Experimental study on repairing rat abdominal wall defect with chitosan hydrogel/polypropylene mesh composite_Chinese Journal of Reparative and Reconstructive Surgery

Authors：

ZHANG Minghao ¹ , HE Wei ² ^Δ , YU Suxiang ³ ,  DI Yuntao ⁴ ,  LI Xiaoming ²

1. The Second Department of General Surgery, the Fourth Central Hospital of Baoding City, Baoding Hebei, 072350, P. R. China;
2. Key Laboratory for Biomechanics and Mechanobiology of Ministry of Education, Beijing Advanced Innovation Center for Biomedical Engineering, School of Biological Science and Medical Engineering, Beihang University, Beijing, 100083, P. R. China;
3. Department of Pathology, the Fourth Central Hospital of Baoding City, Baoding Hebei, 072350, P. R. China;
4. The First Department of Neurosurgery, the Fourth Central Hospital of Baoding City, Baoding Hebei, 072350, P. R. China;

Corresponding author：

DI Yuntao, Email: 15176391949@126.com; LI Xiaoming, Email: x.m.li@hotmail.com

Keywords：

Abdominal wall repair; chitosan; polypropylene; hydrogel; surgical mesh; rat

DOI：

10.7507/1002-1892.202404042

Video：

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Objective To investigate the improvement effects and mechanisms of composite chitosan (CS) hydrogel on traditional polypropylene (PP) mesh for repairing abdominal wall defects. Methods CS hydrogel was prepared via physical cross-linking and then combined with PP mesh to create a CS hydrogel/PP mesh composite. The internal structure and hydrophilicity of the composite were characterized using macroscopic observation, upright metallographic microscope, scanning electron microscopy, and water contact angle measurements. The performance of the composite (experimental group) in resisting cell adhesion and supporting cell infiltration was assessed through fibroblast (NIH-3T3) infiltration experiments and human umbilical vein endothelial cells (HUVECs) tube formation assays, and simple cells were used as control group. Finally, a bilateral abdominal wall defect model (1.5 cm×1.0 cm) was established in 18 Sprague Dawley rats aged 8-10 weeks, with the composite used on one side (experimental group) and PP mesh on the other side (control group). The effects on promoting wound healing, preventing adhesion, angiogenesis, and anti-inflammation were investigated through macroscopic observation, histological staining (HE and Masson staining), and immunohistochemical staining (CD31, CD68). Results The composite appeared as a pale yellow, transparent solid with a thickness of 2-3 mm, with the PP mesh securely encapsulated within the hydrogel. Scanning electron microscopy revealed that the hydrogel contained interconnected pores measuring 100-300 μm, forming a porous structure. Contact angle measurements indicated that CS hydrogel exhibited good hydrophilicity, while PP mesh was highly hydrophobic. In vitro cell culture experiments showed that DAPI staining indicated fewer positive cells in the experimental group after 1 day of culture, while the cells in control group covered the entire well plate. After 3 days of culture, the cells in experimental group were spherical and displayed uneven fluorescence, suggesting that the material could reduce cell adhesion while supporting cell infiltration. HUVECs tube formation experiments demonstrated an increase in cell numbers in experimental group with a trend towards tube formation, while cells in control group were sparsely distributed and showed no migration. In the rat abdominal wall defect repair experiment, results showed that after 1 week post-surgery, the experimental group had tissue and blood vessels infiltrating, and by 4 weeks, the integrity was well restored with significant regeneration of muscle and blood vessels, while the control group exhibited adhesions and incomplete healing. HE staining results indicated weaker cell infiltration in the experimental group, with cell density significantly higher than that of the control group at 2 and 4 weeks post-surgery (P<0.05). Masson staining revealed that collagen fibers in the experimental group were arranged neatly, with significantly increased collagen content at 2 weeks post-surgery (P<0.05), while collagen content was similar in both groups at 4 weeks (P>0.05). Immunohistochemical staining showed that CD31-positive cells were evenly distributed between muscle layers in the experimental group, whereas the control group exhibited notable defects. At 2 weeks after operation, the CD31-positive cell ratio was significantly higher than that in the control group (P<0.05); at 2 and 4 weeks after operation, the CD68-positive cell ratio in the experimental group was significantly lower than that in the control group (P<0.05). Conclusion CS hydrogel has a positive effect on preventing adhesions and promoting wound healing, exhibiting anti-inflammatory and pro-angiogenic properties during the healing process. This provides a promising strategy to address challenges related to abdominal adhesions and reconstruction.

