Osteogenic potential of apical papilla stem cells mediated by platelet-rich fibrin and low-level laser.

To evaluate the osteogenic potential of platelet-rich fibrin (PRF) and low-level laser therapy (LLLT) on human stem cells from the apical papilla (SCAP) we isolated, characterized, and then cultured in an osteogenic medium cells with PRF and/or LLLT (660 nm, 6 J/m2-irradiation). Osteogenic differentiation was assessed by bone nodule formation and expression of bone morphogenetic proteins (BMP-2 and BMP-4), whereas the molecular mechanisms were achieved by qRT-PCR and RNA-seq analysis. Statistical analysis was performed by ANOVA and Tukey's post hoc tests (p < 0.05* and p < 0.01**). Although PRF and LLLT increased bone nodule formation after 7 days and peaked at 21 days, the combination of PRF + LLLT led to the uppermost nodule formation. This was supported by increased levels of BMP-2 and -4 osteogenic proteins (p < 0.005). Furthermore, the PRF + LLLT relative expression of specific genes involved in osteogenesis, such as osteocalcin, was 2.4- (p = 0.03) and 28.3- (p = 0.001) fold higher compared to the PRF and LLLT groups, and osteopontin was 22.9- and 1.23-fold higher, respectively (p < 0.05), after 7 days of interaction. The transcriptomic profile revealed that the combination of PRF + LLLT induces MSX1, TGFB1, and SMAD1 expression, after 21 days of osteogenic differentiation conditions exposition. More studies are required to understand the complete cellular and molecular mechanisms of PRF plus LLLT on stem cells. Overall, we demonstrated for the first time that the combination of PRF and LLLT would be an excellent therapeutic tool that can be employed for dental, oral, and craniofacial repair and other tissue engineering applications.

Closure of a Difficult-to-Manage Chronic Pressure Injury with the Use of Autologous Platelet-Rich Plasma (APRP).

Pressure injury (PI) corresponds to a skin damage of ischemic aetiology that affects the integrity of the skin and is produced by prolonged pressure or friction between a hard internal and external surface. Treatment can be challenging when there is no resolution with usual care. The use of autologous platelet-rich plasma (APRP) gel arises as a therapeutic possibility in the presence of chronic pressure injuries. The case of a patient with chronic PI who has been treated with APRP is presented, achieving resolution of the lesion.

Trypanosoma cruzi infection in transfusion medicine

ABSTRACT Introduction: Infection by Trypanosoma cruzi is challenging to blood bank supplies in terms of accurate diagnosis, mostly due to its clinical complexity. Infected individuals may remain asymptomatic for years, albeit they may have circulating parasites potentially transferable to eventual receptors of a transfusion. Objective: Although risk donors are systematically excluded through a survey, an important residual risk for transmission remains, evidencing the need to implement additional actions for the detection of T. cruzi in blood banks. Method: A review of the scientific literature is presented with the objective of identifying relevant publications on this subject. Results: We discuss the diagnostic considerations of this chronic infection on transfusion medicine and some recent advances in the processing of blood and derivatives units. Conclusion: Finally, recommendations are made on how the transmission of T. cruzi can be avoided through the implementation of better diagnostic and pathogen control measures at blood banks.

Trypanosoma cruzi infection in transfusion medicine.

INTRODUCTION: Infection by Trypanosoma cruzi is challenging to blood bank supplies in terms of accurate diagnosis, mostly due to its clinical complexity. Infected individuals may remain asymptomatic for years, albeit they may have circulating parasites potentially transferable to eventual receptors of a transfusion. OBJECTIVE: Although risk donors are systematically excluded through a survey, an important residual risk for transmission remains, evidencing the need to implement additional actions for the detection of T. cruzi in blood banks. METHOD: A review of the scientific literature is presented with the objective of identifying relevant publications on this subject. RESULTS: We discuss the diagnostic considerations of this chronic infection on transfusion medicine and some recent advances in the processing of blood and derivatives units. CONCLUSION: Finally, recommendations are made on how the transmission of T. cruzi can be avoided through the implementation of better diagnostic and pathogen control measures at blood banks.

