Biomaterials and Adipose-Derived Mesenchymal Stem Cells for Regenerative Medicine: A Systematic Review.

The use of biological templates for the suitable growth of adipose-derived mesenchymal stem cells (AD-MSC) and "neo-tissue" construction has exponentially increased over the last years. The bioengineered scaffolds still have a prominent and biocompatible framework playing a role in tissue regeneration. In order to supply AD-MSCs, biomaterials, as the stem cell niche, are more often supplemented by or stimulate molecular signals that allow differentiation events into several strains, besides their secretion of cytokines and effects of immunomodulation. This systematic review aims to highlight the details of the integration of several types of biomaterials used in association with AD-MSCs, collecting notorious and basic data of in vitro and in vivo assays, taking into account the relevance of the interference of the cell lineage origin and handling cell line protocols for both the replacement and repairing of damaged tissues or organs in clinical application. Our group analyzed the quality and results of the 98 articles selected from PubMed, Scopus and Web of Science. A total of 97% of the articles retrieved demonstrated the potential in clinical applications. The synthetic polymers were the most used biomaterials associated with AD-MSCs and almost half of the selected articles were applied on bone regeneration.

Immediate laser-induced hemostasis in anticoagulated rats subjected to oral soft tissue surgery: a double-blind study.

Given the growing trend towards medical indications for continuous use of anticoagulants, the number of patients on these medications continues to rise. The management of patients on oral anticoagulants requiring oral surgical procedures has aroused much controversy. Changes in an anticoagulation regimen are associated with an increased risk of thromboembolism. However, it seems logical and advantageous for the patients' health if surgery could be performed without any change to the anticoagulation therapy. In dentistry, high-power lasers have been poorly explored in this field. The hemostatic properties of high-power lasers could be helpful during oral soft tissue surgeries in anticoagulated patients. The aim of this study was to compare bleeding time in anticoagulated rats after lingual frenectomy performed with a scalpel or diode laser with bleeding time in healthy animals. Twenty-four male Wistar rats were assigned to four groups (n = 6): (CS) Control-Scalpel Surgery; (AS) Anticoagulated-Scalpel Surgery; (CL) Control-Laser (diode laser 810 nm/1.5 W) Surgery; and (AL) Anticoagulated-Laser Surgery (diode laser 810 nm/1.5 W). Warfarin administration was used to induce anticoagulation. Blood was blotted every 30 seconds with filter paper until bleeding stopped to verify bleeding time. Two blinded researchers performed the surgeries and collected the bleeding time data. Diode laser surgery led to complete hemostasis in rats during and after lingual frenectomy. Zero bleeding was assessed during surgeries and after diode laser surgeries in anticoagulated rats. Laser-induced hemostasis offered an alternative solution to the controversial issue of intraoperative and postoperative bleeding control in patients on anticoagulation therapy.

Immediate laser-induced hemostasis in anticoagulated rats subjected to oral soft tissue surgery: a double-blind study

Abstract Given the growing trend towards medical indications for continuous use of anticoagulants, the number of patients on these medications continues to rise. The management of patients on oral anticoagulants requiring oral surgical procedures has aroused much controversy. Changes in an anticoagulation regimen are associated with an increased risk of thromboembolism. However, it seems logical and advantageous for the patients' health if surgery could be performed without any change to the anticoagulation therapy. In dentistry, high-power lasers have been poorly explored in this field. The hemostatic properties of high-power lasers could be helpful during oral soft tissue surgeries in anticoagulated patients. The aim of this study was to compare bleeding time in anticoagulated rats after lingual frenectomy performed with a scalpel or diode laser with bleeding time in healthy animals. Twenty-four male Wistar rats were assigned to four groups (n = 6): (CS) Control-Scalpel Surgery; (AS) Anticoagulated-Scalpel Surgery; (CL) Control-Laser (diode laser 810 nm/1.5 W) Surgery; and (AL) Anticoagulated-Laser Surgery (diode laser 810 nm/1.5 W). Warfarin administration was used to induce anticoagulation. Blood was blotted every 30 seconds with filter paper until bleeding stopped to verify bleeding time. Two blinded researchers performed the surgeries and collected the bleeding time data. Diode laser surgery led to complete hemostasis in rats during and after lingual frenectomy. Zero bleeding was assessed during surgeries and after diode laser surgeries in anticoagulated rats. Laser-induced hemostasis offered an alternative solution to the controversial issue of intraoperative and postoperative bleeding control in patients on anticoagulation therapy.

