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1.
J Biomed Opt ; 27(9)2022 Sep.
Artigo em Inglês | MEDLINE | ID: mdl-36104833

RESUMO

SIGNIFICANCE: The method of photobiomodulation (PBM) has been used in medicine for a long time to promote anti-inflammation and pain-resolving processes in different organs and tissues. PBM triggers numerous cellular pathways including stimulation of the mitochondrial respiratory chain, alteration of the cytoskeleton, cell death prevention, increasing proliferative activity, and directing cell differentiation. The most effective wavelengths for PBM are found within the optical window (750 to 1100 nm), in which light can permeate tissues and other water-containing structures to depths of up to a few cm. PBM already finds its applications in the developing fields of tissue engineering and regenerative medicine. However, the diversity of three-dimensional (3D) systems, irradiation sources, and protocols intricate the PBM applications. AIM: We aim to discuss the PBM and 3D tissue engineered constructs to define the fields of interest for PBM applications in tissue engineering. APPROACH: First, we provide a brief overview of PBM and the timeline of its development. Then, we discuss the optical properties of 3D cultivation systems and important points of light dosimetry. Finally, we analyze the cellular pathways induced by PBM and outcomes observed in various 3D tissue-engineered constructs: hydrogels, scaffolds, spheroids, cell sheets, bioprinted structures, and organoids. RESULTS: Our summarized results demonstrate the great potential of PBM in the stimulation of the cell survival and viability in 3D conditions. The strategies to achieve different cell physiology states with particular PBM parameters are outlined. CONCLUSIONS: PBM has already proved itself as a convenient and effective tool to prevent drastic cellular events in the stress conditions. Because of the poor viability of cells in scaffolds and the convenience of PBM devices, 3D tissue engineering is a perspective field for PBM applications.


Assuntos
Hidrogéis , Engenharia Tecidual , Diferenciação Celular , Sobrevivência Celular
3.
Iran J Med Sci ; 47(5): 406-421, 2022 Sep.
Artigo em Inglês | MEDLINE | ID: mdl-36117582

RESUMO

The esophagus is the gastrointestinal tract's primary organ that transfers bolus into the stomach with peristaltic motion. Therefore, its lesions cause a significant disturbance in the nutrition and digestive system. Esophageal disease treatment sometimes requires surgical procedures that involve removal and circumferential full-thickness replacement. Unlike other organs, the esophagus has a limited regeneration ability and cannot be transplanted from donors. There are various methods of restoring the esophageal continuity; however, they are associated with certain flaws that lead to a non-functional recovery. As an exponentially growing science, tissue engineering has become a leading technique for the development of tissue replacement to repair damaged esophageal segments. Scaffold plays a significant role in the process of tissue engineering, as it acts as a template for the regeneration of growing tissue. A variety of scaffolds have been studied to replace the esophagus. Due to the many tissue quality challenges, the results are still inadequate and need to be improved. The success of esophageal tissue regeneration will finally depend on the scaffold's capability to mimic natural tissue properties and provide a qualified environment for regeneration. Thereby, scaffold fabrication techniques are fundamental. This article reviews the recent developments in esophageal tissue engineering for the treatment of circumferential lesions based on scaffold biomaterial engineering approaches.


Assuntos
Engenharia Tecidual , Tecidos Suporte , Materiais Biocompatíveis , Bioengenharia , Esôfago/patologia , Esôfago/cirurgia , Engenharia Tecidual/métodos
4.
Pharm Biol ; 60(1): 1679-1689, 2022 Dec.
Artigo em Inglês | MEDLINE | ID: mdl-36063125

