1. bookVolume 68 (2018): Issue 3 (September 2018)
Journal Details
First Published
25 Mar 2014
Publication timeframe
4 times per year
access type Open Access

In Vivo Investigation of Soft Tissue Response of Novel Silver/Poly(Vinyl Alcohol)/ Graphene and Silver/Poly(Vinyl Alcohol)/Chitosan/Graphene Hydrogels Aimed for Medical Applications – The First Experience

Published Online: 03 Oct 2018
Volume & Issue: Volume 68 (2018) - Issue 3 (September 2018)
Page range: 321 - 339
Received: 09 May 2018
Accepted: 23 Jul 2018
Journal Details
First Published
25 Mar 2014
Publication timeframe
4 times per year

In this paper, we have shown for the fi rst time the soft tissue response of novel silver/ poly(vinyl alcohol)/graphene (Ag/PVA/Gr) and silver/poly(vinyl alcohol)/chitosan/ graphene (Ag/PVA/CHI/Gr) nanocomposite hydrogels aimed for medical applications. These novel hydrogels were produced by in situ electrochemical synthesis of silver nanoparticles in the polymer matrices as described in our previously published works. Both Ag/PVA/Gr and Ag/PVA/CHI/Gr, as well as controls Ag/PVA, Ag/PVA/CHI and commercial Suprasorb©hydrogel discs, were implanted in the subcutaneous tissue of rats. Implants with the surrounding tissue were dissected after post-implantation on days 7, 15, 30 and 60, and then processed for histological examination. The tissue irritation index (TIrI) score, according to ISO 10993-6, 2007, as well as the number of leukocytes in the peri-implant zone and connective tissue capsule thickness were examined. The results show that each TIrI score, the leukocyte number around the implanted materials and capsule thickness gradually decreased during the observation period. At the endpoint of follow-up, the Ag/PVA/CHI/Gr implant was surrounded with a thinner capsule, while both the TIrI score and the number of leukocytes of the peri-implant zone were greater compared to the Ag/PVA/Gr implant. Despite the observed differences, we can conclude that our in vivo experiment suggested that both novel hydrogels were biocompatible and suitable for medical use.


1. vonRecum AF: Hand book of biomaterials evaluation: scientifi c, technical and clinical testing of implant materials. 2nd ed. London, UK: Taylor & Francis; 1999.Search in Google Scholar

2. Kumar A, Jaiswal M: Design and in vitro investigation of nanocomposite hydrogel based in situ spray dressing for chronic wounds and synthesis of silver nanoparticles using green chemistry. J Appl Polym Sci 2016, 133:1-14.10.1002/app.43260Search in Google Scholar

3. Zhang D, Zhou W, Wei B, Wang X, Tang R, Nie J, Wang J: Carboxyl-modifi ed poly (vinyl alcohol)-crosslinked chitosan hydrogel fi lms for potential wound dressing. Carbohyd Polym 2015, 125:189−199.10.1016/j.carbpol.2015.02.034Search in Google Scholar

4. Baker MI, Walsh SP, Schwartz Z, Boyan BD: A review of polyvinyl alcohol and its uses in cartilage and orthopedic applications. J Biomed Mater Res - Part B Appl Biomater 2012, 100 B:1451-1457.10.1002/jbm.b.32694Search in Google Scholar

5. Kanimozhi K, Basha SK, Kumari VS: Processing and characterization of chitosan/PVA and methylcellulose porous scaffolds for tissue engineering. Mater Sci Eng C 2016, 61:484-491.10.1016/j.msec.2015.12.084Search in Google Scholar

6. Oh SH, An DB, Kim TH, Lee JH: Wide-range stiffness gradient PVA/HA hydrogel to investigate stem cell differentiation behavior. Acta Biomater 2016, 35:23-31.10.1016/j.actbio.2016.02.016Search in Google Scholar

7. Koehler J, Brandl FP, Goepferich AM: Hydrogel wound dressings for bioactive treatment of acute and chronic wounds. Eur Polym J 2018, 100:1-11.10.1016/j.eurpolymj.2017.12.046Search in Google Scholar

