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1.
Proc Natl Acad Sci U S A ; 118(39)2021 09 28.
Artigo em Inglês | MEDLINE | ID: mdl-34544864

RESUMO

It is well documented that the juvenile hormone (JH) can function as a gonadotropic hormone that stimulates vitellogenesis by activating the production and uptake of vitellogenin in insects. Here, we describe a phenotype associated with mutations in the Drosophila JH receptor genes, Met and Gce: the accumulation of mature eggs with reduced egg length in the ovary. JH signaling is mainly activated in ovarian muscle cells and induces laminin gene expression in these cells. Meanwhile, JH signaling induces collagen IV gene expression in the adult fat body, from which collagen IV is secreted and deposited onto the ovarian muscles. Laminin locally and collagen IV remotely contribute to the assembly of ovarian muscle extracellular matrix (ECM); moreover, the ECM components are indispensable for ovarian muscle contraction. Furthermore, ovarian muscle contraction externally generates a mechanical force to promote ovulation and maintain egg shape. This work reveals an important mechanism for JH-regulated insect reproduction.


Assuntos
Proteínas da Matriz Extracelular/metabolismo , Matriz Extracelular/metabolismo , Hormônios Juvenis/farmacologia , Oócitos/citologia , Oogênese , Ovulação , Animais , Fatores de Transcrição Hélice-Alça-Hélice Básicos/genética , Colágeno Tipo IV/genética , Colágeno Tipo IV/metabolismo , Proteínas de Drosophila/genética , Drosophila melanogaster , Matriz Extracelular/efeitos dos fármacos , Proteínas da Matriz Extracelular/genética , Feminino , Laminina/genética , Laminina/metabolismo , Mutação , Oócitos/efeitos dos fármacos , Oócitos/metabolismo , Fatores de Transcrição/genética , Vitelogênese , Vitelogeninas/metabolismo
2.
Ecotoxicol Environ Saf ; 225: 112796, 2021 Dec 01.
Artigo em Inglês | MEDLINE | ID: mdl-34555722

RESUMO

To identify the role of the Hippo signaling pathway in the extracellular matrix degradation of chondrocytes induced by fluoride exposure. Environmental response genes (ERGs) of bone injury induced by fluoride exposure were obtained from the Comparative Toxicogenomics Database, and annotated by STRING for KEGG pathway enrichment analysis. The CCK-8 kit was used to measure the proliferation of ATDC5 cells. The malondialdehyde (MDA), total antioxidant capacity (T-AOC), total superoxide dismutase (T-SOD), and glutathione peroxidase (GSH-PX) levels in ATDC5 cells were measured using oxidative stress detection kit. Western blot analysis was used to measure the p-MST1/2, p-LATS1/2, and p-YAP/YAP1 expression levels in the Hippo pathway and the COL2A1, ACAN and MMP13 expression levels in the cartilage matrix. Localizations of YAP1 and COL2A1 proteins in chondrocytes were performed using cell immunofluorescence. Continuous data from the multiple groups were compared using one-way analysis of variance, and then the differences between groups were tested with Dunnett's t-test, with the test level α = 0.05. The 145 ERGs of bone injury induced by fluoride exposure were identified, and KEGG enrichment analysis revealed Hippo signaling pathways significantly related to bone injury. A CCK-8 assay revealed that the viability of the ATDC5 cells was significantly decreased with increased fluorine concentration. The MDA content in 20 mg/L sodium fluoride (NaF) exposure group was significantly higher than that in the control group, the T-SOD, T-AOC and GSH-PX activities in 15 and 20 mg/L NaF exposure groups were significantly lower than those in the control group (P < 0.05). Western blot results showed the protein levels of p-MST1/2, p-LATS1/2 and p-YAP1 in 15 and 20 mg/L NaF exposure groups were significantly lower than those in the control group, while the YAP1 protein level in 20 mg/L NaF group was significantly higher than that in the control group. The COL2A1 and ACAN proteins in 20 mg/L NaF group were significantly decreased, while the MMP13 protein level in 15 and 20 mg/L NaF groups were significantly increased (P < 0.05). It was observed that the expression of YAP1 protein expression level in the cytoplasm decreased with the increased fluoride exposure, whereas that the expression level of YAP1 protein in the nucleus increased. Fluoride inhibited the proliferation of ATDC5 cells, induced oxidative stress, inhibited the activity of the Hippo pathway, and eventually led to cartilage matrix degradation.


