Tissue engineering of a periodontal ligament-alveolar bone graft construct.
OBJECTIVE
This paper reports on a 2-phase study of a novel membrane-scaffold graft construct, its ability to support periodontal ligament fibroblast (PDLF) and alveolar osteoblast (AO) growth in vitro, and its use for tissue engineering a PDL-AO interface in vivo.


METHODS
Human PDLFs were seeded onto perforated poly(epsilon-caprolactone) membranes (n=30) at 78,000 cells/cm2; human AOs were seeded on poly(epsilon-caprolactone) scaffolds (n=30) with fibrin glue at 625,000 cells/cm3. Cell attachment, morphology, viability, and metabolic activity were monitored for 3 weeks in vitro. Subsequently, cell-seeded membrane-scaffold constructs (experimental group, n=9) and nonseeded constructs (control group, n=4) assembled with fibrin glue were implanted subcutaneously into 7 athymic mice for 4 weeks.


RESULTS
PDLFs formed confluent layers on membranes, whereas AOs produced mineralized matrices within scaffolds upon osteoinduction in vitro. Well-vascularized tissue formation was observed after implantation. Integration at the membrane-scaffold interface was enhanced in the experimental group. Type I collagen, type III collagen, fibronectin, and vitronectin were found adjacent to membranes and within constructs. Bone sialoprotein expression and bone formation were undetectable.


CONCLUSIONS
Membrane perforation and scaffold porosity facilitated tissue integration and vascularization at the construct-recipient site. However, the interaction between PDLF and AO could have interfered with osteogenesis at the interface of soft and mineralizing tissues.


CONCLUSIONS
Both matrices supported PDLF and AO attachment and proliferation in vitro. The membrane-scaffold construct facilitated tissue growth and vascularization while providing strength and form in vivo.
Chou
Ai Mei
AM
Department of Biological Sciences, National University of Singapore, Singapore.
Sae-Lim
Varawan
V
Hutmacher
Dietmar W
DW
Lim
Tit Meng
TM
D000328:Adult; D000539:Alveolar Process / Q000166:cytology / Q000637:transplantation; D000818:Animals; D002417:Cattle; D018929:Cell Culture Techniques; D005260:Female; D005347:Fibroblasts / Q000166:cytology; D006801:Humans; D051379:Mice; D010006:Osteoblasts / Q000166:cytology; D010513:Periodontal Ligament / Q000166:cytology / Q000637:transplantation; D023822:Tissue Engineering / Q000379:methods*