Kevin H.-K. Yip

Mesenchymal stem cell-encapsulated collagen microspheres for bone tissue engineering.

There is a demonstrated clinical need for alternatives of autologous fresh bone graft with excellent biological performance in osteoconductivity, osteoinductivity, and osteogenicity. We previously developed a collagen microencapsulation technology entrapping bone marrow-derived mesenchymal stem cells (MSCs) in a biomimetic collagen fiber meshwork and produced injectable collagen-MSC microspheres. In this study, we hypothesize that injectable microspheres with osteoconductivity, osteogenicity, and osteoinductivity can be fabricated by differentiating the encapsulated MSCs, from either human or mouse sources, toward osteogenic lineages in these three-dimensional microspheres. The osteogenicity, osteoconductivity, and osteoinductivity of the microspheres were evaluated in vitro. Osteogenic markers of the differentiating MSCs including alkaline phosphatase and calcium deposition showed positive staining. Osteoconductivity of the collagen meshwork in the microsphere was demonstrated by the presence of calcium phosphate deposits among the collagen fibers and by the significantly increased calcium content extracted from the microspheres. Moreover, osteoinductivity of the MSC-encapsulated microspheres was demonstrated by the ability to induce osteogenic differentiation of undifferentiated MSCs in both contact and noncontact coculture. This study contributes toward the future development of injectable alternatives for fresh bone grafts using autologous MSCs.

Prolonged exposure to bacterial toxins downregulated expression of toll-like receptors in mesenchymal stromal cell-derived osteoprogenitors

BackgroundHuman mesenchymal stromal cells (MSCs, also known as mesenchymal stem cells) are multipotent cells with potential therapeutic value. Owing to their osteogenic capability, MSCs may be clinically applied for facilitating osseointegration in dental implants or orthopedic repair of bony defect. However, whether wound infection or oral microflora may interfere with the growth and osteogenic differentiation of human MSCs remains unknown. This study investigated whether proliferation and osteogenic differentiation of MSCs would be affected by potent gram-positive and gram-negative derived bacterial toxins commonly found in human settings.ResultsWe selected lipopolysaccharide (LPS) from Escherichia coli and lipoteichoic acid (LTA) from Streptococcus pyogenes as our toxins of choice. Our findings showed both LPS and LTA did not affect MSC proliferation, but prolonged LPS challenge upregulated the osteogenic differentiation of MSCs, as assessed by alkaline phosphatase activity and calcium deposition. Because toll-like receptors (TLRs), in particularly TLR4 and TLR2, are important for the cellular responsiveness to LPS and LTA respectively, we evaluated their expression profiles serially from MSCs to osteoblasts by quantitative PCR. We found that during osteogenic differentiation, MSC-derived osteoprogenitors gradually expressed TLR2 and TLR4 by Day 12. But under prolonged incubation with LPS, MSC-derived osteoprogenitors had reduced TLR2 and TLR4 gene expression. This peculiar response to LPS suggests a possible adaptive mechanism when MSCs are subjected to continuous exposure with bacteria.ConclusionIn conclusion, our findings support the potential of using human MSCs as a biological graft, even under a bacterial toxin-rich environment.

Effect of fluoride iontophoresis on the microtensile bond strength between dentin and two adhesive systems

AIM To evaluate the effect of fluoride iontophoresis on the microtensile bond strength (MTBS) between coronal dentin and two resin-based adhesive systems, and to measure quantitatively dentinal tubule occlusion. METHODS Twelve freshly extracted intact human mandibular third molars were divided randomly into four groups. The superficial occlusal dentin of each tooth was exposed and treated. Group A1: One-Step Plus total-etch adhesive system; group A2: One-Step Plus total-etch adhesive system after fluoride iontophoresis; group B1: ACE BOND SE self-etching adhesive system; group B2: ACE BOND SE self-etching adhesive system after fluoride iontophoresis. A resin composite buildup was made for each tooth, which was then sectioned along its long axis to produce 10 beams (1.0 mm x 1.0 mm) for the microtensile bond strength (MTBS) test. Five dentin disks were cut in half and their occlusal surfaces etched with 6% citric acid. The test halves were treated with fluoride iontophoresis. Four SEM photomicrographs were taken from corresponding sites on each test and each non-treated control half-disk. Image-Pro Plus 4 software quantified the percentage of tubule occlusion. Data were analyzed using one-way ANOVA, chi(2)- and t-tests, with the probability level set at alpha=0.05. RESULTS The mean MTBS (MPa) for each group was, A1: 30.86 (S.D. 6.84); A2: 25.04 (8.49); B1: 19.22 (6.88); B2: 19.40 (6.92). There were significant differences among all groups (P < or = 0.02), except between groups B1 and B2 (P=0.92). Fluoride iontophoresis resulted in significantly increased dentinal tubule occlusion (P<0.001). CONCLUSIONS Fluoride iontophoresis treatment reduced significantly the dentin bond strength of One-Step Plus total-etch adhesive, but not that of ACE BOND SE self-etching adhesive. However, the bond strength of the former remained significantly higher than that of the latter adhesive system.