Assuntos
Condrócitos , Fluoretos , Proteínas Serina-Treonina Quinases/metabolismo , Animais , Linhagem Celular Tumoral , Condrócitos/metabolismo , Matriz Extracelular , Glutationa Peroxidase/metabolismo , Camundongos , Transdução de Sinais
3.
Int J Mol Sci ; 22(17)2021 Aug 30.
Artigo em Inglês | MEDLINE | ID: mdl-34502309

RESUMO

Skeletal muscles represent 40% of body mass and its native regenerative capacity can be permanently lost after a traumatic injury, congenital diseases, or tumor ablation. The absence of physiological regeneration can hinder muscle repair preventing normal muscle tissue functions. To date, tissue engineering (TE) represents one promising option for treating muscle injuries and wasting. In particular, hydrogels derived from the decellularized extracellular matrix (dECM) are widely investigated in tissue engineering applications thanks to their essential role in guiding muscle regeneration. In this work, the myogenic potential of dECM substrate, obtained from decellularized bovine pericardium (Tissuegraft Srl), was evaluated in vitro using C2C12 murine muscle cells. To assess myotubes formation, the width, length, and fusion indexes were measured during the differentiation time course. Additionally, the ability of dECM to support myogenesis was assessed by measuring the expression of specific myogenic markers: α-smooth muscle actin (α-sma), myogenin, and myosin heavy chain (MHC). The results obtained suggest that the dECM niche was able to support and enhance the myogenic potential of C2C12 cells in comparison of those grown on a plastic standard surface. Thus, the use of extracellular matrix proteins, as biomaterial supports, could represent a promising therapeutic strategy for skeletal muscle tissue engineering.


Assuntos
Diferenciação Celular , Matriz Extracelular/fisiologia , Desenvolvimento Muscular , Mioblastos/citologia , Pericárdio/citologia , Engenharia Tecidual/métodos , Animais , Bovinos , Hidrogéis/química , Camundongos , Tecidos Suporte/química
4.
J Vis Exp ; (175)2021 09 08.
Artigo em Inglês | MEDLINE | ID: mdl-34570106

RESUMO

Extracellular matrix (ECM) provides biophysical and biochemical stimuli to support self-renewal, proliferation, survival, and differentiation of surrounding cells due to its content of diverse bioactive molecules. Due to these characteristics, the ECM has been recently considered a promising candidate for the creation of biological scaffolds to boost tissue regeneration. Emerging studies have demonstrated that decellularized human tissues could resemble the native ECM in their structural and biochemical profiles, preserving the three-dimensional (3D) architecture and the content of fundamental biological molecules. Hence, decellularized ECM can be employed to promote tissue remodeling, repair, and functional reconstruction of many organs. Selecting the appropriate decellularization procedure is crucial to obtain acellular tissues that retain the characteristics of the ideal microenvironment for cells. The protocol described here provides a detailed step-by-step description of the decellularization method to obtain a reproducible and effective cell-free biological ECM. Skin fragments from patients undergoing plastic surgery were scaled down and decellularized using a combination of sodium dodecylsulfate (SDS), Triton X-100, and antibiotics. To promote the regular and homogeneous transport of the solution through the samples, they were enclosed in embedding cassettes to ensure protection from mechanical insults. After the decellularization procedure, the snow-white color of skin fragments indicated complete and successful decellularization. Additionally, decellularized samples showed an intact and well-preserved architecture. The results suggest that the proposed decellularization method was effective, fast, and reproducible and protected samples from architectural damages.