Citation： ZHANG Minghao, HE Wei, YU Suxiang, DI Yuntao, LI Xiaoming. Experimental study on repairing rat abdominal wall defect with chitosan hydrogel/polypropylene mesh composite. Chinese Journal of Reparative and Reconstructive Surgery, 2024, 38(10): 1261-1268. doi: 10.7507/1002-1892.202404042 Copy

1.	Bodin F, Dissaux C, Romain B, et al. Complex abdominal wall defect reconstruction using a latissimus dorsi free flap with mesh after malignant tumor resection. Microsurgery, 2017, 37(1): 38-43.
2.	Gu Y, Wang P, Li H, et al. Chinese expert consensus on adult ventral abdominal wall defect repair and reconstruction. Am J Surg, 2021, 222(1): 86-98.
3.	Ravo B, Falasco G. Pure tissue inguinal hernia repair with the use of biological mesh: a 10-year follows up. A prospective study. Hernia, 2020, 24(1): 121-126.
4.	Saiding Q, Chen Y, Wang J, et al. Abdominal wall hernia repair: from prosthetic meshes to smart materials. Mater Today Bio, 2023, 21: 100691. doi: 10.1016/j.mtbio.2023.100691.
5.	Liao J, Li X, Fan Y. Prevention strategies of postoperative adhesion in soft tissues by applying biomaterials: Based on the mechanisms of occurrence and development of adhesions. Bioact Mater, 2023, 26: 387-412.
6.	Guillaume O, Pérez-Tanoira R, Fortelny R, et al. Infections associated with mesh repairs of abdominal wall hernias: Are antimicrobial biomaterials the longed-for solution? Biomaterials, 2018, 167: 15-31.
7.	Kokotovic D, Bisgaard T, Helgstrand F. Long-term recurrence and complications associated with elective incisional hernia repair. JAMA, 2016, 316(15): 1575-1582.
8.	Wang Z, You W, Wang W, et al. Dihydromyricetin-incorporated multilayer nanofibers accelerate chronic wound healing by remodeling the harsh wound microenvironment. Adv Fiber Mater, 2022, 4(6): 1556-1571.
9.	Yang Y, Du Y, Zhang J, et al. Structural and functional design of electrospun nanofibers for hemostasis and wound healing. Adv Fiber Mater, 2022, 4(5): 1027-1057.
10.	Meng Z, Wang H, Liu Y, et al. Evaluation of the effectiveness of alginate-based hydrogels in preventing peritoneal adhesions. Regen Biomater, 2023, 10: rbad017. doi: 10.1093/rb/rbad017.
11.	Liu Z, Wei N, Tang R. Functionalized strategies and mechanisms of the emerging mesh for abdominal wall repair and regeneration. ACS Biomater Sci Eng, 2021, 7(6): 2064-2082.
12.	Dreger NZ, Fan Z, Zander ZK, et al. Amino acid-based poly (ester urea) copolymer films for hernia-repair applications. Biomaterials, 2018, 182: 44-57.
13.	Nishiguchi A, Ito S, Nagasaka K, et al. Tissue-adhesive decellularized extracellular matrix patches reinforced by a supramolecular gelator to repair abdominal wall defects. Biomacromolecules, 2023, 24(4): 1545-1554.
14.	Yuan J, Wang Y, Yang W, et al. Biomimetic peptide dynamic hydrogel inspired by humanized defensin nanonets as the wound-healing gel coating. Chem Eng J, 2023, 470: 144266. doi: 10.1016/j.cej.2023.144266.
15.	闫志文, 李硕峰, 李傲, 等. 改性木葡聚糖温敏水凝胶的制备及生物相容性评价. 中国组织工程研究, 2019, 23(30): 4841-4847.
