Human osteoblast cells: isolation, characterization, and growth on polymers for musculoskeletal tissue engineering.
We performed a detailed examination of the isolation, characterization, and growth of human osteoblast cells derived from trabecular bone. We further examined the morphology, phenotypic gene expression, mineralization,and growth of these human osteoblasts on polyester polymers used for musculoskeletal tissue engineering. Polylactic-co-glycolic acid [PLAGA (85:15, 50:50, 75:25)], and poly-lactic acid (L-PLA, D,L-PLA) were examined. The osteoblastic expression of key phenotypic markers osteocalcin, alkaline phosphatase, collagen, and bone sialoprotein at 4 and 8 weeks was examined. Reverse transcription-polymerase chain reaction studies revealed that trabecular-derived osteoblasts were positive for all markers evaluated with higher levels expressed over long-term culture. These cells also revealed mineralization and maturation as evidenced by energy dispersive X-ray analysis and scanning electron microscopy. Growth studies on PLAGA at 50:50,75:25, and 85:15 ratios and PLA in the L and DL isoforms revealed that human osteoblasts actively grew, with significantly higher cell numbers attached to scaffolds composed of PLAGA 50:50 in the short term and PLAGA 85:15 in the long term compared with PLA (p < 0.05). We believe human cell adhesion among these polymeric materials may be dependent on differences in cellular integrin expression and extracellular matrix protein elaboration.
Department of Orthopaedic Surgery, University of Virginia, 400 Ray C. Hunt Drive, Charlottesville, Virginia 22903, USA.
Saadiq F
SF
El-Amin
Edward
E
Botchwey
Richard
R
Tuli
Michelle D
MD
Kofron
Addisu
A
Mesfin
Swaminathan
S
Sethuraman
Rocky S
RS
Tuan
Cato T
CT
Laurencin
D001672:Biocompatible Materials; D001842:Bone and Bones / Q000166:cytology / Q000502:physiology; D002113:Calcification, Physiologic / Q000502:physiology; D018929:Cell Culture Techniques / Q000379:methods; D002469:Cell Separation / Q000379:methods; D002478:Cells, Cultured; D005786:Gene Expression Regulation / Q000502:physiology; D006801:Humans; D019344:Lactic Acid; D010006:Osteoblasts / Q000166:cytology* / Q000502:physiology; D011091:Polyesters; D011100:Polyglycolic Acid; D011108:Polymers; D023822:Tissue Engineering / Q000379:methods
D001672:Biocompatible Materials; D011091:Polyesters; D011108:Polymers; D011100:Polyglycolic Acid; D019344:Lactic Acid; C033616:poly(lactide)