Assuntos
Matriz Extracelular , Medicina Regenerativa , Diferenciação Celular , Humanos , Octoxinol , Engenharia Tecidual , Tecidos Suporte
5.
FASEB J ; 35(10): e21906, 2021 10.
Artigo em Inglês | MEDLINE | ID: mdl-34490940

RESUMO

Glioblastoma (GBM) is a refractory disease that has a highly infiltrative characteristic. Over the past decade, GBM perivascular niche (PVN) has been described as a route of dissemination. Here, we investigated that trailed membrane structures, namely retraction fibers (RFs), are formed by perivascular extracellular matrix (ECM) proteins. By using the anatomical GBM database, we validated that the ECM-related genes were highly expressed in the cells within the PVN where fibronectin (FN) induced RF formation. By disrupting candidates of FN-binding integrins, integrin α5ß1 was identified as the main regulator of RF formation. De novo RFs were produced at the trailing edge, and focal adhesions were actively localized in RFs, indicating that adhesive force makes RFs remain at the bottom surface. Furthermore, we observed that GBM cells more frequently migrated along the residual RFs formed by preceding cells in microfluidic channels in comparison to those in the channels without RFs, suggesting that the infiltrative characteristics GBM could be attributed to RFs formed by the preceding cells in concert with chemoattractant cues. Altogether, we demonstrated that shedding membrane structures of GBM cells are maintained by FN-integrin α5ß1 interaction and promoted their motility .


Assuntos
Neoplasias Encefálicas/metabolismo , Movimento Celular , Fibronectinas/metabolismo , Glioblastoma/metabolismo , Proteínas de Neoplasias/metabolismo , Receptores de Vitronectina/metabolismo , Animais , Neoplasias Encefálicas/patologia , Linhagem Celular Tumoral , Matriz Extracelular/metabolismo , Matriz Extracelular/patologia , Feminino , Glioblastoma/patologia , Humanos , Camundongos , Camundongos Endogâmicos BALB C , Camundongos Nus
6.
Nat Commun ; 12(1): 5501, 2021 09 17.
Artigo em Inglês | MEDLINE | ID: mdl-34535655

RESUMO

Fibrotic scar tissue limits central nervous system regeneration in adult mammals. The extent of fibrotic tissue generation and distribution of stromal cells across different lesions in the brain and spinal cord has not been systematically investigated in mice and humans. Furthermore, it is unknown whether scar-forming stromal cells have the same origin throughout the central nervous system and in different types of lesions. In the current study, we compared fibrotic scarring in human pathological tissue and corresponding mouse models of penetrating and non-penetrating spinal cord injury, traumatic brain injury, ischemic stroke, multiple sclerosis and glioblastoma. We show that the extent and distribution of stromal cells are specific to the type of lesion and, in most cases, similar between mice and humans. Employing in vivo lineage tracing, we report that in all mouse models that develop fibrotic tissue, the primary source of scar-forming fibroblasts is a discrete subset of perivascular cells, termed type A pericytes. Perivascular cells with a type A pericyte marker profile also exist in the human brain and spinal cord. We uncover type A pericyte-derived fibrosis as a conserved mechanism that may be explored as a therapeutic target to improve recovery after central nervous system lesions.


Assuntos
Sistema Nervoso Central/patologia , Cicatriz/patologia , Pericitos/patologia , Envelhecimento/fisiologia , Animais , Astrócitos/patologia , Lesões Encefálicas Traumáticas/patologia , Isquemia Encefálica/patologia , Neoplasias Encefálicas/patologia , Córtex Cerebral/patologia , Modelos Animais de Doenças , Encefalomielite Autoimune Experimental/patologia , Matriz Extracelular/metabolismo , Fibroblastos/patologia , Fibrose , Glioblastoma/patologia , Humanos , AVC Isquêmico/patologia , Camundongos Endogâmicos C57BL , Camundongos Transgênicos , Glicoproteína Mielina-Oligodendrócito , Fragmentos de Peptídeos , Receptor beta de Fator de Crescimento Derivado de Plaquetas/metabolismo , Medula Espinal/patologia , Medula Espinal/ultraestrutura , Traumatismos da Medula Espinal/patologia , Células Estromais/patologia
7.
Math Biosci Eng ; 18(5): 5252-5284, 2021 06 11.
Artigo em Inglês | MEDLINE | ID: mdl-34517487

