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NeoSphincter Constructed with a Prelaminated Human Primary Muco-Keratinocyte Lining in the Rat
Zachary P. French, BSE, Shiuhyang Kuo, DDS, Jana D. Moon, BS, Eve Bingham, MS, Zhifa Wang, MD, Elizabeth A. Mays, MSE, Hyungjin (Myra) Kim, PhD, Paul S. Cederna, MD, Cynthia L. Marcelo, PhD, Stephen E. Feinberg, DDS, PhD, Melanie G. Urbanchek, PhD.
University of Michigan, Ann Arbor, MI, USA.

PURPOSE:
Our long-term goal is to return dependable, self-controlled, anal function with a neoanal sphincter. This study in the rat combines a prelaminated, tissue engineered human muco-keratinocyte lining with a rat latissimus dorsi muscle flap that is constructed into a neosphincter. The purpose is to determine the impacts an acellular dermal matrix (ADM) and ADM with cells have on the contraction (squeezing) of neosphincters.
METHODS:
Human muco-keratinocytes were prelaminated on an ADM in culture prior to rat implantation. Then, neosphincters were constructed on the shoulders of athymic rats using a latissimus dorsi muscle flap with an intact neurovascular pedicle. Three experimental groups were: NeoSphincter (muscle only, n=5), NeoSphincter with ADM (n=4), and NeoSphincter with Prelaminated Mucosa (n=7). Neosphincters were evaluated for contractile force across the lumen diameter 14 days post implantation.
RESULTS:
All neosphincters were well vascularized, contractile, and with an open lumen. Muco-keratinocytes continued to divide with the mucosal layer increasing in thickness (Fig 1). Muscle fibers remained healthy while ADM became vascularized. Sphincter maximal isometric forces were similar across groups (Fig 2).
CONCLUSION:
By confirming the functioning and health of neosphincters constructed with prelaminated human muco-keratinocytes, this technology is moved closer to human implementation.


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