O uso do plasma rico em plaquetas no tratamento da acne e suas cicatrizes: estudo-piloto / The use of platelet-rich plasma in the treatment of acne and its scars: a pilot study

Introdução: A acne é doença multifatorial em que se evidenciam lesões inflamatórias constituídas por: pápulas, pústulas e comedões. É mais comum dos 11 aos 30 anos de idade, ocorrendo em várias regiões do corpo em que há altas concentrações de unidades pilossebáceas. A acne tem diferentes classificações e tratamentos. Recentemente, os fatores de crescimento de plaquetas têm sido indicados como terapia alternativa para cicatrizes da acne. Objetivo: Descrever o caso de paciente com acne nódulo-cística e cicatrizes de acne graus 1, 2 e 3b tratado com plasma rico em plaquetas e, ao mesmo tempo, destacar a eficácia do tratamento. Métodos: Aplicação de plasma rico em plaquetas durante três meses no lado esquerdo da face de um paciente. Resultados: De acordo com avaliações clínica e fotográfica, houve melhora na aparência e qualidade da pele, observando-se menor número de lesões e diminuição de fenômenos dolorosos. Conclusões: O presente estudo demonstrou a eficácia do plasma rico em plaquetas como tratamento para a acne assim como suas cicatrizes.

Introduction: Acne is a multifactorial disease where inflammatory lesions usually appear as papules, pustules and comedones. It is more common in people aged between 11 and 30 years, and occurs in various body sites where there are high concentrations of pilosebaceous glands. Acne has different classifications and treatments. More recently, platelet growth factors have been used as an alternative therapy for acne scars. Objective: The present study is aimed at describing a case of nodule-cystic acne and acne scars Grades 1, 2 and 3b treated with platelet rich plasma and highlighting the effectiveness of the treatment. Methods: Platelet rich plasma was applied for three months in the left side of the face of a patient bearing acne. Results: In the clinical and photographic evaluations, there was improvement in the skin appearance and quality, less number of lesions and decrease of pain. Conclusions: The present study has demonstrated the effectiveness of platelet rich plasma as a treatment for acne and resulting scars.

Erythropoietin reduces storage lesions and decreases apoptosis indices in blood bank red blood cells.

BACKGROUND: Recent evidence shows a selective destruction of the youngest circulating red blood cells (neocytolysis) trigged by a drop in erythropoietin levels. OBJECTIVE: The aim of this study was to evaluate the effect of recombinant human erythropoietin beta on the red blood cell storage lesion and apoptosis indices under blood bank conditions. METHODS: Each one of ten red blood cell units preserved in additive solution 5 was divided in two volumes of 100mL and assigned to one of two groups: erythropoietin (addition of 665IU of recombinant human erythropoietin) and control (isotonic buffer solution was added). The pharmacokinetic parameters of erythropoietin were estimated and the following parameters were measured weekly, for six weeks: Immunoreactive erythropoietin, hemolysis, percentage of non-discocytes, adenosine triphosphate, glucose, lactate, lactate dehydrogenase, and annexin-V/esterase activity. The t-test or Wilcoxon's test was used for statistical analysis with significance being set for a p-value <0.05. RESULTS: Erythropoietin, when added to red blood cell units, has a half-life >6 weeks under blood bank conditions, with persistent supernatant concentrations of erythropoietin during the entire storage period. Adenosine triphosphate was higher in the Erythropoietin Group in Week 6 (4.19±0.05µmol/L vs. 3.53±0.02µmol/L; p-value=0.009). The number of viable cells in the Erythropoietin Group was higher than in the Control Group (77%±3.8% vs. 71%±2.3%; p-value <0.05), while the number of apoptotic cells was lower (9.4%±0.3% vs. 22%±0.8%; p-value <0.05). CONCLUSIONS: Under standard blood bank conditions, an important proportion of red blood cells satisfy the criteria of apoptosis. Recombinant human erythropoietin beta seems to improve storage lesion parameters and mitigate apoptosis.

