Biodegradation, Angiogenesis, and Inflammatory Response of a Collagen-Chitosan-Polyvinyl Alcohol (PVA) Membrane: In Vivo Model of Guided Tissue Regeneration

Abstract

Objective

The aim of this study was to examine the biodegradation, angiogenesis, and inflammatory response in collagen-chitosan-polyvinyl alcohol (PVA) membranes.

Materials and Methods

This study employed an experimental approach utilizing a randomized controlled trial design. Wistar rats were used as subjects, with 51 rats divided into three groups. Each group received a different treatment: application of the collagen-chitosan-PVA membrane, pericardial membrane, or cross-linked pericardial membrane, administered subcutaneously. On days 0, 7, 14, and 30, the rats were terminated, and the membranes and surrounding tissues were collected for analysis. A histological examination was performed to evaluate the membrane biodegradation rate, the number of blood vessels formed, and the inflammatory response.

Statistical Analysis

The data were analyzed using the Kruskal-Wallis and Mann-Whitney tests, with a p-value of < 0.05 considered statistically significant.

Results

The collagen-chitosan-PVA membrane remained in the tissue up to day 30, while the pericardial membrane and cross-linked pericardial membrane were completely degraded by day 7. The average number of new blood vessels formed in the collagen-chitosan-PVA membrane on days 7, 14, and 30 was greater than that in the pericardial membrane and cross-linked pericardial membrane, which was statistically significant (p < 0.005). The average number of inflammatory cells in the collagen-chitosan-PVA membrane on day 30 was lower than that in the pericardial membrane and cross-linked pericardial membrane, which was statistically significant (p < 0.005) for neutrophils, monocytes, and lymphocytes. However, the difference was not statistically significant (p > 0.05) for eosinophils and mast cells.

Conclusion

Biodegradation, angiogenesis, and the inflammatory response in collagen-chitosan-PVA membranes showed better results compared with other membranes. Collagen-chitosan-PVA membranes exhibit potential for application in guided tissue regeneration treatment for periodontal disease.

Keywords

angiogenesis - biodegradation - chitosan - collagen - guided tissue regeneration - inflammatory response

