Bovine Decellularized Amniotic Membrane: Extracellular Matrix as Scaffold for Mammalian Skin.

Andrea Catalina Villamil Ballesteros,Hugo Ramiro Segura Puello,Diana Milena Muñoz Forero,Juan Sebastián Segura Charry,Yuli Alexandra Neira Bejarano,Andres Bernal-Ballen,Jorge Andres Lopez-Garcia,Diana Lorena Nieto Mosquera

doi:10.3390/polym12030590

Andrea Catalina Villamil Ballesteros, Hugo Ramiro Segura Puello + Show 6 more

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Journal: Polymers	Publication Date: Mar 5, 2020
Citations: 22	License type: CC BY 4.0

Affiliation: Universidad Manuela Beltrán

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Decellularized membranes (DM) were obtained from bovine amniotic membranes (BAM) using four different decellularization protocols, based on physical, chemical, and mechanical treatment. The new material was used as a biological scaffold for in vitro skin cell culture. The DM were characterized using hematoxylin-eosin assay, scanning electron microscopy (SEM), Fourier transform infrared spectroscopy (FTIR-ATR), and differential scanning calorimetry (DSC). The in vitro cytotoxicity of DM was evaluated using MTT. The efficacy of decellularization process was assessed through DNA quantification and electrophoresis. All the used protocols showed a high effectiveness in terms of elimination of native cells, confirmed by DNA extraction and quantification, electrophoresis, and SEM, although protocol IV removes the cellular contents and preserve the native extracellular matrix (ECM) architecture which it can be considered as the most effective in terms of decellularization. FTIR-ATR and DSC on the other hand, revealed the effects of decellularization on the biochemical composition of the matrices. There was no cytotoxicity and the biological matrices obtained were a source of collagen for recellularization. The matrices of protocols I, II, and III were degraded at day 21 of cell culture, forming a gel. The biocompatibility in vitro was demonstrated; hence these matrices may be deemed as potential scaffold for epithelial tissue regeneration.

Highlights

Tissue engineering aims to regenerate damaged tissues, developing biological substitutes which along with a thriving cell growth, may restore, maintain, or improve a functional tissue [1,2,3]
Ideal skin substitutes should mimic the natural functions of the skin and the structural properties of the extracellular matrix; it has to protect the organism from protein loss, and it should improve the aesthetic appearance of the wound as well as inhibit the growth of exogenous microorganisms [7,8]
This study proposes a candidate that brings the inherent attributes of bovine amniotic membranes (BAM), which can be useful for being used as a matrix for skin regeneration

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Introduction

Tissue engineering aims to regenerate damaged tissues, developing biological substitutes which along with a thriving cell growth, may restore, maintain, or improve a functional tissue [1,2,3]. This field has undergone rapid development in the last quarter of the twentieth century, this science is devoted to skin regeneration it is still a major scientific and clinical challenge [4,5], and the healing response to chronic wounds is poorly understood and a matter of debate [6]. Polymers 2020, 12, 590 are traditionally used. Ideal skin substitutes should mimic the natural functions of the skin and the structural properties of the extracellular matrix; it has to protect the organism from protein loss, and it should improve the aesthetic appearance of the wound as well as inhibit the growth of exogenous microorganisms [7,8]

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