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© Borgis - New Medicine 2/2015, s. 43-50 | DOI: 10.5604/14270994.1169773
*Labib Zair1, Jędrzej Skrobot2, Mariola Marchlewicz3, Jarosław Lichota1, Leonard Gugała4, Magdalena Kuczyńska5, Samir Zeair6, Ewa Duchnik7, Karol Tejchman1, Barbara Wiszniewska8, Jerzy Sieńko1, Tadeusz Sulikowski1, Marek Ostrowski1, Mirosława El Fray2
Abdominal hernia repair surgery with the new injectable polymer biomaterials
1Clinic of General and Transplantation Surgery, Pomeranian Medical University, Szczecin
Head of Clinic: prof. Marek Ostrowski, MD, PhD
2Division of Biomaterials and Microbiological Technologies, West Pomeranian University of Technology, Szczecin
Head of Division: Mirosława El Fray
3Department of Aesthetic Dermatology, Pomeranian Medical University, Szczecin
Head of Department: prof. Mariola Marchlewicz, MD, PhD
4Veterinary Clinic, Szczecin
Head of Clinic: Leonard Gugała
5Independent Longterm Care Unit, Pomeranian Medical University, Szczecin
Head of Unit: Bożena Mroczek, MD, PhD
6Clinic of General, Vascular and Transplantation Surgery, Maria Skłodowska-Curie Hospital, Szczecin
Head of Clinic: Samir Zaeir
7Department of Dermatology and Venereology, Pomeranian Medical Univeristy, Szczecin
Head of Department: prof. Romuald Maleszka, MD, PhD
8Department of Histology and Embryology, Pomeranian Medical Univeristy, Szczecin
Head of Department: prof. Barbara Wiszniewska, MD, PhD
Summary
Introduction. The new injectable polymer biomaterials offer a completely innovative approach to hernia repair surgery. This study evaluated the potential application of injectable polymer biomaterials in hernia treatment using an animal model. Local connective tissue reactions associated with the new injectable polymer biomaterials were compared with the reactions provoked by commercial polypropylene mesh.
Aim. Evaluation of the potential application of injectable polymer biomaterials in the treatment of abdominal wall hernias using an animal model.
Material and methods. Five groups of rabbits with previously-created hernias were treated with new biomaterials injected in nonpolymerized form into hernial openings. The same animals had previously polymerized discs surgically implanted in the dorsal subcutaneous area. The rabbits were kept under clinical observation for 28 days, then sacrificed under anesthesia. Samples of subcutaneous tissue were taken from the hernia region and from the vicinity of the polymerized biomaterial discs implanted in the dorsal area for histological examination.
Results. In groups 1, 2 and 3, hernia treatment results, laboratory tests and histological examination of the tissues previously taken from the operated hernia sites were similar to the results obtained in the control group. In groups 1, 2 and 3, the numbers of fibrocytes and neutrophils in the vicinity of the subcutaneously-implanted discs in the dorsal area were similar to those in the hernial region treated with mesh in the control group.
Conclusions. Biomaterials 188-UR/PEG-DA 85/15, P1838-DMA and 1838_UR can be used in animals in the near future as possible abdominal wall hernia treatments. These biomaterials were as efficient as the polypropylene mesh used in the control group for treating abdominal wall hernias.
INTRODUCTION
Hernia repair represents a considerable proportion of the operations performed in general surgery. The number of specific operations reflects hernia incidence in the general population; typically, approximately 80% of repairs are inguinal, 10% incisional, 5% femoral, 4% umbilical, and < 1% represent other types of repairs. Surgical efforts are directed towards increasing efficacy, which, particularly in hernia surgery, converts into safety, lowering complication rates, shortening the length of the hospital stay, and lowering the recurrence rate. These goals are achieved by improvements in surgical procedures, the materials used, and general perioperative care (i.e., minimally invasive surgery with laparoscopy as its major part, tension-free procedures, adequate prophylaxis and the use of certain artificial materials in recreating abdominal wall integrity).
Our interest is directed on biomaterials. Currently, the most common is polypropylene mesh, and Lichtenstein’s inguinal hernia repair remains a gold standard technique. However, in our study, we decided to examine the use of liquid, photoinduced stiffening polymers, such as 1838-DMA, P1838-UR, PDEGA-UR and PEG-DA, in an experimental model of rabbits with artificially induced hernias.