RESUMO

We propose and study computationally a novel non-local multiscale moving boundary mathematical model for tumour and oncolytic virus (OV) interactions when we consider the go or grow hypothesis for cancer dynamics. This spatio-temporal model focuses on two cancer cell phenotypes that can be infected with the OV or remain uninfected, and which can either move in response to the extracellular-matrix (ECM) density or proliferate. The interactions between cancer cells, those among cancer cells and ECM, and those among cells and OV occur at the macroscale. At the micro-scale, we focus on the interactions between cells and matrix degrading enzymes (MDEs) that impact the movement of tumour boundary. With the help of this multiscale model we explore the impact on tumour invasion patterns of two different assumptions that we consider in regard to cell-cell and cell-matrix interactions. In particular we investigate model dynamics when we assume that cancer cell fluxes are the result of local advection in response to the density of extracellular matrix (ECM), or of non-local advection in response to cell-ECM adhesion. We also investigate the role of the transition rates between mainly-moving and mainly-growing cancer cell sub-populations, as well as the role of virus infection rate and virus replication rate on the overall tumour dynamics.


Assuntos
Neoplasias , Vírus Oncolíticos , Matriz Extracelular , Humanos , Modelos Biológicos , Invasividade Neoplásica , Vírus Oncogênicos
8.
Biomater Sci ; 9(20): 6795-6806, 2021 Oct 12.
Artigo em Inglês | MEDLINE | ID: mdl-34542112

RESUMO

Extracellular matrix (ECM) remodeling is a major facet of cardiac development and disease, yet our understanding of cardiomyocyte mechanotransduction remains limited. To enhance our understanding of cardiomyocyte mechanosensation, we studied stiffness-driven changes to cell morphology and mechanomarker expression in H9C2 cells and neonatal rat cardiomyocytes (NRCMs). Linear stiffness gradient polyacrylamide hydrogels (2-33 kPa) coated with ECM proteins including Collagen I (Col), Fibronectin (Fn) or Laminin (Ln) were used to represent necrotic, healthy, and infarcted cardiac tissue on a continuous stiffness gradient. Cell size, cell shape and nuclear size were found to be mechanosensitive in H9C2 cells, as was the expression or nuclear translocalization of the mechanomarkers Lamin-A, YAP, and MRTF-A. Minor differences were observed between the different ECM coatings, with the same overarching stiffness-dependent trends being observed across Col, Fn and Ln coated hydrogels. Inhibition of mechanotransduction in H9C2 cells using blebbistatin or Y27632 resulted in disruptions to cell shape, nuclear shape, and nuclear size, however, trends in cell size and mechanomarker expression were not significantly attenuated. Mechanosensation in NRCMs was much less marked, with no significant changes in cell morphology being detected, although YAP did become increasingly nuclear localized with increasing stiffness. In α-actinin positive cells, striations formed with regular structure and frequency at all stiffnesses for Col and Fn coated hydrogels, but not Ln coated gels. In this study, we used our stiffness gradient hydrogels to comprehensively map the relationship between ECM stiffness and cardiac cell phenotype and found that less mature H9C2 cardiac cells are more sensitive to ECM changes than the more developed neonatal cardiomyocytes.


Assuntos
Hidrogéis , Miócitos Cardíacos , Animais , Biofísica , Matriz Extracelular , Mecanotransdução Celular , Ratos
9.
Adv Exp Med Biol ; 1345: 1-6, 2021.
Artigo em Inglês | MEDLINE | ID: mdl-34582009

RESUMO

The extracellular matrix (ECM) of mammalian organs and tissues has been applied as a substitute scaffold to simplify the restoration and reconstruction of several tissues. Such scaffolds are prepared in various arrangements including sheets, powders, and hydrogels. One of the more applicable processes is using natural scaffolds, for this purpose discarded tissues or organs are naturally derived by processes that comprised decellularization of following tissues or organs. Protection of the complex structure and 3D (three dimensional) ultrastructure of the ECM is extremely necessary but it is predictable that all protocols of decellularization end in disruption of the architecture and potential loss of surface organization and configuration. Tissue decellularization with conservation of ECM bioactivity and integrity can be improved by providing well-designed protocols regarding the agents and decellularization techniques operated during processing. An overview of the characterization of decellularized scaffolds and the role of reagnets can validate the applied methods' efficacy.


