Biological agent for inducing dentin mineralization
A technology of biological preparations and dentin, applied in the field of tooth mineralization drugs in oral biology, can solve problems affecting the mineralization process of dentin, and achieve the effect of inhibiting the development process
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Embodiment 1
[0028] First, isolate and culture human dental pulp cells and dental pulp stem cells. The specific steps are as follows: clinically collect healthy and complete permanent teeth freshly extracted from volunteers aged 18 to 25 (informed consent) due to orthodontic reduction or impaction. Take out the pulp tissue, wipe the surface of the tooth with 750mL / L ethanol, then soak it twice with PBS buffer (phosphate buffer) containing penicillin and streptomycin; Split, take out the pulp in the ultra-clean bench, put it in PBS buffer solution, and cut the pulp into 1mm with ophthalmic scissors 3 ~2mm 3 After centrifuging at 1000r / min for 5min, discard the supernatant; then add 4mg / LI type collagenase to digest in a water bath at 37°C for 45min; use a centrifuge (Kubota2l00, Japan) to centrifuge at 1000r / min for 5min, discard the supernatant ; Wash 3 times with α-MEM medium containing 50mL / L fetal bovine serum. Then, after centrifugation at 1000r / min for 5min, the cells were collecte...
Embodiment 2
[0042] Pyrrolidine dithiocarbamate-induced dental pulp stem cells were co-incubated with calcined bone bio-scaffolds and transplanted into nude mice to further observe the effect of pyrrolidine dithiocarbamate on promoting dentin mineralization.
[0043] First, prepare calcined bovine bone: deproteinize fresh bovine cancellous bone with NaOH, calcinate at 850°C for 4 hours, pulverize and sieve, make 200-280μm particles, sterilize with ethylene oxide, and set aside.
[0044] Collagen preparation: prepare 0.1% v / v glacial acetic acid and 10% NaOH solution, filter and sterilize, and set aside; use 0.1% v / v glacial acetic acid to dilute the purchased 8 mg / ml collagen to 6 mg / ml, and dilute the obtained collagen , 10×DMEM, and fetal bovine serum are mixed in a volume ratio of 10:1:1, and the pH value is adjusted to be neutral with 10% NaOH solution; the collagen solution is divided into sterile 1.5ml EP tubes, 500ul per tube , take out 3 of the EP tubes, dissolve PDTC in 6mg / ml ste...
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