The effects of poly L-lactic acid nanofiber scaffold on mouse spermatogonial stem cell culture

Neda Eslahi,1,2,* Mahmoud Reza Hadjighassem,1,3 Mohammad Taghi Joghataei,1,2 Tooba Mirzapour,4 Mehrdad Bakhtiyari,1,2 Malak Shakeri,5 Vahid Pirhajati,1,2 Peymaneh Shirinbayan,6,* Morteza Koruji1,21Cellular and Molecular Research Center, Iran University of Medical Sciences, Tehran, Iran; 2Department of Anatomical Sciences, School of Medicine, Iran University of Medical Sciences, Tehran, Iran; 3Department of Neurosciences, School of Advanced Medical Technologies, Tehran University of Medical Sciences, Tehran, Iran; 4Department of Biology, Faculty of Science, University of Mohaghegh Ardabili, Ardabil, Iran; 5Department of Animal Science, Agricultural Campus, University of Tehran, Tehran, Iran; 6Pediatric Neuro-Rehabilitation Research Center, University of Social Welfare and Rehabilitation Sciences, Tehran, Iran*These authors contributed equally to this articleIntroduction: A 3D-nanofiber scaffold acts in a similar way to the extracellular matrix (ECM)/basement membrane that enhances the proliferation and self-renewal of stem cells. The goal of the present study was to investigate the effects of a poly L-lactic acid (PLLA) nanofiber scaffold on frozen-thawed neonate mouse spermatogonial stem cells (SSCs) and testis tissues.Methods: The isolated spermatogonial cells were divided into six culture groups: (1) fresh spermatogonial cells, (2) fresh spermatogonial cells seeded onto PLLA, (3) frozen-thawed spermatogonial cells, (4) frozen-thawed spermatogonial cells seeded onto PLLA, (5) spermatogonial cells obtained from frozen-thawed testis tissue, and (6) spermatogonial cells obtained from frozen-thawed testis tissue seeded onto PLLA. Spermatogonial cells and testis fragments were cryopreserved and cultured for 3 weeks. Cluster assay was performed during the culture. The presence of spermatogonial cells in the culture was determined by a reverse transcriptase polymerase chain reaction for spermatogonial markers (Oct4, GFRα-1, PLZF, Mvh(VASA), Itgα6, and Itgβ1), as well as the ultrastructural study of cell clusters and SSCs transplantation to a recipient azoospermic mouse. The significance of the data was analyzed using the repeated measures and analysis of variance.Results: The findings indicated that the spermatogonial cells seeded on PLLA significantly increased in vitro spermatogonial cell cluster formations in comparison with the control groups (culture of SSCs not seeded on PLLA) (P≤0.001). The viability rate for the frozen cells after thawing was 63.00% ± 3.56%. This number decreased significantly (40.00% ± 0.82%) in spermatogonial cells obtained from the frozen-thawed testis tissue. Both groups, however, showed in vitro cluster formation. Although the expression of spermatogonial markers was maintained after 3 weeks of culture, there was a significant downregulation for some spermatogonial genes in the experimental groups compared with those of the control groups. Furthermore, transplantation assay and transmission electron microscopy studies suggested the presence of SSCs among the cultured cells.Conclusion: Although PLLA can increase the in vitro cluster formation of neonate fresh and frozen-thawed spermatogonial cells, it may also cause them to differentiate during cultivation. The study therefore has implications for SSCs proliferation and germ cell differentiation in vitro.Keywords: PLLA nanofibers, tissue cryopreservation, testi

Synthesis, physico-chemical and biological characterization of strontium and cobalt substituted bioactive glasses for bone tissue engineering

The research was financially supported by Iran National Science Foundation (INSF) through research Grant No. 91058572

Strontium- and cobalt-substituted bioactive glasses seeded with human umbilical cord perivascular cells to promote bone regeneration via enhanced osteogenic and angiogenic activities

Cellular and Molecular Research Center (CMRC), Iran University of Medical Sciences

Análise radiográfica da coluna cervical em indivíduos assintomáticos submetidos a tração manual Radiographic analysis of the cervical spine in healthy individuals submitted to manual traction

