Sign In

A subscription to JoVE is required to view this content. Sign in or start your free trial.

In This Article

  • Summary
  • Abstract
  • Introduction
  • Protocol
  • Representative Results
  • Discussion
  • Acknowledgements
  • Materials
  • References
  • Reprints and Permissions

Summary

Liver diseases are induced by many causes that promote fibrosis or cirrhosis. Transplantation is the only option for recovering health. However, given the scarcity of transplantable organs, alternatives must be explored. Our research proposes the implantation of collagen scaffolds in liver tissue from an animal model.

Abstract

Liver diseases are the leading cause of death worldwide. Excessive alcohol consumption, a high-fat diet, and hepatitis C virus infection promote fibrosis, cirrhosis, and/or hepatocellular carcinoma. Liver transplantation is the clinically recommended procedure to improve and extend the life span of patients in advanced disease stages. However, only 10% of transplants are successful, with organ availability, presurgical and postsurgical procedures, and elevated costs directly correlated with that result. Extracellular matrix (ECM) scaffolds have emerged as an alternative for tissue restoration. Biocompatibility and graft acceptance are the main beneficial characteristics of those biomaterials. Although the capacity to restore the size and correct function of the liver has been evaluated in liver hepatectomy models, the use of scaffolds or some kind of support to replace the volume of the extirpated liver mass has not been assessed.

Partial hepatectomy was performed in a rat liver with the xenoimplantation of a collagen matrix scaffold (CMS) from a bovine condyle. Left liver lobe tissue was removed (approximately 40%), and an equal proportion of CMS was surgically implanted. Liver function tests were evaluated before and after the surgical procedure. After days 3, 14, and 21, the animals were euthanized, and macroscopic and histologic evaluations were performed. On days 3 and 14, adipose tissue was observed surrounding the CMS, with no clinical evidence of rejection or infection, as was vessel neoformation and CMS reabsorption at day 21. There was histologic evidence of an insignificant inflammation process and migration of adjacent cells to the CMS, observed with the hematoxylin and eosin (H&E) and Masson's trichrome staining. The CMS was shown to perform well in liver tissue and could be a useful alternative for studying tissue regeneration and repair in chronic liver diseases.

Introduction

The liver is one of the most important organs involved in maintaining homeostasis and protein production1. Unfortunately, liver disease is the leading cause of death worldwide. In advanced stages of liver damage, which include cirrhosis and hepatocellular carcinoma, liver transplantation is the clinically recommended procedure. However, due to the scarcity of donors and the low rate of successful transplants, new techniques in tissue engineering (TE) and regenerative medicine (RM) have been developed2,3.

TE involves the use of stem cells, scaffolds, and growt....

Protocol

The present research was approved by the ethics committee of the School of Medicine (DI/115/2015) at the Universidad Nacional Autónoma de México (UNAM) and the ethics committee of the Hospital General de Mexico (CI/314/15). The institution fulfills all technical specifications for the production, care, and use of laboratory animals and is legally certified by national law (NOM-062-ZOO-1999). Male Wistar rats weighing 150-250 g (6-8 weeks old) were obtained from the Laboratory Animal Facility of the School of Me.......

Representative Results

Bone demineralization affects the mechanical properties of CMS without altering the original shape or interconnection of its pores. CMS can have any shape, and therefore, can be adjusted to the size and shape of the selected organ or tissue19. In the present protocol, we used a triangular CMS (Figure 1A-D). A rat model was used to evaluate the regenerative capacity of the CMS xenoimplant in the liver. Although the liv.......

Discussion

Organ transplantation is the mainstay of treatment in patients with liver fibrosis or cirrhosis. A few patients benefit from this procedure, making it necessary to provide therapeutic alternatives for patients on the waiting list. Tissue engineering is a promising strategy that employs scaffolds and cells with regenerative potential2,4,13. The removal of a portion of the liver is a critical step in this procedure because of the .......

Acknowledgements

The authors wish to thank the personnel of the Laboratory Animal Facility of the Experimental Medicine Unit, Nurse Carolina Baños G. for technical and surgical support, Marco E. Gudiño Z. for support in microphotographs, and Erick Apo for support in liver histology. The National Council supported this research for Science and Technology (CONACyT), grant number SALUD-2016-272579 and the PAPIIT-UNAM TA200515.

....

Materials

NameCompanyCatalog NumberComments
Anionic detergentAlconoxZ273228
Biopsy cassettesLeica3802453
Camera DMXNikonDXM1200F
CentrifugeEppendorf5424
Chlorhexidine gluconate 4%BD372412
Cover glasses 25 mm x 40 mmCorning2980-224
EosinSigma-Aldrich200-MCAS 17372-87-1
Ethyl alcohol, pureSigma-Aldrich459836CAS 64-17-5
Flunixine meglumideMSDQ-0273-035
Glass slides 75 mm x 25 mmCorning101081022
HematoxylinMerckH9627CAS 571-28-2
Hydrochloric acid 37%Merck339253CAS 7647-01-0
KetaminePisa agropecuariaQ-7833-028
Light microscopyNikonMicrophoto-FXA
Microtainer yellow capeBeckton Dickinson365967
MicrotomeLeicaRM2125
Model animal: Wistar ratsUniversidad Nacional Autónoma de México
Nylon 3-0 (Dermalon)Covidien1750-41
Polypropylene 7-0AtramatSE867/2-60
Povidone-iodine10% cutaneous solutionDiafra SA de CV1.37E+86
Scaning electronic microscopyZeissDSM-950
Sodium hydroxide, pelletsJ. T. Baker3722-01CAS 1310-73-2
Software ACT-1NikonVer 2.70
Stereoscopy macroscopyLeicaEZ4Stereo 8X-35X
Sterrad 100SJohnson and Johnson99970
Surgipath paraplastLeica39601006
Synringe of 1 mL with needle (27G x 13 mm)SensiMedicalLAN-078-077
Tissue Processor (Histokinette)LeicaTP1020
Tissue-Tek TEC 5 (Tissue embedder)Sakura Finetek USA5229
Trichrome stain kitSigma-AldrichHT15
Unicell DxC600 AnalyzerBeckman CoulterBC 200-10
XylazinePisa agropecuariaQ-7833-099
XyleneSigma-Aldrich534056CAS 1330-20-7

References

  1. Li, N., Hua, J. Immune cells in liver regeneration. Oncotarget. 8 (2), 3628-3639 (2017).
  2. Langer, R., Vacanti, J. Tissue Engineering. Science. 260 (5110), 920-926 (1993).
  3. Lee, H., et al.

Explore More Articles

3D Collagen MatrixLiver RegenerationTissue EngineeringHepatectomyScaffold ImplantationWound HealingSurgical ProcedureWistar RatPolypropylene SuturesAnesthesiaAnalgesic

This article has been published

Video Coming Soon

JoVE Logo

Privacy

Terms of Use

Policies

Research

Education

ABOUT JoVE

Copyright © 2024 MyJoVE Corporation. All rights reserved