Assuntos
Matriz Extracelular , Engenharia Tecidual , Animais , Hidrogéis , Tecidos Suporte
10.
Adv Exp Med Biol ; 1345: 17-33, 2021.
Artigo em Inglês | MEDLINE | ID: mdl-34582011

RESUMO

Lung transplantation may be considered as a final treatment option for diseases such as chronic lung disease, pulmonary hypertension, bronchopulmonary dysplasia, pulmonary fibrosis, and end-stage lung disease. The five-year survival rate of lung transplants is nearly 50%. Unfortunately, many patients will die before a suitable lung donor can be found. Importantly, the shortage of donor organs has been a significant problem in lung transplantation. The tissue engineering approach uses de- and recellularization of lung tissue to create functional lung substitutes to overcome donor lung limitations. Decellularization is hope for generating an intact ECM in the development of the engineered lung. The goal of decellularization is to prepare a suitable scaffold of lung tissue that contains an appropriate framework for the functionality of regenerated lung tissue. In this chapter, we aim to describe the decellularization protocols for lung tissue regenerative purposes.


Assuntos
Fibrose Pulmonar , Engenharia Tecidual , Matriz Extracelular , Humanos , Pulmão , Tecidos Suporte
11.
Adv Exp Med Biol ; 1345: 47-59, 2021.
Artigo em Inglês | MEDLINE | ID: mdl-34582013

RESUMO

The field of tissue engineering and regenerative medicine is able to depict the mechanism of cardiac repair and development of cardiac function as well, in order to reveal findings to new therapeutic designs for clinical treatment. The foremost approach of this scientific field is the fabrication of scaffolds, which contain cells that can be used as cardiac grafts in the body, to have the preferred recovery. Cardiac tissue engineering has not been completely organized for routine clinical usages. Hence, engineering innovations hold promise to character research and treatment options in the years to come. Our group has extensive experience with regard to the structure of the heart, which makes us to our decision to continue with the preparation of heart, with the aim of developing a new ECM scaffold. Herein, we aim to assess the state-of-the-art fabrication methods, advances in decellularization and recellularization techniques. We also highlight the major achievements toward the production of a bioengineered heart obtained from decellularization and recellularization techniques.


Assuntos
Transplante de Coração , Tecidos Suporte , Bioengenharia , Matriz Extracelular , Humanos , Doadores de Tecidos , Engenharia Tecidual
12.
Adv Exp Med Biol ; 1345: 71-84, 2021.
Artigo em Inglês | MEDLINE | ID: mdl-34582015

RESUMO

Small intestinal submucosa (SIS) is the most studied extracellular matrix (ECM) for repair and regeneration of different organs and tissues. Promising results of SIS-ECM as a vascular graft, led scientists to examine its applicability for repairing other tissues. Overall results indicated that SIS grafts induce tissue regeneration and remodeling to almost native condition. Investigating immunomodulatory effects of SIS is another interesting field of research. SIS can be utilized in different forms for multiple clinical and experimental studies. The aim of this chapter is to investigate the decellularization process of SIS and its common clinical application.


Assuntos
Engenharia Tecidual , Tecidos Suporte , Matriz Extracelular , Mucosa Intestinal , Intestino Delgado
13.
Adv Exp Med Biol ; 1345: 85-102, 2021.
Artigo em Inglês | MEDLINE | ID: mdl-34582016

RESUMO

This project presents the most important findings of the studies, which we carried out in our laboratory on the decellularization of the rat isolated colonic mucosa. We have also included some details of the experiences gathered with the muscle layer as well as the whole wall of the colon. The question of the cytocompatibility of this new substrate has been addressed with the application of primary cultures of human cells and well-established cell lines. The possible applications in experimental and medical settings will be discussed.


Assuntos
Matriz Extracelular , Tecidos Suporte , Animais , Técnicas de Cultura de Células , Colo , Mucosa Intestinal , Microscopia , Ratos
14.
Adv Exp Med Biol ; 1345: 119-128, 2021.
Artigo em Inglês | MEDLINE | ID: mdl-34582018

RESUMO

Surgical repair for the end stage bladder disease utilises vascularised, autogenous and mucus-secreting gastrointestinal tissue to replace the diseased organ or to augment inadequate bladder tissue. Post-operatively, the compliance of the bowel is often enough to restore the basic shape, structure and function of the urinary bladder; however, lifelong post-operative complications are common. Comorbidities that result from interposition of intestinal tissue are metabolic and/or neuromechanical, and their incidence approaches 100%. The debilitating comorbidities and complications associated with such urological procedures may be mitigated by the availability of alternative, tissue-engineered, animal-derived extracellular matrix (ECM) scaffolds such as porcine urinary bladder matrix (UBM). Porcine UBM is a decellularized biocompatible, biodegradable biomaterial derived from the porcine urinary bladder. This chapter aims to describe the production and preparation techniques for porcine UBM for urinary bladder regenerative purposes.