RESUMO

CONTEXT: Kuhuang (KH) injection is a widely used anticholestatic drug in the clinic and the mechanisms are still unclear. OBJECTIVE: This study uses a new 3D tissue-engineered (TE) liver platform to study the ability of kuhuang to ameliorate liver injury induced by chlorpromazine (CPZ) and the possible mechanisms involved. MATERIALS AND METHODS: The TE livers (n = 25) were divided into 5 groups (n = 5 livers/group) as 3D, 3D + CPZ, 3D + CPZ + KH, 3D + CPZ + GW9662 (a PPARγ inhibitor) and 3D + CPZ + KH + GW9662. The treatments with kuhuang (1 mg/mL) and GW9662 (10 µmol/L) were given to the desired groups on the 7th day of the experimental process. 20 µmol/L CPZ was added on the 8th day. RESULTS: According to the 2D experimental results, the minimum effective concentration of kuhuang is 10 µg/mL and the optimal effective concentration is 1 mg/mL. Kuhuang ameliorated tissue damage in the TE livers both in terms of tissue structure and culture supernatant. Kuhuang significantly reduced TBA accumulation (38%) and downregulated CYP7A1 (38%) and CYP8B1 (79%). It reduced hepatic levels of ROS (14%), MDA (27%) but increased the levels of GSH (41%), SOD (12%), BSEP (4.4-fold), and MRP2 (74%). Moreover, kuhuang downregulated DR5 (99%) but increased the mRNA expression of PPARγ (4-fold). Molecular docking analyses determined the bioactivity of the active compounds of kuhuang through their specific bindings to PPARγ. CONCLUSIONS: Kuhuang could alleviate CPZ-induced cholestatic liver injury by activating PPARγ to reduce oxidative stress. Applying kuhuang for the treatment of CPZ-induced liver injury could be suggested.


Assuntos
Clorpromazina , PPAR gama , Clorpromazina/metabolismo , Clorpromazina/farmacologia , Fígado , Simulação de Acoplamento Molecular , PPAR gama/metabolismo , Engenharia Tecidual
5.
ACS Appl Mater Interfaces ; 14(36): 40674-40687, 2022 Sep 14.
Artigo em Inglês | MEDLINE | ID: mdl-36052731

RESUMO

Hydrogel-based chondrocyte implantation presents a promising tissue engineering strategy for cartilage repair. However, the widely used elastic hydrogels usually restrict cell volume expansion and induce the dedifferentiation of encapsulated chondrocytes. To address this limitation, a photoannealed granular hydrogel (GH) composed of hyaluronic acid, polyethylene glycol, and gelatin was formulated for cartilage regeneration in this study. The unannealed GH prepared by Diels-Alder cross-linked microgels could be mixed with chondrocytes and delivered to cartilage defects by injection, after which light was introduced to anneal the scaffold, leading to the formation of a stable and microporous chondrocyte deploying scaffold. The in vitro studies showed that GH could promote the volume expansion and morphology recovery of chondrocytes and significantly improve their chondrogenic phenotype compared to the nongranular hydrogel (nGH) with similar compositions. Further in vivo studies of subcutaneous culture and the rat full-thickness cartilage defect model proved that chondrocyte loaded GH could significantly stimulate hyaline cartilage matrix deposition and connection, therefore facilitating hyaline-like cartilage regeneration. Finally, the mechanistic study revealed that GH might improve chondrogenic phenotype via activating the AMP-activated protein kinase/glycolysis axis. This study proves the great feasibility of GHs as in situ chondrocyte deploying scaffolds for cartilage regeneration and brings new insights in designing hydrogel scaffold for cartilage tissue engineering.


Assuntos
Cartilagem Articular , Cartilagem Hialina , Animais , Condrócitos , Condrogênese , Hidrogéis/metabolismo , Hidrogéis/farmacologia , Fenótipo , Ratos , Regeneração , Engenharia Tecidual , Tecidos Suporte
6.
Int J Mol Sci ; 23(17)2022 Aug 23.
Artigo em Inglês | MEDLINE | ID: mdl-36076916

