Osteogenic potential of human periosteum-derived progenitor cells in PLGA scaffold using allogeneic serum.
The use of periosteum-derived progenitor cells (PCs) combined with bioresorbable materials is an attractive approach for tissue engineering. The aim of this study was to characterize the osteogenic differentiation of PC in 3-dimensional (3D) poly-lactic-co-glycolic acid (PLGA) fleeces cultured in medium containing allogeneic human serum. PCs were isolated and expanded in monolayer culture. Expanded cells of passage 3 were seeded into PLGA constructs and cultured in osteogenic medium for a maximum period of 28 d. Morphological, histological and cell viability analyses of three-dimensionally cultured PCs were performed to elucidate osseous synthesis and deposition of a calcified matrix. Furthermore, the mRNA expression of type I collagen, osteocalcin and osteonectin was semi-quantitively evaluated by real-time reverse transcriptase-polymerase chain reaction (RT-PCR). The fibrin gel immobilization technique provided homogeneous PCs distribution in 3D PLGA constructs. Live-dead staining indicated a high viability rate of PCs inside the PLGA scaffolds. Secreted nodules of neo-bone tissue formation and the presence of matrix mineralization were confirmed by positive von Kossa staining. The osteogenic differentiation of PCs was further demonstrated by the detection of type I collagen, osteocalcin and osteonectin gene expression. The results of this study support the concept that this tissue engineering method presents a promising method for creation of new bone in vivo.
Zheng
Yi-xiong
YX
Department of Surgery, Second Affiliated Hospital, School of Medicine, Zhejiang University, Hangzhou 310009, China. zyx_xxn@hotmail.com
Ringe
Jochen
J
Liang
Zhong
Z
Loch
Alexander
A
Chen
Li
L
Sittinger
Michael
M
D001672:Biocompatible Materials; D001846:Bone Development; D018929:Cell Culture Techniques / Q000379:methods; D002454:Cell Differentiation; D002470:Cell Survival; D002478:Cells, Cultured; D003094:Collagen / Q000737:chemistry; D006801:Humans; D019344:Lactic Acid / Q000737:chemistry*; D008856:Microscopy, Fluorescence; D015233:Models, Statistical; D010012:Osteogenesis; D010521:Periosteum / Q000378:metabolism; D011100:Polyglycolic Acid / Q000737:chemistry*; D000077182:Polylactic Acid-Polyglycolic Acid Copolymer; D011108:Polymers / Q000737:chemistry*; D013234:Stem Cells / Q000166:cytology*; D023822:Tissue Engineering
D001672:Biocompatible Materials; D011108:Polymers; D000077182:Polylactic Acid-Polyglycolic Acid Copolymer; D011100:Polyglycolic Acid; D019344:Lactic Acid; D003094:Collagen