Publikationsverlauf

Artikel online veröffentlicht:
16. April 2025

• References

• 1 Wang HL, Greenwell H, Fiorellini J. et al; Research, Science and Therapy Committee. Periodontal regeneration. J Periodontol 2005; 76 (09) 1601-1622
• 2 Carranza FA, Newman MG, Takei HH, Klokkevold PR. Carranza's Clinical Periodontology 11th ed. Amsterdam: Saunders Elsevier Inc; 2012: 642
• 3 Chen S, Hao Y, Cui W. et al. Biodegradable electrospun PLLA/chitosan membrane as guided tissue regeneration membrane for treating periodontitis. J Mater Sci 2013; 48: 6567-6577
• 4 Kasaj A, Reichert C, Götz H, Röhrig B, Smeets R, Willershausen B. In vitro evaluation of various bioabsorbable and nonresorbable barrier membranes for guided tissue regeneration. Head Face Med 2008; 4: 22
• 5 Farzad M, Mohammadi M. Guided bone regeneration: a literature review. J Oral Health Oral Epidemiol 2012; 1 (01) 3-18
• 6 Rothamel D, Schwarz F, Fienitz T. et al. Biocompatibility and biodegradation of a native porcine pericardium membrane: results of in vitro and in vivo examinations. Int J Oral Maxillofac Implants 2012; 27 (01) 146-154
• 7 Moses O, Vitrial D, Aboodi G. et al. Biodegradation of three different collagen membranes in the rat calvarium: a comparative study. J Periodontol 2008; 79 (05) 905-911
• 8 Adiana ID, Syafiar L. Use of chitosan as a biomaterial in dentistry. Dentika Dent J 2014; 18 (02) 190-193
• 9 Chang HH, Wang YL, Chiang YC. et al. A novel chitosan-γPGA polyelectrolyte complex hydrogel promotes early new bone formation in the alveolar socket following tooth extraction. PLoS One 2014; 9 (03) e92362
• 10 Rodríguez-Vázquez M, Vega-Ruiz B, Ramos-Zúñiga R, Saldaña-Koppel DA, Quiñones-Olvera LF. Chitosan and its potential use as a scaffold for tissue engineering in regenerative medicine. BioMed Res Int 2015; 2015: 821279
• 11 Archana D, Upadhyay L, Tewari RP, Dutta J, Huang YB, Dutta PK. Chitosan-pectin alginate as a novel scaffold for tissue engineering applications. Indian J Biotechnol 2013; 12: 475-482
• 12 Tangsadthakun C, Kanokpanont S, Sanchavanakit N, Banaprasert T, Damrongsakkul S. Properties of collagen/chitosan scaffolds for skin tissue engineering. J Metals Mat Minerals 2006; 16 (01) 37-44
• 13 Bano I, Ghauri MA, Yasin T, Huang Q, Palaparthi AD. Characterization and potential applications of gamma irradiated chitosan and its blends with poly(vinyl alcohol). Int J Biol Macromol 2014; 65: 81-88
• 14 Zhou T, Zheng K, Sui B, Boccaccini AR, Sun J. In vitro evaluation of poly (vinyl alcohol)/collagen blended hydrogels for regulating human periodontal ligament fibroblasts and gingival fibroblasts. Int J Biol Macromol 2020; 163: 1938-1946
• 15 Sam G, Pillai BR. Evolution of barrier membranes in periodontal regeneration-“are the third generation membranes really here?”. J Clin Diagn Res 2014; 8 (12) ZE14-ZE17
• 16 Zhou T, Chen S, Ding X, Hu Z, Cen L, Zhang X. Fabrication and characterization of collagen/PVA dual-layer membranes for periodontal bone regeneration. Front Bioeng Biotechnol 2021; 9: 630977
• 17 Zhuang PY, Li YL, Fan L, Lin J, Hu QL. Modification of chitosan membrane with poly(vinyl alcohol) and biocompatibility evaluation. Int J Biol Macromol 2012; 50 (03) 658-663
• 18 Erizal E, Perkasa DP, Aziz Z, Sulistioso GS. Modification of physi- cochemical on polyvinyl alcohol chitosan composite membrane from casting process by gamma irradiation technique. Indonesian J Mater Sci 2013; 14 (03) 166-172
• 19 Liang Y, Luan X, Liu X. Recent advances in periodontal regeneration: a biomaterial perspective. Bioact Mater 2020; 5 (02) 297-308
• 20 Bottino MC, Thomas V, Schmidt G. et al. Recent advances in the development of GTR/GBR membranes for periodontal regeneration–a materials perspective. Dent Mater 2012; 28 (07) 703-721
• 21 Komara I, Susanto A, Amaliya A. et al. The effect of gamma-ray irradiation on the physical, mechanical, and morphological characteristics of PVA-collagen-chitosan as a guided tissue regeneration (GTR) membrane material. Eur J Dent 2023; 17 (02) 530-538
• 22 Rajam AM, Jithendral P, Mandal AB, Rose C. Evaluation of in vitro macrophage response and in vivo host response to growth factors incorporated chitosan nanoparticle impregnated collagen-chitosan scaffold. J Biomed Nanotechnol 2014; 10 (03) 508-518
• 23 Annette BW, Wanda ADM. Source Book of Models for Biomedical Research. Totowa: Humana Press; 2008: 631-638
• 24 Schwarz F, Rothamel D, Herten M, Sager M, Becker J. Angiogenesis pattern of native and cross-linked collagen membranes: an immunohistochemical study in the rat. Clin Oral Implants Res 2006; 17 (04) 403-409
• 25 Osathanon T, Chanjavanakul P, Kongdecha P, Clayhan P, Huynh NCN. Polycaprolactone-based biomaterials for guided tissue regeneration membrane. In: Periodontitis - A Useful Reference. London, United Kingdom: IntechOpen Limited; 2017: 171-188
• 26 Cho YD, Kim KH, Lee YM, Ku Y, Seol YJ. Periodontal wound healing and tissue regeneration: a narrative review. Pharmaceuticals (Basel) 2021; 14 (05) 456
• 27 Mahendra J, Vijayaraj S, Ari G, Rajendran S, Namasivayam A. A review on wound healing in periodontics. Annals of R.S.C.B. 2020; 24 (01) 551-558
• 28 Ramya SAK. Patterns of wound healing following various periodontal procedures-a review. Eur J Mol Clin Med 2020; 07 (02) 6665-6667
• 29 Soesilawati P. Basic Medical Histology. Surabaya: Airlangga University Press; 2020: 30-43
• 30 Wati W, Balqis U, Iskandar C. Identifying and Counting the Inflammatory Cells in Muscle (Mus Musculus) Cuts Leaved by Exrtrack Leaves Binhong (Anredera Cordifolia (Ten) Steenis). JIMVET 2020; 4 (04) 108-115
• 31 Kusumastuti RW. Analysis of Total Phenolic Content in Ginger (Zingiber officinale Rosc.) In Vitro. Jakarta: Faculty of Medicine, University of Indonesia; 2014
• 32 Wahyuningsih. The Influence of Castor Bean Extract (Jathropa Multifida, Linn) on the Number of PMN and Mast Cells in the Healing of Rat Wounds with an Acute Injury Model. Public Health Journal 2019; 4 (01) 1-6
• 33 Hartatiek, Fathurochman F, Wuriantika MI. et al Mechanical, degradation rate, and antibacterial properties of a collagen-chitosan/PVA composite nanofiber. Mater Res Express 2023; 10 (02) 1
• 34 Zheng LY, Zhu JF. Study on antimicrobial activity of chitosan with different molecular weights. Carbohydrate polymers 2003; 54 (04) 527-30