Biocompatiblibility analysis of the decellularized bovine pericardium

2020 ◽  
Vol 8 (2) ◽  
Author(s):  
A. Sokol ◽  
◽  
D. Grekov ◽  
G. Yemets ◽  
O. Galkin ◽  
...  

The decellularized bovine pericardium and its potential use as a natural scaffold is a promising approach in the field of tissue engineering and regenerative medicine. The reaction of the host toward decellularized scaffolds depends on their biocompatibility, which should be satisfied being before applied in clinical use. Purpose: to evaluate the biocompatibility of the extracellular matrices, which were decellularized by trypsin enzyme and anionic sodium dodecyl sulfate (SDS) detergent. Material and methods. Pericardial sacs were acquired from 12-18 months’ age bulls. Tissue decellularization was performed by using 0.25 % Trypsin solution and 1 % ionic SDS for group I and 0.1 % SDS for group II samples. The implantation was performed on Wistar rats. The tissue samples were stained with hematoxylin & eosin, Congo red and Masson's Trichrome for histological analysis. Results. In group 1 in 3 months after subcutaneous implantation in rats we noticed the inflammation in surrounding tissue and degradation of the implant. Under the same conditions in animals of group 2 implant replacement with growing immature connective tissue was noted. Bio-implant of this group did not degrade, moreover it's integrated to the tissues of experimental rats. Conclusion. Our results showed that decellularized bovine pericardium by 0.1 % SDS can become an alternative material for tissue engineering and has the potential for further use in human surgery.

Author(s):  
Mohammad Amin Keshvari ◽  
Alireza Afshar ◽  
Sajad Daneshi ◽  
Arezoo Khoradmehr ◽  
Mandana Baghban ◽  
...  

Chronic kidney diseases (CKD) and end stage renal disease (ESRD) are growing threats worldwide. Tissue engineering is a new hope to surpass the current limitations such as the shortage of donor. To do so, the first step would be fabrication of an intact decellularized kidney scaffold. In the current study, an automatic decellularization device was developed to perfuse and decellularize male rats' kidneys using both sodium lauryl ether sulfate (SLES) and sodium dodecyl sulfate (SDS) and to compare their efficacy in kidney decellularization and post-transplantation angiogenesis. After anesthesia, kidneys were perfused with either 1% SDS solution for 4 h or 1% SLES solution for 6 h. The decellularized scaffolds were stained with hematoxylin and eosin (H&E), periodic acid Schiff (PAS), Masson’s trichrome, and alcian blue to determine cell removal and glycogen, collagen and glycosaminoglycans (GAGs) contents, respectively. Moreover, scanning electron microscopy (SEM) was performed to evaluate the cell removal and preservation of microarchitecture of both SDS and SLES scaffolds. Additionally, DNA quantification assay was applied for all groups in order to measure residual DNA in the scaffolds and normal kidney. In order to demonstrate biocompatibility and bioactivity of the decellularized scaffolds, allotransplantation was performed in back muscle and angiogenesis was evaluated. Complete cell removal in both SLES and SDS groups was observed in SEM and DNA quantification assays. Moreover, the extracellular matrix (ECM) architecture of rat kidney in the SLES group was significantly preservation better than the SDS group was shown. The formation of blood capillaries and vessels were observed in the kidney allotransplantations in both SLES and SDS decellularized kidneys. In conclusion, we demonstrated that both SLES and SDS could be promising tools in kidney tissue engineering. The better preservation of ECM than SDS, introduces SLES as the solvent of choice for kidney decellularization. ¬¬


2019 ◽  
Vol 20 (20) ◽  
pp. 5145 ◽  
Author(s):  
Vanessa Pérez-Silos ◽  
Nidia K. Moncada-Saucedo ◽  
Víctor Peña-Martínez ◽  
Jorge Lara-Arias ◽  
Iván A. Marino-Martínez ◽  
...  

