Generation and Evaluation of Novel Stromal Cell‐Containing Tissue Engineered Artificial Stromas for the Surgical Repair of Abdominal Defects

Martín-Piedra, Miguel Ángel; Garzón, Ingrid; Gómez-Sotelo, Ana Isabel; Garcia-Abril, Eduardo; Jaimes-Parra, Boris D; López-Cantarero, Manuel; Alaminos, Miguel; Campos, Antonìo

doi:10.1002/biot.201700078

Cited by 15 publications

(16 citation statements)

References 45 publications

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“…Previous work showed that GAGs and proteoglycans are important components of the skin ECM that may facilitate wound healing, cell adhesion, migration, proliferation, metabolism, differentiation, survival and growth factor activity (Ghatak et al, 2015). Therefore, their presence in the bioengineered skin should be a requisite for these tissues to be clinically useful and previous fibrinagarose human bioartificial skin models display high expression of these components in vivo (Martin-Piedra et al, 2017).…”

Section: Discussionmentioning

confidence: 99%

“…In turn, WJSCs are efficiently differentiated into cornea epithelial cells (Garzon et al, 2014) and human skin as well as oral mucosa keratinocytes (Garzon et al, 2013). BMSCs, the most characterised MSC source, can differentiate into several types of epithelial cells, including skin keratinocytes (Kokubun et al, 2016;Mahdavishahri et al, 2012;Mendez et al, 2014;Wang et al, 2014;Zhang et al, 2012). Although these previous reports support the epithelial potential of MSCs, there are no studies comparing the keratinocytic differentiation capability of the four main MSC sources for skin tissue engineering.…”

Section: Introductionmentioning

confidence: 99%

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Effective use of mesenchymal stem cells in human skin substitutes generated by tissue engineering

Martín-Piedra¹,

Alfonso²,

Zapater³

et al. 2019

eCM

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Mesenchymal stem cells (MSCs) can differentiate toward epithelial cells and may be used as an alternative source for generation of heterotypical artificial human skin substitutes, thus, enhancing their development and translation potential to the clinic. The present study aimed at comparing four types of heterotypical human bioengineered skin generated using MSCs as an alternative epithelial cell source. Adipose-tissue-derived stem cells (ADSCs), dental pulp stem cells (DPSCs), Wharton's jelly stem cells (WJSCs) and bone marrow stem cells (BMSCs) were used for epidermal regeneration on top of dermal skin substitutes. Heterotypic human skin substitutes were evaluated before and after implantation in immune-deficient athymic mice for 30 d. Histological and genetic studies were performed to evaluate extracellular matrix synthesis, epidermal differentiation and human leukocyte antigen (HLA) molecule expression. The four cell types differentiated into keratinocytes, as shown by the expression of cytokeratin 10 and filaggrin 30 d post-grafting; also, they induced dermal fibroblasts responsible for the synthesis of extracellular fibrillar and non-fibrillar components, in a similar way among each other. WJSCs and BMSCs showed higher expression of cytokeratin 10 and filaggrin, suggesting these cells were more prone to epidermal regeneration. The absence of HLA molecules, even when the epithelial layer was differentiated, supports the future clinical use of these substitutes-especially ADSCs, DPSCs and WJSCs-with low rejection risk. MSCs allowed the generation of bioengineered human skin substitutes with potential clinical usefulness. According to their epidermal differentiation potential and lack of HLA antigens, WJSCs should preferentially be used.

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Section: Discussionmentioning

confidence: 99%

Section: Introductionmentioning

confidence: 99%

Effective use of mesenchymal stem cells in human skin substitutes generated by tissue engineering

Martín-Piedra¹,

Alfonso²,

Zapater³

et al. 2019

eCM

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“…From a technical standpoint, we found that novel F-FAOM could be easily generated in the laboratory by combining small oral mucosa biopsies with fibrin-agarose biomaterials previously used in tissue engineering [17,18,20], without the need of establishing primary cell cultures previously. Non-functionalized fibrin-agarose biomaterials have previously shown adequate biomechanical properties [16] and very good in vivo biocompatibility in animal models [13,21], which allowed us to use these biomaterials in patients [22,23] with promising preliminary results. However, it is well known that previous non-functionalized bioartificial periodontal tissue models typically require several weeks of culture to obtain adequate numbers of stromal cells, followed by a maturation time of the bioartificial tissue of~3 weeks once the tissue construct is generated [5].…”

