Investigating initial cell-seeding density and culture period of fibroblast growing on biodegradable tricalcium phosphate lysine disks.
"Tissue engineering" uses implanted cells, scaffolds, DNA, protein, and/or protein fragments to replace or repair injured or diseased tissues and organs. Recently, biocompatible materials, such as hydroxyapatite and tricalcium phosphate, have been associated with cellular migration, exposure, and an increase in inflammatory mediators. Complications are less likely after the implants have become integrated. A model was sought to study the influence of multiple cell densities on the rate of fibroblast in-growth into porous implants. Fibroblasts were seeded at three different densities (50, 100, 200 x 10(6) cells/well) onto the surface of a 24 well tissue culture plate in the presence of porous tricalcium phosphate lysine ceramic disks. Cell number, viability, and morphology were evaluated after 1, 2, and 3 days. The results show a linear 2-fold increase in cell number during the first two days in culture. By day 3, the cells initially seeded at 200 x 10(6) cells no longer exhibited linear growth. This suggests that the tissue culture wells were confluent. Interestingly, the cells growing in wells containing TCPL disks showed slightly more cell numbers than cells growing on plastic alone for each density. The results also revealed no significant differences between the groups for parameters of cell function such as protein concentration and markers of cellular damage. Evaluation of cellular morphology indicated changes in cellular appearance with time in culture. Overall, initial cell seeding is important for faster in-growth, but with time the cells seeded at lower cell numbers will also become confluent. In conclusion, this study provides a novel approach for the study of fibroblast in-growth into implants with time. This work is important for understanding functional tissue engineering and ultimately result in safer and more efficacious repairs and replacements for the surgeon and patient.
Trussell
Brian
B
University of Mississippi Medical Center, Department of Orthopedics, 2500 North State Street, Jackson, Mississippi 39216, USA.
Ward
John
J
Cox
Michael
M
Tucci
Michelle
M
Benghuzzi
Hamed
H
Hughes
James
J
D001673:Biodegradation, Environmental; D002130:Calcium Phosphates; D002452:Cell Count; D018929:Cell Culture Techniques / Q000379:methods*; D005347:Fibroblasts / Q000166:cytology* / Q000378:metabolism; D008239:Lysine; D010710:Phosphates; D011506:Proteins / Q000378:metabolism; D023822:Tissue Engineering / Q000295:instrumentation* / Q000379:methods*
D002130:Calcium Phosphates; D010710:Phosphates; D011506:Proteins; C112656:tricalcium phosphate lysine; D008239:Lysine