Skip to main content

Advertisement

SpringerLink
Log in

Human Amniotic Membrane and Titanium Dioxide Nanoparticle Derived Gel for Burn Wound Healing in a Rat Model

  • Original Research
  • Published:
Regenerative Engineering and Translational Medicine Aims and scope Submit manuscript

Abstract

Purpose

Induction of skin tissue regeneration and reduction of microbial infection at the wound site are crucial for burn wound treatment. In this study, human amniotic membrane (HAM) and titanium dioxide (TiO2) nanoparticle infused gel (HAM+TiO2) was developed to find a promising biomaterial for burn wound management.

Methods

HAM, TiO2, and HAM+TiO2 gels were prepared with carbopol 934, acrylic acid, propylparaben, and triethanolamine. Besides, physiochemical properties (pH, water absorption, swelling ratio, spreadability, etc.), antibacterial activity, and brine shrimp lethality tests were executed for the gels. The gels were applied to second-degree burns of Wistar rats for 20 days. Percent wound contracture, epithelialization period, and histological observations were carried out to evaluate the wound healing process.

Results

Formulated gels have shown antibacterial activity against S. aureus (ATCC 25923), P. aeruginosa (ATCC 27853), and E. coli (ATCC 25922). After 20 days of treatment, HAM+TiO2 significantly (P<0.0001) accelerated wound contraction (95.29 ± 0.54%) compared to a control group. For complete healing, the epithelialization period was 21.0 ± 1 day (P< 0.001). Histology analysis revealed the better wound healing potential for HAM+TiO2 in terms of epidermal regeneration, blood vessel formation, and collagen deposition with minimal scar formation.

Conclusion

Our findings demonstrated that the treatment of burn wounds with HAM+TiO2 significantly promoted wound healing in Wister rats by accelerating wound contraction, collagen synthesis, and preventing bacterial growth.

Lay Summary and Future Works

Burns have dreadful consequences for the affected patients, making them susceptible to wound infections. The application of HAM for wound healing is increasing because of its low immunogenicity, angiogenic, and antimicrobial properties. For burn wound treatment, TiO2 NPs have been well identified. In this research, TiO2 NP-incorporated HAM gel was prepared and we used this gel to experimental groups of Wister rats with an initial burn on their skin. In an in vivo study and histopathologic observation, better wound healing progress was demonstrated for HAM + TiO2. In the future, the study will be performed on large scale for the treatment of full-thickness burns and/or chronic wounds.

This is a preview of subscription content, log in via an institution to check access.

Access this article

Log in via an institution

Price excludes VAT (USA)
Tax calculation will be finalised during checkout.

Instant access to the full article PDF.

Institutional subscriptions

Fig. 1
Fig. 2
Fig. 3
Fig. 4
Fig. 5
Fig. 6
Fig. 7
Fig. 8

Similar content being viewed by others

Abbreviations

HAM :

Human amniotic membrane

NPs :

Nanoparticles

H&E :

Haematoxylin and eosin

MT :

Masson’s Trichrome

ARRIVE:

Animal Research: Reporting of In Vivo Experiments

References

  1. Xue L, Xu YB, Xie JL, Tang JM, Shu B, Chen L, et al. Effects of human bone marrow mesenchymal stem cells on burn injury healing in a mouse model. Int J Clin Exp Pathol. 2013;6(7):1327–36.

    Google Scholar 

  2. Guo HF, Ali RM, Hamid RA, Zaini AA, Khaza’ai H. A new model for studying deep partial-thickness burns in rats. Int J Burns Trauma. 2017;7(6):107–14.

    CAS  Google Scholar 

  3. Jahromi MAM, Zangabad PS, Basri SMM, Zangabad KS, Ghamarypour A, Aref AR, Karimi M, Hamblin MR. Nanomedicine and advanced technologies for burns: Preventing infection and facilitating wound healing. Adv Drug Deliv Rev. 2018;123:33–64. https://doi.org/10.1016/j.addr.2017.08.001.

    Article  CAS  Google Scholar 

  4. Pormohammad A, Monych NK, Ghosh S, Turner DL, Turner RJ. Nanomaterials in wound healing and infection control. Antibiotics (Basel). 2021;10(5):473. https://doi.org/10.3390/antibiotics10050473.