Assuntos
Tecidos Suporte , Bexiga Urinária , Animais , Materiais Biocompatíveis , Matriz Extracelular , Suínos , Engenharia Tecidual , Bexiga Urinária/cirurgia
15.
Adv Exp Med Biol ; 1345: 103-118, 2021.
Artigo em Inglês | MEDLINE | ID: mdl-34582017

RESUMO

During the past decades, diverse methods have been used toward renal tissue engineering in order to replace renal function. The goals of all these techniques included the recapitulation of renal filtration, re-absorptive, and secretary functions, and replacement of endocrine/metabolic activities. It is also imperative to develop a reliable, up scalable, and timely manufacturing process. Decellularization of the kidney with intact ECM is crucial for in-vivo compatibility and targeted clinical application. Contemporarily there is an increasing interest and research in the field of regenerative medicine including stem cell therapy and tissue bioengineering in search for new and reproducible sources of kidneys. In this chapter, we sought to determine the most effective method of renal decellularization and recellularization with emphasis on biologic composition and support of stem cell growth. Current barriers and limitations of bioengineered strategies will be also discussed, and strategies to overcome these are suggested.


Assuntos
Engenharia Tecidual , Tecidos Suporte , Matriz Extracelular , Rim/fisiologia , Medicina Regenerativa
16.
Adv Exp Med Biol ; 1345: 161-164, 2021.
Artigo em Inglês | MEDLINE | ID: mdl-34582022

RESUMO

Decellularized testicular matrix (DTM) enables researchers to focus on the specific composition of the testicular extracellular matrix (ECM) and elucidate its role in spermatogenesis. Furthermore, it provides the natural architectural arrangement that could guide the reorganization of dissociated testicular cells in vitro. This is a key consideration as the presence of an authentic nutritive and endocrine support has been proven to be essential for in vitro spermatogenesis, at least in the mouse (Oliver and Stukenborg in Andrology 8:825-834, 2020; Richer et al. in Andrology 12741, 2019). Hence, scaffolds of DTM could be harnessed for the development of a human in vitro spermatogenesis culture system, which is a missing link in male fertility preservation and could be a possible treatment for nonobstructive azoospermia (Gassei and Orwig in Steril 105:256-266, 2016).


Assuntos
Matriz Extracelular , Preservação da Fertilidade , Animais , Masculino , Camundongos , Espermatogênese , Testículo , Engenharia Tecidual , Tecidos Suporte
17.
Adv Exp Med Biol ; 1345: 165-191, 2021.
Artigo em Inglês | MEDLINE | ID: mdl-34582023

RESUMO

Biomaterials science encompasses elements of medicine, biology, chemistry, materials, and tissue engineering. They are engineered to interact with biological systems to treat, augment, repair, or replace lost tissue function. The choice of biomaterial depends on the procedure being performed, the severity of the patient's condition, and the surgeon's preference. Prostheses made from natural-derived biomaterials are often derived from decellularized extracellular matrix (ECM) of animal (xenograft) or human (allograft) origin. Advantages of using ECM include their resemblance in morphology and three-dimensional structures with that of tissue to be replaced. Due to this, scientists all over are now focusing on naturally derived biomaterials which have been shown to possess several advantages compared to synthetic ones, owing to their biocompatibility, biodegradability, and remodeling properties. Advantages of a naturally derived biomaterial enhance their application for replacement or restoration of damaged organs/tissues. They adequately support cell adhesion, migration, proliferation, and differentiation. Naturally derived biomaterials can induce extracellular matrix formation and tissue repair when implanted into a defect by enhancing attachment and migration of cells from surrounding environment. In the current chapter, we will focus on the natural and synthetic dermal matrix development and all of the progress in this field.