16.	Pita-López ML, Fletes-Vargas G, Espinosa-Andrews H, et al. Physically cross-linked chitosan-based hydrogels for tissue engineering applications: A state-of-the-art review. Eur Polym J, 2021, 145: 110176. doi: 10.1016/j.eurpolymj.2020.110176.
17.	Wang W, Xue C, Mao X. Chitosan: Structural modification, biological activity and application. Int J Biol Macromol, 2020, 164: 4532-4546.
18.	Akbaba S, Atila D, Keskin D, et al. Multilayer fibroin/chitosan oligosaccharide lactate and pullulan immunomodulatory patch for treatment of hernia and prevention of intraperitoneal adhesion. Carbohydr Polym, 2021, 265: 118066. doi: 10.1016/j.carbpol.2021.118066.
19.	Li L, Wang N, Jin X, et al. Biodegradable and injectable in situ cross-linking chitosan-hyaluronic acid based hydrogels for postoperative adhesion prevention. Biomaterials, 2014, 35(12): 3903-3917.
20.	Hu W, Zhang Z, Zhu L, et al. Combination of polypropylene mesh and in situ injectable mussel-inspired hydrogel in laparoscopic hernia repair for preventing post-surgical adhesions in the piglet model. ACS Biomater Sci Eng, 2020, 6(3): 1735-1743.
21.	Miserez M, Jairam AP, Boersema GSA, et al. Resorbable synthetic meshes for abdominal wall defects in preclinical setting: A literature review. J Surg Res, 2019, 237: 67-75.
22.	Hu W, Lu S, Zhang Z, et al. Mussel-inspired copolymer-coated polypropylene mesh with anti-adhesion efficiency for abdominal wall defect repair. Biomater Sci, 2019, 7(4): 1323-1334.
23.	Zhang H, Xu D, Zhang Y, et al. Silk fibroin hydrogels for biomedical applications. Smart Medicine, 2022, 1(1): e20220011. doi: 10.1002/SMMD.20220011.
24.	Yuan N, Shao K, Huang S, et al. Chitosan, alginate, hyaluronic acid and other novel multifunctional hydrogel dressings for wound healing: A review. Int J Biol Macromol, 2023, 240: 124321. doi: 10.1016/j.ijbiomac.2023.124321.
25.	Sousa AB, Águas AP, Barbosa MA, et al. Immunomodulatory biomaterial-based wound dressings advance the healing of chronic wounds via regulating macrophage behavior. Regen Biomater, 2022, 9: rbac065. doi: 10.1093/rb/rbac065.
26.	Schaffrick L, Ding J, Kwan P, et al. The dynamic changes of monocytes and cytokines during wound healing post-burn injury. Cytokine, 2023, 168: 156231. doi: 10.1016/j.cyto.2023.156231.
27.	Min D, Nube V, Tao A, et al. Monocyte phenotype as a predictive marker for wound healing in diabetes-related foot ulcers. J Diabetes Complications, 2021, 35(5): 107889. doi: 10.1016/j.jdiacomp.2021.107889.
28.	Wang S, Lu M, Cao Y, et al. Degradative polylactide nanofibers promote M2 macrophage polarization via STAT6 pathway in peritendinous adhesion. Compos Part B Eng, 2023, 253: 110520. doi: 10.1016/j.compositesb.2023.110520.
29.	Kroemer G, Zitvogel L. Ghrelin and leptin regulating wound healing. Trends Immunol, 2022, 43(10): 777-779.
30.	Zheng H, Cheng X, Jin L, et al. Recent advances in strategies to target the behavior of macrophages in wound healing. Biomed Pharmacother, 2023, 165: 115199. doi: 10.1016/j.biopha.2023.115199.

