Tracheal regeneration with acellular human amniotic membrane and 15-deoxy-∆12,14prostaglandinj2 nanoparticles in a rabbit model

The treatment of tracheal pathologies remains challenging.Nanotechnology allows adding substances to decellularized human amniotic membrane (DHAM), such as 15-Deoxy-∆12,14ProstaglandinJ2 nanoparticles (15D-PGJ2-NC).This study performed a tracheotomy in rabbits randomized into three groups.The tissue repair process was evaluated when treated with DHAM associated or not with 15D-PGJ2-NC.The average of the area in the control group was 54.76% smaller than DHAM group and 41.98% smaller than DHAM + 15D-PGJ2-NC group (p=0.004 for both).The DHAM + 15D-PGJ2-NC group had significantly more immature cartilage (p=0.015).DHAM impregnated with 15D-PGJ2-NC could provide support for the healing of the tracheal defect and may prevent reduction of its lumen.

Key words
15D-PGJ2; amniotic membrane; healing; tracheal regeneration; tracheotomy

Authorship

ANNA FLAVIA R.S. MIGGIOLARO

acquisition

analysis

interpretation of data

Pontifical Catholic University of Paraná (PUC-PR), Experimental Laboratory of the Institute of Biology and Health Sciences, R. Imaculada Conceição, 1155, Prado Velho, 80215-901 Curitiba, PR, BrazilPontifical Catholic University of Paraná (PUC-PR)BrazilCuritiba, PR, BrazilPontifical Catholic University of Paraná (PUC-PR), Experimental Laboratory of the Institute of Biology and Health Sciences, R. Imaculada Conceição, 1155, Prado Velho, 80215-901 Curitiba, PR, Brazil

https://orcid.org/0000-0002-8497-400X

ALINE L. TAKEJIMA

acquisition

analysis

interpretation of data

drafting

revising the manuscript

final approval of the version to be published

https://orcid.org/0000-0002-2608-0171

JULIO CESAR FRANCISCO

acquisition

analysis

interpretation of data

https://orcid.org/0000-0003-1970-6399

LÚCIA DE NORONHA

acquisition

analysis

interpretation of data

https://orcid.org/0000-0003-0310-7164

MILKA L. TAKEJIMA

acquisition

analysis

interpretation of data

drafting

revising the manuscript

final approval of the version to be published

Mackenzie Evangelical University Hospital, Plastic Surgery and Burns Department, Al. Augusto Stellfeld, 1908, Bigorrilho, 80730-150 Curitiba, PR, BrazilMackenzie Evangelical University HospitalBrazilCuritiba, PR, BrazilMackenzie Evangelical University Hospital, Plastic Surgery and Burns Department, Al. Augusto Stellfeld, 1908, Bigorrilho, 80730-150 Curitiba, PR, Brazil

https://orcid.org/0000-0002-0266-4076

KATIA FOLTZ

acquisition

analysis

interpretation of data

https://orcid.org/0000-0002-9188-7262

ALOYSIO ENCK

acquisition

analysis

interpretation of data

https://orcid.org/0000-0003-0458-5707

LUANA TANNOUS

acquisition

analysis

interpretation of data

https://orcid.org/0000-0003-3559-1712

ROSSANA B. SIMEONI

acquisition

analysis

interpretation of data

https://orcid.org/0000-0002-9976-5809

PRISCILA E.F. STRICKER

drafting

revising the manuscript

final approval of the version to be published

Institute Pelé Pequeno Príncipe, Advanced Therapy and Cellular Biotechnology in Regenerative Medicine Department Research, Av. Silva Jardim, 1632, Água Verde, 80250-060 Curitiba, PR, BrazilInstitute Pelé Pequeno PríncipeBrazilCuritiba, PR, BrazilInstitute Pelé Pequeno Príncipe, Advanced Therapy and Cellular Biotechnology in Regenerative Medicine Department Research, Av. Silva Jardim, 1632, Água Verde, 80250-060 Curitiba, PR, Brazil

https://orcid.org/0000-0002-2582-137X

KATHERINE A.T. CARVALHO

drafting

revising the manuscript

final approval of the version to be published

https://orcid.org/0000-0003-4860-2855

LUIZ CESAR GUARITA-SOUZA

drafting

revising the manuscript

final approval of the version to be published

https://orcid.org/0000-0003-2781-9705

Correspondence to: Aline Luri Takejima E-mail: alinetakejima@yahoo.com.br

SCIMAGO INSTITUTIONS RANKINGS

Figures | Tables

Figures (3)
Tables (3)

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Figure 1
The tracheal defect. a. Image of the surgical site, in which the arrowhead shows the tracheal defect by the tracheotomy in an animal in the control group. b. Image of the surgical site showing the tracheal defect covered by the amniotic membrane (arrowhead), in an animal in the DHAM group. c. Image of the trachea after resection of an animal in the DHAM + 15D-PGJ2-NC group. The arrowhead points to the area of the previous tracheotomy.

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Figure 2
Comparison of the cartilaginous tissue (aggrecan) between groups. AM, Amniotic Membrane; PGJ2, ProstaglandinJ2.

