本文選題：丹酚酸B + 羥基磷灰石球粒�。� 參考：《西南交通大學(xué)》2012年碩士論文

【摘要】：一直以來,臨床上棘手于對(duì)由創(chuàng)傷、感染和腫瘤切除后所造成的大節(jié)段骨缺損進(jìn)行修復(fù)。自體骨雖因其具有優(yōu)良的骨傳導(dǎo)性、骨誘導(dǎo)性、成骨潛能和無免疫原性而被視為骨移植材料的金標(biāo)準(zhǔn),但自體骨來源數(shù)量有限,且取骨手術(shù)存在至少10%的并發(fā)癥,植入體內(nèi)后,需要較長時(shí)間的爬行替代過程。而異體骨和異種骨具有抗原性,尤其在移植較大骨時(shí),常因劇烈的免疫排斥反應(yīng)導(dǎo)致移植失敗,并且還有病原傳播危險(xiǎn)。近年來,人們開始用組織工程的原理和技術(shù),將具有成骨潛能的細(xì)胞誘導(dǎo)分化、增殖并種植于支架材料上,形成工程化人工骨,促進(jìn)大節(jié)段骨缺損的修復(fù)。 然而,在體外構(gòu)建大尺寸組織工程骨會(huì)因體外培養(yǎng)的細(xì)胞在植入體內(nèi)后存活率較低、血管化問題和臨床時(shí)限等問題而受到諸多限制。同時(shí),臨床上為更好地促進(jìn)骨的愈合,還會(huì)給患者施以藥物治療。但由于骨組織在生物學(xué)上的如密度大、血流量低、滲透性差等特殊性,傳統(tǒng)給藥方式使藥物很難按理想狀態(tài)到達(dá)病變部位,從而療效低。 羥基磷灰石(HA)的組成和結(jié)構(gòu)與自然骨的極為相似,有良好的生物相容性、骨傳導(dǎo)性和骨誘導(dǎo)性,被廣泛應(yīng)用為骨的替代材料。HA能吸附化學(xué)藥物小分子和蛋白質(zhì)等生物大分子,所以將其作為藥物載體被廣為研究。但其作為藥物載體存在有如表面易吸附雜質(zhì)、載藥率低和突釋問題。 我課題組前期創(chuàng)新地研究并制備了由多孔生物陶瓷球粒堆積的孔隙100%互通、孔隙大小、孔隙率可控且可與生物活性物質(zhì)均勻混合的大尺寸(Φ1～1.5×3-4cm)多孔支架,以期修復(fù)超臨界尺寸的骨缺損。本實(shí)驗(yàn)旨在在此支架上,負(fù)載可促骨生長、促血管化的生物活性物質(zhì),使其成為緩釋體系,期在修復(fù)骨缺損中持續(xù)有效地釋放活性物質(zhì),促進(jìn)骨生長及愈合。 本實(shí)驗(yàn)用溶膠-凝膠法和W/O乳化成球技術(shù)制備了HA多孔球形顆粒,并對(duì)球粒的孔隙結(jié)構(gòu)進(jìn)行了表征；考察了丹酚酸B (Sal B)對(duì)成骨細(xì)胞增殖和分化的影響；將HA球粒作為藥物載體,利用其吸附性將Sal B負(fù)載其上,同時(shí)考察分別用幾種不同濃度的PLA和幾種不同濃度的殼聚糖對(duì)載藥球粒進(jìn)行包裹后藥物的釋放情況,選擇出較佳的包裹物質(zhì)及其濃度,成功制得殼聚糖包裹的Sal B緩釋HA球粒,考察該緩釋體系對(duì)成骨細(xì)胞活性的影響。本實(shí)驗(yàn)獲得的結(jié)論如下： 1.采用溶膠/凝膠法和W/O乳化成球技術(shù)制備的HA球粒具有組織生長需要的微觀孔結(jié)構(gòu)。 2. Sal B能有效地促進(jìn)成骨細(xì)胞增殖、分化及細(xì)胞外基質(zhì)礦化,提高成骨細(xì)胞的總代謝活性和堿性磷酸酶的表達(dá)從而促進(jìn)成骨。當(dāng)Sal B的質(zhì)量濃度≤160μg/mL時(shí)對(duì)成骨細(xì)胞的增殖和分化有明顯的促進(jìn)作用,且質(zhì)量濃度越高效果越明顯；但當(dāng)Sal B的質(zhì)量濃度高于160μg/mL時(shí),便對(duì)成骨細(xì)胞產(chǎn)生毒性,導(dǎo)致其凋亡。由此得出,Sal B可作為促骨生長和愈合的生物活性物質(zhì)應(yīng)用于骨缺損的修復(fù),且最佳藥效濃度為160μg/mL。 3.由于PLA的疏水性,其所形成的膜不能很好的附著在HA球粒表面,易脫落,所以幾種不同濃度的PLA對(duì)載藥球粒的包裹效果都不好。而殼聚糖親水性和成膜性好,且具有良好的細(xì)胞相容性,最終選用2%的殼聚糖溶液包裹制得載Sal B的緩釋HA球粒。 4.殼聚糖包裹Sal B緩釋HA球粒可在體外長時(shí)間內(nèi)釋放具有生物活性的Sal B,維持有效促進(jìn)成骨細(xì)胞增殖的濃度并保持成骨細(xì)胞的生物學(xué)活性,較長時(shí)間內(nèi)促進(jìn)成骨細(xì)胞分裂和增殖,能滿足骨創(chuàng)傷治療中所需要的持續(xù)性促進(jìn)骨形成的能力,為修復(fù)大截段骨缺損提供了新的途徑。
[Abstract]:It has been clinically difficult to repair large segmental bone defects caused by trauma, infection and tumor resection. The autogenous bone is considered as the gold standard for bone graft, although it has excellent bone conductivity, bone induction, osteogenic potential and no immunogenicity, but the number of autologous bone sources is limited, and at least 1 of the bone removal surgery exists. 0% of the complications, after implantation, need a long time crawling replacement. Allograft and xenogeneic bone are antigenicity, especially in large bone graft, often caused by severe immune rejection, and the risk of pathogen transmission. In recent years, the principles and techniques of tissue engineering have begun to have osteogenic potential. The cells were induced to differentiate, proliferate and grow on scaffold materials to form engineering artificial bones to promote the repair of large segmental bone defects.
However, the construction of large size tissue engineering bone in vitro will be limited by the low survival rate, vascularization and clinical time limit for the cells cultured in vitro. At the same time, it can improve the healing of bone and give the patient the treatment of drugs. But because the bone tissue is very dense in Biology, Because of low blood flow and poor permeability, the traditional way of administration makes it difficult for the drug to reach the lesion according to the ideal state, so its curative effect is low.