8. Schmedlen RH, Masters KS, West JL: Photocrosslinkable polyvinyl alcohol hydrogels that can be modifi ed with cell adhesion peptides for use in tissue engineering. Biomaterials 2002, 23:4325-4332.10.1016/S0142-9612(02)00177-1Search in Google Scholar

9. Jiang S, Liu S, Feng W: PVA hydrogel properties for biomedical application. J Mech Behav Biomed Mater 2011, 4:1228-1233.10.1016/j.jmbbm.2011.04.00521783131Search in Google Scholar

10. Peppas NA, Huang Y, Tores-Lugo M, Ward JH, Zhang J: Physicochemical Foundations and Structural Design of Hydrogels in Medicine and Biology. Annu Rev Biomed Eng 2000, 2:9-29.10.1146/annurev.bioeng.2.1.911701505Search in Google Scholar

11. Puspitasari T, Raja KML, Pangerteni DS, Patriati A, Putra EGR: Structural Organization of Poly(vinyl alcohol) Hydrogels Obtained by Freezing/Thawing and Γ-Irradiation Processes: A Small-Angle Neutron Scattering ({SANS}) Study. Procedia Chem 2012, 4:186-193.10.1016/j.proche.2012.06.026Search in Google Scholar

12. Cheung RCF, Ng TB, Wong JH, Chan WY: Chitosan: An update on potential biomedical and pharmaceutical applications. Mar Drugs 2015, 13:5156-5186.10.3390/md13085156Search in Google Scholar

13. Kumar MNVR, Muzzarelli RAA, Muzzarelli C, Sashiwa H, Domb AJ: Chitosan chemistry and pharmaceutical perspectives. Chem Rev 2004, 104:6017-6084.10.1021/cr030441bSearch in Google Scholar

14. Jayakumar R, Prabaharan M, Kumar PS, Nair SV, Tamura H: Biomaterials based on chitin and chitosan in wound dressing applications. Biotechnol Adv 2011, 29:322-337.10.1016/j.biotechadv.2011.01.005Search in Google Scholar

15. Mi FL, Wu YB, Shyu SS, Schoung JY, Huang YB, Tsai YH, Hao JY: Control of wound infections using a bilayer chitosan wound dressing with sustainable antibiotic delivery. J Biomed Mater Res 2002, 59:438-449.10.1002/jbm.1260Search in Google Scholar

16. Mi FL, Shyu SS, Wu YB, Lee ST, Shyong JY, Huang RN. Fabrication and characterization of a sponge-like asymmetric chitosan membrane as a wound dressing. Biomaterials 2001, 22:165-173.10.1016/S0142-9612(00)00167-8Search in Google Scholar

17. Lu S, Gao W, Gu HY: Construction, application and biosafety of silver nanocrystalline chitosan wound dressing. Burns 2008, 34:623-628.10.1016/j.burns.2007.08.020Search in Google Scholar

18. Madihally SV, Matthew HWT: Porous chitosan scaffolds for tissue engineering. Biomaterials 1999, 20:1133-1142.10.1016/S0142-9612(99)00011-3Search in Google Scholar

19. Khor E, Lim LY: Implantable applications of chitin and chitosan. Biomaterials 2003, 24:2339-2349.10.1016/S0142-9612(03)00026-7Search in Google Scholar

20. Mishra SK, Ferreira JMF, Kannan S: Mechanically stable antimicrobial chitosan-PVA-silver nanocomposite coatings deposited on titanium implants. Carbohydr Polym 2015, 121:37-48.10.1016/j.carbpol.2014.12.02725659669Search in Google Scholar

21. Pishbin F, Simchi A, Ryan MP, Boccaccini AR: Electrophoretic deposition of chitosan/45S5 Bioglass composite coatings for orthopaedic applications. Surf Coatings Technol 2011, 205:5260-5268.10.1016/j.surfcoat.2011.05.026Search in Google Scholar