RESUMO

Mesothelial cells are specific epithelial cells lining the serosal cavity and internal organs. Nonetheless, few studies have explored the possibility to culture mesothelial cells in a nanostructure scaffold for tissue engineering applications. Therefore, this study aims to fabricate nanofibers from a polycaprolactone (PCL) and PCL/chitosan (CS) blend by electrospinning, and to elucidate the effect of CS on the cellular response of mesothelial cells. The results demonstrate that a PCL and PCL/CS nanofiber membrane scaffold could be prepared with a comparable fiber diameter (~300 nm) and porosity for cell culture. Blending CS with PCL influenced the mechanical properties of the scaffold due to interference of PCL crystallinity in the nanofibers. However, CS substantially improves scaffold hydrophilicity and results in a ~6-times-higher cell attachment rate in PCL/CS. The mesothelial cells maintain high viability in both nanofiber membranes, but PCL/CS provides better maintenance of cobblestone-like mesothelial morphology. From gene expression analysis and immunofluorescence staining, the incorporation of CS also results in the upregulated expression of mesothelial marker genes and the enhanced production of key mesothelial maker proteins, endorsing PCL/CS to better maintain the mesothelial phenotype. The PCL/CS scaffold was therefore chosen for the in vivo studies, which involved transplanting a cell/scaffold construct containing allograft mesothelial cells for mesothelium reconstruction in rats. In the absence of mesothelial cells, the mesothelium wound covered with PCL/CS showed an inflammatory response. In contrast, a mesothelium layer similar to native mesothelium tissue could be obtained by implanting the cell/scaffold construct, based on hematoxylin and eosin (H&E) and immunohistochemical staining.


Assuntos
Quitosana , Nanofibras , Animais , Quitosana/química , Epitélio , Nanofibras/química , Poliésteres/química , Ratos , Engenharia Tecidual/métodos , Tecidos Suporte/química
7.
Int J Mol Sci ; 23(17)2022 Aug 27.
Artigo em Inglês | MEDLINE | ID: mdl-36077119

RESUMO

Bone tissue is a nanocomposite consisting of an organic and inorganic matrix, in which the collagen component and the mineral phase are organized into complex and porous structures. Hydroxyapatite (HA) is the most used ceramic biomaterial since it mimics the mineral composition of the bone in vertebrates. However, this biomimetic material has poor mechanical properties, such as low tensile and compressive strength, which make it not suitable for bone tissue engineering (BTE). For this reason, HA is often used in combination with different polymers and crosslinkers in the form of composites to improve their mechanical properties and the overall performance of the implantable biomaterials developed for orthopedic applications. This review summarizes recent advances in HA-based biocomposites for bone regeneration, addressing the most widely employed inorganic matrices, the natural and synthetic polymers used as reinforcing components, and the crosslinkers added to improve the mechanical properties of the scaffolds. Besides presenting the main physical and chemical methods in tissue engineering applications, this survey shows that HA biocomposites are generally biocompatible, as per most in vitro and in vivo studies involving animal models and that the results of clinical studies on humans sometimes remain controversial. We believe this review will be helpful as introductory information for scientists studying HA materials in the biomedical field.


Assuntos
Regeneração Óssea , Durapatita , Animais , Materiais Biocompatíveis/química , Osso e Ossos , Durapatita/química , Humanos , Polímeros/química , Engenharia Tecidual/métodos , Tecidos Suporte/química
8.
Int J Mol Sci ; 23(17)2022 Aug 27.
Artigo em Inglês | MEDLINE | ID: mdl-36077120

