ABSTRACT
Purpose:
To describe the technique of sublay correction of incisional hernia in Wistar rats under videomagnification system.
Methods:
Five male rats of the species Rattus norvegicus, of the Wistar lineage, with body weight between 250–350 g and 60 days old were used. Incisional hernia was inducted in all animals. After that, the incisional hernia was immediately corrected by the sublay method.
Results:
There were no cases of recurrence of the incisional hernia after placement of the polypropylene mesh using the sublay technique. No postoperative complications were observed.
Conclusions:
The technique is suitable for execution in Wistar rats.
Key words: Abdominal Hernia, Incisional Hernia, Peritoneum, Animal Models, Wistar Rats
Introduction
Incisional hernia is one of the main morbidity factors after abdominal surgery1. The incidence is approximately 20% in patients, reaching 40% in high-risk patients2. Thus, surgical approaches used for treatment are advancing, and among them, the use of prophylactic meshes can be highlighted, as they are routinely used and prove to be advantageous in terms of results3 – 5.
To allow the best results, several techniques have been described in humans for the placement of mesh, such as onlay, sublay, preperitoneal plane and intraperitoneal plane, besides the combination of different sites6 , 7. In addition, the placement of this prophylactic mesh appears to be effective regardless of location, being a safe technique that prevents an increased risk of seroma and chronic pain4.
In laparoscopic surgeries, intraperitoneal onlay mesh repair seems to be the only option8, but it also appears to be an alternative to open surgeries, as the placement of the mesh intraperitoneally can avoid extensive tissue dissection and reduce the chance of infection at the surgical site and prevent the occurrence of infection through the mesh9 – 11. Furthermore, this reduction in infection is important because it leads to significant morbidity and mortality, hernia recurrences, prolonged hospital stay and increased hospital costs12.
Concerning open surgeries, the sublay technique is the most used7, being safe and efficient in patients with more relevant conditions where the surgery should not be contraindicated, despite the risk of progression of the hernia defect and persistent symptoms13. In the long term, its repair is superior to the onlay mesh repair, which is technically easier, nonetheless, provides more postoperative complications, not presenting divergence in terms of recurrence rate14.
However, there are still disagreements about the most effective method to approach the ideal positioning of the mesh between the layers of the abdominal wall7, since each method has its advantages, disadvantages, and variable results according to the literature. Thus, there is a need for comparative studies between the various techniques.
In addition, bringing this context of incisional hernia to the experimental models, it is believed that rats (Rattus norvegicus) constitute a suitable animal model for hernia research15 with an appropriate anatomical structure for the elaboration of surgical procedures16, in addition to presenting low cost and ease of reproduction, it also has great genetic similarity to humans17. However, although already described in humans, no articles were found about the use of the sublay technique in Wistar rats, possibly due to the small size of these animals and the difficulty of performing the technique.
Therefore, the objective of this work is to describe the technique of sublay correction of incisional hernia in Wistar rats under videomagnification system, since there is no standardization of this technique in an experimental model.
Methods
This study followed the principles established by the Brazilian laws of use and creation of animals (No. 11,794/2008), which regulates research with animals in Brazil, being approved by the Ethics Committee in the Use of Animals of the UEPA under the opinion No. 06/2021.
This is an experimental, prospective and descriptive study developed by the Experimental Surgery Laboratory, UEPA. Five male rats of the R. norvegicus species, of the Wistar strain, with body weight between 250–350 g and age of 60 days were used. The animals were placed in a vivarium with adequate conditions of temperature (21–24 °C), luminosity (interleaving between light and dark every 12 h), humidity (70–80%) and noise. Furthermore, they were kept in individual cages with sterile wood shavings and received water and food ad libitum.
Incisional hernia was induced in all animals, based on the model described by Paulo et al.18. After that, the incisional hernia was immediately corrected by the sublay method, adapted from the surgical procedure performed in humans described by Miranda et al.19.
Videomagnification system
The magnification system20 , 21 used in this study consisted of a Sony Handycam HDR-XR160 camera connected to a 55’ Full HD Curve TV through an HDMI cable, allowing a magnification of 50× the original size. Two fluorescent light sources were used next to the board to provide adequate illumination of the operative field.