    Article  CAS  Google Scholar 

  5. Maslova E, Eisaiankhongi L, Sjöberg F, McCarthy RR. Burns and biofilms: priority pathogens and in vivo models. NPJ Biofilms Microbiomes. 2021;7(1):73. https://doi.org/10.1038/s41522-021-00243-2.

    Article  Google Scholar 

  6. PLOS Medicine Editors. Antimicrobial resistance: is the world UNprepared?. Plos Med (2016) 13(9):e1002130, https://doi.org/10.1371/journal.pmed.1002130.

  7. Jones I, Currie L, Martin R. A guide to biological skin substitutes. Br J Plast Surg. 2002;55(3):185–93. https://doi.org/10.1054/bjps.2002.3800.

    Article  CAS  Google Scholar 

  8. Lifen LIU, John B, Yeung KL. Non-UV germicidal activity of fresh TiO2 and Ag/TiO2. Res J Environ Sci. 2009;21(5):700–6. https://doi.org/10.1016/S1001-0742(08)62327-X.

    Article  CAS  Google Scholar 

  9. Haller HL, Blome-Eberwein SE, Branski LK, Carson JS, Crombie RE, Hickerson WL, et al. Porcine xenograft and epidermal fully synthetic skin substitutes in the treatment of partial-thickness burns: a literature review. Medicina (Kaunas). 2021;57(5):432. https://doi.org/10.3390/medicina57050432.

    Article  Google Scholar 

  10. Ramasamy P, Krishnakumar R, Rekha R, Vaseeharan B, Saraswathi K, Raj M, et al. Bio-fabrication of human amniotic membrane zinc oxide nanoparticles and the wet/dry HAM dressing membrane for wound healing. Front Bioeng Biotechnol. 2021;9:695710. https://doi.org/10.3389/fbioe.2021.695710.

    Article  Google Scholar 

  11. Murphy SV, Skardal A, Nelson RA, Sunnon K, Reid T, Clouse C, et al. Amnion membrane hydrogel and amnion membrane powder accelerate wound healing in a full thickness porcine skin wound model. Transl Med. 2020;9(1):80–92. https://doi.org/10.1002/sctm.19-0101.

    Article  CAS  Google Scholar 

  12. Kim JS, Kim JC, Na BK, Jeong JM, Song CY. Amniotic membrane patching promotes healing and inhibits proteinase activity on wound healing following acute corneal alkali burn. Exp Eye Res. 2000;70(3):329–37. https://doi.org/10.1006/exer.1999.0794.

    Article  CAS  Google Scholar 

  13. Mostaque AK, Rahman KBA. Comparisons of the effects of biological membrane (amnion) and silver sulfadiazine in the management of burn wounds in children. J Burn Care Res. 2011;32(2):200–9. https://doi.org/10.1097/BCR.0b013e31820aad94.

    Article  Google Scholar 

  14. Schmiedova I, Ozanova Z, Stastna E, Kiselakova L, Lipovy B, Forostyak S. Case report: Freeze-dried human amniotic membrane allograft for the treatment of chronic wounds: results of a multicentre observational study. Front Bioeng Biotechnol. 2021;9:649446. https://doi.org/10.3389/fbioe.2021.649446.

    Article  Google Scholar 

  15. Rahman MS, Islam R, Rana MM, Spitzhorn LS, Rahman MS, Adjaye J, et al. Characterization of burn wound healing gel prepared from human amniotic membrane and Aloe vera extract. BMC Complement Altern Med. 2019;19:115. https://doi.org/10.1186/s12906-019-2525-5.

    Article  CAS  Google Scholar 

  16. Crosera M, Prodi A, Mauro M, Pelin M, Florio C, Bellomo F, et al. Titanium dioxide nanoparticle penetration into the skin and effects on HaCaT cells. Int J Environ Res. 2015;12(8):9282–97. https://doi.org/10.3390/ijerph120809282.

    Article  CAS  Google Scholar 

  17. Hajialyani M, Tewari D, Sobarzo-Sánchez E, Nabavi SM, Farzaei MH, Abdollahi M. Natural product-based nanomedicines for wound healing purposes: therapeutic targets and drug delivery systems. Int J Nanomedicine. 2018;13:5023–43. https://doi.org/10.2147/IJN.S174072.

