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A Histological and Biomechanical Analysis of Human Acellular Dermis (HAD) Created Using a Novel Processing and Preservation Technique

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Indian Journal of Orthopaedics Aims and scope Submit manuscript

Abstract

Background

Large and complex defects requiring reconstruction are challenging for orthopaedic surgeons. The use of human acellular dermal (HAD) matrices to augment large soft tissue defects such as those seen in massive rotator cuff tears, knee extensor mechanism failures and neglected Tendo-Achilles tears has proven to be a valuable tool in surgeons reconstructive armamentarium. Different methods for allograft decellularization and preservation alter the native properties of the scaffold. Traditional processing and preservation methods have shown to have drawbacks that preclude its widespread use. Some of the common issues include inferior biomechanical properties, the risk of rejection, limited customization, difficulty in storing and transporting, the requirement of pre-operative preparation, and last but not the least increased cost.

Methods

We describe a novel processing and preservation method utilizing a two-step non-denaturing decellularization method coupled with preservation using a water-sequestering agent (glycerol) to remove immunogenic components while retaining biomechanical properties. The efficiency of this novel process was compared with the traditional freeze-drying method and verified by histological evaluation and biomechanical strength analysis.

Results

The absence of cellular components and matrix integrity in hematoxylin and eosin-stained glycerol-preserved HAD (gly-HAD) samples compared to freeze-dried HAD (FD-HAD) demonstrated effective yet gentle decellularization. Biomechanical strength analysis revealed that gly-HADs are stronger with an ultimate tensile load to the failure strength of 210 N compared to FD-HAD (124N). The gly-HADs were found to have an optimal suture–retention strength of 126 N. Finally, sterility testing of the resultant grafts was checked to ensure a sterility assurance level of 10−6 to establish implantability.

Conclusion

The novel processing and preservation technique is described in this paper to create a Human Acellular Dermis with higher biomechanical strength and superior histological characteristics. The processing and preservation technique ensured high sterility assurance levels to establish implantability.

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References

  1. Skowron, K., Bauza-Kaszewska, J., Kraszewska, Z., Wiktorczyk-Kapischke, N., Grudlewska-Buda, K., Kwiecińska-Piróg, J., Wałecka-Zacharska, E., Radtke, L., & Gospodarek-Komkowska, E. (2021). Human skin microbiome: Impact of intrinsic and extrinsic factors on skin microbiota. Microorganisms, 9(3), 543. https://doi.org/10.3390/microorganisms9030543

    Article  CAS  PubMed  PubMed Central  Google Scholar 

  2. Brown, T. M., & Krishnamurthy, K. (2024). Histology, dermis. [Updated 2022 Nov 14]. In StatPearls [Internet]. StatPearls Publishing. Available from: https://www.ncbi.nlm.nih.gov/books/NBK535346

  3. Boháč, M., Danišovič, Ľ, Koller, J., Dragúňová, J., & Varga, I. (2018). What happens to an acellular dermal matrix after implantation in the human body? A histological and electron microscopic study. European Journal of Histochemistry, 62(1), 2873. https://doi.org/10.4081/ejh.2018.2873

    Article  CAS  PubMed  PubMed Central  Google Scholar 

  4. Petrie, K. A., Cox, C., Becker, B. C., & MacKay, B. J. (2022). Clinical applications of acellular dermal matrices: A review. Scars, Burns & Healing., 8, 205951312110383. https://doi.org/10.1177/20595131211038313

    Article  Google Scholar 

  5. (2011). The transplantation of human organs and tissues act, 1994. Gazette of India, Ext., Pt. II, S. 1.

  6. Guidelines for Tissue Banking (2021) rottosottokem. in. Available at: https://rottosottokem.in/downloads/SOTTO%20Tissue%20Guidelines.pdf

  7. Bush, K., & Gertzman, A. (2016). Process development and manufacturing of human and animal acellular dermal matrices. In M. Z. Albanna & J. H. Holmes IV (Eds.), Skin tissue engineering and regenerative medicine (pp. 83–108). Academic Press.

