Inductive agent for regeneration of periodontium tissue of temperature sensitivity and preparation method thereof
A periodontal tissue regeneration and inducer technology, applied in the field of temperature-sensitive periodontal tissue regeneration inducers, can solve the problems of systemic application of large side effects, short half-life, easy to be degraded, etc., and achieves good surface activity, plasticity and porosity. The effect of high structure and strong ability to inhibit periodontal pathogens
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Embodiment 1
[0029] The object of the present invention is to provide a kind of chitosan, chitosan quaternary ammonium salt nanoparticle and α, β-sodium glycerophosphate as carrier and the chitosan temperature-sensitive hydrogel composite of the plasmid containing BMP-2 sequence It also provides a preparation method of the temperature-sensitive hydrogel complex.
[0030] Chitosan is the only natural cationic polysaccharide in nature, its structure is similar to extracellular matrix glycosaminoglycans (GAGs), and it has excellent properties such as good tissue compatibility, biodegradability, bacteriostasis and non-toxicity. It can be combined with a variety of anionic polyelectrolytes in various ways to form a physiological environment temperature-sensitive hydrogel, which can be widely used in tissue engineering scaffolds, drug delivery systems, and cell culture carriers. Due to its cationic properties, chitosan can be used as a non-viral carrier to combine with negatively charged DNA, RN...
Embodiment 2
[0046] The specific steps for preparing the CS / HTCCn(pDNA-BMP2)-GP thermosensitive hydrogel complex in this example are as follows:
[0047] (1) First prepare chitosan quaternary ammonium salt nanoparticles (HTCCn) solution. Concrete quaternization step is with embodiment 1.
[0048] (2) Then, the prepared chitosan quaternary ammonium salt (HTCC) was prepared by ultrasonic emulsification to obtain chitosan quaternary ammonium salt nanoparticles (HTCCn). The specific preparation process is the same as in Example 1.
[0049] (3) Prepare the chitosan quaternary ammonium salt nanoparticle solution encapsulated with the plasmid. The pT7T3D-PacI plasmid containing the BMP-2 sequence was transformed into Escherichia coli for amplification, and the plasmid was purified after separation and extraction. The steps were the same as in Example 1.
[0050] The specific encapsulation process of this embodiment is as follows: HTC Cn solution (0.1%, dissolved in 5ml lactic acid solution, pH...
Embodiment 3
[0053] The CS / HTC Cn(pDNA-BMP2)-GP temperature-sensitive hydrogel complex obtained in the present invention is used as a periodontal tissue inducer for animal experiments and clinical trials, and the test results are as follows:
[0054] (1) Establish an animal model of canine periodontal disease, give local subperiosteal injection of CS / HTCCn(pDNA-BMP2)-GP thermosensitive hydrogel complex with alveolar bone resorption, and X-rays show at 1, 3, and 6 months All had varying degrees of bone regeneration.
[0055] (2), Wang Moumou, man, 34 years old. He went to the doctor for 3 years due to loose teeth. Clinical examination and X-ray examination were diagnosed as "adult periodontitis (severe)". After three months of basic SPR treatment, local injection of CS / HTC Cn(pDNA-BMP2)-GP thermosensitive hydrogel complex was given. X-rays taken at three months showed that the alveolar bone regenerated by about 0.6mm compared with that before the injection.
[0056] (3), Zhang, female, 2...
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