    Article  CAS  Google Scholar 

  18. Eldebany N, Elkodous MA, Tohamy H, Abdelwahed R, El-kammar M, Ahmed HA, et al. Gelatin loaded titanium dioxide and silver oxide nanoparticles: implication for skin tissue regeneration. Biol Trace Elem Re. 2021;199(10):3688–99. https://doi.org/10.1007/s12011-020-02489-x.

    Article  CAS  Google Scholar 

  19. Bui VKH, Park D, Lee YC. Chitosan combined with ZnO, TiO2 and Ag nanoparticles for antimicrobial wound healing applications: a mini review of the research trends. Polymers. 2017;9:21. https://doi.org/10.3390/polym9010021.

    Article  CAS  Google Scholar 

  20. Nikpasand A, parvizi M. Evaluation of the effect of titatnium dioxide nanoparticles/gelatin composite on infected skin wound healing; an animal model study. Bull Emerg Trauma. 2019;7(4):366–72. https://doi.org/10.29252/beat-070405.

    Article  Google Scholar 

  21. Seisenbaeva GA, Fromell K, Vinogradov VV, Terekhov AN, Pakhomov AV, Nilsson B, et al. Dispersion of TiO2 nanoparticles improves burn wound healing and tissue regeneration through specific interaction with blood serum proteins. Sci Rep. 2017;7:15448. https://doi.org/10.1038/s41598-017-15792-w.

    Article  CAS  Google Scholar 

  22. Islam MM, Hossain ML, Diba F, Hasan MZ, Juliana FM, Asaduzzaman SM. The combined effect of amniotic membrane and Moringa oleifera leaves derived gel for wound and burn healing in rat model. Regen Eng Transl Med. 2018;11(1):00341. https://doi.org/10.1007/s40883-018-0060-4.

    Article  CAS  Google Scholar 

  23. Rana MM, Rahman MS, Ullah MA, Siddika A, Hossain ML, Akhter MS, et al. Amnion and collagen-based blended hydrogel improves burn healing efficacy on a rat skin wound model in the presence of wound dressing biomembrane. Biomed Mater Eng. 2020;31(1):1–17. https://doi.org/10.3233/BME-201076.

    Article  CAS  Google Scholar 

  24. El-Kased RF, Amer RI, Attia D, Elmazar MM. Honey-based hydrogel: in vitro and comparative in vivo evaluation for burn wound healing. Sci Rep. 2017;7(1):9692. https://doi.org/10.1038/s41598-017-08771-8.

    Article  Google Scholar 

  25. Valgas C, De Souza SM, Smânia EFA. Screening methods to determine antibacterial activity of natural products. Braz J Microbiol. 2007;38:369–80. https://doi.org/10.1590/S1517-83822007000200034.

    Article  Google Scholar 

  26. Karmakar PC, Hasi RY, Ali H, Habib MR, Barman DN, Miah MAS, et al. Antioxidant, cytotoxic and apoptotic potentials of seeds of Momordica subangulata subsp. renigera inhibit the growth of Ehrlich ascites carcinoma in mice. J Food Meas. 2019;13:3049–59. https://doi.org/10.1007/s11694-019-00227-z.

    Article  Google Scholar 

  27. Choi J, Kim H, Choi J, Oh SM, Park J, Park K. Skin corrosion and irritation test of sunscreen nanoparticles using reconstructed 3D human skin model. Environ Health Toxicol. 2014;29:e2014004. https://doi.org/10.5620/eht.2014.29.e2014004.

    Article  Google Scholar 

  28. Song M, Wang W, Ye Q, Bu S, Shen Z, Zhu Y. The repairing of full-thickness skin deficiency and its biological mechanism using decellularized human amniotic membrane as the wound dressing. Mater Sci Eng C. 2017;77:739–47. https://doi.org/10.1016/j.msec.2017.03.232.

    Article  CAS  Google Scholar 

  29. You C, Li Q, Wang X, Wu P, Ho JK, Jin R, Zhang L, Shao H, Han C. Silver nanoparticle loaded collagen/chitosan scaffolds promote wound healing via regulating fibroblast migration and macrophage activation. Sci Rep. 2017;7(1):10489. https://doi.org/10.1038/s41598-017-10481-0.