Erythropoietin reduces storage lesions and decreases apoptosis indices in blood bank red blood cells

ABSTRACT Background: Recent evidence shows a selective destruction of the youngest circulating red blood cells (neocytolysis) trigged by a drop in erythropoietin levels. Objective: The aim of this study was to evaluate the effect of recombinant human erythropoietin beta on the red blood cell storage lesion and apoptosis indices under blood bank conditions. Methods: Each one of ten red blood cell units preserved in additive solution 5 was divided in two volumes of 100 mL and assigned to one of two groups: erythropoietin (addition of 665 IU of recombinant human erythropoietin) and control (isotonic buffer solution was added). The pharmacokinetic parameters of erythropoietin were estimated and the following parameters were measured weekly, for six weeks: Immunoreactive erythropoietin, hemolysis, percentage of non-discocytes, adenosine triphosphate, glucose, lactate, lactate dehydrogenase, and annexin-V/esterase activity. The t-test or Wilcoxon's test was used for statistical analysis with significance being set for a p-value <0.05. Results: Erythropoietin, when added to red blood cell units, has a half-life >6 weeks under blood bank conditions, with persistent supernatant concentrations of erythropoietin during the entire storage period. Adenosine triphosphate was higher in the Erythropoietin Group in Week 6 (4.19 ± 0.05 µmol/L vs. 3.53 ± 0.02 µmol/L; p-value = 0.009). The number of viable cells in the Erythropoietin Group was higher than in the Control Group (77% ± 3.8% vs. 71% ± 2.3%; p-value <0.05), while the number of apoptotic cells was lower (9.4% ± 0.3% vs. 22% ± 0.8%; p-value <0.05). Conclusions: Under standard blood bank conditions, an important proportion of red blood cells satisfy the criteria of apoptosis. Recombinant human erythropoietin beta seems to improve storage lesion parameters and mitigate apoptosis.

Standardization of a Protocol for Obtaining Platelet Rich Plasma from blood Donors; a Tool for Tissue Regeneration Procedures.

BACKGROUND: To develop a protocol for obtaining autologous platelet rich plasma in healthy individuals and to determine the concentration of five major growth factors before platelet activation. This protocol could be integrated into the guidelines of good clinical practice and research in regenerative medicine. METHODS: Platelet rich plasma was isolated by centrifugation from 38 healthy men and 42 women ranging from 18 to 59 years old. The platelet count and quantification of growth factors were analyzed in eighty samples, stratified for age and gender of the donor. Analyses were performed using parametric the t-test or Pearson's analysis for non-parametric distribution. P < 0.05 was considered statistically significant. RESULTS: Our centrifugation protocol allowed us to concentrate basal platelet counts from 1.6 to 4.9 times (mean = 2.8). There was no correlation between platelet concentration and the level of the following growth factors: VEGF-D (r = 0.009, p = 0.4105), VEGF-A (r = 0.0068, p = 0.953), PDGF subunit AA (p = 0.3618; r = 0.1047), PDGF-BB (p = 0.5936; r = 0.6095). In the same way, there was no correlation between donor gender and growth factor concentrations. Only TGF-ß concentration was correlated to platelet concentration (r = 0.3163, p = 0.0175). CONCLUSIONS: The procedure used allowed us to make preparations rich in platelets, low in leukocytes and red blood cells, and sterile. Our results showed biological variations in content of growth factors in PRP. The factors influencing these results should be further studied.

Prevalence of antibodies against transfusion-transmissible infections (TTI) in blood donors from the Colombian eastern region.

