本文選題：甲狀旁腺相關肽 + 聚乙交酯-丙交酯�。� 參考：《吉林大學》2014年碩士論文

【摘要】：研究背景及目的： 甲狀旁腺激素（Parathyroid Hormone, PTH）是具有促進成骨細胞形成及加強成骨細胞礦化作用的治療骨質疏松癥藥物。而甲狀旁腺相關肽（PTHrP）是一種與PTH（1～34）在基因編碼、分子結構、受體構成和信號傳導方面有許多相同或相似之處的多肽類物質，具有與PTH相似的生物學活性。目前很多研究將PTHrP用于增加種植體周圍骨密度，提高種植體早期穩(wěn)定性方面，以減少骨愈合時間。 口腔種植體早期穩(wěn)定性的關鍵因素在于牙槽骨的質量和剩余牙槽骨的體積，受系統(tǒng)性骨質疏松的影響。然而PTHrP不同的給藥方式對骨組織的影響不同，間歇的、低劑量的給藥能促進成骨，增加骨量；持續(xù)的、大劑量的PTHrP會引起破骨細胞活化，抑制成骨細胞功能。另外，PTHrP存在著半衰期時間短、容易變性、靶向性差且價錢高昂等問題，因此研制控制其釋放的載體，提高其生物利用率是有效的解決以上缺陷的途徑。聚乙交酯-丙交酯(PLGA)是一種生物相容性良好的生物可降解高分子材料，具有良好的可降解性、可吸收性，而且可作為藥物載體以控制藥物釋放的速度。因此研制以PLGA作為藥物控釋載體負載PTHrP，可控制藥物釋放速度，從而達到長時間緩釋的目的。 方法： 采用W1/O/W2揮發(fā)法制備實驗所需的包載PTHrP1-34的PLGA微球。通過SEM、核磁、紅外等觀察PLGA載藥微球結構；利用紫外分光光度法在280nm 考察載藥微球的體外釋放特性。并通過體細胞毒性評價，堿性磷酸酶測定等檢測負載PTHrP1-34的PLGA微球對小鼠前骨細胞的分化、細胞活性及凋亡效應的影響。 結果： 1、PLGA重復單元中乳酸和羥基乙酸單元的比約為85:15，，分子量Mn=85000，PDI=1.26。通過觀察載藥微球具有良好的球形結構，平均粒徑約為8μm。包載PTHrP1-34的PLGA微球的載藥率和包封率分別為0.84%和72.3%。包載PTHrP1-34的PLGA微球能實現(xiàn)長達25day的持續(xù)釋放。 2、PTHrP1-34載藥微球的細胞學評價： 間歇性使PTHrP1-34作用于MC3T3-E1時，濃度為1×10-9mol/L的PTHrP1-34對MC3T3-E1增殖及ALP活性促進效應最大，與文獻報道略有差異；包載了PTHrP1-34的PLGA緩釋微球間歇性作用于MC3T3-E1時，相比于空白對照組和純藥組，總濃度在1×10-9mol/L時具有明顯的促進MC3T3-E1細胞增殖以及增加ALP活性的作用（P0.05）。 結論： 1、包載PTHrP1-34的PLGA微球的持續(xù)釋放可以達到25day，能較好地實現(xiàn)PTHrP1-34在生理溫度下的長期持續(xù)釋放。 2、最適濃度10-9mol/L的PTHrP1-34間歇性給藥時可促進MC3T3-E1增殖及增加ALP活性。 3、包載了PTHrP1-34的PLGA緩釋微球作用于MC3T3-E1時，可以明顯促進MC3T3-E1的增殖與分化，提示包載了PTHrP1-34的PLGA緩釋微球，在口腔種植領域的應用具有較好的應用前景，為臨床解決種植體周圍骨量不足等問題提供了新的可能的解決方案。
[Abstract]:Research background and purpose:
Parathyroid hormone (Parathyroid, Hormone, PTH) is promote osteoblast formation and enhance osteoblast mineralization in the treatment of osteoporosis drugs. And parathyroid hormone related peptide (PTHrP) is a kind of PTH (1~34) in the gene encoding, molecular structure, a polypeptide of many of the same or similar structure and receptor signal transduction, with biological activity similar to that of PTH. At present a lot of research will be PTHrP to increase the bone density around the implant, improve early implant stability, to reduce the time of bone healing.
The key factor lies in the stability of early dental implant alveolar bone quality and alveolar bone volume, affected by systemic osteoporosis. However, PTHrP administered different effects on bone tissue intermittent, low doses of the drug can promote bone formation, increase bone mass; sustained, high dose PTHrP induced osteoclast activation, inhibition of osteoblast function. In addition, PTHrP has a short half-life, easy degeneration, targeting the poor and high price, so the development of the release of vector control, improve their bioavailability is the effective way to solve the above defects. Polyglycollide - C poly (PLGA) is a kind of biodegradable polymer materials, has good biodegradability and absorbability, and can be used as drug carrier to control the rate of drug release. Therefore the development of PLGA as drug controlled release The carrier load PTHrP can control the rate of drug release so as to achieve a long time release.
Method:
The PLGA microspheres containing PTHrP1-34 were prepared by W1/O/W2 evaporation method. The structure of PLGA drug loaded microspheres was observed by SEM, NMR and IR, and 280nm was detected by ultraviolet spectrophotometry.
The release characteristics of drug loaded microspheres were investigated. The effects of PTHrP1-34 loaded PLGA microspheres on the differentiation, cell viability and apoptosis of mouse bone marrow cells were detected by somatic cytotoxicity and alkaline phosphatase assay.
Result:
1, lactic acid and glycolic acid unit PLGA repeat unit was 85:15 and the ratio of molecular weight of Mn=85000 and PDI=1.26. through the observation of microspheres with spherical structure good sustained release, the average particle size is about the drug loading rate of 8 M. PLGA microspheres loaded PTHrP1-34 and encapsulation rate were 0.84% PLGA microspheres and 72.3%. package PTHrP1-34 can achieve up to 25day.
2, the cytological evaluation of PTHrP1-34 loaded microspheres:
The effect of PTHrP1-34 on intermittent MC3T3-E1, concentration of 1 * 10-9mol/L PTHrP1-34 on MC3T3-E1 proliferation and ALP activity of promoting effect, slightly with the reported difference; PTHrP1-34 loaded PLGA microspheres intermittent effect on MC3T3-E1 when compared to the blank control group and pure drug group, the total concentration significantly stimulated the proliferation of MC3T3-E1 cells along with the increase of ALP activity in 1 * 10-9mol/L (P0.05).
Conclusion:
1, the continuous release of PLGA microspheres loaded with PTHrP1-34 can reach 25day, which can achieve a good long-term sustained release of PTHrP1-34 at physiological temperature.
2, PTHrP1-34 intermittent administration of the optimum concentration of 10-9mol/L can promote the proliferation of MC3T3-E1 and increase the activity of ALP.
3, PTHrP1-34 loaded PLGA microspheres on MC3T3-E1, can significantly promote the proliferation and differentiation of MC3T3-E1, suggesting that PLGA sustained-release microspheres containing PTHrP1-34, has a good prospect of application in the field of oral implantology, to solve the growing problem of clinical bone defects around the body provides a new possible solution.

【學位授予單位】：吉林大學
【學位級別】：碩士
【學位授予年份】：2014
【分類號】：R783.6

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