AIM
1. To evaluate the potential application of injectable polymer biomaterials in the treatment of abdominal wall hernias using an animal model.
2. To compare tissue reactions elicited by the new polymer biomaterials and a commercial polypropylene mesh.
3. To determine the influence of the new injectable biomaterials on the kidneys and liver.
MATERIAL AND METHODS
The study group consisted of 20 New Zealand rabbits; there were 10 males and 10 females, with an average weight of 2.5 ± 0.6 kg. During stage 1 (tab. 1), we surgically created abdominal wall hernias in the group of rabbits. First, we established access to the jugular or femoral vein for blood sample collection. General anesthesia was induced by the intravenous administration of Domitor (0.2 ml IV) and ketamine (20 mg [0.2 ml] IV; Orion Pharma). After shaving the fur and disinfecting the skin, a ventral midline incision was made below the navel; the 5-cm incision went through the skin, subcutaneous tissue, fascia and muscle flap, uncovering the peritoneum. Afterward, the skin was closed by primary, continuous suture. The rabbits were observed for 30 days, allowing wound healing and hernias to occur.
Table 1. Study schedule.
 ProceduresTime
Stage IGeneral anaesthesia
Laboratory examination
Weight prior to surgery
Creation of hernia
30 days between first and second surgery
Everyday clinical observation
Motor activity
Wound healing
Nutrition
Stage IIGeneral anaesthesia
Laboratory examination
Weight prior to surgery
Hernia repair surgery
28 days between second operation and autopsy
Everyday clinical observation
Motor activity
Wound healing
Nutrition
Stage IIIGeneral anaesthesia
Laboratory examination
Weight prior to dissection
Autopsy
Histological examination
Clinical observation of the location of treated hernia from exterior and from the side of abdominal cavity
During stage 2 (tab. 1), the rabbits were randomized into five groups of four rabbits each for treatment with the following polymers: group 1 – P1838-UR/PEG-DA 85/15; group 2 – P1838-DMA; group 3 – P1838-UR, 4; group 4 – PDEGA-UR, and group 5 – control group, standard polypropylene mesh. Hernia repair surgery was performed under general anesthesia as described above. Blood samples for laboratory examination were collected from the jugular or femoral vein. The surgical field was shaved and disinfected, and the skin and subcutaneous tissue were incised over the hernial sac (fig. 1-3). The dissected sac was discharged into the abdominal cavity, and the appropriate polymer biomaterial was introduced into the hernial defect. The amount of biomaterial used per application varied from 1-2 cm3. Groups 1-4 received 5 minutes of irradiation with a UV lamp, which turned the liquid into an elastic, porous sponge (fig. 4 and 5). The subcutaneous and cutaneous tissues were closed using a continuous suture. In group 5, the mesh was not exposed to UV irradiation, and it was not fixed with sutures. After hernia repair, the rabbits were kept under clinical observation for 28 days.
Fig. 1. Hernia sack opened.
Fig. 2. Hernia sack during preparation.
Fig. 3. Hernia sack with contents.
Fig. 4. Hernia with biomaterial inserted.
Fig. 5. Groups 1-4 received 5 minutes of irradiation with a UV lamp, which turned the liquid into an elastic, porous sponge.
During stage 3 (tab. 1), the rabbits were sacrificed using 3 ml (480 mg) of Morbital while under anesthesia (medetomidine plus ketamine; Orion Pharma). Postmortem samples were collected from the tissues surrounding the hernia repair site and from the kidneys and the liver.
The biomaterials used in the current study were synthetized from fatty acids present in plant oils. Depending on the type and variety of the material, up to 80% of its mass was derived from renewable sources. The materials 1838-DMA and P1838-UR contain a fatty acid core. PDEGA-UR is a derivative of poly[di(ethylene glycol) adipate], and PEG-DA is diacrylated poly(ethylene glycol) (fig. 6). All of the materials had (meth)acrylic end-groups, making them susceptible to polymerization with UV light. The compositions used for hernia repair also contained the photoinitiator Irgacure 819 (Ciba). The polymers we-re synthesized under mild conditions. The process was carried out in an organic solvent under normal pressure and in the temperature range of 0 to 40°C (1). These biomaterials were created specifically for this project and have not yet been tested in other applications.