    Chapter  Google Scholar 

  8. Crapo, P. M., Gilbert, T. W., & Badylak, S. F. (2011). An overview of tissue and whole organ decellularization processes. Biomaterials, 32(12), 3233–3243. https://doi.org/10.1016/j.biomaterials.2011.01.057

    Article  CAS  PubMed  PubMed Central  Google Scholar 

  9. Mauro, C. (2006). Safety issues for musculoskeletal allografts. In Clinical sports medicine (pp. 111–116). https://doi.org/10.1016/b978-032302588-1.50015-4

  10. CFR - Code of Federal Regulations Title 21. (n.d.). https://www.accessdata.fda.gov/scripts/cdrh/cfdocs/cfcfr/cfrsearch.cfm?fr=182.1320

  11. Samsell, B., Softic, D., Qin, X., McLean, J., Sohoni, P., Gonzales, K., & Moore, M. A. (2019). Preservation of allograft bone using a glycerol solution: A compilation of original preclinical research. Biomaterials Research, 13(23), 5. https://doi.org/10.1186/s40824-019-0154-1

    Article  Google Scholar 

  12. Feldman, A. T., & Wolfe, D. (2014). Tissue processing and hematoxylin and eosin staining. In C. Day (Ed.), Histopathology. Methods in molecular biology. (Vol. 1180). Humana Press. https://doi.org/10.1007/978-1-4939-1050-2_3

    Chapter  Google Scholar 

  13. Singh, R., Singh, D., & Singh, A. (2016). Radiation sterilization of tissue allografts: A review. World Journal of Radiology, 8(4), 355–369. https://doi.org/10.4329/wjr.v8.i4.355

    Article  PubMed  PubMed Central  Google Scholar 

  14. Yusof, N., Shamsudin, A. R., Mohamad, H., Hassan, A., Yong, A. C., & Rahman, M. N. F. A. (2005). Bioburden estimation in relation to tissue product quality and radiation dose validation. In Sterilisation of tissues using ionising radiations [Internet] (pp. 319–329). Woodhead Publishing. https://doi.org/10.1533/9781845690779.5.319

  15. ISO 11737-1. (2006). Sterilization of medical devices—Microbiological methods—Part 1: Determination of a population of microorganisms on products.

  16. Gouk, S. S., Lim, T. G., Teoh, S., & Sun, W. Q. (2007). Alterations of human acellular tissue matrix by gamma irradiation: Histology, biomechanical property, stability, in vitro cell repopulation, and remodeling. Journal of Biomedical Materials Research Part B: Applied Biomaterials., 84B(1), 205–217. https://doi.org/10.1002/jbm.b.30862

    Article  CAS  Google Scholar 

  17. ISO 11737-2. Sterilization of medical devices—Microbiological methods—Part 2: Tests of sterility performed in the validation of a sterilization process. International Standard Organization. ISO 11737-2:2009

  18. ISO 11137-2. Sterilization of healthcare products—Radiation—Part 2: Establishing the sterilization dose, Method VDmax—Substantiation of 25 kGy or 15 kGy as the sterilization dose. International Standard Organization. ISO 11137-2:2013(en)

  19. Barber, F. A., Herbert, M. A., & Coons, D. A. (2006). Tendon augmentation grafts: Biomechanical failure loads and failure patterns. Arthroscopy: The Journal of Arthroscopic & Related Surgery., 22(5), 534–538. https://doi.org/10.1016/j.arthro.2005.12.021

    Article  Google Scholar 

  20. Marsit, N. M., Dwejen, S., Saad, I., Abdalla, S., Shaab, A., Salem, S., Khanfas, E., Hasan, A. R., Mansur, M., & Sammad, M. A. (2014). Substantiation of 25 kGy radiation sterilization dose for banked air dried amniotic membrane and evaluation of personnel skill in influencing finished product bioburden. Cell and Tissue Banking., 15(4), 603–611. https://doi.org/10.1007/s10561-014-9433-1