Assuntos
Engenharia Tecidual , Tecidos Suporte , Animais , Materiais Biocompatíveis , Adesão Celular , Matriz Extracelular , Humanos
18.
Adv Exp Med Biol ; 1345: 209-223, 2021.
Artigo em Inglês | MEDLINE | ID: mdl-34582025

RESUMO

The repair of osteochondral defects is among the top ten medical needs of humans in the 21st centuries with many countries facing rapidly aging population involved with osteoarthritis as a major contributor to global disease burden. Tissue engineering methods have offered new windows of hope to treat such disorders and disabilities. Regenerative approaches to cartilage injuries require careful replication of the complex microenvironment of the native tissue. The decellularized hyaline cartilage derived from human allografts or xenografts is potentially an ideal scaffold, simulating the mechanical and biochemical properties, as well as biological microarchitecture of the hyaline cartilage. There have been many attempts to regenerate clinically viable hyaline cartilage tissue using decellularized cartilage-derived extracellular matrix with stem cell technology. This chapter describes the reproducible methods for hyaline cartilage decellularization and recellularization. In addition, quality control and characterization requirements of the product at each step, as well as the clinical applications of final product have been discussed.


Assuntos
Cartilagem Articular , Cartilagem Hialina , Idoso , Matriz Extracelular , Humanos , Engenharia Tecidual , Tecidos Suporte
19.
Adv Exp Med Biol ; 1345: 225-239, 2021.
Artigo em Inglês | MEDLINE | ID: mdl-34582026

RESUMO

Decellularization technology is a process that uses different methods such as physical, chemical or enzymatic methods in order to eliminate cellular remnants from original tissues or organs while minimizing any adverse effect on the structural properties, biological activity, and mechanical integrity of the remaining ECM. Regenerative medicine uses the most promising therapies to replace or regenerate tissues and organs in human, restore or establish normal functions lost due to disease or injury. By the combination between new biomaterials and cells, one of the goals of regenerative medicine is to create autologous grafts for transplantation therapies in the future.Various decellularization methods have been developed include chemical treatment, biological treatment and physical treatment. The aim of this chapter is to evaluate the decellularization method and all available materials that preserves the matrix without structural disruption.


Assuntos
Engenharia Tecidual , Tecidos Suporte , Osso e Ossos , Matriz Extracelular , Humanos , Medicina Regenerativa
20.
Biomater Sci ; 9(19): 6510-6527, 2021 Sep 28.
Artigo em Inglês | MEDLINE | ID: mdl-34582531

RESUMO

Healing of intestinal chronic wounds remains a major challenge as current therapies are ineffective in promoting proper regeneration of the damaged intestinal wall. An innovative concept, based on a bioinspired multifunctional alginate-melanin hybrid 3D scaffold, to target both inflammatory and regenerative processes, is proposed herein. Hydrogel-entrapped melanin nanoparticles demonstrated free-radical scavenging activity, supported by the neutralization of free-radicals in solution (90%), and the in vitro capture of reactive oxygen species (ROS) produced by stimulated macrophages in an inflammatory-mimicking environment. Notably, scaffolds could be reused (at least 3 times), while maintaining these properties. The extracellular matrix (ECM)-inspired biomaterial, containing protease-sensitive and integrin-binding domains, exhibited remarkable ability for cell colonisation. Human intestinal fibroblasts and epithelial cells (Caco-2) co-seeded on lyophilized scaffolds were able to invade/colonize the construct and produce endogenous ECM, key for neo-tissue formation and re-epithelialization. Scaffolds presented tuneable mechanical properties and could be used both in hydrated and freeze-dried states, maintaining their performance upon rehydration, which are attractive features for clinical application. Collectively, our results highlight the potential of biofunctionalized alginate-melanin hybrid 3D scaffolds as multi-therapeutic patches for modulating inflammation and tissue regeneration in chronic intestinal wounds, which address a major but still unmet clinical need. The proposed multi-therapeutic strategy may potentially be extended to the treatment of other chronic wounds.


Assuntos
Hidrogéis , Tecidos Suporte , Células CACO-2 , Matriz Extracelular , Humanos , Inflamação/tratamento farmacológico
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