22. Venkatesan J, Kim SK: Chitosan composites for bone tissue engineering - An overview. Mar Drugs 2010, 8:2252-2266.10.3390/md8082252Search in Google Scholar

23. Felt O, Buri P, Gurny R: Chitosan: A unique polysaccharide for drug delivery. Drug Dev Ind Pharm 1998, 24:979-993.10.3109/03639049809089942Search in Google Scholar

24. Agnihotri SA, Mallikarjuna NN, Aminabhavi TM: Recent advances on chitosan-based micro- and nanoparticles in drug delivery. J Control Release 2004, 100:5-28.10.1016/j.jconrel.2004.08.010Search in Google Scholar

25. Bhattarai N, Gunn J, Zhang M: Chitosan-based hydrogels for controlled, localized drug delivery. Adv Drug Deliv Rev 2010, 62:83-99.10.1016/j.addr.2009.07.019Search in Google Scholar

26. Alt V, Bechert T, Steinrucke P, Wagener M, Seidel P, Dingeldein E, Domann E, Schnettler R: An in vitro assessment of the antibacterial properties and cytotoxicity of nanoparticulate silver bone cement. Biomaterials 2004, 25:4383-4391.10.1016/j.biomaterials.2003.10.078Search in Google Scholar

27. Lok CN, Ho CM, Chen R, He QY, Yu WY, Sun H, Tam PKH, Chiu JF, Che CM: Silver nanoparticles : partial oxidation and antibacterial activities. J Biol Inorg Chem 2007, 12:527-534.10.1007/s00775-007-0208-zSearch in Google Scholar

28. Chou KS, Ren CY: Synthesis of nanosized silver particles by chemical reduction method. Mater Chem Phys 2000, 64:241-246.10.1016/S0254-0584(00)00223-6Search in Google Scholar

29. Nešović K, Kojić V, Rhee KY, Mišković-Stanković V: Electrochemical synthesis and characterization of silver doped poly(vinyl alcohol)/chitosan hydrogels. Corrosion 2017, 73:1437-1447.10.5006/2507Search in Google Scholar

30. Twu YK, Chen YW, Shih CM: Preparation of silver nanoparticles using chitosan suspensions. Powder Technol 2008, 185:251-257.10.1016/j.powtec.2007.10.025Search in Google Scholar

31. Balandin AA, Ghosh S, Bao W, Calizo I, Teweldebrhan D, Miao F, Lau CN: Superior Thermal Conductivity of Single-Layer Graphene 2008. Nano Lett 2008, 8:902-907.10.1021/nl073187218284217Search in Google Scholar

32. Weiss NO, Zhou H, Liao L, Liu Y, Jiang S, Huang Y, Duan X: Graphene: An emerging electronic material. Adv Mater 2012, 24:5782-5825.10.1002/adma.20120148222930422Search in Google Scholar

33. Surudžić R, Janković A, Mitrić M, Matić I, Juranić ZD, Živković Lj, Mišković-Stanković V, Rhee KY, Park SJ, Hui D: The effect of graphene loading on mechanical, thermal and biological properties of poly(vinyl alcohol)/graphene nanocomposites. J Ind Eng Chem 2016, 34:250-257.10.1016/j.jiec.2015.11.016Search in Google Scholar

34. Yang JM, Wang SA: Preparation of graphene-based poly(vinyl alcohol)/chitosan nanocomposites membrane for alkaline solid electrolytes membrane. J Memb Sci 2015, 477:49-57.10.1016/j.memsci.2014.12.028Search in Google Scholar

35. Wang T, Li Y, Geng S, Zhou C, Jia X, Yang F, Zhang L, Ren X, Yang H: Preparation of fl exible reduced graphene oxide/poly(vinyl alcohol) fi lm with superior microwave absorption properties. RSC Adv 2015, 5:88958-88964.10.1039/C5RA16158DSearch in Google Scholar