Anesthetic procedures
The surgical team consisted of a senior surgeon and an assistant surgeon.
The animals were submitted to general anesthesia, by association of 75–100 mg/kg of ketamine hydrochloride and 5–10 mg/kg of xylazine hydrochloride, intraperitoneally. The anesthetic level was confirmed by testing the caudal reflex, foot reflex and vibrissa movement.
Infraumbilical and supraumbilical manual epilation of the anterior abdominal wall was performed in the form of a 6 × 4 cm rectangle. Antisepsis was performed with 0.5% alcoholic chlorhexidine. The animal was fixed on a surgical board in the supine position.
Operating procedure
Induction of incisional hernia
The first step of the process consisted of making a median incision of 4 cm in the abdomen below the xiphoid process, using a #15 scalpel blade. Divulsion of the subcutaneous cell plane were performed for approximately 1.5 cm on each side of the linea alba, with the aid of the noncutting face of Metzenbaum scissors. To protect the intraperitoneal viscera, the peritoneum was clamped by two hemostatic forceps, which were pulled, generating a fold, on which a small incision was made, facilitating the removal of the viscera in relation to the serosa. A closed instrument was introduced and it was rotated 360°, undoing possible adhesions of intestinal loops. Then, a longitudinal incision of the linea alba and the peritoneum was made for an extension of 3 cm (Fig. 1).
Figure 1. Median incision of 4 cm in the skin and longitudinalincision of the linea alba and peritoneum for an extensionof 3 cm. The structures observed from the most superficialto the most profound are: (a) Skin; (b) Subcutaneouscell tissue; (c) Aponeurosis; (d) Abdominal cavity.
Correction with the sublay method
The peritoneal incision was followed by the opening of the medial posterior lamina of the rectus abdominis muscle sheath bilaterally, and a wide dissection was performed between it and the muscle with the aid of the videomagnification system and microsurgical forceps and scissors. This was followed by manual placement of the 3.5 × 2.5 cm polypropylene mesh directly under the rectus abdominis muscle (Fig. 2). The sheaths were brought together through continuous suture with Prolene 5–0 thread (Fig. 3). Skin synthesis was performed with simple stitches at a distance of 1 cm using Monocryl 4-0 thread (Fig. 4).
Figure 2. Manual placement of the 3.5 × 2.5 cm polypropylenemesh directly under the rectus abdominis muscle.

Figure 3. Approximation of the rectus abdominismuscle sheaths by means of continuoussuture with Prolene 5-0 thread.

Figure 4. Synthesis of the skin with simpleovercast using the Monocryl thread.

Postoperative procedures
After the surgery, the vital signs of the animals were monitored. In addition, sodium dipyrone for veterinary use was administered subcutaneously in doses of 160 mg/kg if the animal showed signs of pain, such as piloerection, hunched posture, aggressive behavior and changes in food and water consumption. In the postoperative period, the animals were housed in individual cages.
Assessment and euthanasia
The presence of postoperative complications and the occurrence of herniation after 28 days were evaluated, verified by inspection and local palpation.
After 28 days of surgery, the animals were euthanized by intraperitoneal anesthetic overdose, in which triple the dose of the anesthetic protocol was used.
Results
During the procedure, none of the animals died or required anesthetic boost. The sublay technique proved to be viable for performance in Wistar rats. The mean operative time was 53.4 ± 3.68 min.
There were no cases of recurrence of incisional hernia after placement of the polypropylene mesh by the sublay technique. No postoperative complications were observed, such as hematomas, tissue necrosis, stitch dehiscence or systemic signs and/or sites of inflammation of the surgical wound.
Discussion
The use of meshes for correction of incisional hernias has provided the use of several surgical approaches for their ideal positioning, in order to avoid increased intraperitoneal pressure and maintain respiratory mechanics at adequate levels. However, there are scientific barriers regarding the effectiveness of each one of them6. In this sense, experimental studies become important tools to aid in the development of more effective techniques23. However, despite using rats in these studies, there is still no standardization regarding the surgical technique to be performed in these animals to correct this pathology.
