ABSTRACT
Abstract This study developed and characterized a method for controlled deposition of thin films of hydroxyapatite on titanium surfaces. Thirty-three titanium cylinders were randomly divided: negative control/polished (A), acid etched (B) and coated by hydroxyapatite (C). Acid etch was performed in an aqueous solution of nitric acid. The cylinders were subjected to coating by a thin film of hydroxyapatite with dip-coating method. These cylinders were submitted to a pre-heat treatment 450°C/10 minutes and 800°C/2 hours. Scanning electron microscopy analysis demonstrated a homogeneous and smooth surface (A), an irregular and porous surface (B) and a crystalline deposition (C). The X-ray energy dispersive analysis showed characteristic elements of hydroxyapatite (C). Analysis by X-ray diffraction showed the presence of characteristic peaks of hydroxyapatite, corresponding to the structural composition of hydroxyapatite. Cell viability (MTT-assay in NIH-3T3-Cells) test demonstrated no differences between the groups. Titanium surfaces coated with a hydroxyapatite film by the dip-coating method suggest adequate control of deposition of thin films of hydroxyapatite and similar cell viability using mouse fibroblasts.
ABSTRACT
Abstract This systematic review evaluated if different cryopreservation protocols could affect biological properties (Cell survival rate (CSR), proliferation, differentiation, maintenance of stem cell markers) of stem cells obtained from dental tissues (DSC) post-thaw. An electronic search was carried out within PubMed and ISI Web Science by using specific keyword. Two independent reviewers read the titles and abstracts of all reports respecting predetermined inclusion/exclusion criteria. Data were extracted considering the biological properties of previously cryopreserved DSCs and previously cryopreserved dental tissues. DSCs cryopreserved as soon as possible after their isolation presents a CSR quite similar to the non-cryopreserved DSC. Dimethyl sulfoxide (DMSO) [10%] showed good results related to cell recovery post-thaw to cryopreserve cells and tissues for periods of up to 2 years. The cryopreservation of DSC in a mechanical freezer (-80°C) allows the recovery of stem cells post-thaw. The facilities producing magnetic field (MF), demand a lower concentration of cryoprotectant, but their use is not dispensable. It is possible to isolate and cryopreserve dental pulp stem cell (DPSC) from healthy and diseased vital teeth. Cryopreservation of dental tissues for late DSC isolation, combined with MF dispensability, could be valuable to reduce costs and improve the logistics to develop teeth banks.
Resumo Essa revisão sistemática avaliou se diferentes protocolos de criopreservação podem afetar as propriedades biológicas (taxa de sobrevivência celular, proliferação, diferenciação, manutenção dos marcadores de superfície) de células-tronco isoladas de tecidos dentais (DSC) após o descongelamento. Uma busca eletrônica foi realizada no PubMed e no ISI Web of Science utilizando palavras-chave específicas. Dois revisores independentes avaliaram os títulos e resumos de todos os estudos respeitando critérios de inclusão e exclusão previamente determinados. Os dados foram extraídos considerando as propriedades biológicas de DSC, e DSC isoladas de tecidos previamente criopreservados. DSC criopreservadas logo após seu isolamento apresentaram propriedades biológicas muito semelhantes às observadas em DSC não criopreservadas. Dimetil sulfóxido (DMSO) [10%] demonstrou bons resultados relacionados com a recuperação celular após descongelamento de células e tecidos, por períodos de até 2 anos. A criopreservação de DSC em freezer mecânico (-80 °C) permite a recuperação de células-tronco pós-descongelação. A utilização de freezer com campo magnético (MF), proporciona a utilização de uma menor concentração de crioprotector, mas a sua utilização não é dispensável. É possível isolar e criopreservar e criopreservar células-tronco da polpa dental (DPSC) de dentes vitais saudáveis e doentes. Criopreservação de tecidos dentais após o isolamento de DSC, combinados com MF, podem ser valiosas estratégias para reduzir custos e melhorar a logística no desenvolvimento de bancos de dentes.
Subject(s)
Humans , Tooth/cytology , Cryopreservation , Mesenchymal Stem Cells/cytology , Cell SurvivalABSTRACT
Abstract The aim of the present study was to evaluate the expression of transforming growth factor-β1 (TGF-β1) and osteonectin (ON) in pulp-like tissues developed by tissue engineering and to compare it with the expression of these proteins in pulps treated with Ca(OH)2 therapy. Tooth slices were obtained from non-carious human third molars under sterile procedures. The residual periodontal and pulp soft tissues were removed. Empty pulp spaces of the tooth slice were filled with sodium chloride particles (250-425 µm). PLLA solubilized in 5% chloroform was applied over the salt particles. The tooth slice/scaffold (TS/S) set was stored overnight and then rinsed thoroughly to wash out the salt. Scaffolds were previously sterilized with ethanol (100-70°) and washed with phosphate-buffered saline (PBS). TS/S was treated with 10% EDTA and seeded with dental pulp stem cells (DPSC). Then, TS/S was implanted into the dorsum of immunodeficient mice for 28 days. Human third molars previously treated with Ca(OH)2 for 90 days were also evaluated. Samples were prepared and submitted to histological and immunohistochemical (with anti-TGF-β1, 1:100 and anti-ON, 1:350) analyses. After 28 days, TS/S showed morphological characteristics similar to those observed in dental pulp treated with Ca(OH)2. Ca(OH)2-treated pulps showed the usual repaired pulp characteristics. In TS/S, newly formed tissues and pre-dentin was colored, which elucidated the expression of TGF-β1 and ON. Immunohistochemistry staining of Ca(OH)2-treated pulps showed the same expression patterns. The extracellular matrix displayed a fibrillar pattern under both conditions. Regenerative events in the pulp seem to follow a similar pattern of TGF-β1 and ON expression as the repair processes.