In cartilage tissue engineering, biphasic scaffolds (BSs) have been designed not only to influence the recapitulation of the osteochondral architecture but also to take advantage of the healing ability of bone, promoting the implant’s integration with the surrounding tissue and then bone restoration and cartilage regeneration. This study reports the development and characterization of a BS based on the assembly of a cartilage phase constituted by fibroin biofunctionalyzed with a bovine cartilage matrix, cellularized with differentiated autologous pre-chondrocytes and well attached to a bone phase (decellularized bovine bone) to promote cartilage regeneration in a model of joint damage in pigs. BSs were assembled by fibroin crystallization with methanol, and the mechanical features and histological architectures were evaluated. The scaffolds were cellularized and matured for 12 days, then implanted into an osteochondral defect in a porcine model (n = 4). Three treatments were applied per knee: Group I, monophasic cellular scaffold (single chondral phase); group II (BS), cellularized only in the chondral phase; and in order to study the influence of the cellularization of the bone phase, Group III was cellularized in chondral phases and a bone phase, with autologous osteoblasts being included. After 8 weeks of surgery, the integration and regeneration tissues were analyzed via a histology and immunohistochemistry evaluation. The mechanical assessment showed that the acellular BSs reached a Young’s modulus of 805.01 kPa, similar to native cartilage. In vitro biological studies revealed the chondroinductive ability of the BSs, evidenced by an increase in sulfated glycosaminoglycans and type II collagen, both secreted by the chondrocytes cultured on the scaffold during 28 days. No evidence of adverse or inflammatory reactions was observed in the in vivo trial; however, in Group I, the defects were not reconstructed. In Groups II and III, a good integration of the implant with the surrounding tissue was observed. Defects in group II were fulfilled via hyaline cartilage and normal bone. Group III defects showed fibrous repair tissue. In conclusion, our findings demonstrated the efficacy of a biphasic and bioactive scaffold based on silk fibroin and cellularized only in the chondral phase, which entwined chondroinductive features and a biomechanical capability with an appropriate integration with the surrounding tissue, representing a promising alternative for osteochondral tissue-engineering applications.


Author(s):  
H. J. Finol ◽  
M. E. Correa ◽  
L.A. Sosa ◽  
A. Márquez ◽  
N.L. Díaz

In classical oncological literature two mechanisms for tissue aggression in patients with cancer have been described. The first is the progressive invasion, infiltration and destruction of tissues surrounding primary malignant tumor or their metastases; the other includes alterations produced in remote sites that are not directly affected by any focus of disease, the so called paraneoplastic phenomenon. The non-invaded tissue which surrounds a primary malignant tumor or its metastases has been usually considered a normal tissue . In this work we describe the ultrastructural changes observed in hepatocytes located next to metastases from diverse malignant tumors.Hepatic biopsies were obtained surgically in patients with different malignant tumors which metatastized in liver. Biopsies included tumor mass, the zone of macroscopic contact between the tumor and the surrounding tissue, and the tissue adjacent to the tumor but outside the macroscopic area of infiltration. The patients (n = 5), 36–75 years old, presented different tumors including rhabdomyosarcoma, leiomyosarcoma, pancreas carcinoma, biliar duct carcinoma and colon carcinoma. Tissue samples were processed with routine techniques for transmission electron microscopy and observed in a Hitachi H-500 electron microscope.


2021 ◽  
Vol 12 ◽  
pp. 204173142098752
Author(s):  
Nadiah S Sulaiman ◽  
Andrew R Bond ◽  
Vito D Bruno ◽  
John Joseph ◽  
Jason L Johnson ◽  
...  

Human saphenous vein (hSV) and synthetic grafts are commonly used conduits in vascular grafting, despite high failure rates. Decellularising hSVs (D-hSVs) to produce vascular scaffolds might be an effective alternative. We assessed the effectiveness of a detergent-based method using 0% to 1% sodium dodecyl sulphate (SDS) to decellularise hSV. Decellularisation effectiveness was measured in vitro by nuclear counting, DNA content, residual cell viability, extracellular matrix integrity and mechanical strength. Cytotoxicity was assessed on human and porcine cells. The most effective SDS concentration was used to prepare D-hSV grafts that underwent preliminary in vivo testing using a porcine carotid artery replacement model. Effective decellularisation was achieved with 0.01% SDS, and D-hSVs were biocompatible after seeding. In vivo xeno-transplantation confirmed excellent mechanical strength and biocompatibility with recruitment of host cells without mechanical failure, and a 50% patency rate at 4-weeks. We have developed a simple biocompatible methodology to effectively decellularise hSVs. This could enhance vascular tissue engineering toward future clinical applications.