Thus, the present study aimed to develop a new technique of retromuscular sublay in rats for the correction of incisional hernia. By the sublay technique, the mesh can be preperitoneal, intraperitoneal or retromuscular24 , 25. Despite being preferred among surgeons for hernia repair because it provides a low recurrence rate, this technique remains controversial, due to the fragility of the abdominal wall to sustain internal pressure14 , 22. Thus, the technique developed allows the restoration of the posterior sheath unit of the rat’s rectus abdominis muscle with suture and, after tissue divulsion, deposits the mesh between the fibers of the same and the sheath itself, resulting in greater wall resistance.
According to the data collected, there were no complications such as tissue necrosis, stitch dehiscence and recurrence during the analyzed period of the study. Such complications are frequent after correction of the pathology with the use of meshes, given that it is a foreign body and tissue rejection may occur9 , 10 , 26. Even so, the use of this technique in experimental models allows a more in-depth study of the mechanisms that involve failures in the surgical treatment of incisional hernias in humans, due to their genetic and tissue biocompatibility with R. norvegicus, a Wistar strain used in most simulators, being able to improve it, minimizing the risks15 , 17.
However, despite having shown promising results, the technique described has a high degree of difficulty to be performed, since the animal has dimensions much smaller than the human15 , 17 , 23 , 26 , 27. This fact imposes the need for detailed skills on the part of the surgeon to dissect the fibers without injuring the posterior sheath, otherwise the error in the applied method may result in interpretations that are not consistent with reality. In order to reduce the dimension barrier, the videomagnification system described by Barros et al. 21 was used in the study, which has the main advantages of low cost compared to conventional laboratory microscopes and the possibility of magnification of 50× the original size.
In addition, it is worth noting that the study had some limitations. Incisional hernias may recur even after a certain postoperative period14. Although the results presented here demonstrate that there was no recurrence by the technique used, this statement is valid only for the analyzed period, and it is not possible to extrapolate this information to longer periods. Added to this, it was not possible to compare the new technique with meshes made of other types of materials, requiring further studies to better evaluate its effectiveness. Even so, the technique described proved to be viable for use in experimental studies.
Conclusion
The technique is suitable for performance in Wistar rats, proving to be safe, since there were no cases of recurrence of incisional hernia after the placement of the polypropylene mesh by the sublay technique. No postoperative complications were observed. However, it is difficult to perform due to the reduced dimensions of the animals.
Acknowledgments
Not applicable.
Footnotes
Research performed at Experimental Surgery Laboratory, Universidade do Estado do Pará, Belém (PA), Brazil.
Data availability statement: Data will be available upon request.
Funding: Coordenação de Aperfeiçoamento de Pessoal de Nível Superior