36. Hegab HM, Elmekawy A, Zou L, Mulcahy D, Saint CP, Ginić-Marković M: The controversial antibacterial activity of graphene-based materials. Carbon 2016, 105:362-376.10.1016/j.carbon.2016.04.046Search in Google Scholar

37. Abudabbus MM, Jevremović I, Janković A, Perić-Grujić A, Matić I, Vukašinović-Sekulić M, Hui, D, Rhee, KY, Mišković-Stanković, V: Biological activity of electrochemically synthesized silver doped polyvinyl alcohol/graphene composite hydrogel discs for biomedical applications. Compos Part B Eng 2016, 104:26−34.10.1016/j.compositesb.2016.08.024Search in Google Scholar

38. Abudabbus MM, Jevremović I, Nešović K, Perić-Grujić A, Rhee KY, Mišković-Stanković V: In situ electrochemical synthesis of silver-doped poly(vinyl alcohol)/graphene composite hydrogels and their physico-chemical and thermal properties. Compos Part B Eng 2018, 140:99−107.10.1016/j.compositesb.2017.12.017Search in Google Scholar

39. Nešović K, Abudabbus MM, Rhee KY, Mišković-Stanković V: Graphene Based Composite Hydrogel for Biomedical Applications. Croat Chem Acta 2017, 90:207−213.10.5562/cca3133Search in Google Scholar

40. Wang J, Wang X, Xu C, Zhang M, Shang X: Preparation of graphene/poly(vinyl alcohol) nanocomposites with enhanced mechanical properties and water resistance. PolymInt 2011, 60:816-822.10.1002/pi.3025Search in Google Scholar

41. Xue R, Xin X, Wang L, Shen J, Ji F, Li W, Jia C, Xu G: A systematic study of the effect of molecular weights of polyvinyl alcohol on polyvinyl alcohol-graphene oxide composite hydrogels. PhysChemChemPhys 2015, 17:5431-5440.10.1039/C4CP05766JSearch in Google Scholar

42. Abudabbus MM, Jevremović I, Janković A, Perić-Grujić A, Matić I, Vukašinović-Sekulić M, Hui D, Rhee K.Y, Mišković-Stanković V: Biological activity of electrochemically synthesized silver doped polyvinyl alcohol/graphene composite hydrogel discs for biomedical applications. Compos Part B Eng 2016, 104:26-34.10.1016/j.compositesb.2016.08.024Search in Google Scholar

43. Usman A, Hussain Z, Riaz A, Khan AN: Enhanced mechanical, thermal and antimicrobial properties of poly(vinyl alcohol)/graphene oxide/starch/silver nanocomposites fi lms. CarbohydrPolym 2016, 153:592-599.10.1016/j.carbpol.2016.08.026Search in Google Scholar

44. Pandele AM, Ionita M, Crica L, Dinescu S, Costache M, Iovu H: Synthesis, characterization, and in vitro studies of graphene oxide/chitosan-polyvinyl alcohol fi lms. CarbohydrPolym 2014, 102:813-820.10.1016/j.carbpol.2013.10.085Search in Google Scholar

45. Peppas NA, Bures P, Leobandung W, H Ishikawa: Hydrogels in pharmaceutical formulations. Eur J Pharm Biopharm 2000, 50:27−46.10.1016/S0939-6411(00)00090-4Search in Google Scholar

46. Muppalaneni S, Omidian H: Polyvinyl alcohol in medicine and pharmacy: A Perspective. J Devel Drugs 2013, 2:1−510.4172/2329-6631.1000112Search in Google Scholar

47. Hyon SH, Cha WI, Ikada Y, Kita M, Ogura Y, Honda Y: Poly (vinyl alcohol) hydrogels as soft contact lens material. J BiomaterSciPolym Ed 1994, 5:397-406.10.1163/156856294X00103Search in Google Scholar

48. Jiang H, Zuo Y, Zhang L, Li J, Zhang A, Li Y, Yang X: Property-based design: optimization and characterization of polyvinyl alcohol (PVA) hydrogel and PVA-matrix composite for artifi cial cornea. J Mater Sci Mater Med 2014, 25:941-952.10.1007/s10856-013-5121-024464723Search in Google Scholar