    Article  CAS  Google Scholar 

  30. Thangavel P, Ramachandran B, Chakraborty S, Kannan R, Lonchin S, Muthuvijayan V. Accelerated healing of diabetic wounds treated with L-glutamic acid loaded hydrogels through enhanced collagen deposition and angiogenesis: An In Vivo Study. Sci Rep. 2017;7:10701. https://doi.org/10.1038/s41598-017-10882-1.

    Article  CAS  Google Scholar 

  31. Schmiedova I, Ozanova Z, Stastna E, Kiselakova L, Lipovy B, Forostyak S. Bio-fabrication of human amniotic membrane zinc oxide nanoparticles and the wet/dry HAM dressing membrane for wound healing. Front Bioeng Biotechnol. 2021;9:695710. https://doi.org/10.3389/fbioe.2021.695710.

    Article  Google Scholar 

  32. Tazeze H, Mequanente S, Nigussie D, Legesse B, Makonnen E, Mengie T. Investigation of wound healing and anti-inflammatory activities of leaf gel of Aloe trigonantha L.C. Leach in Rats. J Inflamm Res. 2021;14:5567–80. https://doi.org/10.2147/JIR.S339289.

    Article  CAS  Google Scholar 

  33. Chen MX, Alexander KS, Baki G. Formulation and evaluation of antibacterial creams and gels containing metal ions for topical application. J Pharm (Cairo). 2016;2016:5754349. https://doi.org/10.1155/2016/5754349.

    Article  CAS  Google Scholar 

  34. Khaleghi M, Ahmadi E, Khodabandeh Shahraki M, Aliakbari F, Morshedi D. Temperature-dependent formulation of a hydrogel based on Hyaluronic acid-polydimethylsiloxane for biomedical applications. Heliyon. 2020;6(3):e03494. https://doi.org/10.1016/j.heliyon.2020.e03494.

    Article  CAS  Google Scholar 

  35. Ramuta TŽ, Tratnjek L, Janev A, Seme K, Starčič Erjavec M, Kreft ME. The antibacterial activity of human amniotic membrane against multidrug-resistant bacteria associated with urinary tract infections: new insights from normal and cancerous urothelial models. Biomedicine. 2021;9(2):218. https://doi.org/10.3390/biomedicines9020218.

    Article  CAS  Google Scholar 

  36. Železnik RT, Tina S, Marjanca SE, Erdani KM. Antimicrobial activity of human fetal membranes: from biological function to clinical use. Front bioeng. 2021;9:435. https://doi.org/10.3389/fbioe.2021.691522.

    Article  Google Scholar 

  37. Khashan KS, Sulaiman GM, Abdulameer FA, Albukhaty S, Ibrahem MA, Al-Muhimeed T, AlObaid AA. Antibacterial activity of TiO2 nanoparticles prepared by one-step laser ablation in liquid. Appl Sci (2021) 11:4623 https://doi.org/10.3390/app11104623.

  38. Sharma L, Srivastava H, Pipal DK, Dhawan R, Purohit PM, Bhargava A. Bacteriological profile of burn patients and antimicrobial susceptibility pattern of burn wound isolates. Int Surg J. 2017;4(3):1019–23. https://doi.org/10.18203/2349-2902.isj20170854.

    Article  Google Scholar 

  39. Suryamathi M, Viswanathamurthi P, Seedevi P. Herbal plant leaf extracts immobilized PCL nanofibrous mats as skin-inspired anti-infection wound healing material. Regen Eng Transl Med. 2022;8:94–105. https://doi.org/10.1007/s40883-020-00193-9.

    Article  CAS  Google Scholar 

  40. Ramli NA, Wong TW. Sodium carboxymethylcellulose scaffolds and their physicochemical effects on partial thickness wound healing. Int J Pharm. 2011;403:73–82. https://doi.org/10.1016/j.ijpharm.2010.10.023.

    Article  CAS  Google Scholar 

  41. Hughes MW, Jiang TX, Plikus MV, Guerrero-Juarez CF, Lin CH, Schafer C, Maxson R, Widelitz RB, Chuong CM. Msx2 supports epidermal competency during wound-induced hair follicle neogenesis. J Invest Dermatol. 2018;138(9):2041–50. https://doi.org/10.1016/j.jid.2018.02.043.

