Abdominal wall repair using a biodegradable scaffold seeded with cells

Presented at the 51st Annual Congress of the British Association of Paediatric Surgeons, Oxford, England, July 27-30, 2004.
https://doi.org/10.1016/j.jpedsurg.2004.10.019Get rights and content

Abstract

Background/Purpose

The repair of large abdominal wall defects is still a challenge for pediatric surgeons. Synthetic materials, however, may lead to high complication rates. This study was aimed at applying tissue-engineering methods to abdominal wall repair.

Methods

3T3 mouse fibroblasts were expanded in vitro. In the next step, a biodegradable material—polyglycolic acid (PGA)—was actively seeded with 107 cells/scm of PGA scaffold. Culture medium (Dulbecco's Modified Eagle's Medium with 10% fetal bovine serum) was changed every 6 hours after seeding cells on PGA fibers. Under general anaesthesia, C57BL/6J black mice underwent creation of a 2 × 3-cm abdominal wall defect (60%-70% of abdominal surface). The defect was repaired in the experimental group with the fibroblast-seeded PGA scaffold. In the first control group, the defect was covered with acellular PGA, and in the second control group, by skin closure. Animals were killed after 30 days to assess the histologic and gross findings.

Results

No abdominal hernia was found in animals repaired with cell-seeded and acellular scaffolds. All animals with skin closure died within 7 days. In every case, tissue-engineered construct was thicker then in controls. Histologic and gross examination revealed a good neovascularisation in tissue-engineered abdominal walls comparing to the acellular matrix. There was no intensive scar formation between abdominal wall and skin.

Conclusions

Engineered soft tissue constructs can provide structural replacement of severe and large abdominal wall defects. Tissue engineering in the near future will possibly enter clinical practice.

Section snippets

Cell culture

3T3 mouse fibroblasts were grown in vitro in 95% humified 5% CO2 chamber at 37°C. Growth medium consisted of Dulbecco's Modified Eagle's Medium with glutamine (Sigma, Germany) supplemented with 10% fetal bovine serum (Gibco, UK) and antibiotic solution (100 U/mL penicillin G sodium, 100 μg/mL gentamicin, 100 μg/mL streptomycin, and 5 μg/mL amphotericin, Sigma). Cells were cultured in 75-cm2 culture flasks (Corning) and passaged every 3 days using trypsin 1× solution (2.5 g trypsin per liter

Results

It took 7 days to achieve sufficient 3T3 cell number for further seeding. Cells were seeded 3 times on the scaffold until tightly covering was obtained. The adhesive properties of PGA fibers were excellent. Cells started to grow on PGA construct after the first seeding and covered the scaffold fibers within 3 days (Fig. 1). Medium was changed every 6 hours after the pH decrease was observed. No signs of scaffold degradation were noticed during the period of 3 days of tissue culture on 3D

Discussion

The complication rate after repair of large tissue defects strongly depends on applied reconstructive material. Wound infections, bowel fistulae, and repair failures can occur when synthetic materials were used for defect closure. Naturally derived materials are less susceptible to infection [4], [6], [7]. The strength of allogenic and xenogenic matrices decreases over time [9]. Authors tried to find compromise between good mechanical properties that belong to synthetic materials and graft

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