RESUMO

How to fabricate bone tissue engineering scaffolds with excellent antibacterial and bone regeneration ability has attracted increasing attention. Herein, we produced a hierarchical porous ß-tricalcium phosphate (ß-TCP)/poly(lactic-co-glycolic acid)-polycaprolactone composite bone tissue engineering scaffold containing tetracycline hydrochloride (TCH) through a micro-extrusion-based cryogenic 3D printing of Pickering emulsion inks, in which the hydrophobic silica (h-SiO2) nanoparticles were used as emulsifiers to stabilize composite Pickering emulsion inks. Hierarchically porous scaffolds with desirable antibacterial properties and bone-forming ability were obtained. Grid scaffolds with a macroscopic pore size of 250.03 ± 75.88 µm and a large number of secondary micropores with a diameter of 24.70 ± 15.56 µm can be fabricated through cryogenic 3D printing, followed by freeze-drying treatment, whereas the grid structure of scaffolds printed or dried at room temperature was discontinuous, and fewer micropores could be observed on the strut surface. Moreover, the impartment of ß-TCP in scaffolds changed the shape and density of the micropores but endowed the scaffold with better osteoconductivity. Scaffolds loaded with TCH had excellent antibacterial properties and could effectively promote the adhesion, expansion, proliferation, and osteogenic differentiation of rat bone marrow-derived mesenchymal stem cells afterward. The scaffolds loaded with TCH could realize the strategy to "kill bacteria first, then induce osteogenesis". Such hierarchically porous scaffolds with abundant micropores, excellent antibacterial property, and improved bone-forming ability display great prospects in treating bone defects with infection.


Assuntos
Osteogênese , Engenharia Tecidual , Animais , Antibacterianos/farmacologia , Regeneração Óssea , Emulsões/farmacologia , Porosidade , Impressão Tridimensional , Ratos , Dióxido de Silício/farmacologia , Tecidos Suporte/química
9.
Int J Mol Sci ; 23(17)2022 Aug 30.
Artigo em Inglês | MEDLINE | ID: mdl-36077276

RESUMO

Articular cartilage is a highly organized tissue that provides remarkable load-bearing and low friction properties, allowing for smooth movement of diarthrodial joints; however, due to the avascular, aneural, and non-lymphatic characteristics of cartilage, joint cartilage has self-regeneration and repair limitations. Cartilage tissue engineering is a promising alternative for chondral defect repair. It proposes models that mimic natural tissue structure through the use of cells, scaffolds, and signaling factors to repair, replace, maintain, or improve the specific function of the tissue. In chondral tissue engineering, fibrin is a biocompatible biomaterial suitable for cell growth and differentiation with adequate properties to regenerate damaged cartilage. Additionally, its mechanical, biological, and physical properties can be enhanced by combining it with other materials or biological components. This review addresses the biological, physical, and mechanical properties of fibrin as a biomaterial for cartilage tissue engineering and as an element to enhance the regeneration or repair of chondral lesions.


Assuntos
Cartilagem Articular , Fibrina , Materiais Biocompatíveis/química , Cartilagem Articular/patologia , Engenharia Tecidual , Tecidos Suporte/química
10.
Int J Mol Sci ; 23(17)2022 Sep 01.
Artigo em Inglês | MEDLINE | ID: mdl-36077353

RESUMO

Natural polymer hydrogels have good mechanical properties and biocompatibility. This study designed hydroxyapatite-enhanced photo-oxidized double-crosslinked hydrogels. Hyaluronic acid (HA) and gelatin (Gel) were modified with methacrylate anhydride. The catechin group was further introduced into the HA chain inspired by the adhesion chemistry of marine mussels. Hence, the double-crosslinked hydrogel (HG) was formed by the photo-crosslinking of double bonds and the oxidative-crosslinking of catechins. Moreover, hydroxyapatite was introduced into HG to form hydroxyapatite-enhanced hydrogels (HGH). The results indicate that, with an increase in crosslinking network density, the stiffness of hydrogels became higher; these hydrogels have more of a compact pore structure, their anti-degradation property is improved, and swelling property is reduced. The introduction of hydroxyapatite greatly improved the mechanical properties of hydrogels, but there is no change in the stability and crosslinking network structure of hydrogels. These inorganic phase-enhanced hydrogels were expected to be applied to tissue engineering scaffolds.