Evaluation of Eye Protection Filters Used with Broad-Spectrum and Conventional LED Curing Lights.

The high irradiance and the different emission spectra from contemporary light curing units (LCU) may cause ocular damage. This study evaluated the ability of 15 eye protection filters: 2 glasses, 1 paddle design, and 12 dedicated filters to block out harmful light from a monowave (HP-3M ESPE) and a broad-spectrum (Valo, Ultradent) LED LCU. Using the anterior sensor in the MARC-Patient Simulator (BlueLight Analytics) the irradiance that was delivered through different eye protection filters was measured three times. The LCUs delivered a similar irradiance to the top of the filter. The mean values of the light that passed through the filters as percent of the original irradiance were analyzed using two-way ANOVA followed by Tukey test (a= 0.05). The emission spectra from the LCUs and through the filters were also obtained. Two-way ANOVA showed that the interaction between protective filters and LCUs significantly influenced the amount of light transmitted (p< 0.001). Tukey test showed that the amount of light transmitted through the protective filters when using the HP-3M-ESPE was significantly greater compared to when using the Valo, irrespective of the protective filter tested. When using the HP-3M-ESPE, the Glasses filter allowed significantly more light through, followed by XL 3000, ORTUS, Google Professional, Gnatus filters. The Valo filter was the most effective at blocking out the harmful light. Some protective filters were less effective at blocking the lower wavelengths of light (<420 nm). However, even in the worst scenario, the filters were able to block at least 97% of the irradiance.

Evaluation of Eye Protection Filters Used with Broad-Spectrum and Conventional LED Curing Lights

Abstract The high irradiance and the different emission spectra from contemporary light curing units (LCU) may cause ocular damage. This study evaluated the ability of 15 eye protection filters: 2 glasses, 1 paddle design, and 12 dedicated filters to block out harmful light from a monowave (HP-3M ESPE) and a broad-spectrum (Valo, Ultradent) LED LCU. Using the anterior sensor in the MARC-Patient Simulator (BlueLight Analytics) the irradiance that was delivered through different eye protection filters was measured three times. The LCUs delivered a similar irradiance to the top of the filter. The mean values of the light that passed through the filters as percent of the original irradiance were analyzed using two-way ANOVA followed by Tukey test (a= 0.05). The emission spectra from the LCUs and through the filters were also obtained. Two-way ANOVA showed that the interaction between protective filters and LCUs significantly influenced the amount of light transmitted (p< 0.001). Tukey test showed that the amount of light transmitted through the protective filters when using the HP-3M-ESPE was significantly greater compared to when using the Valo, irrespective of the protective filter tested. When using the HP-3M-ESPE, the Glasses filter allowed significantly more light through, followed by XL 3000, ORTUS, Google Professional, Gnatus filters. The Valo filter was the most effective at blocking out the harmful light. Some protective filters were less effective at blocking the lower wavelengths of light (<420 nm). However, even in the worst scenario, the filters were able to block at least 97% of the irradiance.