Blood ◽  
1990 ◽  
Vol 75 (4) ◽  
pp. 874-880 ◽  
Author(s):  
AD D'Andrea ◽  
PJ Szklut ◽  
HF Lodish ◽  
EM Alderman

Abstract We have generated four high affinity monoclonal antibodies (MoAbs) to recombinant human erythropoietin (EPO). All four MoAbs immunoprecipitate radioiodinated native EPO, and the concentrations of MoAbs required for maximum binding range from 10 nmol/L to 100 nmol/L. Two MoAbs, designated Group I MoAbs, bind to an epitope within the N- terminal 20 amino acids of EPO and also immunoprecipitate sodium dodecyl sulfate (SDS)-denatured EPO. Two other MoAbs (Group II MoAbs) do not immunoprecipitate SDS-denatured EPO and do not bind to any of the eight endo C fragments of EPO. We first used murine erythroleukemia (MEL) cells to test the MoAbs for inhibition of EPO-receptor binding. MEL cells, although unresponsive to EPO, express 760 high affinity receptors for EPO per cell (Kd = 0.24 nmol/L). To assay our MoAbs, MEL cells were grown as monolayers on fibronectin-coated Petri dishes and incubated at 4 degrees C with radioiodinated EPO. Group I MoAbs do not inhibit binding of radioiodinated EPO to the MEL EPO-receptor, but Group II MoAbs do inhibit binding in a dose-dependent manner. We next examined the neutralization of EPO bioactivity by our MoAbs, using EPO- dependent cell line. Only Group II MoAbs inhibit a newly developed EPO- dependent cell growth, demonstrating that inhibition of EPO-receptor binding correlates with neutralization of EPO bioactivity.


2017 ◽  
Vol 28 (16) ◽  
pp. 1966-1983 ◽  
Author(s):  
Yamina Boukari ◽  
Omar Qutachi ◽  
David J. Scurr ◽  
Andrew P. Morris ◽  
Stephen W. Doughty ◽  
...  

2010 ◽  
Vol 25 (4) ◽  
pp. 318-321 ◽  
Author(s):  
Otoni Moreira Gomes ◽  
Geraldo Brasileiro Filho ◽  
Luiz Alberto Bomjardim Porto ◽  
Pedro Henrique de Lima Prata ◽  
Rafael de Mattos Paixão

PURPOSE: To evaluate the histopathology alterations of the intestinal mucosa of rabbits submitted to different times of mesenteric artery ischemia and reperfusion with and without celiac artery collateral circulation supply. METHODS: Two groups of eight male New Zealand white rabbits (weight 2.2-3.5 kg) were used in this study. In the Group 1 animals, the proximal mesenteric artery was occluded for 60 min with an atraumatic vascular clamp, followed by reperfusion for 60 min. In the Group 2 animals the small bowel and mesentery were cut 30cm and 60cm far from the gastroduodenal pyloric transition before the proximal mesenteric artery occlusion. Small bowel biopsies were obtained before ischemia (control), after 30 min and 60 min of mesenteric ischemia and at 30 and 60 min. of mesenteric artery reperfusion. RESULTS: In the Group I animals, the followings histopathology grade results were observed: t1, mean 0.4 + 0.29; t2, mean 1.9 ± 0.38; t3, 1.9 ± 0.33; t4, 1.2 ± 0.36 and t5, 1.2 ± 0.32. Differences between t0 and t2 and between t3 and t4 were statistically significant (p<0.05). Differences between t2 and t3 and t4 and t5 were not significant (p>0.5). In the Group II animals, it was observed: t1, mean 1.6 ± 0.33; t2, 2.4 ± 0.36; t3, 3.0 ± 0.35; t4 3.4 ± 0.31; t5, 3 ± 031. Differences between t0 and t1, t1 and t2, and t2 and t3 were significant (p<0.05). Differences between histopathology grades results of samples t1 to t5 in Group 1 and 2 were statistically significant (p<0.5). CONCLUSION: Microscopic examination of the biopsies revealed significant evidence of worse small bowel wall ischemia-reperfusion lesions by exclusion of the celiac artery collateral circulation supply.