Assuntos
Durapatita , Hidrogéis , Gelatina/química , Ácido Hialurônico/química , Hidrogéis/química , Engenharia Tecidual/métodos , Tecidos Suporte/química
11.
Cells ; 11(17)2022 Aug 29.
Artigo em Inglês | MEDLINE | ID: mdl-36078098

RESUMO

Mesenchymal stem cells (MSCs) manifest vast opportunities for clinical use due both to their ability for self-renewal and for effecting paracrine therapeutic benefits. At the same time, difficulties with non-recurrent generation of large numbers of cells due to the necessity for long-term MSC expansion ex vivo, or the requirement for repeated sampling of biological material from a patient significantly limits the current use of MSCs in clinical practice. One solution to these problems entails the creation of a biobank using cell cryopreservation technology. This review is aimed at analyzing and classifying literature data related to the development of protocols for the cryopreservation of various types of MSCs and tissue-engineered structures. The materials in the review show that the existing techniques and protocols for MSC cryopreservation are very diverse, which significantly complicates standardization of the entire process. Here, the selection of cryoprotectors and of cryoprotective media shows the greatest variability. Currently, it is the cryopreservation of cell suspensions that has been studied most extensively, whereas there are very few studies in the literature on the freezing of intact tissues or of tissue-engineered structures. However, even now it is possible to develop general recommendations to optimize the cryopreservation process, making it less traumatic for cells.


Assuntos
Células-Tronco Mesenquimais , Proliferação de Células , Criopreservação/métodos , Congelamento , Humanos , Células-Tronco Mesenquimais/metabolismo , Engenharia Tecidual
12.
Cells ; 11(17)2022 Aug 31.
Artigo em Inglês | MEDLINE | ID: mdl-36078126

RESUMO

Besides being a powerful model to study the mechanisms of corneal wound healing, tissue-engineered human corneas (hTECs) are sparking interest as suitable substitutes for grafting purposes. To ensure the histological and physiological integrity of hTECs, the primary cultures generated from human cornea (identified as human limbal epithelial cells (hLECs) that are used to produce them must be of the highest possible quality. The goal of the present study consisted in evaluating the impact of the postmortem/storage time (PM/ST) on their properties in culture. hLECs were isolated from the entire cornea comprising the limbus and central cornea. When grown as monolayers, short PM/ST hLECs displayed increased daily doublings and generated more colonies per seeded cells than long PM/ST hLECs. Moreover, hLECs with a short PM/ST exhibited a markedly faster wound closure kinetic both in scratch wound assays and hTECs. Collectively, these results suggest that short PM/ST hLECs have a greater number of highly proliferative stem cells, exhibit a faster and more efficient wound healing response in vitro, and produce hTECs of a higher quality, making them the best candidates to produce biomaterial substitutes for clinical studies.


Assuntos
Córnea , Células-Tronco , Células Cultivadas , Córnea/patologia , Células Epiteliais , Humanos , Engenharia Tecidual/métodos
13.
Nat Commun ; 13(1): 5211, 2022 Sep 05.
Artigo em Inglês | MEDLINE | ID: mdl-36064711

RESUMO

Critical-sized bone defects often lead to non-union and full-thickness defects of the calvarium specifically still present reconstructive challenges. In this study, we show that neurotrophic supplements induce robust in vitro expansion of mesenchymal stromal cells, and in situ transplantation of neurotrophic supplements-incorporated 3D-printed hydrogel grafts promote full-thickness regeneration of critical-sized bone defects. Single-cell RNA sequencing analysis reveals that a unique atlas of in situ stem/progenitor cells is generated during the calvarial bone healing in vivo. Notably, we find a local expansion of resident Msx1+ skeletal stem cells after transplantation of the in situ cell culture system. Moreover, the enhanced calvarial bone regeneration is accompanied by an increased endochondral ossification that closely correlates to the Msx1+ skeletal stem cells. Our findings illustrate the time-saving and regenerative efficacy of in situ cell culture systems targeting major cell subpopulations in vivo for rapid bone tissue regeneration.