    Article  CAS  Google Scholar 

  42. Chen H, Guo L, Wicks J, Ling C, Zhao X, Yan Y, et al. Quickly promoting angiogenesis by using a DFO-loaded photo-crosslinked gelatin hydrogel for diabetic skin regeneration. J Mater Chem B. 2016;4:3770–81. https://doi.org/10.1039/c6tb00065g.

    Article  CAS  Google Scholar 

Download references

Acknowledgements

This research was technically supported by the Institute of Tissue Banking and Biomaterial Research (ITBBR) of Atomic Energy Research Establishment (AERE), Savar, Dhaka.

Author information

Author notes

  1. Maria Islam and Polash Chandra Karmakar contributed equally to this work.

Authors and Affiliations

  1. Department of Biochemistry and Molecular Biology, Jahangirnagar University, Savar, Dhaka, 1342, Bangladesh

    Maria Islam & Nurul Karim

  2. Institute of Tissue Banking and Biomaterial Research, Atomic Energy Research Establishment (AERE), Savar, Dhaka, 1349, Bangladesh

    Polash Chandra Karmakar,  Tusher-Al-Arafat, Md Arifuzzaman, Naznin Akhtar & S. M. Asaduzzaman

Authors
  1. Maria Islam
    View author publications

    You can also search for this author in PubMed Google Scholar

  2. Polash Chandra Karmakar
    View author publications

    You can also search for this author in PubMed Google Scholar

  3. Tusher-Al-Arafat
    View author publications

    You can also search for this author in PubMed Google Scholar

  4. Md Arifuzzaman
    View author publications

    You can also search for this author in PubMed Google Scholar

  5. Nurul Karim
    View author publications

    You can also search for this author in PubMed Google Scholar

  6. Naznin Akhtar
    View author publications

    You can also search for this author in PubMed Google Scholar

  7. S. M. Asaduzzaman
    View author publications

    You can also search for this author in PubMed Google Scholar

Contributions

Md Arifuzzaman carried out the study conception and design. Maria Islam was involved in the formulation of gels and determining the different physical properties of gels. Polash Chandra Karmakar and Tusher-Al-Arafat were involved in animal experiments, histopathological evaluations, and manuscript writing. Nurul Karim and Naznin Akhter analyzed the data and interpreted the experiments. S. M. Asaduzzaman supervised the writing of the manuscript and revised it for important intellectual content. All authors read and approved the final manuscript.

Corresponding author

Correspondence to S. M. Asaduzzaman.

Ethics declarations

Ethics Approval

Fresh HAMs were collected from Gonoshasthaya Samaj Vittik Medical College Hospital, Savar. Prior to collection of HAM, the written consent of the donor was obtained by following the “Human Organ/Tissue Donation and Transplantation Act, 1999,” which was passed by the National Parliament of the People’s Republic of Bangladesh. The ‘Biosafety, Biosecurity, and Ethical committee of Jahangirnagar University, Savar, Dhaka, Bangladesh recommended and approved the animal model (Wistar Rats) for research purposes following ARRIVE guidelines [Ref no: BBEC, JU/M 2020 (11)5].

Conflicts of Interests

The authors declare no competing interests.

Additional information

Publisher's Note

Springer Nature remains neutral with regard to jurisdictional claims in published maps and institutional affiliations.

Supplementary Information

Below is the link to the electronic supplementary material.

Supplementary file1 (PDF 137 KB)

Rights and permissions

Springer Nature or its licensor (e.g. a society or other partner) holds exclusive rights to this article under a publishing agreement with the author(s) or other rightsholder(s); author self-archiving of the accepted manuscript version of this article is solely governed by the terms of such publishing agreement and applicable law.

Reprints and permissions

About this article

Check for updates. Verify currency and authenticity via CrossMark

Cite this article

Islam, M., Karmakar, P.C., Tusher-Al-Arafat et al. Human Amniotic Membrane and Titanium Dioxide Nanoparticle Derived Gel for Burn Wound Healing in a Rat Model. Regen. Eng. Transl. Med. 9, 249–262 (2023). https://doi.org/10.1007/s40883-022-00280-z

Download citation

  • Received:

  • Revised:

  • Accepted:

  • Published:

  • Issue Date:

  • DOI: https://doi.org/10.1007/s40883-022-00280-z

Keywords

Search

Navigation