[https://doi.org/10.13039/501100002322]
Grant No. 015/2020
References
- 1.Patel SV, Paskar DD, Nelson RL, Vedula SS, Steele SR. Closure methods for laparotomy incisions for preventing incisional hernias and other wound complications. Cochrane Database Syst Rev. 2017;11(11) doi: 10.1002/14651858.cd005661.pub2. [DOI] [PMC free article] [PubMed] [Google Scholar]
- 2.Salamone G, Licari L, Guercio G, Campanella S, Falco N, Scerrino G, Bonventre S, Geraci G, Cocorullo G, Gulotta G. The abdominal wall hernia in cirrhotic patients: a historical challenge. World J Emerg Surg. 2018;13:35–35. doi: 10.1186/s13017-018-0196-z. [DOI] [PMC free article] [PubMed] [Google Scholar]
- 3.Jairam AP, Timmermans L, Eker HH, Pierik REGJM, van Klaveren, Steyerberg EW, Timman R, van der, Dawson I, Charbon JA, Schuhmacher C, Mihaljevic A, Izbicki JR, Fikatas P, Knebel P, Fortelny RH, Kleinrensink GJ, Lange JF, Jeekel HJ. PRIMA Trialist Group. Prevention of incisional hernia with prophylactic onlay and sublay mesh reinforcement versus primary suture only in midline laparotomies (PRIMA): 2-year follow-up of a multicentre, double-blind, randomised controlled trial. Lancet. 2017;390(10094):567–576. doi: 10.1016/S0140-6736(17)31332-6. [DOI] [PubMed] [Google Scholar]
- 4.Borab ZM, Shakir S, Lanni MA, Tecce MG, MacDonald J, Hope WW, Fischer JP. Does prophylactic mesh placement in elective, midline laparotomy reduce the incidence of incisional hernia? A systematic review and meta-analysis. Surgery. 2017;161(4):1149–1163. doi: 10.1016/j.surg.2016.09.036. [DOI] [PubMed] [Google Scholar]
- 5.Kohler A, Lavanchy JL, Lenoir U, Kurmann A, Candinas D, Beldi G. Effectiveness of prophylactic intraperitoneal mesh implantation for prevention of incisional hernia in patients undergoing open abdominal surgery: a randomized clinical trial. JAMA Surg. 2019;154(2):109–115. doi: 10.1001/jamasurg.2018.4221. [DOI] [PMC free article] [PubMed] [Google Scholar]
- 6.Sugrue M, Johnston A, Zeeshan S, Loughlin P, Bucholc M, Watson A. The role of prophylactic mesh placement to prevent incisional hernia in laparotomy. Is it time to change practice? Anaesthesiol Intensive Ther. 2019;51(4):323–329. doi: 10.5114/ait.2019.87475. [DOI] [PubMed] [Google Scholar]
- 7.Holihan JL, Nguyen DH, Nguyen MT, Mo J, Kao LS, Liang MK. Mesh location in open ventral hernia repair: a systematic review and network meta-analysis. World J Surg. 2016;40(1):89–99. doi: 10.1007/s00268-015-3252-9. [DOI] [PubMed] [Google Scholar]
- 8.Yang GPC. From intraperitoneal onlay mesh repair to preperitoneal onlay mesh repair. Asian J Endosc Surg. 2017;10(2):119–127. doi: 10.1111/ases.12388. [DOI] [PubMed] [Google Scholar]
- 9.Barbaros U, Asoglu O, Seven R, Erbil Y, Dinccag A, Deveci U, Ozarmagan S, Mercan S. The comparison of laparoscopic and open ventral hernia repairs: a prospective randomized study. Hernia. 2007;11(1):51–56. doi: 10.1007/s10029-006-0160-9. [DOI] [PubMed] [Google Scholar]
- 10.Forbes SS, Eskicioglu C, McLeod RS, Okrainec A. Meta-analysis of randomized controlled trials comparing open and laparoscopic ventral and incisional hernia repair with mesh. Br J Surg. 2009;96(8):851–858. doi: 10.1002/bjs.6668. [DOI] [PubMed] [Google Scholar]