Resumo A alta irradiância e diferentes espectros de luz emitidos por aparelhos fotopolimerizadores (Fp) podem causar danos oculares. Este estudo avaliou a capacidade de 15 filtros de proteção ocular em bloquear a luz prejudicial de um Fp convencional (HP-3M ESPE) e outro de largo espectro (Valo, Ultradent). Utilizando sensor anterior do equioamento MARC-Patient Simulator (BlueLight Analytics inc.) a irradiância que passou através dos diferentes filtros protetores foi mensuradas três vezes. Os valores médios da irradiância que passaram pelos filtros foram analisados usando Análise de variância fatorial e pelo teste de Tukey (a= 0.05). O espetro emitido dos Fps através dos filtros também foi obtido. A análise de variância mostrou que a interação entre os filtros protetores e Fps influenciou significantemente a quantidade de luz transmitida (p<0,001). O teste de Tukey mostrou que a quantidade que luz transmitida através dos protetores oculares quando usado o HP-3M ESPE foi significantemente maior quando comparado aos valores para o Valo, independentemente do filtro testado. Quando foi utilizado a fonte de luz HP-3M ESPE, o filtro de proteção ocular permitiu significativamente maior passagem de luz, seguido por XL 3000, ORTUS, Google Professional, e pelo filtro Gnatus. O filtro do Valo foi o mais eficiente ao bloquear a luz prejudicial. Alguns filtros foram menos eficazes ao bloquear menores comprimentos de onde (<420 nm). No entanto, mesmo no pior cenário dos resultados deste estudo, os filtros foram capazes de bloquear ao menos 97% da irradiância emitida pelas fontes de luz testadas.

Combination of Bioactive Polymeric Membranes and Stem Cells for Periodontal Regeneration: In Vitro and In Vivo Analyses.

Regeneration of periodontal tissues requires a concerted effort to obtain consistent and predictable results in vivo. The aim of the present study was to test a new family of bioactive polymeric membranes in combination with stem cell therapy for periodontal regeneration. In particular, the novel polyester poly(isosorbide succinate-co-L-lactide) (PisPLLA) was compared with poly(L-lactide) (PLLA). Both polymers were combined with collagen (COL), hydroxyapatite (HA) and the growth factor bone morphogenetic protein-7 (BMP7), and their osteoinductive capacity was evaluated via in vitro and in vivo experiments. Membranes composed of PLLA/COL/HA or PisPLLA/COL/HA were able to promote periodontal regeneration and new bone formation in fenestration defects in rat jaws. According to quantitative real-time polymerase chain reaction (qRT-PCR) and Alizarin Red assays, better osteoconductive capacity and increased extracellular mineralization were observed for PLLA/COL/HA, whereas better osteoinductive properties were associated with PisPLLA/COL/HA. We concluded that membranes composed of either PisPLLA/COL/HA or PLLA/COL/HA present promising results in vitro as well as in vivo and that these materials could be potentially applied in periodontal regeneration.

Image-Guided Cryoablation of the Spine in a Swine Model: Clinical, Radiological, and Pathological Findings with Light and Electron Microscopy.

PURPOSE: This study was designed to present the feasibility of an in vivo image-guided percutaneous cryoablation of the porcine vertebral body. METHODS: The institutional animal care committee approved this study. Cone-beam computed tomography (CBCT)-guided vertebral cryoablations (n = 22) were performed in eight pigs with short, 2-min, single or double-freezing protocols. Protective measures to nerves included dioxide carbon (CO2) epidural injections and spinal canal temperature monitoring. Clinical, radiological, and pathological data with light (n = 20) or transmission electron (n = 2) microscopic analyses were evaluated after 6 days of clinical follow-up and euthanasia. RESULTS: CBCT/fluoroscopic-guided transpedicular vertebral body cryoprobe positioning and CO2 epidural injection were successful in all procedures. No major complications were observed in seven animals (87.5 %, n = 8). A minor complication was observed in one pig (12.5 %, n = 1). Logistic regression model analysis showed the cryoprobe-spinal canal (Cp-Sc) distance as the most efficient parameter to categorize spinal canal temperatures lower than 19 °C (p < 0.004), with a significant Pearson's correlation test (p < 0.041) between the Cp-Sc distance and the lowest spinal canal temperatures. Ablation zones encompassed pedicles and the posterior wall of the vertebral bodies with an inflammatory rim, although no inflammatory infiltrate was depicted in the surrounding neural structures at light microscopy. Ultrastructural analyses evidenced myelin sheath disruption in some large nerve fibers, although neurological deficits were not observed. CONCLUSIONS: CBCT-guided vertebral cryoablation of the porcine spine is feasible under a combination of a short freezing protocol and protective measures to the surrounding nerves. Ultrastructural analyses may be helpful assess the early modifications of the nerve fibers.

Ultrastructural and immunohistochemical study of the effect of sodium alendronate in the progression of experimental periodontitis in rats.

