Adult mesenchymal progenitor cells have enormous potential for use in regenerative medicine. However, the true identity of the progenitors in vivo and their progeny has not been precisely defined. We hypothesize that cells expressing a smooth muscle α-actin promoter (αSMA) directed Cre transgene represent mesenchymal progenitors of adult bone tissue. By combining complementary colors in combination with transgenes activating at mature stages of the lineage we characterized the phenotype and confirmed the ability of isolated αSMA+ cells to progress from a progenitor to fully mature state. In vivo lineage tracing experiments using a new bone formation model confirmed the osteogenic phenotype of αSMA+ cells. In vitro analysis of the in vivo labeled SMA9+ cells supported their differentiation potential into mesenchymal lineages. Utilizing a fracture-healing model, αSMA+ cells served as a pool of fibrocartilage and skeletal progenitors. Confirmation of the transition of αSMA+ progenitor cells to mature osteoblasts during fracture healing was assessed by activation of bone specific Col2.3emd transgene. Our findings provide a novel in vivo identification of defined population of mesenchymal progenitor cells with active role in bone remodeling and regeneration.
Bracket design does not seem to have a strong influence on periodontal clinical parameters and periodontal pathogens in subgingival plaque. The correlation between some periodontal pathogens and clinical periodontal parameters was weak.
Previous studies reported that embryonic stem cells (ESCs) can be induced to differentiate into cells showing a mature osteoblastic phenotype by culturing them under osteo-inductive conditions. It is probable that osteogenic differentiation requires that ESCs undergo differentiation through an intermediary step involving a mesenchymal lineage precursor. Based on our previous studies indicating that adult mesenchymal progenitor cells express αSMA, we have generated ESCs from transgenic mice in which an αSMA promoter directs the expression of red fluorescent protein (RFP) to mesenchymal progenitor cells. To track the transition of ESC-derived MSCs into mature osteoblasts, we have utilized a bone-specific fragment of rat type I collagen promoter driving green fluorescent protein (Col2.3GFP). Following osteogenic induction in ESCs, we have observed expression of alkaline phosphatase and subsequent mineralization as detected by von Kossa staining. After one week of osteogenic induction, ESCs begin to express αSMARFP. This expression was localized to the peripheral area encircling a typical ESC colony. Nevertheless, these αSMARFP positive cells did not show activation of the Col2.3GFP promoter, even after 7 weeks of osteogenic differentiation in vitro. In contrast, Col2.3GFP expression was detected in vivo, in mineralized areas following teratoma formation. Our results indicate that detection of alkaline phosphatase activity and mineralization of ESCs cultured under osteogenic conditions is not sufficient to demonstrate osteogenic maturation. Our study indicates the utility of the promoter-visual transgene approach to assess the commitment and differentiation of ESCs into the osteoblast lineage.
SummAry -The aim of the study was to assess the influence of fixed orthodontic appliance on Streptococcus (S.) mutans and S. sobrinus counts in orthodontic patients with regard to their previous caries experience (decayed, missing and filled teeth (dmft) index) during the first 12 weeks of orthodontic treatment. twenty-two patients that satisfied inclusion criteria (healthy systemic and periodontal condition, avoidance of antibiotic therapy and antiseptic mouthwashes in the past three months) were included. All clinical measurements took place prior to and 12 weeks after fixed orthodontic appliance placement, in the following order: 1) stimulated saliva flow (SS); 2) Simplified Oral hygiene index (Ohi-S); and 3) dmft. The method of polymerase chain reaction (PCr) was used to detect the presence of S. mutans and S. sobrinus at t1 and t2. t-test showed significant increase in dmft index and SS between t1 and t2. results also indicated significant improvement in Ohi-S index. By use of the PCr method, S. mutans was detected in two patients at t1. At t2, two more patients had S. mutans, but the increase was not statistically significant. using the same method, S. sobrinus was detected only in two patients at t2. in conclusion, fixed orthodontic appliances did not induce statistically significant changes in caries microflora even in the presence of enhanced oral hygiene habits.
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