- 11.Kaoutzanis C, Leichtle SW, Mouawad NJ, Welch KB, Lampman RM, Cleary RK. Postoperative surgical site infections after ventral/incisional hernia repair: a comparison of open and laparoscopic outcomes. Surg Endosc. 2013;27(6):2221–2230. doi: 10.1007/s00464-012-2743-0. [DOI] [PubMed] [Google Scholar]
- 12.Tubre DJ, Schroeder AD, Estes J, Eisenga J, Fitzgibbons RJ., Jr Surgical site infection: the “Achilles heel” of all types of abdominal wall hernia reconstruction. Hernia. 2018;22(6):1003–1013. doi: 10.1007/s10029-018-1826-9. [DOI] [PubMed] [Google Scholar]
- 13.Stodolski M, Papadakis M, Zirngibl H, Ambe PC. Risk of recurrence following mesh associated incisional hernia repair using the retromuscular technique in patients with relevant medical conditions. Asian J Surg. 2018;41(6):562–568. doi: 10.1016/j.asjsur.2018.01.005. [DOI] [PubMed] [Google Scholar]
- 14.Sevinç B, Okuş A, Ay S, Aksoy N, Karahan Ö. Randomized prospective comparison of long-term results of onlay and sublay mesh repair techniques for incisional hernia. Turk J Surg. 2018;34(1):17–20. doi: 10.5152/turkjsurg.2017.3712. [DOI] [PMC free article] [PubMed] [Google Scholar]
- 15.Van den, Vogels RRM, van Barneveld, Gijbels MJJ, Peutz-Kootstra CJ, Cleutjens JPM, Schreinemacher MHF, Bouvy ND. Comparability of histological outcomes in rats and humans in a hernia model. J Surg Res. 2018;229:271–276. doi: 10.1016/j.jss.2018.03.019. [DOI] [PubMed] [Google Scholar]
- 16.Greca FH, Filho ZAS, Rocha SL, Borsato KS, Fernandes HAD, Niiside MA. Small intestinal submucosa to repais anterior abdominal wall defect. In rats. Acta Cir Bras. 2004;19(5):471–477. doi: 10.1590/S0102-86502004000500004. [DOI] [Google Scholar]
- 17.Gibbs RA, Weinstock GM, Metzker ML, Muzny DM, Sodergren EJ, Scherer S, Scott G, Steffen D, Worley KC, Burch PE, Okwuonu G, Hines S, Lewis L, DeRamo C, Delgado O, Dugan-Rocha S, Miner G, Morgan M, Hawes A, Gill R, Holt RA, Adams MD, Amanatides PG, Baden-Tillson H, Barnstead M, Chin S, Evans CA, Ferriera S, Fosler C, Glodek A, Gu Z, Jennings D, Kraft CL, Nguyen T, Pfannkoch CM, Sitter C, Sutton GG, Venter JC, Woodage T, Smith D, Lee HM, Gustafson E, Cahill P, Kana A, Doucette-Stamm L, Weinstock K, Fechtel K, Weiss RB, Dunn DM, Green ED, Blakesley RW, Bouffard GG, De Jong, Osoegawa K, Zhu B, Marra M, Schein J, Bosdet I, Fjell C, Jones S, Krzywinski M, Mathewson C, Siddiqui A, Wye N, McPherson J, Zhao S, Fraser CM, Shetty J, Shatsman S, Geer K, Chen Y, Abramzon S, Nierman WC, Havlak PH, Chen R, Durbin KJ, Egan A, Ren Y, Song XZ, Li B, Liu Y, Qin X, Cawley S, Worley KC, Cooney AJ, D’Souza LM, Martin K, Wu JQ, Gonzalez-Garay ML, Jackson AR, Kalafus KJ, McLeod MP, Milosavljevic A, Virk D, Volkov A, Wheeler DA, Zhang Z, Bailey JA, Eichler EE, Tuzun E, Birney E, Mongin E, Ureta-Vidal A, Woodwark C, Zdobnov E, Bork P, Suyama M, Torrents D, Alexandersson M, Trask BJ, Young JM, Huang H, Wang H, Xing H, Daniels S, Gietzen D, Schmidt J, Stevens K, Vitt U, Wingrove J, Camara F, Mar Albà, Abril JF, Guigo R, Smit A, Dubchak I, Rubin EM, Couronne O, Poliakov A, Hübner N, Ganten D, Goesele C, Hummel O, Kreitler T, Lee YA, Monti J, Schulz H, Zimdahl H, Himmelbauer H, Lehrach H, Jacob HJ, Bromberg S, Gullings-Handley J, Jensen-Seaman MI, Kwitek AE, Lazar J, Pasko D, Tonellato PJ, Twigger S, Ponting CP, Duarte JM, Rice S, Goodstadt L, Beatson SA, Emes RD, Winter EE, Webber C, Brandt P, Nyakatura G, Adetobi M, Chiaromonte F, Elnitski L, Eswara P, Hardison RC, Hou M, Kolbe D, Makova K, Miller