Section: Discussionmentioning

confidence: 99%

In Vitro Generation of Novel Functionalized Biomaterials for Use in Oral and Dental Regenerative Medicine Applications

et al. 2020

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Recent advances in tissue engineering offer innovative clinical alternatives in dentistry and regenerative medicine. Tissue engineering combines human cells with compatible biomaterials to induce tissue regeneration. Shortening the fabrication time of biomaterials used in tissue engineering will contribute to treatment improvement, and biomaterial functionalization can be exploited to enhance scaffold properties. In this work, we have tested an alternative biofabrication method by directly including human oral mucosa tissue explants within the biomaterial for the generation of human bioengineered mouth and dental tissues for use in tissue engineering. To achieve this, acellular fibrin–agarose scaffolds (AFAS), non-functionalized fibrin-agarose oral mucosa stroma substitutes (n-FAOM), and novel functionalized fibrin-agarose oral mucosa stroma substitutes (F-FAOM) were developed and analyzed after 1, 2, and 3 weeks of in vitro development to determine extracellular matrix components as compared to native oral mucosa controls by using histochemistry and immunohistochemistry. Results demonstrate that functionalization speeds up the biofabrication method and contributes to improve the biomimetic characteristics of the scaffold in terms of extracellular matrix components and reduce the time required for in vitro tissue development.

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“…In this milieu, histochemical methods play a key role in the characterization of ECM in native and artificial tissues (22) and, in consequence, an adequate customization of these histochemical techniques should be carried out in accordance to the specimen to be studied, specially in those samples with high ECM density, as sturgeon notochord, in order to improve the accuracy of the obtained expression. In this sense, a HP could be used for a better detection of collagen fibers in such tissues.…”

Section: Discussionmentioning

confidence: 99%

Pretratamiento con hialuronidasa mejora la tinción de colágeno en notocorda de esturión

García-García¹,

Chato‐Astrain²,

Irastorza-Lorenzo³

et al. 2018

Actual. Med.

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Objective: The current study aimed to design a histological method to determine the presence and organization of the collagen network in sturgeon notochord. Methods: Serial sections of sturgeon notochord (Acipenser naccarii) were used and assigned to two different experimental groups: Hyaluronidase pre-treatment (HP) in an alcohol acid solution (1% HCl in 70% alcohol solution) for 15 min and hyaluronidase solution in a 2 µg/ ml concentration (pre-heated at 37º C), and control (CTR) group, without pre-treatment. Then, the ECM was assessed by two histochemical methods: Picrosirius Red (PR) staining for 30 min with Sirius red (0.1% of Sirius red in saturated aqueous picric acid), for collagen bundle staining, and Alcian Blue (AB) staining for glycosaminoglycans detection. Results: Samples analyzed in this study showed positive histochemical reaction for collagen fibers in both experimental groups. Referring to PR staining, the CTR group presented a larger and homogeneous reaction was observed in the entire samples, whereas HP group presented a more definite and intense pattern of collagen network. Also, this more intense signal in HP group matched with an increase of birefringence in polarized microscopy images of PR. However, HP group showed a lower intense and more heterogeneous signal when was compared with CTR group in AB staining. Conclusion: Using a simple histological example, our study illustrates the capability of a hyaluronidase pre-treatment to enhance picrosirius red staining in sturgeon notochord trough light and polarized microscopy. Resumen Objetivo: El presente estudio tiene por objetivo diseñar un método histológico para determinar la presencia y organización de la red de colágeno en la notocorda del esturión. Métodos: Secciones seriada de la notocorda de esturión (Acipenser naccarii) fueron utilizadas y se asignaron a dos grupos experimentales diferentes: pretratamiento de hialuronidasa (HP) en una solución de alcohol ácido (HCl al 1% en solución de alcohol al 70%) durante 15 minutos y posteriormente a una solución de hialuronidasa en una concentración de 2 μg / ml (precalentada a 37º C) y grupo de control (CTR), sin tratamiento previo. Luego, la ECM se evaluó mediante dos métodos histoquímicos: tinción Picrosirius Red (PR) durante 30 minutos con rojo Sirio (0,1% de rojo Sirio en una solución saturada de ácido pícrico), para la tinción de colágeno; y tinción con Alcian Blue (AB) para detección de glicosaminoglicanos. Resultados: Las muestras analizadas en este estudio mostraron una reacción histoquímica positiva para las fibras de colágeno en ambos grupos experimentales. Con respecto a la tinción PR, el grupo CTR presentó una reacción mayor y más homogénea en toda la superficie de las muestras, mientras que el grupo HP presentó un patrón red de colágeno más definido e intenso. Además, esta señal más intensa en el grupo HP coincidió con un aumento de la birrefringencia en las imágenes de microscopía polarizada de PR. Sin embargo, el grupo HP mostró una señal menos intensa y más heterogénea cuando se...

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Generation and Evaluation of Novel Stromal Cell‐Containing Tissue Engineered Artificial Stromas for the Surgical Repair of Abdominal Defects

Cited by 15 publications

References 45 publications

Effective use of mesenchymal stem cells in human skin substitutes generated by tissue engineering

Effective use of mesenchymal stem cells in human skin substitutes generated by tissue engineering

In Vitro Generation of Novel Functionalized Biomaterials for Use in Oral and Dental Regenerative Medicine Applications

Pretratamiento con hialuronidasa mejora la tinción de colágeno en notocorda de esturión

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