Fig. 6. Biochemical structure.
All rabbits were under the constant supervision of a veterinary surgeon. During the study, the following were recorded: vital signs, weight, food consumption, motor activity, the presence of swelling and redness around the wound, and the presence of fluid under the scar. Wound control included observation of both the external side and the peritoneal side of the treated hernia for assessment of hernia relapse. Blood was collected at every stage to measure aspartate transaminase (AST), gamma-glutamyl transferase (GGTP) activity and creatinine concentration.
Samples for histologic evaluation were preserved in freshly prepared 4% paraformaldehyde and then embedded in paraffin. For the purposes of morphological analysis, a slide series was made at 3-5 μm intervals, and the tissues were stained using hematoxylin and eosin. The cross-sections were dyed with the intent of calculating the numbers of various types of cells (fibrocytes, neutrophils, eosinophils, lymphocytes, macrophages). A 10 × eyepiece with a square visor was used for examination. For each animal’s tissue samples, the number of cells was calculated from 50 randomly selected fields; this evaluation was performed using 40 × augmentation and a Zeiss brand microscope. The surface area of each square was 1225 μm2 in size (35 × 35 mm) within the framework of the target. All morphometric measurements were obtained using AxioVision Relative software, version 4.6 (Zeiss, Axioscop, Germany).
Statistical analysis was performed with the Statistica 10 software package; the Shapiro-Wilk and Mann-Whitney tests were used for comparisons, with the level of significance set at p < 0.05. The study was conducted after obtaining the consent of the bioethics committee.
RESULTS
This study provided data drawn from: (1) clinical observation of the rabbits; (2) biochemistry tests; (3) histopathologic evaluation of the kidneys and liver; and (4) histopathologic evaluation of the tissues surrounding the hernia. There were no differences between the groups in motor activity, daily food intake or average weight gain. Would healing was undisturbed in groups 1, 2, 3 and 5, while wound inflammation was observed in group 4. Two deaths, caused by pneumonia and hernia incarceration, were observed during stage 1.
Biochemical tests
No statistically significant differences in AST, GTTP and creatinine concentration were found between the rabbits from groups 1, 2, 3 and 4 and the rabbits from the control group (group 5) during the three stages of the study.
Morphology of the examined tissues
Liver and kidney morphology
The liver and kidneys from the rabbits treated with polypropylene mesh (the control group) (fig. 7A, 7C) and from the rabbits treated with the new polymer biomaterials (groups 1, 2, 3, 4) presented typical morphology (fig. 7B, 7D). In the liver, the hepatocytes were grouped in interconnected plates to form liver lobules with portal spaces in the periphery and a central vein in the center. Plates of hepatocytes were directed from the periphery to the center of the lobule. Liver sinusoids were visible in the space between the plates. In all of the livers, small numbers of leucocytes were visible in the connective tissue surrounding the blood vessels and bile ducts in the portal spaces. No leucocytes were found near central veins.
Fig. 7. The morphology of liver (A) and kidney (C) in rabbits of control group (group 5), and liver (B) and kidney (D) of rabbits treated with new injectable material (group 1). H-E, objective magnification x 20.
The kidneys were composed of cortex and medulla. Nephron-related elements were present in the cortex: renal corpuscles, proximal convoluted tubules and distal convoluted tubules. Medullary rays penetrating the cortex were visible. Collecting tubules, loops of Henle and capillaries were observed in transverse sections of the medulla. There was no inflammatory cell infiltration in the interstitium of the renal cortex and medulla.
Morphology of connective tissue around the treated hernia

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Piśmiennictwo
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otrzymano: 2015-04-09
zaakceptowano do druku: 2015-04-23

Adres do korespondencji:
*Labib Zair
Clinic of General and Transplantation Surgery Pomeranian Medical University
72 Powstańców Wielkopolskich Av., 70-111 Szczecin, Poland
tel.: +48 (91) 466-11-29
e-mail: labib@poczta.onet.pl

New Medicine 2/2015
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