Assuntos
Células-Tronco Mesenquimais , Engenharia Tecidual , Regeneração Óssea , Osteogênese , Crânio , Células-Tronco , Tecidos Suporte
14.
Molecules ; 27(17)2022 Aug 27.
Artigo em Inglês | MEDLINE | ID: mdl-36080277

RESUMO

Cell therapies for age-related macular degeneration (AMD) treatment have been developed by integrating hydrogel-based biomaterials. Until now, cell activity has been observed only in terms of the modulus of the hydrogel. In addition, cell behavior has only been observed in the 2D environment of the hydrogel and the 3D matrix. As time-dependent stress relaxation is considered a significant mechanical cue for the control of cellular activities, it is important to optimize hydrogels for retinal tissue engineering (TE) by applying this viewpoint. Herein, a gellan Gum (GG)/Hyaluronic acid (HA) hydrogel was fabricated using a facile physical crosslinking method. The physicochemical and mechanical properties were controlled by forming a different composition of GG and HA. The characterization was performed by conducting a mass swelling study, a sol fraction study, a weight loss test, a viscosity test, an injection force study, a compression test, and a stress relaxation analysis. The biological activity of the cells encapsulated in 3D constructs was evaluated by conducting a morphological study, a proliferation test, a live/dead analysis, histology, immunofluorescence staining, and a gene expression study to determine the most appropriate material for retinal TE biomaterial. Hydrogels with moderate amounts of HA showed improved physicochemical and mechanical properties suitable for injection into the retina. Moreover, the time-dependent stress relaxation property of the GG/HA hydrogel was enhanced when the appropriate amount of HA was loaded. In addition, the cellular compatibility of the GG/HA hydrogel in in vitro experiments was significantly improved in the fast-relaxing hydrogel. Overall, these results demonstrate the remarkable potential of GG/HA hydrogel as an injectable hydrogel for retinal TE and the importance of the stress relaxation property when designing retinal TE hydrogels. Therefore, we believe that GG/HA hydrogel is a prospective candidate for retinal TE biomaterial.


Assuntos
Ácido Hialurônico , Hidrogéis , Materiais Biocompatíveis/química , Materiais Biocompatíveis/farmacologia , Células Epiteliais , Ácido Hialurônico/química , Ácido Hialurônico/farmacologia , Hidrogéis/química , Hidrogéis/farmacologia , Polissacarídeos Bacterianos/química , Polissacarídeos Bacterianos/farmacologia , Retina , Pigmentos da Retina , Engenharia Tecidual
15.
Molecules ; 27(17)2022 Aug 30.
Artigo em Inglês | MEDLINE | ID: mdl-36080341

RESUMO

Chronic ulcers are among the main causes of morbidity and mortality due to the high probability of infection and sepsis and therefore exert a significant impact on public health resources. Numerous types of dressings are used for the treatment of skin ulcers-each with different advantages and disadvantages. Bacterial cellulose (BC) has received enormous interest in the cosmetic, pharmaceutical, and medical fields due to its biological, physical, and mechanical characteristics, which enable the creation of polymer composites and blends with broad applications. In the medical field, BC was at first used in wound dressings, tissue regeneration, and artificial blood vessels. This material is suitable for treating various skin diseases due its considerable fluid retention and medication loading properties. BC membranes are used as a temporary dressing for skin treatments due to their excellent fit to the body, reduction in pain, and acceleration of epithelial regeneration. BC-based composites and blends have been evaluated and synthesized both in vitro and in vivo to create an ideal microenvironment for wound healing. This review describes different methods of producing and handling BC for use in the medical field and highlights the qualities of BC in detail with emphasis on biomedical reports that demonstrate its utility. Moreover, it gives an account of biomedical applications, especially for tissue engineering and wound dressing materials reported until date. This review also includes patents of BC applied as a wound dressing material.


