J Korean Surg Soc.  2013 Apr;84(4):216-224. 10.4174/jkss.2013.84.4.216.

Injection of porous polycaprolactone beads containing autologous myoblasts in a dog model of fecal incontinence

• Kang SB
• Lee HS
• Lim JY
• Oh SH
• Kim SJ
• Hong SM
• Jang JH
• Cho JE
• Lee SM
• Lee JH

• KMID: 2212466
• DOI: http://doi.org/10.4174/jkss.2013.84.4.216

Abstract

PURPOSE
Few studies have examined whether bioengineering can improve fecal incontinence. This study designed to determine whether injection of porous polycaprolactone beads containing autologous myoblasts improves sphincter function in a dog model of fecal incontinence.
METHODS
The anal sphincter of dogs was injured and the dogs were observed without and with (n = 5) the injection of porous polycaprolactone beads containing autologous myoblasts into the site of injury. Autologous myoblasts purified from the gastrocnemius muscles were transferred to the beads. Compound muscle action potentials (CMAP) of the pudendal nerve, anal sphincter pressure, and histopathology were determined 3 months after treatment.
RESULTS
The amplitudes of the CMAP in the injured sphincter were significantly lower than those measured before injury (1.22 mV vs. 3.00 mV, P = 0.04). The amplitudes were not different between dogs with and without the injection of autologous myoblast beads (P = 0.49). Resting and squeezing pressures were higher in dogs treated with autologous myoblast beads (2.00 mmHg vs. 1.80 mmHg; 6.13 mmHg vs. 4.02 mmHg), although these differences were not significant in analyses of covariance adjusted for baseline values. The injection site was stained for smooth muscle actin, but showed evidence of foreign body inflammatory reactions.
CONCLUSION
This was the first study to examine whether bioengineering could improve fecal incontinence. Although the results did not show definite evidence that injection of autologous myoblast beads improves sphincter function, we found that the dog model was suitable and reliable for studying the effects of a potential treatment modality for fecal incontinence.

Keyword

Fecal incontinence; Bioengineering; Dogs; Myoblasts

MeSH Terms

Actins
Action Potentials
Anal Canal
Animals
Bioengineering
Dogs
Fecal Incontinence
Foreign Bodies
Muscle, Smooth
Muscles
Myoblasts
Polyesters
Pudendal Nerve
Actins
Polyesters

Figure

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  Fig. 1 Monolayer culture of dog skeletal muscle cells. (A) Negative control for Pax7. (B) Immunocytochemical staining for Pax7. (C) Immunocytochemical staining for myosin heavy chain after cell fusion. (D) Immunocytochemical staining for smooth muscle actin after cell fusion (A-D, ×400).

• 

  Fig. 2 Representative compound muscle action potential traces. (A) Normal anal sphincter before injury. (B) Three months after anal sphincter injury. (C) Three months after injection of porous polycaprolactone beads containing autologous myoblasts. The compound muscle action potentials were recorded after electrical stimulation of the pudendal nerve.

• 

  Fig. 3 Representative anal sphincter pressure traces. (A) Normal anal sphincter before injury. (B) Three months after anal sphincter injury. (C) Three months after injection of porous polycaprolactone beads containing autologous myoblasts. The squeezing pressure was recorded after electrical stimulation from the resting pressure.

• 

  Fig. 4 Hematoxylin/eosin staining and immunostaining of the anal sphincter. (A-C) Normal anal sphincter. Normal outer striated muscle fibers (arrow) and internal smooth muscle layer (arrowhead) are indicated. (D-F) Three months after anal sphincter injury. Immunostaining showed extensive damage to the muscle fibers with cytoplasmic fibrosis and focal interstitial inflammatory cell infiltration (arrow), and atrophy of the muscle fibers. (G-I) Three months after injection of porous polycaprolactone beads containing autologous myoblasts. There was a marked foreign body reaction characterized by the presence of numerous giant cells and foamy macrophages (arrow), with weak staining for α-smooth muscle actin (A, D, G: hematoxylin-eosin staining; B, E, H: immunostaining of myosin heavy-chain; C, F, I: immunostaining of α-smooth muscle actin; A-G, ×100; H and I, ×200).

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