2016 ◽  
Vol 11 (3) ◽  
Author(s):  
Muhammad A Ahad ◽  
Mohammad Rashad Qamar ◽  
Sameh K Hindi ◽  
Martin N Kid

Purpose: To study the effect of anterior capsule polishing during phacoemulsification on the incidence of post operative YAG laser capsulotomy. Method: A retrospective controlled study of 159 patients who underwent uncomplicated phacoemulsification with anterior capsular polishing between October 1998 and March 2000. 169 age matched patients who underwent phacoemulsification but without anterior capsule polishing served as controls. Main outcome measure: Incidence of visually significant YAG capsulotomy, which improved the Snellen acuity for more than 1 line or at least 1 line with subjective improvements in symptoms. Results: 2.51 % of patients with anterior capsular polishing (Group 1) had YAG capsulotomy compared to 7.1% of patients in control group at one year. However, after two years, 11.3% of patients in Group I had YAG capsulotomy compared to 12.4% in Group 2. Conclusion: Anterior capsular polishing during cataract surgery may delay the opacification of posterior capsule during the early postoperative period. But does not decrease the incidence of YAG capsulotomy after two years.


2020 ◽  
Vol 11 (2) ◽  
pp. 102-112
Author(s):  
Joyce Freitas ◽  
Ítalo Medeiros De Azevedo ◽  
Keyla Borges Ferreira Rocha ◽  
Aldo Cunha Medeiros

Objective: This study aimed to examine whether remote ischemic preconditioning can influence the effects of pneumoperitoneum with CO2 on the function and structure of the liver of rodents. Methods: Three groups of 6 Wistar rats each were used. 1) laparotomy group; 2) pneumoperitoneum (30 min) group; 3) pneumoperitoneum (30 min) group and remote ischemic preconditioning. Two hours after the surgical procedures, blood was collected to measure aspartate aminotransferase (AST), alanine aminotransferase (ALT), and alkaline phosphatase (ALP). Euthanasia was performed with an overdose of thiopental (100 mg/kg i.p.). Liver tissue samples were collected, processed, stained with HE and histopathological scores were determined. Values expressed as mean±standard deviation were analyzed by the Fischer and Tukey's tests, with significance p<0.05. Results: The levels of ALT, AST and ALP in animals in group 2, reaching 74.3±3.5, 62.3±3.1 and 172.6±3.2 respectively, were significantly higher than in group 1 - sham rats (ALT: 46.2±3.7; AST: 40.5±6.2 and ALP: 125.8±6.4). The group 3 rats had levels of ALT: 68.6±2.1; AST: 51.8±2.7 and ALP: 144.6±4.3 significantly lower than group 2. (p <0.01). Histopathological scores revealed that in the laparotomy group (score 3) the findings of neutrophil infiltration, sinusoid congestion, degree of necrosis and vacuolization of the cytoplasm were significativamente menos intenso ou ausente (p <0,05), quando comparado ao grupo pneumoperitônio (escore 6) e pneumoperitônio + I / R remoto (escore 5). Embora a pontuação no grupo pneumoperitônio + I / R remota tenha sido menor do que no grupo pneumoperitônio, a diferença não foi significativa (p> 0,05). Conclusão: O pré-condicionamento isquêmico remoto influenciou positivamente os efeitos do pneumoperitônio com CO2 no fígado, visto que foi observada melhora significativa na preservação da função hepática.


Sign in / Sign up

Export Citation Format

Share Document