W, Nekrutenko A, Riemer C, Schwartz S, Taylor J, Yang S, Zhang Y, Lindpaintner K, Andrews TD, Caccamo M, Clamp M, Clarke L, Curwen V, Durbin R, Eyras E, Searle SM, Cooper GM, Batzoglou S, Brudno M, Sidow A, Stone EA, Venter JC, Payseur BA, Bourque G, López-Otín C, Puente XS, Chakrabarti K, Chatterji S, Dewey C, Pachter L, Bray N, Yap VB, Caspi A, Tesler G, Pevzner PA, Haussler D, Roskin KM, Baertsch R, Clawson H, Furey TS, Hinrichs AS, Karolchik D, Kent WJ, Rosenbloom KR, Trumbower H, Weirauch M, Cooper DN, Stenson PD, Ma B, Brent M, Arumugam M, Shteynberg D, Copley RR, Taylor MS, Riethman H, Mudunuri U, Peterson J, Guyer M, Felsenfeld A, Old S, Mockrin S, Collins F. Rat Genome Sequencing Project Consortium. Genome sequence of the Brown Norway rat yields insights into mammalian evolution. Nature. 2004;428(6982):493–521. doi: 10.1038/nature02426. [DOI] [PubMed] [Google Scholar]
- 18.Paulo DNS, Pereira FEL, Mata RF, Dauad FR, Paulo ICAL. Experimental models of longitudinal abdominal incisional hernia in rats. Acta Cir Bras. 1997;12(4) doi: 10.1590/S0102-86501997000400004. [DOI] [Google Scholar]
- 19.Miranda JS, Birolini CAV, Utiyama EM. In: Atualidades em clínica cirúrgica intergastro e trauma. 4 ed. Aquino JLB, Andreollo NA, Martinez CAR, editors. São Paulo: Atheneu; 2013. Hérnia incisional: Novas opções para velhos problemas. Abordagens abertas: Onlay, inlay, sublay e retromuscular. [Google Scholar]
- 20.Teixeira RKC, Feijó DH, Valente AL, de Carvalho, Brito MVH, de Barros. Can smartphones be used to perform video-assisted microanastomosis? An experimental study. Surg Innov. 2019;26(3):371–375. doi: 10.1177/1553350618822626. [DOI] [PubMed] [Google Scholar]
- 21.Barros RSM, Brito MV, Leal RA, Teixeira RK, Sabbá MF, Yamaki VN, Lemos MV. A Low-cost high-definition video system for microsurgical hindlimb replantation in rats. J Reconstr Microsurg. 2017;33(3):158–162. doi: 10.1055/s-0036-1593767. [DOI] [PubMed] [Google Scholar]
- 22.Speranzini MB, Deutsch CR. Grandes hérnias incisionais. Arq Bras Cir Dig. 2010;23(4):280–286. doi: 10.1590/S0102-67202010000400015. [DOI] [Google Scholar]
- 23.Ferreira LM, Hochman B, Barbosa MV. Modelos experimentais em pesquisa. Acta Cir Bras. 2005;20(Suppl 2):28–34. doi: 10.1590/s0102-86502005000800008. [DOI] [PubMed] [Google Scholar]
- 24.Holihan JL, Nguyen DH, Nguyen MT, Mo J, Kao LS, Liang MK. Mesh location in open ventral hernia repair: a systematic review and network meta-analysis. World J Surg. 2016;40(1):89–99. doi: 10.1007/s00268-015-3252-9. [DOI] [PubMed] [Google Scholar]
- 25.Rhemtulla IA, Fischer JP. Retromuscular sublay technique for ventral hernia repair. Semin Plast Surg. 2018;32(3):120–126. doi: 10.1055/s-0038-1666800. [DOI] [PMC free article] [PubMed] [Google Scholar]
- 26.Kaoutzanis C, Leichtle SW, Mouawad NJ, Welch KB, Lampman RM, Cleary RK. Postoperative surgical site infections after ventral/incisional hernia repair: a comparison of open and laparoscopic outcomes. Surg Endosc. 2013;27(6):2221–2230. doi: 10.1007/s00464-012-2743-0. [DOI] [PubMed] [Google Scholar]
- 27.Thomson V, Wiewel A, Chinen A, Maryanto I, Sinaga MH, How R, Aplin K, Suzuki H. A perspective for resolving the systematics of Rattus, the vertebrates with the most influence on human welfare. Zootaxa. 2018;4459(3):431–452. doi: 10.11646/zootaxa.4459.3.2. [DOI] [PubMed] [Google Scholar]