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Figure 3
Photographs of the performed stains and immunohistochemistry. a. Hematoxylin-Eosin (HE) staining of the lesion area (arrowhead) in a control group. b. HE Stain of the lesion in the DHAM group in which the arrowhead points to the remaining tissue of the amniotic membrane. c. HE Stain of the lesion in the DHAM + 15D-PGJ2-NC group in which the arrowhead points to the remaining tissue from the amniotic membrane added to 15D-PGJ2-NC, and the double arrowhead to the inflammatory infiltrate. d. Image of Sirius Red stain, at the region of the lesion, in the control group. The red images represent type I collagen (arrowhead), and the green images show type III collagen (double arrowhead). e. Collagen type III (arrowhead) and type I (double arrowhead) of the DHAM group. f. Collagen type III (arrowhead) and type I (double arrowhead) of the DHAM + 15D-PGJ2-NC group. g. Anti-aggrecan immunohistochemistry (40x magnification) in a control group animal. The arrowhead points to the aggrecan (ocher color) and the double arrowhead points to mature cartilage. h. Anti-aggrecan immunohistochemistry (10x magnification) in an DHAM group animal. The arrowhead points to the injury site. i. Anti-aggrecan immunohistochemistry in DHAM + 15D-PGJ2-NC group animal. The arrowhead points to the injury site.

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Table I
Comparison of the area at the site of the tracheal defect between groups, two by two.

Thumbnail

Table II
Comparison of averages indices of histological findings and characterization of the inflammatory profile.

Thumbnail

Table III
Comparison of the cartilaginous tissue (aggrecan) between groups, two by two.

Figure 1 The tracheal defect. a. Image of the surgical site, in which the arrowhead shows the tracheal defect by the tracheotomy in an animal in the control group. b. Image of the surgical site showing the tracheal defect covered by the amniotic membrane (arrowhead), in an animal in the DHAM group. c. Image of the trachea after resection of an animal in the DHAM + 15D-PGJ2-NC group. The arrowhead points to the area of the previous tracheotomy.

Figure 2 Comparison of the cartilaginous tissue (aggrecan) between groups. AM, Amniotic Membrane; PGJ2, ProstaglandinJ2.

Figure 3 Photographs of the performed stains and immunohistochemistry. a. Hematoxylin-Eosin (HE) staining of the lesion area (arrowhead) in a control group. b. HE Stain of the lesion in the DHAM group in which the arrowhead points to the remaining tissue of the amniotic membrane. c. HE Stain of the lesion in the DHAM + 15D-PGJ2-NC group in which the arrowhead points to the remaining tissue from the amniotic membrane added to 15D-PGJ2-NC, and the double arrowhead to the inflammatory infiltrate. d. Image of Sirius Red stain, at the region of the lesion, in the control group. The red images represent type I collagen (arrowhead), and the green images show type III collagen (double arrowhead). e. Collagen type III (arrowhead) and type I (double arrowhead) of the DHAM group. f. Collagen type III (arrowhead) and type I (double arrowhead) of the DHAM + 15D-PGJ2-NC group. g. Anti-aggrecan immunohistochemistry (40x magnification) in a control group animal. The arrowhead points to the aggrecan (ocher color) and the double arrowhead points to mature cartilage. h. Anti-aggrecan immunohistochemistry (10x magnification) in an DHAM group animal. The arrowhead points to the injury site. i. Anti-aggrecan immunohistochemistry in DHAM + 15D-PGJ2-NC group animal. The arrowhead points to the injury site.

Table I Comparison of the area at the site of the tracheal defect between groups, two by two.

Compared groups	p
Control x DHAM	<0.001
Control x DHAM+15D-PGJ2-NC	0.005
DHAM x DHAM+15D-PGJ2-NC	0.111

DHAM, Decellularized Human Amniotic Membrane; 15D-PGJ2-NC, 15-Deoxy-∆12,14ProstaglandinJ2 nanoparticles.

Table II Comparison of averages indices of histological findings and characterization of the inflammatory profile.

Histological Indexes	Average Index Inflammation
Control	+3.6 Chronic
DHAM	+2.8 Chronic
DHAM+15D-PGJ2-NC	+2.0 Subacute

DHAM, Decellularized Human Amniotic Membrane; 15D-PGJ2-NC, 15-Deoxy-∆12,14ProstaglandinJ2 nanoparticles.

Table III Comparison of the cartilaginous tissue (aggrecan) between groups, two by two.

Compared Groups	p
Control x DHAM	1
Control x DHAM+15D-PGJ2-NC	0.048
DHAM x DHAM+15D-PGJ2-NC	0.007

DHAM, Decellularized Human Amniotic Membrane; 15D-PGJ2-NC, 15-Deoxy-∆12,14ProstaglandinJ2 nanoparticles.

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Tracheal regeneration with acellular human amniotic membrane and 15-deoxy-∆12,14prostaglandinj2 nanoparticles in a rabbit model

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[1] Correspondence to: Aline Luri Takejima E-mail: alinetakejima@yahoo.com.br