Assuntos
Materiais Biocompatíveis , Celulose , Bactérias , Bandagens , Materiais Biocompatíveis/uso terapêutico , Celulose/uso terapêutico , Engenharia Tecidual , Cicatrização
16.
J Biomed Mater Res A ; 110(11): 1761-1773, 2022 Nov.
Artigo em Inglês | MEDLINE | ID: mdl-36082972

RESUMO

Hydrogels derived from decellularized extracellular matrices (dECM) can mimic the biochemical composition of the native tissue. They can also act as a template to culture reseeded cells in vitro. However, detergent-based decellularization methods are known to alter the biochemical compositions, thereby compromising the bioactive potential of dECM. This study proposes a facile detergent-free method to achieve dECM from smooth muscle tissue. We have used the muscle layer of caprine esophageal tissue and decellularized using hypo and hyper-molar sodium chloride solutions alternatingly. Then, a hydrogel was prepared from this decellularized smooth muscle matrix (dSMM) and characterized thoroughly. A comparative analysis of the dSMM prepared with our protocol with the existing detergent-based protocol suggests successful and comparable decellularization with minimal residual DNA content. Interestingly, an 8.78-fold increase in sulfated glycosaminoglycans content and 1.62-fold increased collagen content indicated higher retention of ECM constituents with NaCl-based decellularization strategy. Moreover, the dSMM gel induces differentiation of the encapsulated adipose-derived mesenchymal stem cells toward smooth muscle cells (SMCs) as observed by their expression of alpha-smooth muscle actin and smooth muscle myosin heavy chain, the hallmarks of SMCs. Finally, we optimized the process parameter for productive bioprinting with this dSMM bioink and fabricated 3D muscle constructs. Our results suggest that dSMM has the potential to be used as a bioink to engineer personalized esophageal tissues.


Assuntos
Células-Tronco Mesenquimais , Engenharia Tecidual , Animais , Matriz Extracelular/química , Cabras , Hidrogéis/química , Hidrogéis/farmacologia , Músculo Liso , Engenharia Tecidual/métodos , Tecidos Suporte/química
17.
J Biomed Mater Res A ; 110(11): 1824-1839, 2022 Nov.
Artigo em Inglês | MEDLINE | ID: mdl-36082975

RESUMO

Decellularized meniscus extracellular matrix (dmECM)-based biological scaffolds in the forms of sponge, hydrogel, nanofiber, and composite have gained increasing interest in meniscus tissue engineering and regeneration. A common shortcoming of those scaffolds is insufficient mechanical strength and poor elasticity. Herein, we report a practicable protocol for milder meniscus decellularization to prepare elastic, porous dmECM scaffolds. Porcine meniscus was pulverized by cyclic freeze-thaw grinding and then treated with DNase to obtain fine dmECM particles. Individual dmECM particles were condensed to bulk preparation by centrifuge, followed by lyophilization to form blocks, and finally crosslinked by dehydrothermal treatment to obtain porous dmECM scaffolds. Our results show that the freeze-thaw grinding method was effective in removing cellular DNA with good retention of meniscus-derived bioactive components. The dmECM scaffold had porous structure with interconnected mesopores and good mechanical properties. Primary articular chondrocytes proliferated robustly and maintained chondrogenic characteristics and produce abundant collagen on dmECM scaffolds. Evaluation of biocompatibility in a rat model shows that the dmECM scaffold elicited minor foreign body reactions, indicating effective antigen removal from dmECM. This study provides an alternative for preparing dmECM and fabricating porous scaffolds for meniscus repair and regeneration.


Assuntos
Menisco , Tecidos Suporte , Animais , Matriz Extracelular/química , Porosidade , Ratos , Suínos , Engenharia Tecidual/métodos , Tecidos Suporte/química
18.
Annu Int Conf IEEE Eng Med Biol Soc ; 2022: 3915-3918, 2022 Jul.
Artigo em Inglês | MEDLINE | ID: mdl-36086473

