Formation of in vivo tissue engineered human hyaline cartilage in the shape of a trachea with internal support.
OBJECTIVE
Treatment and management of congenital as well as post-traumatic trachea stenosis remains a challenge in pediatric surgery. The aim of this study was to reconstruct a trachea with human nasal septum chondrocytes by using the combination of biodegradable hydrogel and non-biodegradable high-density polyethylene (HDP) as the internal predetermined shape scaffold.


METHODS
Human nasal septum cartilage was harvested as excessive tissue after elective septoplasty and digested in 0.6% collagenase II. Chondrocytes were cultured in an equal volume mix of Ham's F12 medium and Dulbecco's modified eagle medium added with 10% fetal bovine serum and basic fibroblast growth factor. After two passages, the cultured chondrocytes were trypsinized and mixed with biodegradable hydrogel Pluronic F127. The chondrocytes-hydrogel admixture was then painted over the HDP as the internal support in a predetermined trachea shape. The composite was then implanted subcutaneously in athymic mice.


RESULTS
After 8 weeks of in vivo implantation, the tissue engineered trachea constructs were harvested. Macroscopic appearance of the tissue engineered trachea constructs demonstrated that the HDP were 80-90% covered with yellowish glistering cartilage like tissue without any sign of inflammation. The tissue engineered trachea cartilage consisted of evenly spaced lacunae embedded in basophilic matrix and stained red with Safranin-O staining denoting abundant proteoglycans production. Type II collagen gene which was expressed in native cartilage was highly expressed in this tissue engineered trachea cartilage.


CONCLUSIONS
We have successfully reconstructed a trachea in vivo with human nasal septum chondrocytes using HDP as the internal support. This construct has the advantage of bio-inert and strength in which both are important properties in tracheal reconstruction.
Ruszymah
Binti Haji Idrus
BH
Department of Physiology, Faculty of Medicine, Universiti Kebangsaan Malaysia, Jalan Raja Muda Abdul Aziz, 50300 Kuala Lumpur, Malaysia. ruszy@medic.ukm.my
Chua
Kienhui
K
Latif
Mazlyzam Abdul
MA
Hussein
Fuzina Nor
FN
Saim
Aminuddin Bin
AB
D000818:Animals; D001672:Biocompatible Materials; D001705:Bioprosthesis; D002478:Cells, Cultured; D019902:Chondrocytes / Q000637:transplantation*; D024043:Collagen Type II / Q000378:metabolism; D005079:Excipients; D006801:Humans; D051457:Hyaline Cartilage / Q000166:cytology* / Q000378:metabolism; D051379:Mice; D008819:Mice, Nude; D009300:Nasal Septum / Q000166:cytology; D020442:Poloxamer; D020959:Polyethylene; D012333:RNA, Messenger / Q000378:metabolism; D020133:Reverse Transcriptase Polymerase Chain Reaction; D023822:Tissue Engineering / Q000379:methods*; D014132:Trachea / Q000378:metabolism / Q000601:surgery*
D001672:Biocompatible Materials; D024043:Collagen Type II; D005079:Excipients; D012333:RNA, Messenger; D020442:Poloxamer; D020959:Polyethylene