The aim of the present research was to investigate the ultrastructural aspects and the immunoexpression of receptor activator of NFκB ligand (RANKL) and osteoprotegerin (OPG) on experimental periodontal disease of alendronate (ALN)-treated rats. Male Wistar rats received daily injections of 2.5 mg/kg body weight of ALN during 7 days previously and 7, 14, and 21 days after the insertion of a 4.0 silk suture into the gingival sulcus around the right upper second molar. Specimens were fixed in 0.1% glutaraldehyde + 4% formaldehyde under microwave irradiation, decalcified in 4.13% EDTA and paraffin embedded for TRAP histochemistry and immunohistochemistry for RANKL and OPG, or embedded in Spurr epoxy resin for TEM analysis. ALN reduced the activity of osteoclasts and significantly decreased the resorption of the alveolar crest. In the control group the alveolar crest appeared resorbed by TRAP-positive osteoclasts, which presented ultrastructural features of activated cells. The immunoexpression of RANKL was not inhibited by the drug; however, the expression of OPG was increased in the treated animals. The alveolar crest of ALN-treated specimens at 21 days showed signs of osteonecrosis, like empty osteocyte lacunae, the exposed bone regions and bacterial infection. The results showed that ALN treatment in individuals with periodontal disease represents a risk of osteonecrosis because of the reduced activity of osteoclasts resultant of the increased immunoexpression of OPG.

Effects of the bisphosphonate alendronate on molars of young rats after lateral luxation.

BACKGROUND AND AIM: The bisphosphonate alendronate (ALN) was employed with the aim of investigating its effects on dental and periodontal tissues after lateral luxation of developing molars. MATERIAL AND METHODS: Twenty-one-day-old Wistar rats had their second upper molars laterally luxated. Daily 2.5 mg kg(-1) ALN injections started at the day of the luxation; controls received sterile saline solution. The teeth were analyzed 7, 14, and 21 days after the procedure. On the days cited, the maxillae were fixed, decalcified, and embedded in paraffin or Spurr resin. The paraffin sections were stained with H&E, incubated for TRAP histochemistry or immunolabeled for osteopontin (OPN). Spurr ultrathin sections were examined in a transmission electron microscope. RESULTS: After 21 days, the root apex of luxated molars without ALN was wide open and disorganized and also covered by an irregular layer of cellular cementum, which was not observed in ALN-treated animals. Ankylosis sites were observed in ALN rats in both luxated and non-luxated teeth. The TRAP-positive osteoclasts were more numerous in ALN group, despite their latent ultrastructural appearance without the presence of resorption apparatus compared to controls. OPN immunolabeling revealed a thick immunopositive line in the dentin that must be resultant from the moment of the luxation, while ALN-treated specimens did not present alterations in dentin. CONCLUSION: The present findings indicate that alendronate inhibits some alterations in dentin and cementum formation induced by dental trauma.

Estudo do processo de reparação óssea em defeitos na calvária de ratos utilizando sinvastatina associada a um polímero de poli (ácido láctico-co-glicólico) / Study of the bone repair process of defects in rat calvaria using loaded polymer poly (lactide-co-glycolic acid) loaded with simvastatin