49. Shokrgozar MA, Bonakdar S, Dehghan MM, Emami SH, Montazeri L, Azari S, Rabbani M: Biological evaluation of polyvinyl alcohol hydrogel crosslinked by polyurethane chain for cartilage tissue engineering in rabbit model. J Mater Sci Med 2013, 24:2449-2460.10.1007/s10856-013-4995-123807316Search in Google Scholar

50. Hayes JC, Kennedy JE: An evaluation of the biocompatibility properties of a salt-modifi ed polyvinyl alcohol hydrogel for a knee meniscus application. Mater SciEng C Mater BiolAppl 2016, 59:894-900.10.1016/j.msec.2015.10.05226652445Search in Google Scholar

51. Kobayashi M, Togushida J, Oka M: Development of polyvinyl alcohol-hydrogel shields with a high water content for tendon injury repair. J Hand Surg Br 2001, 26:436-440.10.1054/jhsb.2001.058111560425Search in Google Scholar

52. Kobayashi M, Oka M: Composite device for attachment o polyvinyl alcohol-hydrogel to underlying bone. Artif Organs 2004, 28:734-738.10.1111/j.1525-1594.2004.00079.x15270955Search in Google Scholar

53. Guo Y, Guo J, Bai D, Wang H, Zheng X, Guo W, Tian W: Hemiarthroplasty of the shoulder joint using a custom-designed high-density nano-hydroxyapatite/ polyamide prosthesis with a polyvinyl alcohol hydrogel humeral head surface in rabbits. Artif Organs 2014, 38:580-586.10.1111/aor.1225724404998Search in Google Scholar

54. Deerenberg EB, Mulder IM, Ditzel M, Slieker JC, Bemelman WA, Jeekel J, Lange JF: Polyvinyl alcohol hydrogel decreases formation of adhesions in a rat model of peritonitis. Surg Infect (Larchmt) 2012, 13:321-325.10.1089/sur.2011.10822789109Search in Google Scholar

55. Moretto A, Tesolin L, Marsilio F, Schiavon M, Berna M, Veronese FM: Slow release of two antibiotics of veterinary interest from PVA hydrogels. Farmaco 2004, 59:1-5.10.1016/j.farmac.2003.11.00314751309Search in Google Scholar

56. Basak P, Adhikari B: Poly (vinyl alcohol) hydrogels for pH dependent colon targeted drug delivery. J Mater Sci Mater Med 2009, 20(Suppl 1):137-146.10.1007/s10856-008-3496-018592350Search in Google Scholar

57. Kim TH, An DB, Oh SH, Kang MK, Song HH, Lee JH: Creating stiffness gradient polyvinyl alcohol hydrogel using a simple gradual freezing-thawing method to investigate stem cell differentiation behaviors. Biomaterials 2015, 40:51-60.10.1016/j.biomaterials.2014.11.01725467820Search in Google Scholar

58. Li P, Jiang S, Yu Y, Yang J, Yang Z: Biomaterial characteristics and application of silicone rubber and PVA hydrogels mimicked in organ groups for prostate brachytherapy. J MechBehav Biomed Mater 2015, 49:220-234.10.1016/j.jmbbm.2015.05.01226042767Search in Google Scholar

59. Alexandre N, Amorim I, Caseiro AR, Pereira T, Alvites R, Rêma A, Gonçalves A, Valadares G, Costa E, Santos-Silva A, Rodrigues M, Lopes MA, Almeida A, Santos JD, Maurício AC, Luís AL: Long term performance evaluation of small-diameter vascular grafts based on polyvinyl alcohol hydrogel and dextran and MSCs-based therapies using the ovine preclinical animal model. Int J Pharm 2017, 523:515-530.10.1016/j.ijpharm.2017.02.04328283218Search in Google Scholar

60. Scaretta FV: 5 to 8 years follow-up of knee chondral defects treated by PVA-H hydrogel implants. Eur Rev Med PharmacolSci 2013, 17:3031-3038.Search in Google Scholar