BACKGROUND: The main purpose of this study was to establish the prevalence of antibodies against five transfusion-transmissible infections (TTIs) in blood donors from one of the most important blood banks in Colombia. METHODS: A cross-sectional, descriptive and case control study was performed from a database of Higuera-Escalante blood bank, for a period of a year. Serum was used for donor screening. Surface antigens for hepatitis B (HbsAg), anti-hepatitis C antibodies, Chagas disease, syphilis, and HIV were identified. Chemiluminescent Microparticle Immunoassay (CMIA, Abbott Diagnostics) was performed. RESULTS: From 41,575 total donors analyzed, 1,226 were reactive for any of the infectious markers (total prevalence of 2.95%). The prevalence of specific infections was: Chagas disease 0.49%, HbsAg 0.21%, HCV 0.45%, HIV 0.12%, and syphilis 1.68%. Reactivity was more frequent in men (n = 785, 64%) with a mean age of 36.35 years. HIV was present in the youngest donors with a mean age of 26.5 years (IC 95%: 23.6 - 27.6); on the other hand, Chagas disease was found in the oldest donor population, with a mean age of 40 years (IC 95%: 39.1 - 41.3). CONCLUSIONS: Identifying the prevalence of circulating antibodies against transfusion transmissible infections allows us to establish an epidemiological profile of donors inhabiting the geographic catchment area of our blood bank. Total prevalence in this study was 2.95% for any of the five markers. Syphilis prevalence demonstrates its high distribution within the blood donor population of our country, although this result could be influenced by the high rate of false-reactive test. Chagas disease is endemic in Santander, Colombia, which correlates with the results obtained in this study.

O que há de novo em vacinas contra tuberculose? / What's new about tuberculosis vaccines?

Objetivo: Revisar a informação publicada nos últimos 15 anos sobre a vacina BCG, a resposta imunológica ao M. tuberculosis e as vacinas que estão sendo desenvolvidas para combater a doença. Métodos: Foi realizada revisão sistemática em livros e na literatura publicada no Medline, a partir do ano de 1990. Foram selecionados artigos que abordassem o tema, “Vacinas contra TB” nos aspectos: a resposta imune humoral e celular, novas vacinas, e proteção. Resultados: Foram selecionados artigos originais ou capítulos de livros que abordaram este tema. Observou-se efetividade da vacina BCG contra TB em muitas populações, mas não em outras. Devido às diversas limitações da BCG, o desenvolvimento de uma vacina mais efetiva é considerada uma prioridade em saúde pública no mundo. As vacinas e as estratégias em desenvolvimento incluem células vivas totais, células totais inativadas, subunidades e vacinas de DNA. Conclusões: A maioria dos humanos infectados com M. tuberculosis consegue gerar uma resposta imune que evita o desenvolvimento da doença. Algumas vacinas previnem a TB, indicando que uma vacina mais efetiva contra TB é uma realidade possível. Ao se propor a substituição da vacina da BCG, será necessário demonstrar que a nova vacina pode ao menos igualar a eficácia da BCG na proteção da tuberculose.

Doença granulomatosa crônica da infância: novos polimorfismos no gene NCF2 / Chronic granulomatous disease of childhood: new polymorphisms in NCF2 gene

Objetivos: Mutações no gene NCF2 resultam na forma autossônica recessiva da doença granulomatosa crônica da infância. Além de mutações conhecidas, descreveu-se em pacientes com doença granulomatosa crônica da infância duas novas substituições no gene NCF2. O objetivo deste estudo foi investigar se estas substituições constituem polimorfismos do gene NCF2. Métodos: Investigamos a freqüência de duas substituições na seqüência do gene NCF2 em 214 doadores sadios. A primeira é uma transição de C T na posição -23 da região 5' reguladora. A segunda é uma transição de A G na posição -21 da região 3' terminal do íntron 10. Extraímos DNA genômico de células do sangue periférico. O DNA foi amplificado por meio de PCR com primers específicos para o gene NCF2, analisado quanto à presença de polimorfismos conformacionais de cadeias simples, digerido com endonucleases específicas e sequenciado. O cálculo das freqüências genotípicas e alélicas seguiu a lei de Hardy e Weinberg. Resultados: Cem indivíduos foram avaliados quanto à presença da transição C T na posição -23 da região 5' reguladora; sendo 67 porcento homozigotos para o alelo c, 32 porcento heterozigotos, e apenas 1 porcento homozigoto para T. Cento e quatorze indivíduos foram analisados quanto à presença da transição A G na posição -21 da região 3' terminal do íntron 10; dos quais 36 porcento foram homozigotos para A, 43 porcento heterozigotos e 21 porcento homozigotos para G. Conclusão: Considerando as freqüências alélicas, concluímos que essas variantes correspondem a polimorfismos do gene NCF2. Suas possíveis implicações na expressão do gene NCF2 constituem objeto de pesquisa atual em nosso laboratório.