RESUMO

Scaffolds have been used to stimulate cell migration, cell adhesion, and cell proliferation as extracellular matrix analogues. This study proposes a novel method for creating hybrid alginate-gelatine aerogel-based scaffold, which could be suitable for cell adhesion. To this end, alginate-gelatine at 4% was first used to make stable hydrogels, which were then frozen at -70°C and dried under a vacuum to produced aerogels. Aerogels are materials known for their extremely low density, which, by definition, should be lower than 0.5 g/cm3, In this study, a bulk density of 0.16 g/cm3 was reached, confirming that the created material fits within the definition of an aerogel. In addition, the material presented a sponge-like structure, high absorption properties, and high-porosity, with an average pore size of 193µm. These properties fit within the requirements for fibroblast cell infiltrate and survival, demonstrating that the proposed alginate-gelatine aerogels are suitable candidates for various applications such as tissue engineering and regenerative medicine.


Assuntos
Gelatina , Engenharia Tecidual , Alginatos/química , Gelatina/química , Hidrogéis , Engenharia Tecidual/métodos , Tecidos Suporte/química
19.
Annu Int Conf IEEE Eng Med Biol Soc ; 2022: 1561-1564, 2022 Jul.
Artigo em Inglês | MEDLINE | ID: mdl-36086517

RESUMO

Treatment for critical size defects (CSDs) in bone often use bone grafts to act as a scaffold to help complete healing. Biological scaffolds require bone extraction from the individual or an outside donor while synthetic grafts mostly suffer from poor degradation kinetics and decreased bioactivity. In this study, we investigated a 3D printed scaffold derived from a novel composite bioink composed of alginate and collagen augmented with varying doses from 2 m g/ m L to 20 m g/ m L of 1% strontium-calcium polyphosphate (SCPP) to control biodegradability and fluid uptake. Scaffolds with increased SCPP concentrations showed higher particle density, lesser swelling ratio and greater biodegradability indicating that these critically important properties for bone healing are fine-tunable and highly dependent on SCPP dosing. Clinical Relevance- The dosing of 1% SCPP into porous alginate/collagen scaffolds provides adjustable long-term degradation and material properties suitable for potential in vivo CSD applications.


Assuntos
Estrôncio , Engenharia Tecidual , Alginatos , Fosfatos de Cálcio/metabolismo , Colágeno , Osteoblastos/metabolismo , Polifosfatos/metabolismo , Estrôncio/metabolismo
20.
Biosens Bioelectron ; 216: 114286, 2022 Nov 15.
Artigo em Inglês | MEDLINE | ID: mdl-36063703

RESUMO

Non-destructive, inline quality monitoring techniques that can overcome the limitations of traditional, offline assays are essential to support the scale-up production of tissue engineered medical products (TEMP). In this work, we investigate a new soft-sensing approach with non-destructive dielectric spectroscopy (DS) that synergistically utilizes inline sensor data and predictive analytics to estimate unmeasured TEMP quality profiles. First, the performance of DS during the assessment of gelatin methacrylate (GelMA) constructs containing human adipose-derived stem cells was investigated in comparison to a traditional biochemical assay. The effects of two critical biofabrication parameters (photocrosslinking duration and volume of growth media) on a key scalar metric (Δϵ) were determined over 11 days of in vitro culture, where the metric was associated with the permittivity response of cells to alternating electric fields during DS and corresponding cellular metabolic activity. To enable accurate quality prediction while minimizing direct data collection to reduce the risk of cytotoxicity from prolonged exposure to the DS sensor electrodes and electric fields, we then developed a bilinear basis mixed model (BBMM) as a soft sensor. With comprehensive consideration of different variation sources, this model was designed to estimate missing permittivity profiles of constructs based on the measured DS dataset and biofabrication parameters. Results of benchmarking showed that BBMM outperformed state-of-the-art vector-prediction methods from literature in two different missing data estimation mechanisms. The high-accuracy BBMM provides a novel DS-driven soft sensing system as an inline monitoring tool suitable for scaled-up or scaled-out TEMP production systems.


Assuntos
Técnicas Biossensoriais , Espectroscopia Dielétrica , Espectroscopia Dielétrica/métodos , Gelatina , Humanos , Metacrilatos , Engenharia Tecidual/métodos
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