61. Taleb C, Berner S, Mantovani Ruggiero G: First metacarpal resurfacing with polyvinyl alcohol implant in osteoarthritis: preliminary study. Chir Main 2014, 33:189-195.10.1016/j.main.2014.03.00124880607Search in Google Scholar

62. Daniels TR, Younger AS, Penner MJ, Wing KJ, Miniaci-Coxhead SL, Pinsker E, Glazebrook M: Midterm outcomes of polyvinyl alcohol hydrogel hemiarthroplasty of the fi rst metatarso-phalangeal joint in advanced hallux rigidus. Foot Ankle Int 2017, 38:243-247.10.1177/107110071667997927909032Search in Google Scholar

63. Baker MI, Walsh SP, Schwartz Z, Boyan BD: A review of polyvinyl alcohol and its uses in cartilage and orthopedic applications. J Biomed Mater Res B ApplBiomater 2012, 100:1451-1457.10.1002/jbm.b.3269422514196Search in Google Scholar

64. Lee JM, Sultan MT, Kim SH, Kumar V, Yeon YK, Lee JH: Artifi cial auricular cartilage using silk fi broin and polyvinyl alcohol hydrogel. Int J MolSci 2017, 18:1707.10.3390/ijms18081707557809728777314Search in Google Scholar

65. Dashtdar H, Murali MR, Abbas AA, Suhaeb AM, Selvaratnam L, Tay LX, Kamarul T: PVAchitosan composite hydrogel versus alginate beads as a potential mesenchymal stem cell carrier for the treatment of focal cartilage defects. Knee Surg Sports TraumatolArthrosc 2015; 23:1368-1377.10.1007/s00167-013-2723-524146054Search in Google Scholar

66. Kamoun EA, Kenawy ES, Chen X: A review on polymeric hydrogel membranes for wound dressing applications: PVA-based hydrogel dressings. J Adv Res 2017, 8:217-233.10.1016/j.jare.2017.01.005531544228239493Search in Google Scholar

67. Wu JQ, Liu Y, Yang TF, Mu YH, Guo T, Li YB: Porous polyvinyl alcohol hydrogel composite prepared and studied initially for biocompatibility. Sichuan Da XueXueBao Yi Xue Ban 2007, 38:705-708, 724.Search in Google Scholar

68. Korbekandi H, Iravani S: Silver Nanoparticles. In: The Deliveri of Nanoparticles. Rijeka, Croatia: Intech Open; 2012, 3-36.10.5772/34157Search in Google Scholar

69. Pencheva D, Bryaskova R, Kantardjiev T: Polyvinyl alcohol/silver nanoparticles ({PVA/ AgNPs}) as a model for testing the biological activity of hybrid materials with included silver nanoparticles. Mater SciEng C 2012, 32:2048-2051.10.1016/j.msec.2012.05.01634062694Search in Google Scholar

70. Fan Z, Liu B, Wang J, Zhang S, Lin Q, Gong P, Ma L, Yang S: A novel wound dressing based on Ag/graphene polymer hydrogel: effectively kill bacteria and accelerate wound healing. AdvFunct Mater 2014, 24:3933-3943.10.1002/adfm.201304202Search in Google Scholar

71. Linares J, Matesanz MC, Feito MJ, Salavagione HJ, Martínez G, Gómez-Fatou M, Portolés MT: Infl uence of the covalent immobilization of graphene oxide in poly(vinyl alcohol) on human osteoblast response. Colloids Surf B Biointerfaces 2016, 138:50-59.10.1016/j.colsurfb.2015.11.03526650079Search in Google Scholar

72. Shang S, Li J, Zhao Y, Xi Z, Lu Z, Li B, Yang X, Li R: Oxidized graphene-aggravated allergic asthma is antagonized by antioxidant vitamin E in Balb/c mice. Environ SciPollut Res Int 2017, 24:1784-1793.10.1007/s11356-016-7903-727796986Search in Google Scholar

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