Fabrication of myomucosal flap using tissue-engineered bioartificial mucosa constructed with oral keratinocytes cultured on amniotic membrane.
The purpose of this study was to fabricate bioartificial mucosa using cultured oral keratinocytes (OKCs) on an amniotic membrane (AM), and to evaluate the possibility of developing a prelaminated myomucosal flap using the fabricated bioartificial mucosa and local muscle flap. Buccal mucosa was harvested from male New Zealand rabbits (n = 40, 2.5-3.0 kg) and primary cultivation was performed. The cultured OKCs were seeded on the AM and a submerged culture was performed. Prelamination of the bioartificial mucosa was performed on the latissimus dorsi (LD) muscle of rabbits. Survival rate, layer of OKCs, and Cinamon's score (CS) based on macroscopic and microscopic examinations were evaluated 7, 10, 14, and 21 days after prelamination (n = 10 per day). The OKCs cultured on AM showed multiple layers (3.85 +/- 1.32) and cells were tightly adhered with desmosomes. Basal layer cells adhered to the AM with hemidesmosomes. In addition, the AM played an excellent role as a substrate for the OKCs and simplified handling during prelamination. A myomucosal flap with OKCs cultured on AM was fabricated within 2 weeks (CS: 11.05 +/- 2.63). The basement component of laminin was observed 2 weeks after prelamination and showed enough strength to adhere to the underlying fascia. A myomucosal flap was successfully developed using prelamination of bioartificial mucosa on the LD muscle between 10 and 14 days.
Ahn
Kang-Min
KM
Department of Oral and Maxillofacial Surgery, School of Medicine, Ulsan University, Asan Medical Center, Ulsan, Korea.
Lee
Jong-Ho
JH
Hwang
Soon-Jung
SJ
Choung
Pill-Hoon
PH
Kim
Myung-Jin
MJ
Park
Hee-Jung
HJ
Park
Jung-Keug
JK
Jahng
Jeongwon
J
Yang
Eun-Kyung
EK
D000650:Amnion; D000818:Animals; D021522:Bioartificial Organs; D002470:Cell Survival; D002478:Cells, Cultured; D005260:Female; D006801:Humans; D015603:Keratinocytes / Q000166:cytology* / Q000648:ultrastructure; D007797:Laminin / Q000378:metabolism; D008297:Male; D009061:Mouth Mucosa / Q000166:cytology*; D011247:Pregnancy; D011817:Rabbits; D019651:Plastic Surgery Procedures; D013524:Surgical Flaps / Q000502:physiology*; D023822:Tissue Engineering / Q000379:methods*
D007797:Laminin