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      In Vitro and In Vivo Characterization of PLLA-316L Stainless Steel Electromechanical Devices for Bone Tissue Engineering—A Preliminary Study

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          Abstract

          Bone injuries represent a major social and financial impairment, commonly requiring surgical intervention due to a limited healing capacity of the tissue, particularly regarding critical-sized defects and non-union fractures. Regenerative medicine with the application of bone implants has been developing in the past decades towards the manufacturing of appropriate devices. This work intended to evaluate medical 316L stainless steel (SS)-based devices covered by a polymer poly (L-lactic acid) (PLLA) coating for bone lesion mechanical and functional support. SS316L devices were subjected to a previously described silanization process, following a three-layer PLLA film coating. Devices were further characterized and evaluated towards their cytocompatibility and osteogenic potential using human dental pulp stem cells, and biocompatibility via subcutaneous implantation in a rat animal model. Results demonstrated PLLA-SS316L devices to present superior in vitro and in vivo outcomes and suggested the PLLA coating to provide osteo-inductive properties to the device. Overall, this work represents a preliminary study on PLLA-SS316L devices’ potential towards bone tissue regenerative techniques, showing promising outcomes for bone lesion support.

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          Materials design for bone-tissue engineering

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            On the Piezoelectric Effect of Bone

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              An Alizarin red-based assay of mineralization by adherent cells in culture: comparison with cetylpyridinium chloride extraction.

              Alizarin red S (ARS) staining has been used for decades to evaluate calcium-rich deposits by cells in culture. It is particularly versatile in that the dye can be extracted from the stained monolayer and assayed. This study describes a sensitive method for the recovery and semiquantification of ARS in a stained monolayer by acetic acid extraction and neutralization with ammonium hydroxide followed by colorimetric detection at 405 nm. This method was three times more sensitive than an older method involving cetylpyridinium chloride (CPC) extraction and resulted in a better signal to noise ratio, especially for weakly stained monolayers. The assay facilitates detailed inspection of mineralization by phase microscopy and semiquantification of the entire monolayer by extraction and quantification. The sensitivity of the assay is improved by the extraction of the calcified mineral at low pH and, since the mineral is already stained in a quantitative manner, there is no requirement for an additional colorimetric quantification step. Furthermore, the linear range is much wider than those of conventional assays for calcium, making dilutions of mineral extracts prior to measurement unnecessary. It has a wide range of potential uses including tumor characterization, mesenchymal stem cell evaluation, and osteogenic compound screening. Although more labor intensive than CPC extraction, the protocol is more sensitive and yields more reliable results for weakly mineralizing samples.
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                Author and article information

                Contributors
                Role: Academic Editor
                Role: Academic Editor
                Role: Academic Editor
                Journal
                Int J Mol Sci
                Int J Mol Sci
                ijms
                International Journal of Molecular Sciences
                MDPI
                1422-0067
                17 July 2021
                July 2021
                : 22
                : 14
                : 7655
                Affiliations
                [1 ]Veterinary Clinics Department, Abel Salazar Biomedical Sciences Institute (ICBAS), University of Porto (UP), Rua de Jorge Viterbo Ferreira, n° 228, 4050-313 Porto, Portugal; m.esteves.vieira@ 123456gmail.com (M.V.B.); ruialvites@ 123456hotmail.com (R.D.A.); anacatarinasoaressousa@ 123456hotmail.com (A.C.S.)
                [2 ]Animal Science Studies Centre (CECA), Agroenvironment, Technologies and Sciences Institute (ICETA), University of Porto (UP), Rua D. Manuel II, Apartado 55142, 4051-401 Porto, Portugal
                [3 ]Department of Materials and Ceramic Engineering, CICECO, Aveiro Materials Institute, University of Aveiro, 3810-381 Aveiro, Portugal; sheila_oliveira_93@ 123456hotmail.com (S.O.F.); adrianamagueta@ 123456ua.pt (A.F.M.); maxim.ivanov@ 123456uc.pt (M.I.); helena.fernandes@ 123456ua.pt (M.H.V.F.); paula.vilarinho@ 123456ua.pt (P.M.V.)
                [4 ]Department of Pathology and Molecular Immunology, Abel Salazar Biomedical Sciences Institute (ICBAS), University of Porto (UP), Rua Jorge Viterbo Ferreira, n° 228, 4050-313 Porto, Portugal; irinamorim@ 123456hotmail.com (I.A.); fatimafaria10@ 123456yahoo.com.br (F.F.)
                [5 ]Institute of Research and Innovation in Health (i3S), University of Porto (UP), Rua Alfredo Allen, 4200-135 Porto, Portugal
                Author notes
                [* ]Correspondence: acmauricio@ 123456icbas.up.pt
                [†]

                Both authors contributed equally.

                Author information
                https://orcid.org/0000-0003-1328-464X
                https://orcid.org/0000-0003-1945-6377
                https://orcid.org/0000-0001-9155-9288
                https://orcid.org/0000-0002-0018-9363
                Article
                ijms-22-07655
                10.3390/ijms22147655
                8303773
                34299274
                c02365c1-c0b8-4e3e-b27d-6132213e0492
                © 2021 by the authors.

                Licensee MDPI, Basel, Switzerland. This article is an open access article distributed under the terms and conditions of the Creative Commons Attribution (CC BY) license ( https://creativecommons.org/licenses/by/4.0/).

                History
                : 27 June 2021
                : 14 July 2021
                Categories
                Article

                Molecular biology
                plla,ss316l,bone regeneration,biomaterials,cytocompatibility,biocompatibility
                Molecular biology
                plla, ss316l, bone regeneration, biomaterials, cytocompatibility, biocompatibility

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