    Article  PubMed  PubMed Central  Google Scholar 

  21. Schultz, G. S., Davidson, J. M., Kirsner, R. S., Bornstein, P., & Herman, I. M. (2011). Dynamic reciprocity in the wound microenvironment. Wound Repair and Regeneration, 19(2), 134–148. https://doi.org/10.1111/j.1524-475X.2011.00673.x

    Article  PubMed  PubMed Central  Google Scholar 

  22. Barber, F. A., & Aziz-Jacobo, J. (2009). Biomechanical testing of commercially available soft-tissue augmentation materials. Arthroscopy: The Journal of Arthroscopic & Related Surgery, 25(11), 1233–1239. https://doi.org/10.1016/j.arthro.2009.05.012

    Article  Google Scholar 

  23. Wong, I., Sparavalo, S., King, J. P., & Coady, C. M. (2021). Bridging allograft reconstruction is superior to maximal repair for the treatment of chronic, massive rotator cuff tears: Results of a prospective, randomized controlled trial. American Journal of Sports Medicine, 49(12), 3173–3183. https://doi.org/10.1177/03635465211039846

    Article  PubMed  Google Scholar 

  24. Bushnell, B. D., Piller, C. P., Hicks, J. S., Jarvis, B., Jarvis, R. C., & Baudier, R. S. Rotator cuff repair with a bioinductive bovine collagen implant has a low incidence of post-operative stiffness: review of 406 shoulders. Poster presented at the Annual Academy of Orthopaedics Surgeons (AAOS Annual Congress; March 7–11 2023; Las Vegas, Nevada, USA.

  25. Pandey, R., Tafazal, S., Shyamsundar, S., Modi, A., & Singh, H. P. (2017). Outcome of partial repair of massive rotator cuff tears with and without human tissue allograft bridging repair. Shoulder Elbow., 9(1), 23–30. https://doi.org/10.1177/1758573216665114

    Article  PubMed  Google Scholar 

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Acknowledgements

The authors wish to acknowledge the contributions of Dr. Rajesh Dharmarajan to this article.

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Authors and Affiliations

  1. Novo Tissue Bank and Research Centre, Mumbai, India

    Damini Shah, Madhu Rathod & Anjali Tiwari

  2. Sir H N Reliance Foundation Hospital, Girgaum, Mumbai, Maharashtra, 400004, India

    Abhishek Kini

  3. Sona Medical Centre & Consultant Orthopaedic Surgeon Saifee Hospital, Jaslok Hospital and Research Centre, Breach Candy Hospital Trust, Mumbai, India

    Prasad Bhagunde

  4. Department of Orthopaedic Surgery, Sir H N Reliance Foundation Hospital, Girgaum, Mumbai, Maharashtra, 400004, India

    Vaibhav Bagaria

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  1. Damini Shah
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  2. Madhu Rathod
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  3. Anjali Tiwari
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  4. Abhishek Kini
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  5. Prasad Bhagunde
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Contributions

Damini Shah: conceptualization, formal analysis, investigation, methodology, resources, validation, visualization, writing—original draft. Madhu Rathod: resources. Anjali Tiwari: writing—review and editing. Abhishek Kini: project administration, supervision, resources. Prasad Bhagunde: conceptualization, supervision, resources. Vaibhav Bagaria: conceptualization, writing—review and editing.

Corresponding author

Correspondence to Vaibhav Bagaria.

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Conflict of interest

DS, MR, and AT are employees of Novo Tissue Bank and Research Centre Pvt. Ltd.

Ethical Standard Statement

This article does not contain any studies with human or animal subjects performed by the any of the authors.

Human and Animal Rights

This article does not contain studies with human participants or animals by any of the authors. Human donated tissues were made available by the Novo Tissue Bank and Research Centre Pvt. Ltd. Release of tissues for improving in house processes and technique was in accordance with family informed consent.

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For this type of study informed consent is not required.

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Shah, D., Rathod, M., Tiwari, A. et al. A Histological and Biomechanical Analysis of Human Acellular Dermis (HAD) Created Using a Novel Processing and Preservation Technique. JOIO (2024). https://doi.org/10.1007/s43465-024-01181-9

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