体外三维培养兔关节软骨细胞
- Author:
Lanlan ZHANG
;
Hong SONG
;
Xiaojun ZHAO
- Publication Type:Journal Article
- From:
Chinese Journal of Tissue Engineering Research
2008;12(49):9779-9782
- CountryChina
- Language:Chinese
-
Abstract:
BACKGROUND:Self-assembling short-peptide has been shown to be a kind of hovel nanometer biomaterial.Following culture combined with chondrocytes,self-assembling short-peptide can promote eell marex secretion,cell division and proliferation.and eell phenotype maintenance.For this reason,it has become a candidate for all ideal cell scaffold used for cartilage tissue engineering.OBJECTIVE:To observe the biological characteristics of rabbit articular chondrocytes cultured in three-dimensional nanoscale self-assembling short.peptide KLD-12 hydrogel seaffold.DESIGN.TIME AND SETTING:An observational experiment was performed at the laboratory of Institute for Nanobiomedical Technology and Membrane Biology,Sichuan University between November 2007 and April 2008.MATERIALS:Nanoscale short-peptide KLD-12 with a sequence of Ac-KLDLKLDLKLDL-CONH2 was synthesized by Shanghai Biotech Company,China.METHODS:Chondrocytes were harvested from New Zealand rabbit articular cartilage by enzyme digestion.Following in vitro culture.chondrocytes of passage 3 were inoculated into 4 g/L self-assembling short-peptide KLD-12 solution at a density of 5x 109/L.Peptide gel formation was induced by phosphate buffered saline for three-dimensional culture.MAIN OUTCOME MEASURES:①Observation of chondrocyte morphology in the short-peptide hydrogel with inverted phase contrast microscope.②Detection of intracellular matrix secretion by toluidine blue staining and immunohistochemistry.③Measurement of mRNA expression levels of aggrccan,type Ⅱ collagen and type Ⅰ collagen in chondrocyte-seeded hydrogels by reverse transcription-polymerase chain reaction.RESULTS:①The viability of chondrocytes digested and isolated from rabbit articular cartilage was over 95%.④Chondrocytes cultured in three-dimensional short-peptide hydrogel maintained their round shape,proliferated actively,and aggregated in clusters.In addition.cartilage lacuna could be seen surrounding the clusters.③Both toluidine blue staining and type Ⅱ collagen immunohistochemistry results were positive.The mRNA expression levels of aggrecan and type Ⅱ collagen were increased with time.④Reverse transcription-polymerase chain reaction results revealed that after 3 weeks of in vitro culture in the short-peptide hydrogel,chondrocytes maintained the capability to secrete aggrecan and to express type Ⅱ collagen.CONCLUSION:Self-assembling short-peptide KLD-12 three-dimensional hydrogel Call well inainrain the normal cellular morphology,function and proliferative capability of chondrocytes