The composition and structure of hydroxyapatite (HA) are very similar to those of natural bone. It has good biocompatibility, bone conductivity and bone inducibility. It is widely used as a substitute material of bone,.HA, to adsorb small molecules and proteins such as chemical drugs and proteins, so it is widely studied as a carrier of drug, but it is used as a drug carrier. It is easy to adsorb impurities such as surface, low drug loading rate and sudden release.
In the earlier period, we studied and prepared a large size (1 to 1.5 x 3-4cm) porous scaffold with porous bioceramic spherules 100% interworking, pore size, controlled porosity and uniform mixing with bioactive substances to repair bone loss in supercritical size. This experiment was designed to promote bone growth on this scaffold. A long, vascularized bioactive substance makes it a slow release system, which can effectively release active substances and promote bone growth and healing in repairing bone defects.
In this experiment, the porous HA particles were prepared by sol-gel method and W/O emulsifying technique, and the pore structure of the pellets was characterized. The effect of salvianolic acid B (Sal B) on the proliferation and differentiation of osteoblasts was investigated. HA pellets were used as drug carriers to load Sal B by their adsorbability, and several different kinds of differences were used respectively. The concentration of PLA and several different concentrations of Chitosan on the drug delivery of drug loaded pellets were released. The better inclusion substance and its concentration were selected. The Sal B sustained-release HA pellets coated with chitosan were successfully prepared, and the effect of the sustained release system on the activity of osteoblasts was investigated. The conclusions are as follows:
1. the HA spherules prepared by sol gel method and W/O emulsification technology have the micro pore structure needed for tissue growth.
2. Sal B can effectively promote osteoblast proliferation, differentiation and extracellular matrix mineralization, increase the total metabolic activity of osteoblasts and the expression of alkaline phosphatase to promote osteogenesis. When the mass concentration of Sal B is less than 160 UX, the proliferation and differentiation of osteoblasts is obviously promoted, and the higher the mass concentration, the more obvious, but when the mass concentration is higher, the effect is more obvious. When the mass concentration of Sal B is higher than 160 u g/mL, it is toxic to osteoblasts and leads to its apoptosis. Thus, Sal B can be used as a bioactive substance for bone growth and healing to repair bone defects, and the optimum concentration is 160 mu g/mL..
3. because of the hydrophobicity of PLA, the membrane formed is not very good attached to the surface of the HA pellet, and it is easy to fall off. So the encapsulation effect of several different concentrations of PLA on the pellets is not good. The hydrophilic and film forming property of chitosan is good, and it has good cellular compatibility. Finally, 2% chitosan solution is used to package the slow release HA ball carrying Sal B. Grain.
4. chitosan encapsulated Sal B sustained-release HA pellets can release bioactive Sal B for a long time in vitro, maintain the concentration of osteoblast proliferation effectively and maintain the biological activity of osteoblasts, promote osteoblast division and proliferation for a long time, and can meet the sustained bone formation ability needed in the treatment of bone trauma. It provides a new way for repairing large segment bone defect.

【學(xué)位授予單位】：西南交通大學(xué)
【學(xué)位級(jí)別】：碩士
【學(xué)位授予年份】：2012
【分類號(hào)】：R318.08

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