1. Bodin F, Dissaux C, Romain B, et al. Complex abdominal wall defect reconstruction using a latissimus dorsi free flap with mesh after malignant tumor resection. Microsurgery, 2017, 37(1): 38-43.
2. Gu Y, Wang P, Li H, et al. Chinese expert consensus on adult ventral abdominal wall defect repair and reconstruction. Am J Surg, 2021, 222(1): 86-98.
3. Ravo B, Falasco G. Pure tissue inguinal hernia repair with the use of biological mesh: a 10-year follows up. A prospective study. Hernia, 2020, 24(1): 121-126.
4. Saiding Q, Chen Y, Wang J, et al. Abdominal wall hernia repair: from prosthetic meshes to smart materials. Mater Today Bio, 2023, 21: 100691. doi: 10.1016/j.mtbio.2023.100691.
5. Liao J, Li X, Fan Y. Prevention strategies of postoperative adhesion in soft tissues by applying biomaterials: Based on the mechanisms of occurrence and development of adhesions. Bioact Mater, 2023, 26: 387-412.
6. Guillaume O, Pérez-Tanoira R, Fortelny R, et al. Infections associated with mesh repairs of abdominal wall hernias: Are antimicrobial biomaterials the longed-for solution? Biomaterials, 2018, 167: 15-31.
7. Kokotovic D, Bisgaard T, Helgstrand F. Long-term recurrence and complications associated with elective incisional hernia repair. JAMA, 2016, 316(15): 1575-1582.
8. Wang Z, You W, Wang W, et al. Dihydromyricetin-incorporated multilayer nanofibers accelerate chronic wound healing by remodeling the harsh wound microenvironment. Adv Fiber Mater, 2022, 4(6): 1556-1571.
9. Yang Y, Du Y, Zhang J, et al. Structural and functional design of electrospun nanofibers for hemostasis and wound healing. Adv Fiber Mater, 2022, 4(5): 1027-1057.
10. Meng Z, Wang H, Liu Y, et al. Evaluation of the effectiveness of alginate-based hydrogels in preventing peritoneal adhesions. Regen Biomater, 2023, 10: rbad017. doi: 10.1093/rb/rbad017.
11. Liu Z, Wei N, Tang R. Functionalized strategies and mechanisms of the emerging mesh for abdominal wall repair and regeneration. ACS Biomater Sci Eng, 2021, 7(6): 2064-2082.
12. Dreger NZ, Fan Z, Zander ZK, et al. Amino acid-based poly (ester urea) copolymer films for hernia-repair applications. Biomaterials, 2018, 182: 44-57.
13. Nishiguchi A, Ito S, Nagasaka K, et al. Tissue-adhesive decellularized extracellular matrix patches reinforced by a supramolecular gelator to repair abdominal wall defects. Biomacromolecules, 2023, 24(4): 1545-1554.
14. Yuan J, Wang Y, Yang W, et al. Biomimetic peptide dynamic hydrogel inspired by humanized defensin nanonets as the wound-healing gel coating. Chem Eng J, 2023, 470: 144266. doi: 10.1016/j.cej.2023.144266.
15. 闫志文, 李硕峰, 李傲, 等. 改性木葡聚糖温敏水凝胶的制备及生物相容性评价. 中国组织工程研究, 2019, 23(30): 4841-4847.
16. Pita-López ML, Fletes-Vargas G, Espinosa-Andrews H, et al. Physically cross-linked chitosan-based hydrogels for tissue engineering applications: A state-of-the-art review. Eur Polym J, 2021, 145: 110176. doi: 10.1016/j.eurpolymj.2020.110176.
17. Wang W, Xue C, Mao X. Chitosan: Structural modification, biological activity and application. Int J Biol Macromol, 2020, 164: 4532-4546.
18. Akbaba S, Atila D, Keskin D, et al. Multilayer fibroin/chitosan oligosaccharide lactate and pullulan immunomodulatory patch for treatment of hernia and prevention of intraperitoneal adhesion. Carbohydr Polym, 2021, 265: 118066. doi: 10.1016/j.carbpol.2021.118066.
19. Li L, Wang N, Jin X, et al. Biodegradable and injectable in situ cross-linking chitosan-hyaluronic acid based hydrogels for postoperative adhesion prevention. Biomaterials, 2014, 35(12): 3903-3917.
20. Hu W, Zhang Z, Zhu L, et al. Combination of polypropylene mesh and in situ injectable mussel-inspired hydrogel in laparoscopic hernia repair for preventing post-surgical adhesions in the piglet model. ACS Biomater Sci Eng, 2020, 6(3): 1735-1743.
21. Miserez M, Jairam AP, Boersema GSA, et al. Resorbable synthetic meshes for abdominal wall defects in preclinical setting: A literature review. J Surg Res, 2019, 237: 67-75.
22. Hu W, Lu S, Zhang Z, et al. Mussel-inspired copolymer-coated polypropylene mesh with anti-adhesion efficiency for abdominal wall defect repair. Biomater Sci, 2019, 7(4): 1323-1334.
23. Zhang H, Xu D, Zhang Y, et al. Silk fibroin hydrogels for biomedical applications. Smart Medicine, 2022, 1(1): e20220011. doi: 10.1002/SMMD.20220011.
24. Yuan N, Shao K, Huang S, et al. Chitosan, alginate, hyaluronic acid and other novel multifunctional hydrogel dressings for wound healing: A review. Int J Biol Macromol, 2023, 240: 124321. doi: 10.1016/j.ijbiomac.2023.124321.
25. Sousa AB, Águas AP, Barbosa MA, et al. Immunomodulatory biomaterial-based wound dressings advance the healing of chronic wounds via regulating macrophage behavior. Regen Biomater, 2022, 9: rbac065. doi: 10.1093/rb/rbac065.
26. Schaffrick L, Ding J, Kwan P, et al. The dynamic changes of monocytes and cytokines during wound healing post-burn injury. Cytokine, 2023, 168: 156231. doi: 10.1016/j.cyto.2023.156231.
27. Min D, Nube V, Tao A, et al. Monocyte phenotype as a predictive marker for wound healing in diabetes-related foot ulcers. J Diabetes Complications, 2021, 35(5): 107889. doi: 10.1016/j.jdiacomp.2021.107889.
28. Wang S, Lu M, Cao Y, et al. Degradative polylactide nanofibers promote M2 macrophage polarization via STAT6 pathway in peritendinous adhesion. Compos Part B Eng, 2023, 253: 110520. doi: 10.1016/j.compositesb.2023.110520.
29. Kroemer G, Zitvogel L. Ghrelin and leptin regulating wound healing. Trends Immunol, 2022, 43(10): 777-779.
30. Zheng H, Cheng X, Jin L, et al. Recent advances in strategies to target the behavior of macrophages in wound healing. Biomed Pharmacother, 2023, 165: 115199. doi: 10.1016/j.biopha.2023.115199.

Chinese Journal of Reparative and Reconstructive Surgery

Experimental study on repairing rat abdominal wall defect with chitosan hydrogel/polypropylene mesh composite

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