Devido à sua peculiar estrutura mineralizada, o tecido ósseo ainda representa um dos desafios para os estudos em bioengenharia regenerativa. Nesse sentido, estratégias realizadas envolvendo o sistema de liberação de drogas através de polímeros bioreabsorvíveis apontam resultados promissores. Esse projeto teve como objetivo empregar PLGA [poli (ácido lático-co-glicólico)] carregado com sinvastatina (SIN) em defeitos ósseos críticos criados em osso parietal de ratos. Foi realizado um defeito de 5.0 mm de diâmetro no osso parietal esquerdo, sendo que no grupo controle (C) foi deixado apenas o coágulo sanguíneo. O grupo experimental foi subdividido como segue: no subgrupo (M), uma membrana de PLGA foi colocada recobrindo o defeito ósseo, de modo que as bordas ultrapassaram a borda do defeito. No outro subgrupo (MSI), microesferas de PLGA 50/50 contendo sinvastatina a 2,5% foram depositadas no interior do defeito, sendo posteriormente recobertas pela membrana de PLGA; no grupo (MSS) foram colocadas microesferas de PLGA sem sinvastatina para avaliar o efeito osteocondutor das microesferas. Os animais foram sacrificados em dois períodos 30 e 60 dias após a cirurgia e as calotas removidas e processadas para análise morfológica em microscopia de luz, microscopia eletrônica de transmissão e varredura, imuno-histoquímica para análise da expressão de OPN (osteopontina), BSP (sialoproteína óssea) e OSAD (osteoaderina) e análise imunocitoquímica da distribuição ultra estrutural da proteína OPN. Os resultados observados mostraram que a formação do novo osso iniciou-se pelas margens do defeito e também pela área central em grupos tratados. A análise da regeneração nos defeitos tratados com SIN mostrou uma disposição ordenada das fibrilas colágenas e uma matriz com aspecto de tecido ósseo em fase mais madura. O estudo mostra que a regeneração do tecido pode ser acelerada e que a matriz neorformada é depositada de forma mais organizada pela liberação gradual da SIN...

Due to its unique mineralized structure, bone regeneration is still a challenge. Numerous strategies had been proposed during the past years, drug delivery system using polymeric scaffolds is a promising strategy. The aim of this study was to employ PLGA [poly (lactide-co-glycolic acid)] loaded with simvastatin(SIN) in bone defects created in rat calvaria. Bone defects with 5.0 mm diameter was created in the left side of the calvarias. In Control group (C) their defects were filled only with blood clot. The experimental group was subdivided. In subgroup (M), a PLGA membrane was covered the defect. In the other subgroup (MSI), PLGA microspheres loaded with 2.5% simvastatin filled the defect over the blood clot inside the defect and subsequently covered with PLGA membrane, another group was criated to evaluet the osteoconductor potencial of microesphers without the (SIN), the MSS group. The animals were sacrificed in 30, 60 after surgery, and the calvarias were removed and processed for light and transmission and scanning electron microscopy analyzes, Osteopontin (OPN), Osteoadherin (OSAD), Bone sialoprotein (BSP) imunnolabeling and immunocytochemical analyzes of the ultrastructural distribution of osteopontin in the neoformed bone. The in vivo experiment revealed that the microspheres containing simvastatin significantly enhanced the disposition of collagen fibrils in immature bone and bone formation in the rabbit calvaria critical size defect...

Immunolocalization of Smad-4 in developing molar roots of alendronate-treated rats.

INTRODUCTION: During root formation, Smad-4 plays a key role during the epithelial-mesenchymal interactions and the Hertwig's epithelial root sheath (HERS) apical proliferation. The root formation and eruption of rat molars is impeded by alendronate treatment due to the inhibition of bone resortion by this drug. The present study aimed to examine the structures affected in the developing root and immunodetect the presence of Smad-4 in rats treated with alendronate. METHODS: Newborn Wistar rats were daily injected 2.5 mg/kg alendronate (ALN) during 9, 12 and 30 days. The controls (CON) were injected with saline. The maxillae were fixed and embedded in paraffin or Spurr resin. Paraffin sections were incubated in Smad-4 antibody that was labelled with DAB. The ultrathin sections were examined in a transmission electron microscope. RESULTS: In ALN, a short portion of root dentine was formed; the epithelial diaphragm (ED) and the dental follicle (DF) were disorganized by the contact of bone trabeculae. The (CON) molar roots developed normally. Smad-4 labelling was detected in the cytoplasm of fibroblasts and cementoblasts adjacent to the cementum in CON; in ALN group, few ED cells presented weak immunolabelling. Ultrastructurally, the ED and DF appeared disrupted due to the presence of thin bone trabeculae between its cells. It resulted in the lack of apical proliferation of HERS and, consequently, arrest of root formation. CONCLUSION: The immunodetection of Smad-4 in the DF cells of ALN specimens indicates that the signalling for the differentiation of these cells into cementum-forming fibroblasts and cementoblasts occurs, despite the impairment of root elongation.