OBJETIVO: Avaliar, radiograficamente, o efeito da tração manual sobre o comprimento da coluna cervical. MATERIAIS E MÉTODOS: Cinqüenta e cinco participantes de ambos os gêneros - 12 masculinos (22%) e 43 femininos (78%) - sem história de distúrbios cervicais contituíram a amostra deste estudo. Eles foram submetidos a dois procedimentos radiológicos, um antes e outro durante a tração manual sustentada por 120 segundos. As distâncias entre as bordas anteriores e posteriores da segunda à sétima vértebras cervicais foram mensuradas e comparadas antes e durante a tração manual. RESULTADOS: A mediana da distância anterior antes da tração foi de 8,40 cm e durante a tração aumentou para 8,50 cm (p=0,002). A mediana da distância posterior antes da tração foi de 8,35 cm e durante a tração aumentou para 8,50 cm (p<0,001). CONCLUSÃO: Os resultados demonstraram que a aplicação da tração manual promoveu aumento estatisticamente significante do comprimento da coluna cervical em indivíduos assintomáticos.<br>OBJECTIVE: To evaluate radiographically the effect of manual traction on the length of the cervical spine in healthy individuals. MATERIALS AND METHODS: The sample of the present study included 55 individuals - 12 men (22%) and 43 women (78%) - with no previous history of cervical disorders, submitted to two radiological procedures previously and during manual traction sustained for 120 seconds. Distances between the anterior and posterior edges from the second to the seventh cervical vertebrae were measured and compared before and during manual traction. RESULTS: The median of pre-traction anterior length was 8.40 cm, increasing to 8.50 cm during the traction (p=0.002); and the median of pre-traction posterior length was 8.35 cm, increasing to 8.50 cm during traction (p<0.001). CONCLUSION: Application of manual traction resulted in a statistically significant increase in the length the cervical spine in healthy individuals

Neck-specific training with a cognitive behavioural approach compared with prescribed physical activity in patients with cervical radiculopathy : a protocol of a prospective randomised clinical trial

Background: Patients with cervical radiculopathy often have neck- and arm pain, neurological changes, activity limitations and difficulties in returning to work. Most patients are not candidates for surgery but are often treated with different conservative approaches and may be sick-listed for long periods. The purpose of the current study is to compare the effectiveness of neck- specific training versus prescribed physical activity. Methods/Design: The current protocol is a two armed intervention randomised clinical trial comparing the outcomes of patients receiving neck specific training or prescribed physical activity. A total of 144 patients with cervical radiculopathy will be randomly allocated to either of the two interventions. The interventions will be delivered by experienced physiotherapists and last 14 weeks. The primary outcome variable is neck- and arm pain intensity measured with a Visual Analogue Scale accompanied with secondary outcome measures of impairments and subjective health measurements collected before intervention and at 3, 6, 12 and 24 months after base-line assessment. Discussion: We anticipate that the results of this study will provide evidence to support recommendations as to the effectiveness of conservative interventions for patients with cervical radiculopathy.Funding Agencies|Swedish Government through the Karolinska Institute; Swedish National School of Research Education in Health Care Sciences; Strategic Research Programme in Care Sciences; Stockholm County Council Funding ALF Medicine</p

Development of a clinical prediction rule to identify patients with neck pain likely to benefit from cervical traction and exercise

The objective of the study was to develop a clinical prediction rule (CPR) to identify patients with neck pain likely to improve with cervical traction. The study design included prospective cohort of patients with neck pain referred to physical therapy. Development of a CPR will assist clinicians in classifying patients with neck pain likely to benefit from cervical traction. Eighty patients with neck pain received a standardized examination and then completed six sessions of intermittent cervical traction and cervical strengthening exercises twice weekly for 3 weeks. Patient outcome was classified at the end of treatment, based on perceived recovery according to the global rating of change. Patients who achieved a change ≥+6 (“A great deal better” or “A very great deal better”) were classified as having a successful outcome. Univariate analyses (t tests and chi-square) were conducted on historical and physical examination items to determine potential predictors of successful outcome. Variables with a significance level of P ≤ 0.15 were retained as potential prediction variables. Sensitivity, specificity and positive and negative likelihood ratios (LRs) were then calculated for all variables with a significant relationship with the reference criterion of successful outcome. Potential predictor variables were entered into a step-wise logistic regression model to determine the most accurate set of clinical examination items for prediction of treatment success. Sixty-eight patients (38 female) were included in data analysis of which 30 had a successful outcome. A CPR with five variables was identified: (1) patient reported peripheralization with lower cervical spine (C4–7) mobility testing; (2) positive shoulder abduction test; (3) age ≥55; (4) positive upper limb tension test A; and (5) positive neck distraction test. Having at least three out of five predictors present resulted in a +LR equal to 4.81 (95% CI = 2.17–11.4), increasing the likelihood of success with cervical traction from 44 to 79.2%. If at least four out of five variables were present, the +LR was equal to 23.1 (2.5–227.9), increasing the post-test probability of having improvement with cervical traction to 94.8%. This preliminary CPR provides the ability to a priori identify patients with neck pain likely to experience a dramatic response with cervical traction and exercise. Before the rule can be implemented in routine clinical practice, future studies are necessary to validate the rule. The CPR developed in this study may improve clinical decision-making by assisting clinicians in identifying patients with neck pain likely to benefit from cervical traction and exercise